Sixth International Symposium
on Rose Research and Cultivation
Hannover, Germany, 25th - 30th August 2013
www.rosesymposium2013.uni-hannover.de
Dear Colleagues,
It is a great pleasure to welcome you to the International Symposium on Rose Research and Cultivation
which, this year will be held in Hannover, Germany. It is the 6th Symposium on roses held under the
auspices of the International Society for Horticultural Science (ISHS) and follows a series of rose
symposia that started in Rehovot, Israel in 1985.
Since then rose research has been transformed by technological revolutions in many areas of science
most of all by tremendous developments in plant molecular biology that meanwhile penetrated all
areas of research, not only genetics and breeding but also disciplines as postharvest physiology,
phytomedicine, ecology and taxonomy to name just a few. This year´s symposium will follow the
established structure of the past symposia in that scientific presentations by oral communications
and posters are mixed with a mid-congress professional tour.
The scientific program covers major areas of rose research and provides a large number of very
interesting talks and an even larger number of posters providing up to date information on current
research projects on roses all around the world. The professional tour includes visits to the German
variety protection office involved in a federal program for the conservation of genetic resources
of the rose and visits to two major German rose breeders Kordes´ Söhne and Rosen Tantau. A post
congress tour provides the possibility to visit one of the largest rose collections of the world, the
Europa-Rosarium in Sangerhausen.
I hope that your active participation will lead to intensive and fruitful discussions on rose research
and to new contacts and scientific collaborations and that you enjoy your time in Hannover with its
famous historical gardens.
Thomas Debener
Institute for Plant Genetics
Sixth International Symposium
on Rose Research and Cultivation
25th – 30th August 2013, Hannover Congress Centrum, Germany
Organizing Committee
Thomas Debener, Leibniz Universität Hannover
Jürgen Grunewaldt, Leibniz Universität Hannover
Marcus Linde, Leibniz Universität Hannover
Heiko Mibus, Leibniz Universität Hannover
Traud Winkelmann, Leibniz Universität Hannover
Bettina Winnat, Leibniz Universität Hannover
Scientific Committee
Dave Byrne, USA
Thomas Debener, Germany
Johan van Huylenbroeck, Belgium
QingLin Liu, China
Hilde Nybom, Sweden
H. Brent Pemberton, USA
José Antonio Saraiva Grossi, Brazil
Margrethe Serek, Germany
René Smulders, Netherlands
Yoshihiro Ueda, Japan
We are greatly thankful for the support from the following sources:
Deutsche Forschungsgemeinschaft (DFG)
Niedersächsisches Ministerium für Wissenschaft und Kultur (MWK)
Freundeskreis der Leibniz Universität Hannover e.V.
Kanzlei Würtenberger/Kunze
W. Kordes’ Söhne Rosenschulen
Rosen Tantau KG
Bund Deutscher Baumschulen
Syngenta Agro GmbH
Leibniz Universität Hannover
Scientific Program
Sunday, August 25th, 2013 – Hannover Congress Centrum
15.00 – 19.00
18.00 – 20.00
Registration
Welcome Reception: Wine and Cheese
Monday, August 26th, 2013 – Hannover Congress Centrum
09.00 Opening and Greetings
Thomas Debener
(Leibniz Universität Hannover, Convener)
Margrethe Serek
(Leibniz Universität Hannover, ISHS Chair Section Ornamentals)
Udo Schmitz
(Leibniz Universität Hannover, Dean Faculty of Natural Sciences)
09.30 – 11.35
Session 1: Genetic Resources
Page
Chairperson: Volker Wissemann, Julius-Liebig-University Gießen, Germany
09.30 – 10.15 What´s in a genetic resource?
Volker Wissemann
1
10.15 – 10.35
Scots Roses and related cultivars of Rosa spinosissima –
a review
Peter Boyd
2
10.35 – 10.55
DISTRIBUTION, CHARACTERIZATION AND PROPAGATION OF SICILIAN 3
ROSE SPECIES
Giancarlo Fascella
10.55 – 11.20
Coffee Break
11.20 – 11.40
DIVERSE AND VALUABLE WILD ROSES IN THE HENGUAN
MOUNTAINS, SOUTHWESTERN CHINA
Hongying Jian
4
11.40 – 12.00
GENETIC DIVERSITY ANALYSIS OF OLD GARDEN ROSES FROM THE 19th CENTURY BASED ON SSR MARKERS
Shubin Li
5
12.00 – 13.00
13.00 – 14.00
Postersession
LUNCH
14.00 – 16.25
Session 2: Genetics and Breeding
Page
Chairperson: Dave Byrne, Texas A&M University, USA
14.00 – 14.45
Advances in Rose Breeding and Genetics in North
America
Dave Byrne
6
14.45 – 15.05
Cultivar differences in plant transpiration rate at high relative air humi-dity are not related to genotypic
variation in stomatal responsiveness
Habtamu Giday
7
15.05 – 15.25 THE CONTINUOUS FLOWERING GENE IN ROSE IS A FLORAL
INHIBITOR
Laurence Hibrand-Saint Oyant
8
15.25 – 15.45
GENE-SPECIFIC KNOCKDOWN SYSTEM USING AN APPLE
LATENT SPHERICAL VIRUS VECTOR IN ROSE
Shungo Otagaki
9
15.45 – 16.05
USE OF DIGITAL GENE EXPRESSION TO DISCRIMINATE GENE
EXPRESSION IN DIFFERENT DEVELOPMENTAL STAGE OF ROSA
CHINENSIS ‘PALLIDA’
Huijun Yan
10
16.05 – 16.25
EARTH-KIND® ROSE TRIALING: AN INTERNATIONAL MODEL FOR THE
IDENTIFICATION OF REGIONALLY-ADAPTED LANDSCAPE ROSES
David Zlesak
11
16.25 – 16.50
16.50 – 18.15
Coffee Break
Session 3: Rose Hips
Page
Chairperson: Hilde Nybom, Swedish University of Agricultural Sciences, Sweden
16.50 – 17.35
Rose hips: beauty is as beauty does
Hilde Nybom
12
17.35 – 17.55
FACTORS AFFECTING THE GERMINATION OF HYBRID ROSE ACHENES: A REVIEW
Dik de Vries
13
17.55 – 18.15
LEAF AND FLOWER PHENOLIC PROFILES OF INDIGENOUS
SLOVENIAN ROSE SPECIES
Valentina Schmitzer
14
18.30DEPARTURE TO HERRENHAEUSER GAERTEN
19.00 – 20.30
GUIDED TOUR THROUGH THE HISTORIC BAROQUE GARDENS OF
HERRENHAUSEN
Tuesday, August 27th, 2013 – Hannover Congress Centrum
08.30 – 11.45
Session 4: Rose Genomics
Page
Chairperson: Fabrice Foucher, INRA Angers, France
08.30 – 09.15 Towards the rose genome sequence and its use in
research and breeding
Fabrice Foucher
15
09.15 – 09.35
Genetic analysis of tetraploid F1 rose populations based on the Rose Axiom SNP array
René Smulders
16
09.35 – 09.55
Borrowing from neighbors: leveraging genome sequence 17
of strawberry, apple, and peach to study disease
resistance in rose
James M. Bradeen
09.55 – 10.15
MOLECULAR GENETIC STUDIES ON THE CONTINOUS-FLOWERING ROSES THAT ARE NOT ORIGINATED FROM ROSA CHINENSIS
Koji Kawamura
10.15 – 10.45
18
Coffee Break
10.45 – 11.05
Integration of genetic and physical maps for Rosa wichurana using Tyramide-FISH
Jan De Riek
19
11.05 – 11.25
RNA-SEQ AND DIGITAL GENE EXPRESSION ANALYSIS OF FLOWER
DEVELOPMENT IN RECURRENT AND NON-RECURRENT ROSES
Qinglin Liu
20
11.25 – 11.45
ASSOCIATION MAPPING OF PHENOTYPIC TRAITS IN ROSES
Dietmar Schulz
21
11.45 – 13.00
13.00 – 14.30
Postersession
LUNCH
14.30 – 15.55
Session 5: Postharvest Physiology
Page
Chairperson: Ernst Woltering, Wageningen University, The Netherlands
14.30 – 15.15
Sustainable solutions to preserve rose flower quality 22
and to facilitate long distance transportation
Ernst Woltering
15.15 – 15.35
Morphology, endogenous phytohormones, and related proteins of Bent Peduncle Phenomenon in Cut Rose plants
Wan Soon Kim
15.35 – 16.00
23
Coffee Break
16.00 – 17.35
Session 6: Secondary Metabolites and Biochemistry
Page
Chairperson: Alexander Vainstein, Hebrew University of Jerusalem, Israel
16.00 – 16.45
Sweet smell of rose: navigating the biochemical
network of floral scent production
Alexander Vainstein
24
25
16.45 – 17.05
ROSE SCENT 2-PHENYLETHANOL WAS PRODUCED VIA TWO
BIOSYNTHETIC PATHWAYS IN RESPONSE TO SEASONAL CHANGE IN ENVIRONMENT
Momoyo Kanda
18.30 and 19.00
SHUTTLE SERVICE TO THE RESTAURANT
19.00 Symposium DINNER
12 APOSTEL, PELIKANPLATZ 2; 30177 HANNOVER
22.15 and 22.45
SHUTTLE SERVICE BACK TO THE HOTEL
Wednesday, August 28th, 2013
08.00 – 18.30
Symposium Professional Tour
German variety protection office (Bundessortenamt) in Hannover
Rose breeding companies:
Kordes Rosen in Sparrieshoop
Rosen Tantau in Uetersen
(Lunch Box included)
Thursday, August 29th, 2013 – Hannover Congress Centrum
8.30 – 10.15
Session 7: Growth Physiology and Production/Propagation
Page
Chairperson: Brent Pemberton, A&M University, USA
08.30 – 09.15
A recent history of changing trends in U.S. garden rose plant sales and production methods
Brent Pemberton
26
09.15 – 09.35
GROWERS’ AND CONSUMERS’ KNOWLEDGE, ATTITUDES AND
OPINIONS REGARDING ROSES AVAILABLE FOR PURCHASE
Dave Byrne
27
28
09.35 – 09.55
EFFECT OF FLOWER BUD REMOVAL, STEM AGE, AND LOCATION IN CANOPY OF THE STOCK PLANT ON CUTTING PROPAGATION OF ROSE CULTIVARS
Byoung Ryong Jeong
09.55 – 10.15
DIURNAL RHYTHM OF PETAL GROWTH IN CUT ROSE FLOWER
Takanori Horibe
10.15 – 10.45
10.45 – 12.10
29
Coffee Break
Session 8: Abiotic Stress
Page
Chairperson: Marie Christine van Labeke, Ghent University, Belgium
10.45 – 11.30
Abiotic stress tolerance in roses: physiological and
biochemical changes
Marie-Christine van Labeke
30
11.30 – 11.50
INCREASED SALINITY IMPROVES STOMATAL FUNCTIONING OF
ROSES GROWN UNDER HIGH RELATIVE AIR HUMIDITY
Dália Carvalho
31
11.50 – 12.10
Responses of Selected Garden Roses to Four Different Soil
Moisture Contents
Dave Byrne
32
12.15 – 13.30
LUNCH
13.30 – 16.45
Session 9: Rose Diseases and Pests
Page
Chairperson: Thomas Debener, Leibniz Universität Hannover, Germany
13.30 – 14.15
Genetics and genomics of disease resistance in roses
Thomas Debener
33
14.15 – 14.35
POWDERY MILDEW RESISTANCE IN ROSES IS PATHOTYPE SPECIFIC
Leen Leus
34
14.35 – 14.55
Rose rosette virus is the causal agent of rosette disease
Ioannis Tzanetakis
35
14.55 – 15.15
VOLATILE EMISSIONS FROM ACTIVELY-GROWING GYNANDROPSIS
GYNANDRA AND CLEOME HASSELERIANA THAT PROVIDE EFFECTIVE
CONTROL OF SPIDER MITES IN PROTECTED ROSE CULTIVATION
Brian Grout
36
15.15 – 15.45
Coffee Break
15.45 – 16.05
Symptoms, transmission, and detection of four rose
viruses
Dimitre Mollov
37
16.05 – 16.25
Epidemiology of Blackberry chlorotic ringspot virus
Ioannis Tzanetakis
38
16.25 – 16.45
A bio-test for rose replant problems and investigating
rhizosphere microbial communities using DGGE
fingerprints
Traud Winkelmann
39
16.45 – 17.00
CLOSING REMARKS
Friday, August 30th, 2013
8.00 – 18.30
Post Congress Tour
Europa-Rosarium Sangerhausen
ORAL PRESENTATIONS
Session 1: Genetic Resources
Talk
What´s in a genetic resource?
Wissemann, Volker
Justus-Liebig-Universität Giessen, Institut für Botanik, Professur für Spezielle Botanik &
Botanischer Garten, Heinrich Buff Ring 38, 35392 Giessen, Germany
„Resources“ in general as a scientific term refer to materialistic or structural properties which any
organism needs for its existence, growth and reproduction. In a more narrow definition the trait
of “resources” is limited to environmental factors which are potentially limited and depleted by
competing organism leading to the survival of the fittest. Are genetic resources covered by this
definition? At first glance definitely not. Genetic resources are potentially unlimited, permanently
newly created by evolutionary factors, and they refer to genetic properties of potential or already
applicable value. Thus, in a biological sense genetic resources are unlimited and can not be depleted.
However, we can deplete or exploite the usefulness of a specific genetic resource, e.g. disease
resistance, colour variability and other breeding goals due to limited natural variability. By that
resources become potentially limited. Resources can also become limited if patented or protected by
breeders rights as intellectual property. In that case, the more narrow definition of the resources as
the base for success, e.g. survival of the economic fittest (or fatest?) becomes evident. But there is
one biological mechanism, which separates limited environmental resources from unlimited genetic
resources, and that are the processes of interaction and coevolution. By these processes, genetic
material becomes altered and altered again, sharpened in terms of adaptation to evolutionary
changes and thus potentially unlimited. Therefore I postulate, that the most divergent genetic
resources in roses are found in genetic material related to interaction and coevolution on one hand,
and self-poisoning reactions on the other hand. Disease resistance (e.g. by endophytic reactions),
tolerance to abiotic stress (e.g. by differential salt tolerance mechanisms), differential quenching of
chemical reactions (e.g. lycopene variability), chemical properties of glands (e.g. sesquiterpenoids),
growth forms, differential investment in reproductive modes, recurrent flowering, oil production are
potential targets of further research in resource research. Examples based on interactive characters
and character states will be discussed of yet unexploited functional traits of wild roses for breeding
purposes.
1
Session 1: Genetic Resources
Talk
Scots Roses and related cultivars of
Rosa spinosissima – a review.
Boyd, Peter1
Hill Cottage, Meadowtown, Minsterley, Shrewsbury, Shropshire SY5 0DZ. UK. [Heritage Rose
Committee, World Federation of Rose Societies; Patron, Heritage Roses in Australia (2013-2014);
Holder of Plant Heritage ‘National Collection of Rosa spinosissima (syn. R. pimpinellifolia) Scots
Roses and hybrids’]
1
‘Scots Roses’ and other cultivars of Rosa spinosissima (R. pimpinellifolia) are less well-known today
than other rose groups and an under-exploited genetic resource. However, in 1840, there were
probably more cultivars of ‘Scots Rose’ than cultivars of all the other distinct groups of garden roses
of the time combined. Hundreds of single, semi-double and double Scots Roses were raised in a wide
range of colours in Britain, Europe and North America before 1830. They were less ‘fashionable’
but more widespread by 1840. Immigrants to North America, New Zealand and elsewhere carried
suckers or seeds of Scots Roses to their new homes. Most of the old cultivars originated as variants or
hybrids of the low-growing form of the species that grows wild in north-western Europe. In the 20th
century, Doorenbos in the Netherlands and others raised cultivars with a similar character to the
old Scots Roses. However, Kordes in Germany and others used tall-growing Asian forms with larger
flowers (e.g. ‘Altaica’) to breed large shrubs (e.g. Frühlings hybrids). Such hybrids have a different
habit and character to the old Scots Roses. Depending on latitude and altitude, true Scots Roses
bloom between April and July in the Northern Hemisphere or between October and December in the
Southern Hemisphere. Some are repeat-blooming. They are very floriferous and generally sweetly
scented. Most produce black or dark purple heps. Many exhibit good autumn colour in shades of
yellow, orange, pink, red or purple. Few old cultivars remain in commerce but many old gardenworthy cultivars have been rediscovered and new seedlings raised. There is renewed interest in the
group but their nomenclature is confused. R. spinosissima cultivars are extremely cold-hardy and
resistant to drought and diseases. These charming but tough roses have potential as a source of new
garden roses in a time of climatic uncertainty.
2
Session 1: Genetic Resources
Talk
DISTRIBUTION, CHARACTERIZATION AND
PROPAGATION OF SICILIAN ROSE SPECIES
Fascella Giancarlo, Gaetano Giardina, Antonio Giovino and Silvia Scibetta
Consiglio per la Ricerca e la Sperimentazione in Agricoltura, Unità di Ricerca per il recupero e la
valorizzazione delle specie floricole mediterranee (Research Unit for Mediterranean Flower Species),
S.S.113 - Km 245.500, 90011 Bagheria (Palermo), Italy
In the present work distribution, habitats, characterization and propagative characteristics of wild
Roses native to Sicily were described in order to define their ecological value and ornamental
potentialities.
Among studied species, Rosa canina L. resulted the most widespread in the regional territory (found
from 0 to 1500 m of altitude), followed by R. sempervirens L. (from 100 to 600 m); R. rubiginosa L.
was the least common (only beyond 1600 m).
Sicilian roses showed a high variability on morphological characteristics: R. sempervirens was the
only evergreen species, with creeping habit and long internodes; R. sicula Tratt. showed a dwarf
habit (up to 50 cm). Molecular characterization was accomplished by means of “DNA barcoding”, a
technique of taxonomic identification based on a unique nucleotidic sequence (matK and rbcL) for
each species.
All species were propagated in vivo by placing semihardwood cuttings on rooting substrates
(sphagnum peat, perlite, peat:perlite 1:1 v/v) with or without 0.4% naphthalene acetic acid (NAA)
treatment. Rooting rates were affected by genotypes, substrates and NAA treatment, with higher
values (46.2%) recorded for R. sempervirens cuttings rooted in peat:perlite and treated with NAA.
Roses species were also propagated in vitro by culturing microcuttings in a multiplication medium
with four benzyladenine (BA) concentrations (0, 0.25, 0.5 and 0.75 mg/L). Shoots formation was
influenced by genotype and BA concentration: higher number of shoots (2.2) was obtained from
Rosa canina explants on medium with BA 0.75 mg/L. Microshoots were successively transferred to a
rooting m edium with three indolacetic acid (IAA) concentrations (0, 0.3 and 0.6 mg/L). Rooting rates
were affected by genotype and IAA concentration as R. canina showed higher values (68.5%) with
IAA 0.6 mg/L, whereas R. sempervirens evidenced higher rate (92.1%) with the control.
This research was financed by Italian Ministry of Agriculture through the project “R.G.V./FAO” - D.M.
13763/11.
3
Session 1: Genetic Resources
Talk
DIVERSE AND VALUABLE WILD ROSES IN THE
HENGUAN MOUNTAINS OF SOUTHWESTERN
CHINA
Hongying Jian, Shufa Li, Shubin Li, Qigang Wang, Xianqin Qiu, Lulin Ma, Hao Zhang and Kaixue
Tang
Flower Research Institute, Yunnan Academy of Agricultural Sciences, Kunming, Yunnan, 650205,
China
China is the greatest treasure house of roses with 95 wild species and many varieties and forms of
genus Rosa. The Hengduan Mountains, located on the eastern edge of the Qinghai-Tibetan plateau
in southwestern China, is a core region of the Sino-Himalayan Floristic Region and is characterized
by its richness in biodiversity and endemic species. Based on field expeditions for the search of wild
roses in this area since 2003, the distribution, morphology and other information on wild roses were
recorded and compiled. The results were the following: 1) There were 71 species/varieties and forms
of wild roses mainly from Section Cinnamomeae, Section Synstylae, Section Pimpinellifoliae and
Section Microphyllae. Among them, 44 species/varieties and forms were endemic to the Hengduan
Mountains. 2) Some species were very narrowly distributed and some thrived widely in both elevation
and horizon. The elevational distribution of genus Rosa in the Hengduan Mountains was from
1662m to 4400m. Section Cinnamomeae and Section Pimpinellifoliae occupied the higher areas
with an altitude from 3000m to 4000m, scattering under forests or in alpine scrubs and meadows,
while Section Synstylae densely grew on the sunny barren or along the riversides from 2000m to
3000m except for Rosa soulieana. 3) Many species were diverse in morphology because of natural
hybridization, polyploidization and adaption, resulting in richness and endemism of genus Rosa in this
area. 4) Wild roses embraced desirable traits such as elegant form, vivid flower color, fantastic hips,
pleasant scent, resistant to powdery mildew and tolerant to drought and coldness, etc. The results
suggested that wild roses in the Hengduan Mountains were of great importance for the studies on
systematics and evolution of genus Rosa. It was also necessary to investigate and preserve the wild
roses in this area for directly utilization as ornamentals or as genetic resources for rose breeding.
4
Session 1: Genetic Resources
Talk
GENETIC DIVERSITY ANALYSIS OF OLD GARDEN
ROSES FROM THE 19th CENTURY BASED ON SSR
MARKERS
Shubin Li1,2,3,4, Annie Chastellier1,2,3, Jérémy Clotault1,2,3, Fabrice Foucher1,2,3, Sylvain Gaillard1,2,3,
Laurence Hibrand-Saint Oyant1,2,3, Julien Jeauffre1,2,3, Raymond Loubert5, Thérèse Loubert5, Gilles
Michel1,2,3, Vanessa Soufflet-Freslon1,2,3, Tatiana Thouroude1,2,3, Kaixue Tang4, Alix Pernet1,2,3*
INRA, UMR 1345 Institut de Recherche en Horticulture et Semences , SFR 4207 QUASAV, F-49071
Beaucouzé, France
2
Université d’Angers, UMR 1345 Institut de Recherche en Horticulture et Semences, SFR 4207
QUASAV, PRES L’UNAM, F-49000 Angers, France
3
Agrocampus Ouest, UMR 1345 Institut de Recherche en Horticulture et Semences, SFR 4207 QUASAV, F-49000 Angers, France
4
Flower Research Institute, Yunnan Academy of Agricultural Sciences, Kunming, Yunnan, 650205,
China
5
Roseraie « Roses Loubert », Les Brettes, 49350 Les Rosiers-sur-Loire
* To whom correspondence should be addressed. E-mail: [email protected]
1
The history of rose breeding can be divided into three crucial periods: wild species collection and
domestication, old garden rose cross breeding, and the modern rose production. It is well known
that old garden roses have acted as a bridge for inheritance of several important traits, such as
continuous flowering or scent production. In order to understand the genetic determinism of
continuous flowering trait and select related genes, we propose to develop an association mapping
strategy on an old garden rose collection.
For this purpose, we investigate the genetic diversity and population structure of the old European
and Chinese garden roses using microsatellite markers. Analysis is conducted using 32 microsatellites
primer pairs, either anonymous or from expressed sequenced tags, on 350 old garden roses from
different sections and horticultural groups mainly bred during the 18th-19th centuries.
According to previous studies, alleles are scored in a binary format according to their molecular
weight. Explorative analysis of the variability is performed by calculating distance matrices based on
Dice genetic dissimilarity indices and then visualizing these distance matrices by principal coordinate
analyses. The obtained distance matrices are also subjected to cluster analyses by the “neighbor
joining” method. Inference analysis of the variability is conducted thanks to a model-based clustering
method, using appropriate software package like Structure.
Results concerning the structure of the rose population under study will be presented and the possible
development of association mapping will be discussed.
5
Session 2: Genetics and Breeding
Talk
Advances in Rose Breeding and Genetics in
North America
David H. Byrne
Department of Horticultural Sciences, Texas A&M University, College Station, TX 77843-2133, USA
Rose breeding in the North America is dominated by private breeding programs, both large and small,
whereas the research into rose breeding and genetics is done primarily by public institutions such as
Universities where the new generation of breeders are being trained. Both these efforts are influenced
by the larger trends towards products that are environmentally friendly and convenient to use. Thus
there is an emphasis on the development of well adapted, especially disease resistant, landscape rose
cultivars. This trend is confirmed by a recent survey of the rose industry in the USA which indicated
that disease resistance is the most desired trait for landscape roses. Research in public institutions
also emphasizes disease resistance and other adaptation traits as well as the utilization of molecular
tools in more efficient introgression of these traits into superior horticultural rose germplasm.
6
Session 2: Genetics and Breeding
Talk
Cultivar differences in plant
transpiration rate at high relative air
humidity are not related to genotypic
variation in stomatal responsiveness
*, Katrine H. Kjær1, Dimitrios Fanourakis2 and Carl-Otto Ottosen1
1
AArhus University, Department of Food Science, Kirstinebjergvej 10, DK-5792 Årslev, Denmark
Plant Sciences (IBG-2), Forschungszentrum Jülich, D-52425 Jülich, Germany
* Corresponding author: [email protected]
1
2
Plants grown at high relative air humidity (RH) often show disturbed water relations due to less
responsive stomata. The attenuation of stomatal responsiveness, as a result of high RH during leaf
expansion, depends on the cultivar of pot roses. We hypothesized that tolerant cultivars to high RH
experience less decline in transpiration by high RH, and that the variation in transpiration rate reflect
by differences in leaf temperature (Tleaf). Plant biomass, the stomatal responsiveness to desiccation,
together with transpiration and leaf temperature at growth conditions were analyzed in four pot rose
cultivars grown at moderate (60%) or high (85%) RH. Plants grown at high RH had a larger (9%)
leaf area and transpired less (50%) during the light period. At night time, transpiration decreased
(%) by high RH in one cultivar. Within the cultivars, Tleaf was generally higher (1.53.7 °C) at high as
compared to moderate RH. Following desiccation, leaf weight loss varied (866%) in high RH-grown
plants, indicating a wide variation in RH tolerance. High RH mainly decreased water loss during the
light period, though tolerant cultivars experienced the same decrease in transpiration as sensitive
ones. Cultivars with more responsive stomata transpired less at moderate RH, whereas stomatal
responsiveness was not related to transpiration at high RH. Consequently, plant water loss can be
used as a screening criterion for enhanced stomatal responsiveness only at moderate RH conditions,
while genotypic variation in plant water loss could not be detected by Tleaf differences.
7
Session 2: Genetics and Breeding
Talk
THE CONTINUOUS FLOWERING GENE IN ROSE IS
A FLORAL INHIBITOR
Hibrand-Saint Oyant, Laurence, Marie Randoux, Julien Jeauffre, Tatiana Thouroude, Sandrine Pierre,
Marie-José Jammes, Jean-Paul Reynoird and Fabrice Foucher
INRA, Institut de Recherche en Horticulture et Semences (INRA, Agrocampus-Ouest, Université
d’Angers), SFR 4207 QUASAV, BP 60057, 49071 Beaucouzé Cedex, France
1
In horticulture, control of flowering is an important issue. Rose presents different modes of blooming.
Once-flowering (OF) roses bloom only once a year, whereas recurrent roses have the ability to flower
several times a year. By genetic approach, we have identified that RoKSN, a rose homologue of the
TERMINAL FLOWER 1 (TFL1) floral repressor, is a regulator of continuous flowering (Iwata et al.,
2012). Due to a mutation, continuous-flowering (CF) roses do not accumulate the repressor and
flower continuously. Our objective was to provide functional evidence for RoKSN being a floral
repressor.
In order to functionally validate the gene, we performed complementation of tfl1-11 mutants of
Arabidopsis thaliana, and we over-expressed RoKSNin a CF rose. Transgenic roses were regenerated
and floral behaviour was tested under greenhouse conditions.
In Arabidopsis thaliana, the tfl1 mutants are early flowering and present determinate inflorescence
with a terminal flower. RoKSN over-expression complemented the mutation with suppression of the
late flowering and restoration of the indeterminate inflorescence. Three phenotypic classes were
found: (i) plants that behaved as the WT plants, (ii) plants with a late flowering and some floral
defects and (iii) non flowering plants.
In rose, 6 plants over-expressing RoKSN (OE-RoKSN) and two control plants (transformed with an
empty vector) were regenerated from a CF rose. By Southern blot, the 6 plants were shown to be
independent. After acclimation in greenhouse, whereas control plants presented rapidly terminal
flowers, the 6 OE-RoKSN plants remained vegetative with an indeterminate growth. The OE-RoKSN
plants accumulated RoKSNtranscripts (q-PCR analysis) and RoKSN proteins (Western blot analysis).
We have clearly demonstrated that RoKSN is a floral inhibitor in Arabdidosis thaliana and in rose.
These results opened interesting ways to control flowering in rose. We are presently developing
protocols to transformed once-flowering roses in order to extinguish the floral repressor and see the
effect on flowering.
8
Session 2: Genetics and Breeding
Talk
GENE-SPECIFIC KNOCKDOWN SYSTEM USING
AN APPLE LATENT SPHERICAL VIRUS VECTOR IN
ROSE
Shungo Otagaki, Hiroaki Ito, Masaki Ochiai, Katsuhiro Shiratake and Shogo Matsumoto
Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya 464-8601, Japan
Research objective: Plant virus infection activates an RNA-mediated defense response that targets
viral RNAs in a nucleotide sequence specific manner. When viruses are designed to carry a foreign
sequence homologous to the host gene, the process can also be targeted against the corresponding
endogenous mRNA. This phenomenon is known as virus-induced gene silencing (VIGS). Recent
studies have shown that VIGS is a particularly useful reverse genetic tool for plants that are difficult
to transform. In this study, we present a first VIGS system in rose using Apple latent spherical virus
(ALSV) vector.
Materials and methods: We cloned a partial phytoene desaturase (PDS) coding sequence from Rosa
multiflora and inserted 162 bp of the PDS gene fragment into an ALSV-RNA2 vector. The construct
was co-inoculated with the infectious clone of ALSV-RNA1 into Chenopodium quinoa by mechanical
inoculation. The resulting recombinant virus was designated ALSV_mul PDS. Total RNAs were
extracted from ALSV_mul PDS infected C. quinoa leaves and then inoculated to the seedlings of R.
rugosa and R. multiflora by particle bombardment. Successful induction of VIGS was monitored by
the phenotypic changes and semi-quantitative PCR of the PDS gene.
Results and conclusions: We obtained four R. rugosa plants and one R. multiflora plant showing
a photo-bleached phenotype. Silencing phenotypes in these roses were mainly observed on the
upper leaves of primary shoots. Interestingly, one ALSV_mul PDS-infected R. rugosa plant showed a
completely photo-bleached secondary shoot and this phenotype persisted for more than 1 year. No
obvious symptoms or leaf color change was observed in the roses infected with a wild-type ALSV.
Semi-quantitative PCR showed that the expression level of the PDS gene was drastically decreased
in the photo-bleached leaves. From the results, ALSV vector-mediated VIGS system seemed useful for
determining the function of interesting genes in rose.
9
Session 2: Genetics and Breeding
Talk
USE OF DIGITAL GENE EXPRESSION TO
DISCRIMINATE GENE EXPRESSION IN DIFFERENT
DEVELOPMENTAL STAGE OF ROSA CHINENSIS
‘PALLIDA’
Huijun Yan1, Hao Zhang1, Shubing Li1, Junkai Yang2, Min Chen1, Hongying Jian1, Qigang Wang1,
Kaixue Tang1
Flower Research Institute of Yunnan Academy of Agricultural Sciences, Kunming, Yunnan 650205,
P. R. China
2
College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070,
China
1
Rosa chinensis ‘Pallida’ is one of the most important ancient cultivars originating in China, and
contributes the ‘tea scent’ trait, recurrent blooming, and double flower character to modern roses,
yet little is understood of the genes involved in scent metabolism and flower development. In the
study, we examined the global gene expression patterns of flower buds stage (FB), open flower stage
(OF) and senescent flowers stage (SF) in R. chinensis ‘Pallida’ using digital gene expression (DGE)
analysis. Almost 17 million clean tags were generated using Illumina technology to produce the
expression data for 90757 genes. We compared pairs of DGE profiles of the three libraries. Between
OF and FB libraries, a total of 7145 differentially expressed genes (DEGs) were detected with 4567
up-regulated and 2578 down-regulated genes. Between FB and SF libraries, a total of 3616 DEGs
were found with 1444 up-regulated genes and 2172 down-regulated genes. 3711 DEGs were upregulated and 1751 DEGs were down-regulated in OF compared with SF. The data suggested that
up-regulated DEGs in OF were more than that of FB and SF. To understand the functions of DEGs,
we mapped all the genes to terms in KEGG databases and compared with the whole transcriptome
background, and find that genes involved in metabolic, biosynthesis of secondary metabolites and
plant hormone signal transduction pathways were significantly enriched. To further evaluate the
DGE data, we analyzed the expression level of Orcinol o-methyltransferase (OOMT), Linalool synthase
(Lis), and flower time control protein, which are the genes highly expressed in OF. The results showed
that the genes were expressed at significantly lower levels in the FB and SF, and highly expressed in
OF. The results provide useful information on expressed genes in different developmental stage of
R. chinensis ‘Pallida’, contribute foundation for scent-related and flowering genes discovery in rose.
10
Session 2: Genetics and Breeding
Talk
EARTH-KIND® ROSE TRIALING: AN
INTERNATIONAL MODEL FOR THE IDENTIFICATION
OF REGIONALLY-ADAPTED LANDSCAPE ROSES
Zlesak, David C.1, Derald Harp2, Kathy Zuzek3, John Sloan4, Allen Owings5, and Steve George6
Univeristy of Wisconsin-River Falls, Department of Plant and Earth Science, River Falls, WI USA
Texas A&M University-Commerce, Department of Agricultural Sciences, Commerce, TX USA
3
University of Minnesota Extension, Farmington, MN USA
4
National Great Rivers Research and Education Center, East Alton, IL USA
5
LSU AgCenter, Hammond, LA USA
6
Texas A&M AgriLife Extension Service, Dallas, TX USA
1
2
Landscape roses are exceedingly popular worldwide and have become the flagship flowering shrub
trialed within the Earth-Kind® Environmental Landscape Management System (started at Texas A&M
University to support environmental stewardship). Multi-year, replicated Earth-Kind regional rose
trials are conducted under low-input conditions throughout a geographical region. Cultivars with
superior regional performance are designated as regional Earth-Kind roses providing consumers and
all sectors of the horticulture industry confidence that they can be successful with these cultivars
given basic plant care. Earth-Kind rose trials are now underway in multiple U.S. regions and in
five additional countries. Current trial parameters include: a RCB design with 8’ plant spacing,
incorporating 3-4” of compost preplant and maintaining 3-4” of organic mulch throughout the trials
for nutrition, planting own-root plants (to avoid graft incompatibilities), irrigating regularly during
the first year of establishment then reducing/eliminating irrigation later, and no pruning, pesticides
(unless in extreme circumstances that would otherwise jeopardize the trials) or traditional fertilizers.
Minor modifications are allowed based on regional needs (e.g. closer spacing and pruning allowed to
remove dead wood in colder regions). Plants are evaluated monthly beginning in year two through the
end of the 3 or 4 year study using a 10 point horticultural scale (includes pest tolerance and aesthetic
factors) as well as overall flower number and coverage. The strong experimental design allows for
the performance data to be published in peer-reviewed scientific literature. The trials also support
further studies surveying plants for additional, regionally important traits. Laboratory screens, such
as surveying for black spot resistance to known races, are conducted to help complement the field
trial data. Earth-Kind rose trialing provides a consistent and strong international rose trialing and
outreach model to identify and promote genetically superior cultivars for the benefit of regional
consumers, nursery professionals, landscapers, researchers, and breeders.
11
Session 3: Rose Hips
Talk
BEAUTY IS AS BEAUTY DOES
Hilde Nybom
Department of Plant Breeding – Balsgård, Swedish University of Agricultural Sciences, Fjälkestadsvägen 459, SE-29194 Kristianstad, Sweden
Dogroses, i.e. species belonging to Rosa sect. Caninae, grow wild along roadsides and in disturbed
areas in temperate regions of Europe and W Asia. A minimum of 50 dogrose taxa are usually
recognized, but many hundreds have been described. Most taxa are cross-compatible, and hybrids
often produce fertile offspring, making proper systematic treatment challenging, to say the least.
In mediaeval times, these humble but healthy cousins of the more spectacular ornamental roses,
were highly appreciated as medicinal plants. Nowadays, dogroses are sometimes cultivated or, more
commonly, harvested from wild or naturalized stands to produce juice, dessert soup, jelly and other
tasty products. Recently, dogrose fruits have however enjoyed renewed interest due to the very
high levels of antioxidant compounds, mainly polyphenols, but also carotenoids and vitamins B, C
and E. Extracts of rosehips have anti-inflammatory, anti-nociceptive, anti-diabetic, anti-obesity and
anti-mutagenic effects, and are used to treat, e.g., osteoarthritis and various stomach problems. The
seeds contain high contents of unsaturated fatty acids, which are used for production of oils for skin
treatment and as cosmetics. The amount and composition of bioactive compounds vary greatly with
genotype and environment. Breeding of improved dogrose genotypes or of intersectional hybrids
is, however quite a challenge due to the unique canina meiosis. Regardless of ploidy level (most
species are 5x, but some 4x and 6x taxa/forms also occur), only 7 bivalents are formed in the first
meiotic division. The remaining chromosomes occur exclusively as univalents and are not included
in viable pollen grains, which contain only the 7 divided bivalent chromosomes. All the univalents
are transmitted to one of the daughter cells in the female meiosis, and are eventually included in
the viable egg cells, which therefore contain 21, 28 or 35 chromosomes depending on ploidy levels.
Consequently, reciprocal crosses produce widely different offspring.
12
Session 3: Rose Hips
Talk
FACTORS AFFECTING THE GERMINATION OF
HYBRID ROSE ACHENES: A REVIEW
de Vries, D.P.1 and Dubois, Lidwien A.M.2
Foundation Sub Rosa, P.O. Box 4097, 6710 EB Ede, The Netherlands
Stichting Dienst Landbouwkundig Onderzoek - PPO/PRI, P.O.Box 16, 6700 AA Wageningen, The
Netherlands
1
2
The smooth germination of mature (Hybrid) rose achenes is hampered by (i) hardseededness (HS), (ii)
primary dormancy (PD) and (iii) germination polymorphism (GP). HS is owing to the hard pericarp.
PD is, in principle, a natural phenomenon that protects the seed from precocious germination. For
germination, PD must be lifted by after-ripening. When the process of after-ripening is interrupted
seeds become secondary dormant (SD). When SD, both seeds and naked embryos are highly recalcitrant
to germination. After-ripening of (Hybrid) rose seeds consists of two steps (i) softening the pericarp,
(ii) hydrolysing and/or antagonizing the inhibitor ABA, present in the testa and/or embryo. The ratio
ABA: GA (present or newly synthesized) controls readiness for germination. Imbibing previously coldstratified Hybrid seeds in GA3, enhanced the even emergence and germination percentage. Likely
owing to their unique descent, Hybrid roses bear achenes that are less recalcitrant to breaking PD
than those of many species roses. Different levels of PD, occurring between the species roses, can
be traced back to adaptation to their natural habitat. Comparing the after-ripening requirements
of Hybrid seeds with those of species roses is a false position. GP is caused by varying temperatures
during fruit ripening, is promoted by prolonged pollination periods, effects variation in dormancy
and as such contributes to uneven germination of Hybrid seeds. PD of Hybrid seeds is deeper as
mean ambient temperatures during the last 30 days of fruit ripening were lower. Hence, lower
temperatures in autumn, as commonly set in glasshouses, cause deeper PD. Rose seeds may germinate
without any after-ripening when harvested unripe. Each year, breeders should compose the “best
protocol” for cold stratification/ germination of (Hybrid) rose seeds from the elements: (i) the period
of pollination, (ii) the temperatures during 30 days prior to hip ripening, (iii) the storage of achenes
between extraction from the hip and sowing, (iv) the intended warm period prior to stratification and
(v) the temperatures set for stratification.
13
Session 3: Rose Hips
Talk
LEAF AND FLOWER PHENOLIC PROFILES OF
INDIGENOUS SLOVENIAN ROSE SPECIES
Valentina Schmitzer1, Franci Stampar1
University of Ljubljana, Biotechnical faculty, Department of Agronomy, Jamnikarjeva 101, 1000,
Ljubljana, Slovenia
1
Wild rose species and ecotypes represent a vast genetic pool for further breeding and selection in
particular striving to improve characteristics such as resistance to the major pathogens and stress
tolerance of modern cultivars or enhancing new ornamental values (for example scented leaves).
In Slovenia, several autochthonous roses have been identified and among these, Rosa canina, R.
rubiginosa and R. glauca are prevalent. Wild rose species have been planted in collections at several
locations in central and southwest part of the country and thus represent an ideal research potential
for metabolic studies of different plant parts. The study aims to morphologically describe selected
wild rose species and identify individual compounds from different phenolic classes in rose leaves and
flowers. Anthocyanins and flavonols were quantified and correlated with visual characteristics of rose
flowers, colorimetrically evaluated with a portable colorimeter. Leaf metabolic profiles were studied
in detail and the content level of hydroxycinnamic acid derivatives, flavan-3-ols, dihydrochalcones
and flavonols are discussed from the viewpoint of resistance. All biochemical analyses were performed
with the use of HPLC and compounds further identified with HPLC-MS-ESI. The phenolic profile of
analyzed indigenous rose species differed significantly among species and plant organs: a great
variety of compounds was determined in leaves, whereas flowers prevalently accumulated pigments
and flavonols. Among anthocyanins glycosides of cyanidin and pelargonidin were confirmed in red
and pink flowers and among flavonols, quercetin and kaempferol glycosides have been identified. The
metabolic screening of indigenous rose species provides information on potentially interesting plants
for resistance enhancement via phenolic accumulation.
14
Session 4: Rose Genomics
Talk
Towards the rose genome sequence and
its use in research and breeding
Current “Rose Genome Sequence Initiative” members:
INRA, IRHS, Angers, France (Fabrice Foucher)/ Agrocampus Ouest, IRHS, Angers, France (Soulaiman Sakr)/ Wageningen
UR Plant Breeding, The Netherlands (Marinus JM Smulders)/ Leibniz University Hannover, Germany (Thomas Debener)/ ILVO-Plant, Belgium (Jan De Riek)/ IFAPA, Córdoba, Spain (Ana Torres)/ IFAPA, Málaga, Spain (Iraida Amaya)/ T.
Millán, University of Córdoba, Spain (A Cabrera)/ Hebrew University of Jerusalem, Israël (Dani Zamir)/ East Malling
Research, UK (Dan Sargent)/ SLU Alnarp, Sweden (Hilde Nybom)/ AgroBioinstitute and Joint Genomic Centre, Bulgaria
(A. Attanasov)/ University of Minessota, USA (Stan Hokanson)/ Texas A&M University, USA (Dave Byrne)/ North Carolina State University (NCSU), USA (Bryon Sosinski)/ Washington State University (WSU), USA (Dorrie Main)/ Montreal
Botanical Garden, Canada (Anne Bruneau)/ Agricultural Research Council, Onderstepoort, South Africa (Jasper Rees)/
Nagoya University, Japan (Shogo Matsumoto)/ Chubu University, Japan (Kunio Yamada)/ CEA, Genoscope, Evry (Patrick
Wincker)/ INRA, LIPM, Toulouse (Jérome Gouzy)/ INRA-ENS, RDP, Lyon, France (Mohammed Bendahmane)
Rose is one of the most economically important ornamental crops worldwide. Rosa sp. can become
a model for woody ornamentals. Its genome size is relatively small (500 Mb), its genetic history
with ploïdy events is well documented, and rose has a short life for a woody plant. Furthermore,
different tools are available, including transcriptomic tools, genetic maps and genetic transformation
protocols. Rose represents an original model for studying some ornamental traits that can not be
addressed in other model plant species such as Arabidopsis. Some of these traits, such recurrent
blooming, flower morphogenesis or scent production and emission, are of economic interest.
Different groups involved in rose genetics and genomics gathered to form the ‘Rose Genome Sequence
Initiative’. Our objective is to obtain a high quality rose genome sequence of the diploid R. chinensis
‘Old Blush’. One important issue is the high level of heterozygosity of roses. To tackle this issue,
different strategies are proposed: production of an haploid and development a high density genetic
map to anchor the genome.. This genetic map will be developed from a cross between ‘Old Blush’
and R. wichurana. The genotype R. chinensis ‘Old Blush’ will be sequenced using NGS technologies.
The data will be assembled and arranged using the high density map. In order to increase ESTs and
to facilitate genome annotation, we have recently produced ESTs from various tissues of ‘Old Blush’
under different conditions. Digital expression (RNA Seq) was obtained from the different tissues and
data are available on the following web site (https://iant.toulouse.inra.fr/plants/rosa/FATAL/).
The rose genome sequence will be a great step to help identifying the molecular basis of ornamental
traits and also to study genetic diversity and genome evolution in the genus Rosa and in the Rosaceae
family.
15
Session 4: Rose Genomics
Talk
Genetic analysis of tetraploid F1 rose
populations based on the Rose Axiom SNP
array
M.J.M. Smulders1, Roeland E. Voorrips1, G. Danny Esselink1, Wendy P.C. van ‘t Westende1, Mirjana
Vukosavljev1, Carole F.S. Koning-Boucoiran1, W. Eric van de Weg1, Paul Arens1, Dietmar Schulz2,
Thomas Debener2, L. Bellon3, M. Mittmann3, A. Pirani3, T. Webster3, F. Brew4, Peter Cox5, Chris Maliepaard1
Wageningen UR Plant Breeding, Wageningen, The Netherlands
Institute for Plant Genetics, Leibnitz Universität Hannover, Germany
3
Affymetrix Inc., Santa Clara 95051 California, USA
4
Affymetrix UK Ltd, Wooburn Green, High Wycombe, HP10 OHH, UK
5
Roath, Eindhoven, The Netherlands
1
2
Many ornamentals are polyploid. Genetic analysis using molecular markers is well established for
diploid crops but until recently the tools for mapping and QTL analysis in tetraploids were not readily
available. However, this is rapidly changing: next-generation sequencing enables the identification
of large numbers of SNPs; array hybridisation generates large SNP marker data sets; and software for
dosage scoring (fitTetra, Voorrips et al., BMC Bioinform 12:172, 2011) allows efficient assignment of
the tetraploid SNP genotypes of individuals.
We have generated a SNP data set based on transcriptome sequencing of the flowers and leaves of the
parents of two segregating F1 populations and several garden rose cultivars. This set formed the basis
of the WagRhSNP Axiom array, which contains 68,893 SNPs probed from both directions, totalling
137,786 probes. This array was used to genotype the two F1 populations (a cut rose population
(Koning-Boucoiran et al., TAG 125:591, 2012) and a garden rose population) and a diversity panel of
garden roses. Using the tetraploid SNP dosage scores we will develop linkage maps, and subsequently
work on QTL mapping of traits including frost tolerance for garden roses and flower colour, production
traits and powdery mildew resistance for cut roses. The array is not proprietary but can be used by
all research groups working in rose.
In preparation for this work we have developed software (PedigreeSim, Voorrips and Maliepaard,
BMC Bioinform 13:248, 2012) for simulating marker data in a tetraploid crop, which models both
bivalent and quadrivalent formation during meiosis and therefore also simulates double reduction
(the situation where a gamete receives two copies of the same chromosome segment, which is only
possible in auto-polyploids).
16
Session 4: Rose Genomics
Talk
Borrowing from neighbors: leveraging
genome sequence of strawberry, apple,
and peach to study disease resistance in
rose
James M. Bradeen1, David Zlesak2 and Stan C. Hokanson3
University of Minnesota, Department of Plant Pathology, St. Paul, MN USA
David Zlesak — University of Wisconsin-River Falls, Department of Plant and Earth Science, River
Falls, WI USA
3
Stan C. Hokanson — University of Minnesota, Department ofHorticultural Science, St. Paul, MN
USA
1
2
Roses (genus Rosa) are a member of the Rosaceae family, which also includes many important
fruit crops including apple (genus Malus), peach (genus Prunus), and strawberry (genus Fragaria).
Phylogenetic relationships have been extensively studied in the Rosaceae and indicate that the rose
and the strawberry share a close evolutionary relationship while apple and peach are more distantly
related. Importantly, the genomes of apple, peach, and a diploid wild strawberry (F. vesca) have been
sequenced. Regardless of the plant species of origin or the pathogen against which they are effective,
most plant disease resistance (R) genes encode proteins of conserved structure. One conserved
protein domain, the nucleotide binding site (NBS) has been especially useful for comparative, crossspecies analyses. We have completed analyses of NBS R-genes found in the genome sequences of
apple, peach, and strawberry. Our results indicate tha tmost R-gene lineages are of ancient origin
that predated the evolutionary split of these species. But other R-gene lineages appear to be more
recent and are shared between apple and peach, but missing in strawberry. Our findings confirm
that patterns of R-gene representation across species are consistent with phylogenetic relationships.
Conversely, we hypothesize that phylogenetic relationships between plant species can be leveraged
to study R-genes in species for which complete genome sequences are not yet available. To test
this hypothesis, we are now using our comparative R-gene framework to devise strategies for deep
sequencing of the R-gene content of rose. Based on observations in strawberry, we anticipate
that the rose genome will contain a few hundred NBS R-genes. Our results will provide the first
comprehensive view of R-gene lineages found in the rose genome and are expected to provide allelelevel resolution. This research project is also designed to build upon efforts to develop markers linked
to the rose black spot R-geneRdr3.
17
Session 4: Rose Genomics
Talk
MOLECULAR GENETIC STUDIES ON THE
CONTINOUS-FLOWERING ROSES THAT ARE NOT
ORIGINATED FROM ROSA CHINENSIS
Takanori Horibe1,Kawamura, Koji2, Kunio Yamada1, SyungoOtagaki3, and Shogo Matsumoto3
Department of Environmental Biology, College of Bioscience and Biotechnology, Chubu University,
1200 Matsumoto-cho, Kasugai, Aichi, 487-8501, JAPAN
2
Department of Environmental Engineering, Osaka Institute of Technology, 5-16-1, Ohmiya Asahi-ku, Osaka, 535-8585, JAPAN
3
Laboratory of Horticultural Science, Graduate School of Bioagricultural Sciences, Nagoya University, Furoh-cho, Chikusa, Nagoya, 464-8601, JAPAN
1
The majority of modern rose cultivars are able to flower during all favorable seasons (i.e., continuousflowering). The continuous-flowering character of modern roses was considered to originate from
a Chinese rose, Rosa chinensis in the 18th century. The single origin of continuous-flowering roses
may result in a narrow genetic diversity of modern roses and limit the creation of new continuousflowering cultivars by cross breeding. We aimed to discover roses that can flower continuously but
do not originate from R. chinensis.
Recent studies have identified a molecular basis of the continuous-flowering character in R. chinensis
and and demonstrated that the rose has a mutation of KSNgene, which acts as a repressor of floral
induction. The insertion of a copia-like retrotransposon (9kbp) into the KSN blocks the transcription
of the gene in R. chinensis and allows these genotypes to bloom continuously. The majority of modern
roses are expected to inherit the same mutated allele of KSN (named ksnt) from R. chinensis. We thus
searched continuous-flowering roses “without” the allele ksnt by a genetic screening in order to
discover the continuous-flowering roses that were “not” originated from R. chinensis.
The KSN genotyping of a large number of continuous-flowering roses (>100) identified several
cultivars without the allele ksnt from R. chinensis. All these cultivars were derived from a few wild
ancestral species with the continuous-flowering character, R. moschata, R. rugosa, or R. bracteata. We
proposed that these species are nice genetic resources to expand the genetic background of modern
continuous-flowering roses. We also did further studies on one of the species, R. rugosa to uncover
the molecular basis of its flowering behavior. The seasonal expression of KSN gene was clearly linked
with the alternation of flowering/vegetative stages of shoot apices in R. rugosa, indicating that
continuous-flowering behavior of the species might be also controlled by the KSN.
18
Session 4: Rose Genomics
Talk
Integration of genetic and physical maps
for Rosa wichurana using Tyramide-FISH
Ilya Kirov1,2, Katrijn Van Laere2, Ludmila Khrustalva1, Ellen De Keyser2, Jan De Riek2
Centre of Molecular Biotechnology, Russian State Agricultural University, Moscow Agricultural
Academy named by K.A. Timiryazev, Timiryazev Street 49, 127550 Moscow, Russia
2
Institute for Agricultural and Fisheries Research (ILVO), Plant Sciences Unit, Applied Genetics and
Breeding, Caritasstraat 21, 9090 Melle, Belgium
1
The integration of genetic and chromosomal maps is still very challenging for most plant species.
Physical maps linked to genetically mapped markers are very valuable for plant breeding, especially
when they are integrated with genomic sequence data. The most promising approach to integrate
physical and genetic maps is to directly visualize DNA fragments, derived from genetically mapped
markers, using FISH (Fluorescent In Situ Hybridization). However, because of the sometimes short
length of the DNA fragments, conventional FISH is not always sensitive enough to visualize them.
Therefore, Tyramide-FISH, combining the advantage of an enzymatic and a fluorescence-based
detection, has great potential for physical mapping of short DNA fragments.
In previous research a genetic map based on a diploid population R.wichurana x R. ‘Yesterday’ was
constructed (Moghaddam et al. 2012). As a proof of concept, we physically map Phenylalanine
ammonia-lyase (1700bp), Orcinol O-methyltransferase (1100bp) and Pyrroline-5-carboxylate
synthase (1700bp) using optimized Tyramide-FISH for Rosa wichurana. Optimization of the Tyramide
FISH protocol was necessary as R. wichurana is a difficult species for cytogenetics because its tiny
roots, hard cell wall, low mitotic index, very small chromosomes (size between 0.86 and 1.52 µm)
and high polysaccharide content in the tissues. Also EST-SNP markers were designed for these three
genes and detected by “High Resolution Melting”; their genetic mapping has been performed. The
HRM markers were incorporated into the known genetic maps of Rosa (Moghaddam et al. 2012) and
obtained data allowed us to integrate the genetic and physical maps of chromosomes 1, 4 and 7 of
Rosa wichurana.
19
Session 4: Rose Genomics
Talk
RNA-SEQ AND DIGITAL GENE EXPRESSION
ANALYSIS OF FLOWER DEVELOPMENT IN
RECURRENT AND NON-RECURRENT ROSES
LiJuan Lian, Qianqian Jia, QingLin Liu
Department of Ornamental Horticulture and Landscape Architecture, China Agricultural University,
Beijing, 100193, China
Recurrent flowering habit of modern roses is a very important horticultural trait, yet little is understood
about its genes involved in the regulatory network. The transcriptome of Rosa multiflora ‘Albo-plena’
(non-recurrent flowering) and R. chinensis ‘Old Blush’ (recurrent flowering) were sequenced using
short read sequencing technology (Illumina HiSeqTM2000). In a single run, we obtained 6.96 G and
10.06 G clean reads, respectively. These clean reads were assembled into 158 717 unique sequences
(mean size=921 bp). These assembled sequences were annotated with such bioinformatics methods
as gene descriptions, gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG), coding
sequence prediction and analysis of differential gene expression. Based on sequence homology, 27 714
sequences with GO annotations were classified into 48 categories, belong to three GO terms including
Biological Process, Cellular Component and Molecular Function, in which the three most common
categories are cellular process, cell part and binding respectively. Meanwhile, we investigated the
transcriptome changes on three different flower developmental stages including flower bud stage
(5 April), flowering stage (1 June), re-flowering stage (4 September) using a digital gene expression
(DGE) profiling system. The results showed that there are more than 5 500 differentially expressed
genes between R. multiflora and R. Chinensis during flower development. Among them, about 2/3 of
the genes are expressed at the flower bud stage. There are more than 20 enriched pathways such as
Glycolysis / Gluconeogenesis, Citrate cycle (TCA cycle) and Pentose phosphate, and so on. The DGE
results of RTFL1c (Rosa Teminal Flower 1c) and RLFY (Rosa Leafy) were consistent with our precious
qRT-PCR results. In addition, the expression levels of SPINDLY (SPY) and GIGANTEA(GI) related to
gibberellin signal transduction presented obvious difference in flower development stages between
recurrent and non-recurrent roses.
20
Session 4: Rose Genomics
Talk
Association mapping of phenotypic traits
in roses
Schulz, Dietmar Frank1, Rena Schott1, Ana Priscilla Montenegro1, Dorothee Klostermann1, Marinus J.
M. Smulders2, Roeland E. Voorrips2,Marcus Linde1, Burkhard Spellerberg3 and Thomas Debener1
Gottfried Wilhelm Leibniz Universität Hannover, Department of Molecular Plant Breeding, Herrenhäuser Str. 2, 30419 Hannover, Germany
2
Wageningen UR Plant Breeding, Wageningen, The Netherlands
3
Bundessortenamt, Osterfelddamm 80, 30627 Hannover, Germany
1
In this work we focus on the association of resistance to several important fungal rose pathogens
and different phenotypic traits like scent, anthocyanin and carotinoid content and plant architecture
to molecular markers. A panel of 96 mostly tetraploid rose cultivars were grown in five different field
locations in the north, middle and western part of Germany. Additionally, one trial was conducted
under controlled conditions in a greenhouse in Hannover (Bundessortenamt). The studies included
the generation of AFLP- and SSR- markers for genome wide association associations and markers
for candidate gene associations. Prior to the calculation of associatins molecular markers were used
to estimate population structure (Structure 2.3.4) and genetic diversity of the association panel..
Associations between markers and phenotypic traits were calculated with Tassel 4.1.10. A testcross
progeny was developed to verify the identified marker-trait associations.
21
Session 5: Postharvest Physiology
Talk
Sustainable solutions to preserve
rose flower quality and to facilitate
long-term transport in refrigerated
containers
Ernst J. Woltering1,2, Harmannus Harkema1, Henry A. M. Boerrigter1
Food and Biobased Research, Wageningen University and Research Centre, P.O. Box 17, 6700 AA
Wageningen, The Netherlands
2
Horticultural Supply Chains Group, Wageningen University, P.O. Box 630, 6700 AP Wageningen,
The Netherlands
1
Long-term shipment of flowers in Reefer containers as a replacement of airfreight is the method
of choice with respect to saving on transport costs and decreasing the carbon footprint. Optimal
storage conditions have been developed over the years for a variety of flowers and cultivars with
varying degree of success. In roses, the protocols consist of strict recommendations concerning the
cultivars to be used, the flower development stage at harvest, the type of packaging, the post-harvest
application of fungicides and the use of pretreatment, transport and (when stored dry) rehydration
solutions.
One of the issues hampering long-time storage is the development of Botrytis cinerea. Apart from
fungicide applications during growth, postharvest treatments with regular fungicides may be applied
to tackle this problem during long-term storage. Bacterial growth in the xylem vessels during storage
is another factor causing quality loss. Bacterial growth may lead to blockage of the xylem vessels
and may hamper proper flower development and cause premature flower wilting. Pretreatment,
transport and rehydration solutions therefore often contain biocides. Such compounds often contain
heavy metals and are less desirable from an environmental point of view. Ethylene is another factor
that may cause quality problem during long-term transport. Ethylene may disturb proper flower
opening and may cause leaf abscission in some cultivars. Therefore, pretreatments with silver
thiosulphate solutions are applied toprevent these symptoms but this treatment is less desirable
from an environmental point of view.
We investigated several “sustainable” alternatives to the use of regular fungicides and heavy metalcontaining treatment solutions. The most promising technologies to establish a sustainable flower
distribution chain are discussed.
22
Session 5: Postharvest Physiology
Talk
Morphorogy, endogenous
phytohormones, and related proteins of
Bent Peduncle Phenomenon in Cut Rose
plants
Wan Soon Kim1*, Ji Hyun Seo1, Seul Ki Lee1, and Won Hee Kim2
Department of Environmental Horticulture, University of Seoul, Seoul 130-743, Korea
Floriculture Research Division, National Institute of Horticultural & Herbal Science, Suwon 441440, Korea
1
2
The bent peduncle phenomenon (BPP) known as a kind of physiological disorder in cut rose plants
causes reducing the cut flower yields in rose greenhouse production. In fact, it is not easy to avoid
the BPP in the field as the mechanisms are still unknown. To address this challenge, this study
was conducted to investigate the characteristics of BPP occurrence and related morphorogical
characteristics including floral morphology, shoot growth, and endogenous phytohormone level of
BPP shoots in cut rose plants (Rosa hybrida L. ‘Beast’). Morphological observation confirmed the fact
that BPP was accompanied by the early formation of phyllody of a specific sepal among five sepals,
which gave rise to peduncle bending by fasciation in the upper part of the peduncle.. Year-round BPP
frequency in ‘Beast’ cultivar was in the range of 5 to 20% with seasonal changes, increasing rapidly
with an average temperature rise in summer. Except bent peduncle, the BPP shoots appeared to grow
normally in terms of speed and size of floral development in comparison with normal flowers. However,
carbohydrate distribution to the flower in shoots affected by BPP was significantly reduced. The level
of endogenous IAA (3-Indolylacrylic acid) within the floral part in BPP shoots was maintained at high
levels during floral development, in contrast to normal shoots. GA3 contents were not significantly
different between normal and BPP shoots. Proteins related to BPP were detected by 2-dimensional
polyacrylamide gel electrophoresis (2D-PAGE). This study indicates that BPP is induced by a series of
factors: abnormally early formation of a specific sepal relative to high temperature, phyllody of the
sepal with fasciation, and continuous supply of endogenous IAA by phyllody.
23
Session 6: Secondary Metabolites and Biochemistry
Talk
SWEET SMELL OF ROSE: NAVIGATING THE
NETWORK OF FLORAL SCENT PRODUCTION
Vainstein, Alexander
The Hebrew University of Jerusalem, The Robert H. Smith Institute of Plant Sciences and Genetics
in Agriculture, P.O.Box 12, Rehovot 76100, Israel
Research into flower scent, that of roses in particular, focused for many years on its chemical
elucidation, while the biochemistry of fragrance production and the mechanism of emission
remained sketchy at best. To enable the exploration of metabolic fluxes and the identification of
genes that perform key functions in the transcriptional regulation of floral volatile production,
we employed tobacco rattle virus (TRV)-based gene silencing (VIGS), virus-aided gene expression
(VAGE) and promoter-activation screens for large-scale identification of floral scent genes. A central
regulatory role for the identified myb transcription factors, named EMISSION OF BENZENOIDS (EOBs),
in the biosynthetic cascade leading to the production/emission of unique floral scent bouquets
was revealed by gene-expression profiling, the levels of phenylpropanoid-pathway intermediates,
and the coordinated wide-ranging effect of the identified myb factors on the production of floral
volatiles but not on that of anthocyanin. To evaluate whether scent production in rose flowers, which
originates from diverse metabolic pathways (e.g. phenylpropanoids and isoprenoids) can be affected
by transcriptional regulators, Arabidopsis PRODUCTION OF ANTHOCYANIN PIGMENT1 (PAP1), which
had been shown to exert broad activation of the phenylpropanoid pathway, was introduced into Rosa
hybrida. Olfactory assay revealed that bees and humans could discriminate between the floral scents
of PAP1-transgenic and control flowers. The increase in volatile production in PAP1 transgenes was
not caused solely by transcriptional activation of their respective biosynthetic genes, but probably
also resulted from enhanced metabolic flux in both the phenylpropanoid and isoprenoid pathways.
Integrating knowledge of the structural and regulatory genes involved in floral scent production and
of the mechanism regulating metabolic flow with the ability to modify genome sequences should
enable rational manipulation of the pathway toward enhanced production of floral scent compounds.
24
Session 6: Secondary Metabolites and Biochemistry
Talk
ROSE SCENT 2-PHENYLETHANOL WAS
PRODUCED VIA TWO BIOSYNTHETIC PATHWAYS
IN RESPONSE TO SEASONAL CHANGE IN
ENVIRONMENT
Momoyo Kanda1, Hiroshi Hirata2, Toshiyuki Ohnishi1 and Naoharu Watanabe2,3
Shizuoka University, Graduate School of Agriculture, 836 Ohya, Suruga-Ku, Shizuoka 422-8529,
Japan
2
Shizuoka University, Graduate School of Science and Technology (GSST), 836 Ohya, Suruga-Ku,
Shizuoka 422-8529, Japan
3
Shizuoka University, Graduate School of Engineering, 3-5-1 Johoku, Naka-Ku, Hamamatsu 4328561, Japan
1
2-Phenylethanol (2PE) is one of the prominent scent compounds produced by Damask roses such as
Rosa x damascena, R. ‘Hoh-Jun’, and R. ‘Yves Piaget’. We describe a new season-specific biosynthetic
pathway for the scent compound 2-phenylethanol (2PE) in rose flowers. Over a couple of years
in Shizuoka, Japan, we conducted L-[2H8]phenylalanine (L-[2H8]Phe) feeding experiments with
protoplasts made from flowers of R. ‘Yves Piaget’. We already reported the biosynthetic pathway
for 2PE yielding [2H8]-2PE from L-[2H8]Phe. In this pathway two enzymes, rose aromatic amino acid
decarboxylase: AADC and phenylacetaldehyde reductase:PAR were involved. Contrarily [2H7]-2PE as
a main isotopologue was synthesized by the protoplasts during the period from June to October
in both years, whereas [2H8]-2PE was synthesized as the main product during the rest of the year.
We hypothesize that rose flowers have two different 2PE biosynthetic pathways from L-Phe, and
that each of these pathways contributes differently to 2PE production. Here, we first report the
identification and biochemical characterization of rose aromatic amino acid amino transferase:
RyAAAT and rose α -keto-decaroboxylase:RyKDC for [2H7]-2PE production in planta, demonstrate
season-specific 2PE biosynthetic pathway. Transient RNAi suppression experiments targeting RyPDC
decreased 2PE production from June to October, whereas 2PE production was not affected from
November to April. These results underscored the involvement of two biosynthetic pathway of 2PE in
rose and rose utilizes the novel pathway with season-specific (June-October) to produce 2PE
25
Session 7: Growth Physiology and Production/Propagation
Talk
A recent history of changing trends
in U.S. garden rose plant sales and
production methods
Pemberton, H. Brent1 and John F. Karlik2
Texas A&M AgriLife Research and Extension Center, P.O. Box 200, Overton, TX 75684, USA
University of California Cooperative Extension, 1031 S. Mt. Vernon Ave., Bakersfield, CA 93307,
USA
1
2
Nursery crop sales have steadily increased in the U.S. for the last several decades. Despite this fact,
the number and value of garden rose plants sold in the U.S. has been decreasing since the 1980s
and in particular since 2000. Due to the introduction of landscape rose cultivars during the last two
decades, however, sales of garden roses have stabilized in more recent years. The development of
disease tolerant landscape type rose series such as Meidiland®, Simplicity®, Knock Out® and others
like them have grown in popularity with U.S. landscapers and the consuming public. Concomitant
with the ascent of landscape rose sales has been a change in the method of production. The use of
rooted liners for own-root container nursery and field production of garden rose plants has risen
dramatically as the production of field-grown grafted plants has fallen. Characteristics of landscape
garden roses and changes in production methods for this type of rose vs. a field-grown grafted rose
plant will be discussed.
26
Session 7: Growth Physiology and Production/Propagation
Talk
GROWERS’ AND CONSUMERS’ KNOWLEDGE,
ATTITUDES AND OPINIONS REGARDING ROSES
AVAILABLE FOR PURCHASE
Tina M. Waliczek1, Byrne, Dave2, and Don Holeman3
Texas State University, Department of Agriculture, Texas State University, San Marcos, TX 78666,
United States
2
Texas A&M University, Department of Horticultural Sciences, College Station, TX, 77843, United
States
3
Rose Hybridizer Association, Enfield, Connecticut
1
Rose breeders historically have bred plants based on what they have deemed important, or instead
on variables growers have noted as important for the success in growing the crop. However, enduser opinions have not been formally considered. The purpose of this study was to investigate both
the growers’ and the consumers’ knowledge, attitudes, needs and opinions of roses available on the
market and their experiences in growing them in the nursery and home landscape. A survey tool was
developed to measure what attributes consumers were looking for in purchasing and growing rose
plants, their knowledge of diseases and pests and their hopes for new plants coming to market. The
survey was developed using Limesurvey web-based survey design tool. A link was sent to horticultural
group mailing lists includingthe Rose Hybridizer Association, the American Rose Society, the Texas
Landscape and Nursery Association, Master Gardeners among others as well as distributed through
personal email lists, Facebook, and a news release from Texas A&M University. The survey was posted
for 10 weeks. It included approximately 70 questions and took 30 minutes or more to complete. Over
1800 responses were received from rose growers and consumers nationwide. Data were automatically
downloaded into Microsoft Excel. Data formatting was conducted in Excel and transferred to PASW/
SPSS. Descriptive statistics determined trends among overall growers and consumers. Comparisons
of growers versus consumers, as well as demographic groups were made using ANOVA tests. Results
from the study will help rose breeders determine the needs of both growers and consumers and the
direction of future breeding projects.
27
Session 7: Growth Physiology and Production/Propagation
Talk
EFFECT OF FLOWER BUD REMOVAL, STEM AGE,
AND LOCATION IN CANOPY OF THE STOCK
PLANTON CUTTING PROPAGATION OF ROSE
CULTIVARS
Hwang, Chul Hwan1,Ji Eun Park1,Moon Sook Son2 and Byoung Ryong Jeong1,2,3
Department of Horticulture, Division of Applied Life Science (BK21 Program), Graduate School of
Gyeongsang National University, Jinju 660-701, Republic of Korea
2
Institute of Agriculture & Life Science, Gyeongsang National University, Jinju 660-701, Republic of
Korea
3
Research Institute of Life Science, Gyeongsang National University, Jinju 660-701, Republic of
Korea
Corresponding author: T. +82-55-772-1913; F. +82-55-757-7542; [email protected]
1
Experiments were carried out to investigate rooting and growth of rose cuttings as affected by flower
bud removal from the flowering stem and the age of the flowering stem (i.e., flowering stage of
flower bud). In a second experiment, the effect of the location in canopy of the flowering stem was
investigated. Test plants used were Rosa hybrida ‘Sherbet’ and ‘Lover Letter’in the first experiment and
‘Pink Mimi’ and ‘Free Sun’ in the second experiment. Single node cuttings, each with a five leaflet leaf
and an anaxillary bud, were stuck into pre-wet rockwool cubes (5 cm x5 cm x5 cm, Grodan, Denmark)
to a depth of about 2cm. The results in the first experiment for ‘Sherbet’ showed no significant
differences among treatments. However, in the ‘Lover Letter’ rooting percentage, root length, no. of
roots, and root fresh and dry weights increased, while shoot length, and shoot fresh and dry weights
decreased as the flowering stage decreased. Flower bud removal from the flowering stem promoted
the overall shoot and root growth, except chlorophyll content. Both cultivars showed 100% rooting
percentage in the second experiment, and there were no significant differences in overall shoot and
root growth as affected by the treatment. Scanning electron microscopic examination of the stems
revealed that stems harvested from the center location of the canopy were thicker than those from
the side. The photosynthetic rate, transpiration rate, and stomatal conductance of the leaves grown
in the center of the canopy were lower than those grown in the side, and were also affected by the
cultivar. The results would be worth to consider for the management of rose stock plants for cutting
propagation.
This study was carried out with the support of “On-Site Cooperative Agriculture Research Project
(Project No. PJ906980062012)”, RDA, Republic of Korea.Chul Hwan Hwang and Ji Eun Park were
supported by a scholarship from the BK21 Program, the Ministry of Education and Human Resources
Development, Republic of Korea.
28
Session 7: Growth Physiology and Production/Propagation
Talk
DIURNAL RHYTHM OF PETAL GROWTH IN CUT
ROSE FLOWERS
Takanori Horibe1 and Kunio Yamada1
1
College of Bioscience and Biotechnology, Chubu University, Kasugai, Aichi 487-8501, Japan
In our research, we found that the petal growth of rose flowers placed under 12h-light/12h-dark
photoperiod condition showed a diurnal rhythm. However, little is known about the relationship
between petal growth and photoperiod. Thus, we investigated the effect of photoperiod on petal
growth rhythm of cut rose flowers. We have shown that cell wall loosening proteins, expansin and
xyloglucan endotransglucosylase/hydrolase (XTH), play an important role in petal growth. And we
presume that aquaporin is related to mass water flow during petal growth. Therefore, we conducted
a gene expression analysis to clarify relationship between these genes and petal growth rhythm.
Cut rose flowers (Rosa ‘Pretty Woman’) were held without leaves at various photoperiod conditions
and their growth was observed at regular intervals. For expression analysis, the petals were sampled
twice a day (1 hour after beginning of light-/dark- period) for 2 days from cut rose flowers held at
12h-light/12h-dark photoperiod conditions.
Petal growth rhythm was observed under various photoperiods, but the speed and the period of
flower opening changed depending on particular photoperiod. It seemed that light stimulus is an
important signal to keep a particular growth rhythm. In addition, we found even a petal detached
from the bud showed the same diurnal rhythm of growth. In expression analysis, the amount of
aquaporin mRNA in the light period was higher than that in the dark period, but the amounts of XTH
and expansin mRNA did not show notable differences between light and dark periods. We are now
analyzing the relationship between petal growth and photoperiod in more detail.
29
Session 8: Abiotic Stress
Talk
Abiotic stress tolerance in roses:
physiological and biochemical changes
Marie-Christine Van Labeke
Department of Plant Production, Ghent University, Belgium
Roses are important ornamentals in landscaping and gardens, this in different climate zones. In
climate change scenarios, occurrences of dry and hot summers, now already present in South and
East-Europe, are expected to occur in other parts of Europe as well. Water shortage is a critical
challenge to gardening and landscaping in many regions of the world. Climate change can also affect
winter survival due to irregular temperature patterns leading to insufficient cold acclimation.
Plants perceive and respond to abiotic stress in a way that is both affected by the environment and
their genetic background. Several abiotic stresses affect plants through disruption of the plant water
status, whether it is through decreased availability of water in the environment during drought or
through cellular dehydration caused by formation of extracellular ice during cold stress. Under field
conditions stresses also often occur in combination.
Despite the economic importance of garden roses knowledge about their tolerance and adaptation
to abiotic stresses are still scarce. In roses reduced biomass and flowering is observed under drought
stress. Yet, a good and reproducible approach to invest the drought response is quite challenging.
Drought stress studies often use water withholding which leads to a severe drought stress while
mild to moderate stress might better reflect the actual field conditions. Monitoring the soil water
status is often used as well as physiological approaches such as leaf water potential and leaf gas
exchanges to measure the early effects of drought stress in roses. With increasing drought stress
osmotic adjustment is observed leading to higher proline and soluble carbohydrate contents. Data on
anti-oxidative defence mechanisms are however still scarce.
30
Session 8: Abiotic Stress
Talk
INCREASED SALINITY IMPROVES STOMATAL
FUNCTIONING OF ROSES GROWN UNDER HIGH
RELATIVE AIR HUMIDITY
Dália R.A. Carvalho1, Ep Heuvelink2, Marta W. Vasconcelos1, Susana M.P. Carvalho1,2
CBQF - Centro de Biotecnologia e Química Fina – Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa/Porto, Rua Dr. António Bernardino Almeida, 4200072 Porto, Portugal
2
Department of Plant Sciences, Horticultural Supply Chains Group, Wageningen University, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands
1
Leaves developed under high relative air humidity (RH) have a poor stomatal functioning, leading
to excessive water loss. This study aims at investigating if increasing moderately the salinity level
throughout plant development would enhance stomatal responsiveness to closing stimuli in high
RH-grown plants.
Cut rose ‘Prophyta’ were grown as a single shoot in controlled growth chambers under moderate
(60%) and high (90%) RH, combined with three levels of electrical conductivity (EC) in the irrigation
solution: 2, 4 and 6 dS m-1. Stomatal responsiveness was accessed in fully expanded leaflets subjected
to desiccation or to darkness. It was found that in leaflets developed at high RH, the relative water
content (RWC) after 4 h of desiccation increased from 18% (EC 2, control) to 42% (EC 6). In addition,
it was shown that the nocturnal stomatal conductance (gs) of intact leaflets from high RH-grown
plants was linearly reduced within increased salinity levels (being 320 mmol m-2s-1 at EC 2 and 130
mmol m-2s-1 at EC 6). Nonetheless, the levels achieved of RWC and nocturnal gs were still significantly
different from the ones observed in moderate RH-grown plants grown at EC 2. For instance, leaflets
grown at moderate RH and EC 2 showed a RWC of 70% after 4 h of desiccation and the nocturnal
gs was 86% lower than the gs of high-RH grown plants under EC 6. Concerning stomatal index and
stomatal dimensions, increasing EC only induced very slight changes in these parameters in high
RH leaflets. It is concluded that increased salinity within the studied range can partly counteract
the negative effect of high RH on stomatal functioning. However, this strategy should be carefully
applied to roses as although no leaf chlorosis (due to salt toxicity) was observed in the first flush,
after harvesting the flower stalk, severe symptoms appeared on older leaves, especially at EC 6.
31
Session 8: Abiotic Stress
Talk
Responses of Selected Garden Roses to
Four Different Soil Moisture Contents
Cai, Xiaoya1, Terri Starman1, Genhua Niu2, Charles Hall1, and David Byrne1
Department of Horticultural Science, Texas A&M University, 2133 TAMU, College Station, TX
77843
2
Department of Horticultural Science, Texas AgriLife Research and Extension Center, Texas A&M
System, 1380 A&M Circle, El Paso, TX 79927
1
A greenhouse study was conducted to evaluate the response of two garden roses (Rosa × hybrid L.),
‘Belinda’s Dream’ and ‘RADrazz’ to four soil moisture contents (SMC). An automatic irrigation system
was set up to maintain the four constant SMC levels of 10, 20, 30, and 40%. Before the experiment,
the Acclima sensors were calibrated and the water application rate was tested to measure the total
amount of water applied during the treatment period. Two cultivars were grown in 16 L containers
filled with peat moss-pine park based substrate. The study was a two-factor experimental design
with 4 replications. During the two-month treatment period, total water applied in each container
was 5.7 L, 16.6 L, 39.3 L, and 57.2 L at 10, 20, 30, and 40% SMC, respectively. There was a 90%
reduction in water application at 10% SMC compared to 40% SMC. In both cultivars, there were
no significant differences in growth and physiological responses between 30% and 40% SMC. In
‘RADrazz’, shoot DW was reduced by 20.7% and 87.3%, root DW was reduced by 34.1% and 82.1%,
while flower number was reduced by 27.5% and 87.8% at 20 and 10% SMC, respectively, compared
to 30% and 40% SMC. Photosynthesis was highest at 30 and 40% SMC, and it was reduced by 28.1%
and 62.4% at 20 and 10% SMC, respectively. In ‘Belinda’s Dream’, shoot DW was reduced by 30.7%
and 87.6%, root DW was reduced by 46.8% and 83.5%, while flower number was reduced by 41.9%
and 77.1% at 20 and 10% SMC, respectively, compared to 30% and 40% SMC. Midday leaf water
potential was highest at 40% SMC, and there was no significant difference between 20 and 30%
SMC. It was reduced by 54.5% at 10% SMC, compared to 40% SMC. Photosynthesis was highest at
30 and 40% SMC and lowest at 10% SMC. In summary, plants at 30 and 40% SMC maintained the
highest shoot and root DW, flower number, midday leaf water potential, and photosynthesis. Water
applied at 30% and 20% SMC was reduced by 31% and 70%, compared to 40% SMC, with excellent
performance at 30% SMC and acceptable growth and quality at 20% SMC. The 10% SMC led to
significant growth reduction, poor quality, and 25% mortality.
32
Session 9: Rose Diseases and Pests
Talk
Genetics and genomics of disease
resistance in roses
Debener, Thomas; Linde, Marcus; Schulz, Dietmar; Kaufmann, Helgard; Terefe-Ayana, Diro; Wehmeyer, Juliane; Klostermann, Dorothea; Schott, Rena; Montenegro, Ana-Priscilla,
Leibniz Universität Hannover, Institute for Plant Genetics, Herrenhäuser Str. 2, 30419, Hannover,
Germany
Diseases and pests of roses are an annoyance for the private gardener but their control is a major
cost factor for rose production, especially for cut and pot rose production. As plant protection by
chemicals is both costly and imposes potential risks to the environment resistance breeding would
be one alternative to chemical disease and pest control. Resistance breeding is most efficient if the
genetic basis of resistance factors on the host side and the genetic diversity on the pathogen side
are known. For roses, knowledge about both resistance factors and genetic diversity of some major
pathogens have accumulated over the last years. For black spot, powdery mildew and downy mildew
information about single dominant resistance genes as well as QTLs has been obtained by analyzing
both bi-parental populations segregating for resistance as well as association panels. In addition,
insight into the structure of populations of the pathogen has been gained by several research groups.
Recently, research to develop approaches for MLO based broad spectrum disease resistance to
powdery mildew has been started and first results have been published. The presentation will give an
overview on all of these topics and will discuss the potential of these research results for commercial
resistance breeding.
33
Session 9: Rose Diseases and Pests
Talk
POWDERY MILDEW RESISTANCE IN ROSES IS
PATHOTYPE SPECIFIC
Leen Leus, Hossein Hosseini Moghaddam, Jan De Riek and Johan Van Huylenbroeck
Institute for Agricultural and Fisheries Research (ILVO), Plant Sciences Unit, Caritasstraat 21, 9090
Melle, Belgium
Powdery mildew in roses causes economic losses and is problematic in pot, cut and garden roses.
Pathotypes of rose powdery mildew are well described and various resistance mechanisms are
observed in different rose genotypes. In the present study, Rosa wichurana and Rosa ‘Yesterday’
were found to react differently towards two powdery mildew pathotypes (R-E and R-P) by pathotype
specific responses in the rose leaves. On ‘Yesterday’ conidia of R-E could germinate but no further
development of mycelium was observed. On R. wichurana pathotype R-E developed but cellular
reactions in the leaves were abundant. Both rose genotypes showed susceptibility to R-P. Fungal
development of R-P was observed along with cellular reactions in the rose leaves. For all rosepathotype combinations responses of the host plant were described and quantified.
An F1 population (90 genotypes) obtained from ‘Yesterday’ and R. wichurana was used to develop a
genetic map. Fungal development in the F1 population was scored for the pathotypes R-E and R-P.
For both pathotypes the scores segregated differently resulting in the development of pathotype
specific Quantitative Trait Loci (QTL) on the genetic map.
Our results indicate that resistance reactions to powdery mildew do not only depend on the different
resistance mechanisms present in roses but also on the specific pathotype. These results show that
breeders who aim to select for better resistant roses need to take into account the interaction
between plant and fungus. The best strategy to obtain roses with a durable resistance is to combine
different resistance genes and subsequent resistance mechanisms thereby increasing the number of
pathotypes withstood.
34
Session 9: Rose Diseases and Pests
Talk
Rose rosette virus is the causal agent of
rosette disease
Patrick Di Bello, Thien Ho and Ioannis E. Tzanetakis
Department of Plant Pathology, Division of Agriculture, University of Arkansas System, Fayetteville
72701.
Rose rosette virus (RRV), a member of the genus Emaravirus (negative sense multi-segmented RNA
viruses), was discovered in 2011 in perfect association with Rose rosette disease (RRD). The type
member of the genus, European mountain ash ringspot-associated virus, has been reported having
four RNAs but RRV appears to have more than six segments. Immunological, RT- and qRT-PCR
detection protocols were developed based on data obtained from a population structure study of RRV
allowing for quick detection of the virus. RRV was detected in Phyllocoptes fructiphilus mites which
have previously been shown to transmit the RRD agent. The mites were serially transferred from
RRV-infected to RRV-free roses, and the latter tested positive for RRV two months post infestation.
Typical symptoms of RRD subsequently developed on the material including mosaic and mottling,
leaf distortion, and shoot proliferation. No other viruses were identified in diseased plants indicating
that RRV is indeed the causal agent of RRD.
35
Session 9: Rose Diseases and Pests
Talk
VOLATILE EMISSIONS FROM ACTIVELY-GROWING
GYNANDROPSIS GYNANDRA AND CLEOME
HASSELERIANA THAT PROVIDE EFFECTIVE
CONTROL OF SPIDER MITES IN PROTECTED ROSE
CULTIVATION
Nyalala, Samuel1 and Brian Grout2
Egerton University, Department of Horticulture, P.O. Box 536, Njoro, Kenya
University of Copenhagen, Department of Plant and Environmental Sciences, Højbakkegård Allé13,
2630 Taastrup, Denmark
1
2
Foliage of the African Spider Plant, Gynandropsis gynandra L. (Briq) is traditionally used as a repellent
against livestock ticks in rural Eastern Africa, and our earlier work that shows this characteristic
extends to another acarid, the two-spotted spider mite (Tetranychus urticae Koch), has been briefly
reviewed. As a companion plant to cultivated roses G. gynandra has a highly significant, repellent
effect on spider mites and is similarly effective when used as a barrier to infestation. Analysis by
GC-MS shows that the repellent effect is due to foliar emissions of acetonitrile, which is toxic to
the mites if they cannot escape it. Additionally, the leaves contain a number of other substances
e.g. methyl isothiocyanate, toxic at low vapour concentrations to spider mites. The current GC-MS
study demonstrates that acetonitrile emissions from G. gynandra increase significantly at higher
temperatures and high growing degree days under glasshouse conditions, which is beneficial with
regard to the repellent properties of the plant, as spider mite increase is similarly temperature
dependent. To expand the potential of these findings more clearly towards rose production in
temperate regions, a taxonomically close relative of G. gynandra, also from the family Cleomaceae,
has been investigated. The widely-grown Cleome hassleriana Chod. has been shown by GC-MS to
have comparable acetonitrile emissions and foliar content of volatile miticidal compounds to G.
gynandra, which highlights its potential as a contributory element in spider mite control strategies.
Either of these plants might be used for spider mite control in its own right or as complementary to
other strategies e.g. the use of biopredators. They may also be a natural and sustainable source of
effective miticides.
36
Session 9: Rose Diseases and Pests
Talk
Symptoms, transmission, and detection of
four rose viruses
Mollov, Dimitre1, Ben Lockhart1, and David Zlesak2
Department of Plant Pathology, University of Minnesota, Saint Paul, MN 55108 USA
Department of Plant and Earth Science, University of Wisconsin-River Falls, River Falls, WI 55022.
1
2
Four previously undescribed viruses infecting cultivated roses were identified and fully characterized
in Minnesota. These four viruses were transmitted by grafting from infected to healthy roses and
found to be the likely causal agents of the diseases that they were associated with. Viruses were
provisionally named after the characteristic symptoms in infected plants as follows: Rose yellow
vein virus (RYVV), Rose yellow mosaic virus (RoYMV), Rosa rugosa leaf distortion virus (RrLDV), and
Rose yellow leaf virus (RoYLV). Unlike the currently known viruses that affect rose, the ilarviruses
and the nepoviruses, that only show symptoms and are detected early in the growing season, these
new viruses exhibit symptoms throughout the season and can be detected readily during the entire
year. Based on virion and genome properties it was determined that RYVV is a member of the family
Caulimoviridae, RoYMV is a member of the family Potyviridae, and RrLDV and RoYLV are members
of the family Tombusviridae. Phylogenetic analyses suggest that these four viruses belong to distinct
new genera in their respective taxonomic families. The whole genomic sequence of each virus was
deposited in GenBank under the accession numbers: RYVV JX028536; RoYMV NC_019031; RrLDV
KC166238; and RoYLV, KC166239. Reliable diagnostic protocols were developed for each virus by
PCR for RYVV detection, RT-PCR, immunosorbent electron microscopy (ISEM), and indirect enzymelinked immunosorbent assay (ELISA) for RoYMV detection, and RT-PCR for both RrLDV and RoYMV
detection.
37
Session 9: Rose Diseases and Pests
Talk
Epidemiology of Blackberry chlorotic
ringspot virus
Bindu Poudel, Thien Ho, Archana Khadgi, and Ioannis E. Tzanetakis
Department of Plant Pathology, Division of Agriculture, University of Arkansas System, Fayetteville
72701.
Blackberry chlorotic ringspot virus (BCRV) is an ilarvirus that infects several rosaceous hosts including
rose. The high occurrence of BCRV in wild rose in the United States and the lack of knowledge
on the epidemiology of the virus make it a concern for the ornamental industry. The objective of
this study was to acquire information on different aspects of the virus ecology and epidemiology
including population structure, detection, alternative hosts, and transmission. BCRV appears to have
a homogeneous population in the United States but this population is quite different from the
European isolate. Information on the population structure was used to develop immunological and
molecular tests that can readily detect the virus with high sensitivity. Several plant species were
evaluated and were confirmed as alternative hosts for the virus, including apple, suggesting that
BCRV may be infecting additional rosaceous plants. Seed transmission was tested on seedlings of
BCRV-infected Rosa multiflora as well as Chenopodium quinoa with transmission rates reaching
50%, a fact that should be taken into account in breeding efforts ] where the virus is endemic.
38
Session 9: Rose Diseases and Pests
Talk
Rose replant problems – detection by
bio-tests and investigations of bulk soil
and rhizosphere microbial communities
using DGGE fingerprints
Yim, Bunlong1, Kornelia Smalla2 and Traud Winkelmann1
Gottfried Wilhelm Leibniz Universität Hannover, Institute of Horticultural Production Systems,
Herrenhäuser Str. 2, D-30419 Hannover, Germany
2
Julius Kühn-Institut – Federal Research Centre for Cultivated Plants (JKI), Institute for Epidemiology and Pathogen Diagnostics, Messeweg 11-12, D-38104 Braunschweig, Germany
1
Replant problems or soil sickness is a still unsolved problem in nurseries. Declines in growth and yield
often occur after removal of old plants and replanting the same species in the same field. Nematodes,
bacteria, fungi, toxins and nutrient deficiencies are discussed as possible causes of replant problems
not only in roses but also in other rosaceous plants. The aim of this study was (i) to detect the replant
problems of soils from three nurseries using Rosa corymbifera ‘Laxa’ seedlings under greenhouse
conditions based on different soil disinfection treatments, and (ii) to explore soil and rhizosphere
microbial community structure applying denaturing gradient gel electrophoresis (DGGE) fingerprints
of these soils before and at the end of the bio-tests.
The bio-test in 2013 involved determining plant growth in nine soil variants collected in three
different nurseries. In 2012, each nursery had applied three treatments on the replant problems
sites: (a) growing grass to maintain the replant problems status, (b) & (c) growing ‘Laxa’ and apple
rootstocks to intensify the replant problems, respectively. These soils were used in 3L containers
either untreated, heat-treated (1 h at 50 °C), or gamma-irradiated (10 Gy min. dose).
Significant differences in height were observed earliest after 2 weeks and latest after 4 weeks of
culture in the bio-tests, except for soil from one nursery where no significant differences were
found between untreated and heat-treated variants. After 8 weeks, shoot dry mass was closely
correlated to plant height with significant differences between treated and untreated soil variants.
The DGGE fingerprints of 16S rRNA gene fragments representing dominant bacteria and ITS fragments
representing dominant fungi are currently carried out with DNA from bulk and rhizosphere soil
collected in the nurseries and at the final evaluation of the bio-test. The identification of differentially
abundant bacteria and fungi by sequencing will follow.
39
POSTER PRESENTATIONS
Poster
Poster Session 1: Genetic Resources
ROSES OF VAN AND CULTURE OF ROSE
Şevket Alp1 Mehmet Koyuncu2
Yüzüncü Yıl University, Faculty of Agriculture, Department of Landscape Architecture Van Turkey
Ankara Uni., Faculty of Pharmacy, Department of Pharmaceutical Botany, Ankara, Turkey
1
2
According to some historians, the 18th century is the beginning of changing the established culture
and the term of proliferation of European culture in the Ottoman community. During that period, a
great evolution has been observed in the kind of plants which were used for designing the outdoor
plant cultivation for daily use.
For example, the rose which had been growing for many years in palace gardens of Istanbul, lost
its value after European roses were imported which caused Ottoman roses to lose their traditional
admire.
However, those old original Ottoman roses were continued to be cultivated in some Anatolian cities
without being effected by these trends. One of the regions which had not been affected by this
conversion is the Lake Van region in East Anatolia. As the result of some research, six different types
of original roses were detected in the gardens and old graveyards in this region. Research are also
shows that; these roses have a great reflection on Van’s traditional handicrafts and cuisine.
This rescript contains an establishment for the old original roses for representing its value in
traditional culture.
41
Poster Session 1: Genetic Resources
Poster
19. CENTURY OTTOMAN’S ROSES and VARIETIES
Şevket Alp1 and Karaca Zafer2
Yüzüncü Yıl University, Faculty of Agriculture, Department of Landscape Architecture Van Turkey
Directory of BağbahceMagazineIstanbul, Turkey
1
2
During the Ottoman period, an agricultural inspector, İzzetliZakaryanEfendi, in 1841, wrote a book
named “GülveMahsulatı- Rose and Crops”. In this book, the rose has been explained as a member
of the “Rosa” group a spiky, quite smelly, acceptably flowery plant or small bushy tree. In the book,
roses were classified into ten different groups and each of them has been identified one by one.
In this study first the information about these groups is given, then it also inform about rose cultivation and about the cities in which roses were cultivated during the Ottoman period in Anatolia.
42
Poster
Poster Session 1: Genetic Resources
CELLULOSE AND LIGNIN CONTENT RATIOS IN THE
PERICARP OF SOME SPECIES OF ROSA GROWING
IN THE EAST ANADOLIA REGION
Şevket Alp1 and Aldemir Reşit2
1Yüzüncü Yıl University, Faculty of Agriculture, Department of
Landscape Architecture Van Turkey
2YüzüncüYılUniversity, Gevaş Vocational School, 65080 Van Turkey
The dormancy of rose achene germination may be delayed due to a hard woody pericarp. This barrier
in the form of a hard pericarp contributes to dormancy in the rose. In this study, cellulose and lignin
ratios which cause pericarp hardness has been determined for seed of some Rosa species that grow
in the Van region. Also the mentioned polymers were detected in the pericarp of Rosa canina seed in
different harvest periods. As a result, the percentage of ligninification that influences hardness has
been determined, in Rosa dumalis (30.95 %); in Rosa canina (27.49 %); in “Hoşapgülü” (20.18 %) and
in Rosa pisiformis (16.22 %). The ratio of lignin in R. canina has been found to be variable depending
on the time of harvest. As a result of the study, the seed of some rose spevies could be used as animal
nutrition due to high doses of lignin.
43
Poster Session 1: Genetic Resources
Poster
KARYOTYPE ANALYSIS OF WILD ROSA SPECIES IN
XINJIANG, NORTHWESTERN CHINA
ChaoYu1, Le Luo1,2, Huitang Pan1,2, Runhua Guo3 Yunji Sui3 and Qixiang Zhang1,2,*
College of Landscape Architecture, Beijing Forestry University, 35 Tsinghua East Road, Haidian
District, 100083, Beijing, China
2
National Engineering Research Center for Floriculture, 100083, Beijing, China
3
Xinjiang Career Technical College, Xinjiang, China
* Corresponding Author: e-mail: [email protected]
1
Wild Rosa species, which have strong adaptability and numerous variations, are widely distributed in
the Xinjiang Uygur autonomous region of China. These species display many important horticultural
traits which were not found in the gene pool of modern cultivated roses. However, little is known
about the cytological characteristics and few of them were karyologically analyzed. In this research,
karyological data of 13 samples of 7 wild Rosa taxa, including Rosa berberifolia, R. spinosissima, R.
platyacantha, R. beggeriana and R. laxa etc., were investigated via the squash method. The results
showed that 7 samples were diploid (2n=2x=14) and the other 6 samples were tetraploid (2n=4x=28).
The karyotypes of all the test samples were made of m and sm chromosomes, which include 1A, 2A,
1B and 2B. The asymmetry index, centromere index and constitution of relative length etc. were
different between samples. The karyotype of the tetraploid R. laxa sample from Aksu showed obvious
differences with other R. laxa samples tested, namely the highest asymmetry index and the most
evolved karyotype, which should be placed as a new variety of R. laxa. The karyological data, most
of which are reported for the first time, provided a comprehensive cytogenetic resource that can
be used to better understand the taxonomy, evolution and speciation in genus Rosa and to identify
candidate species for breeding strategies.
44
Poster
Poster Session 2: Genetics and Breeding
ASSESSMENT OF THE ROSE BUSH FLORIBUNDITY
AND ARCHITECTURE
Boumaza Rachid1, Nathalie Leduc2, Lydie Huché-Thélier3, Sabine Demotes-Mainard3, Hervé Autret3,
François Barbier1, Jessica Bertheloot3, Nathalie Brouard2, Mathieu Caradeuc3, Laurent Crespel1,
Odile Douillet3, Bénédicte Dubuc2, Fabrice Foucher2, Pierre-Maxime Furet2, Gilles Galopin1, Morgan Garbez1, Laurence Hibrand Saint-Oyant2, Anita Lebrec2, Marion Lecerf1, José Le Gourrierec2,
Camille Li-Marchetti1, Jérémy Lothier2, Philippe Morel3, Sandrine Pelleschi-Travier2, Maria-Dolores
Perez-Garcia1, Alix Pernet2, Thomas Peron2, Daniel Relion3, Hanaé Roman2, Pierre Santagostini4,
Monique Sigogne3, Gérard Sintès3, Alain Vian2, Vincent Guérin3, Soulaiman Sakr1
Agrocampus-Ouest, UMR 1345 Institut de Recherche en Horticulture et Semences, SFR 4207 QUASAV, Angers, France.
2
Université d’Angers, UMR 1345 Institut de Recherche en Horticulture et Semences, SFR 4207
QUASAV, PRES L’UNAM, Angers, France.
3
INRA, UMR 1345 Institut de Recherche en Horticulture et Semences, SFR 4207 QUASAV, Beaucouzé, France.
4
Agrocampus-Ouest, Angers, France.
1
Floribundity defined as “the capacity of a plant to produce abundant flowers at high density on each
of its branches”, is a key element of the aesthetic quality of ornamental plants. In many species,
including the rosebush, this feature is strongly related to the plant architecture which is determined
by the genotype, the environment and the crop management.
To address these questions, the Architecture and Environment (ARCH-E) team of the Research
Institute on Horticulture and Seeds (http://www6.angers-nantes.inra.fr/irhs) in collaboration with
the Genetic determinism & Diversity of Ornamental plants (GDO) team of the same institute, is
developing an integrative research program whereby environmental effects on the establishment
of the plant architecture are examined from the molecular to the whole plant levels using plant
sciences (physiology, genetics, molecular biology, biochemistry…), sensory analysis and modelling
studies.
The aim of the poster is to present a review of the studies (listed below) managed by the team,
focusing on methods to assess and compare the floribundity of some rose cultivars, considered as
example varieties by the International Union for the Protection of New Varieties of Plants (UPOV).
45
Poster Session 2: Genetics and Breeding
Poster
Pollen grain preservation at low
temperatures in valuable Rosa hybrida L.
commercial cultivars
Annalisa Giovannini1*, Anca Macovei2, Mattia Donà2, Alberto Valassi2, Matteo Caser3, Valentina
Scariot3, Andrea Mansuino4, Gian Guido Ghione4, Daniela Carbonera2 and Alma Balestrazzi2
Consiglio per la Ricerca e la Sperimentazione in Agricoltura-Unità di Ricerca per la Floricoltura e le
Specie Ornamentali, Corso Degli Inglesi 508, 18038 Sanremo (Imperia), Italy;
2
Dipartimento di Biologia e Biotecnologie “L. Spallanzani”, via Ferrata 1, 27100 Pavia, Italy;
3
Dipartimento di Scienze Agrarie, Forestali ed Alimentari, Via Leonardo da Vinci 44, 10095,
Grugliasco, Torino, Italy;
4
NIRP International, Azienda Agricola di Ghione L. & Figli, Via San Rocco,1 - Fraz. Bevera - 18039
Ventimiglia (Imperia), Italy
*Corresponding author E-mail: [email protected]
1
Rosa hybrida L. modern cultivars require hand pollination for rosehip production and collection of
seeds. Breeding programs are often focused on the quality of rose pollen which is genotype dependent
and affected by the conditions used for its conservation. Decrease in pollen viability during storage
condition could be due to dehydration, which results in loss of pollen colloidal properties. In addition,
H2O2 and NO overaccumulation can inhibit stored pollen germination (Macovei et al., submitted). In
this study the in vitro pollen viability of 6 different rose cultivars, provided by NIRP International, was
evaluated under different storage conditions in order to implement preservation protocols. Flowers
of the cultivars ‘Alba’, ‘Anastasia’, ‘Encanto’, ‘Marvelle’, ‘Swan’ and ‘Touch of Class’ were collected
from plants cultivated in the NIRP greenhouses (Bevera, Ventimiglia, Northwest Italy). Anthers were
removed from flowers and dried on Petri dishes for 24 hours (T= 24.0°C). About one gram of pollen
was recovered from 12 flowers of each cultivar. To assess the storage period the pollen grains were
divided and placed into polyethylene tubes in the freezer (T= - 20.0°C) and in the deep freezer (T= 80.0°C). Pollen viability was measured soon after 24 hours from flower collection and after 40, 120
and 240 days of storage, respectively. The pollen grains were put to germinate on a culture medium
containing agar (0,7%), sucrose (15%), calcium chloride (152 mg L-1) and boric acid (40 mg L-1).
Our results confirmed that the viable fraction in fresh pollen varied among genotypes, and also that
pollen preservation at low temperatures is cultivar dependent.
46
Poster
Poster Session 2: Genetics and Breeding
MEAN DAILY MAXIMUM TEMPERATURE FOR
8 – 14 DAYS BEFORE HARVEST AS A GOOD
PREDICTOR FOR CHANGE IN LANDSCAPE ROSE
FLOWER DRY WEIGHT
Greyvenstein, Ockert1, Terri Starman1, Brent Pemberton2, Genhua Niu3 and David Byrne1
Department of Horticultural Science, Texas A&M University, 2133 TAMU, College Station, TX
77843
2
Texas AgriLife Research and Extension Center, P.O. Box 200, Overton, TX
3
Department of Horticultural Science, Agricultural Research and Extension Center at El Paso, 1380
A&M Circle, El Paso, TX 79927
1
Heat stress is a major limiting factor for growing crops and can severely affect rose flower quality and
post harvest quality. Flower dry weight and the number of vegetative nodes to flowering of fourteen
adapted landscape roses were studied during 2010 and 2011. A simple linear model based on the
mean daily maximum temperature for 8 – 14 days (2WkMaxºC) before the flowers open groups the
14 cultivars into two groups: cluster 1 with a predicted 4.28 % and cluster 2 with a 6.45 % decrease
in average flower dry weight for a 1 ºC increase in 2WkMaxºC. The average mean daily temperature
recorded during the growth period for plants was 24.7 ºC, with the maximum and minimum daily
average of 34.4 and 2.22 ºC. The number of vegetative nodes to the inflorescence is affected by the
growing season but the change and magnitude were cultivar dependent. The number of nodes of
some cultivars was not affected by the growing season and could be an indicator of heat tolerance.
The change in number of nodes and predicted change in flower size appears to be separate traits and
could aid breeders in selecting better adapted material.
47
Poster Session 2: Genetics and Breeding
Poster
Mechanism of Self-Incompatibility in
Diploid Roses
Paudel, Dinesh Babu1 and Debener, Thomas1
Gottfried Wilhelm Leibniz Universität Hannover, Department of Molecular Plant Breeding, Herrenhäuser Str. 2, 30419 Hannover, Germany
1
Self-incompatibility is natural mechanism by which self-pollination is arrested and out-crossing
is facilitated. Many plants adopted this mechanism to reduce the chance of inbreeding depression
during the course of evolution. Self-incompatibility is widely present in diploid Rosa species but
studies related to self-incompatibility on this genus are very limited. As other members of the
Rosaceae family, rose is believed to have gametophytic self-incompatibility controlled by single
locus genes with multiple alleles; where an S-RNase ribonuclease inhibits the growth of pollen in
the style. This study focuses principally on 1. confirmation of self-incompatibility in rose, 2. analyzing
the number of alleles present in the diploid populations available at the Institute for Plant Genetics,
3. finding the possible mechanism of self-incompatibility and 4. locating the genes governing the
self-incompatibility reaction. For this study, Rosa hybrid population 94/1 and its two parents 93/1117 and 93/1-119 were selected. Back-cross pollination on the segregating population 94/1 showed
a segregation ratio consistent with the presence of a single gametophytic SI locus for which the
two parents share one allele and differ in the second allele. Tetraploid plants have been induced
from the selected diploid genotypes and self-pollination of the tetraploid plants in comparison to
diploids is being done to elucidate whether the SI system breaks down in tetraploids by competitive
interaction of the pollen S-function. Inter-crosses between the available diploid genotypes of the
institute are being carried out to find out the number of different alleles present in the collection.
In addition, DNA sequences from the orthologous genes of related species as well as partial Illumina
sequenced genomic DNA of genotype 88/124-46 and cDNA obtained from the pistil have been used
for developing molecular markers in order to position these markers on the integrated consensus
map of diploid roses, based on the polymorphism detected.
48
Poster
Poster Session 2: Genetics and Breeding
New strategies for improving varietal
creation in Rosa
Valentina Scariot1*, Luca Pipino1,2,3, Annalisa Giovannini2, Matteo Caser1, Fulvio Dente2, Andrea
Mansuino4, Gian Guido Ghione4, Leen Leus5, Marie-Chiristine Van Labeke3
Department of Agricultural, Forest and Food Sciences, University of Turin, Via Leonardo da Vinci
44, I-10095 Grugliasco (TO), Italy, [email protected]
2
Consiglio per la Ricerca e la Sperimentazione in Agricoltura-Unità di Ricerca per la Floricoltura e le
Specie Ornamentali, Corso Degli Inglesi 508, 18038 Sanremo (Imperia), Italy;
3
Department of Plant Production, Ghent University, Coupure links 653, 9000, Gent, Belgium
4
NIRP International, Az. Agricola di Ghione L. & Figli, via San Rocco 1, Fraz. Bevera, I-18039, Ventimiglia (IM)
5
Institute for Agricultural and Fisheries Research (ILVO), Plant Sciences Unit, Caritasstraat 21, 9090,
Melle, Belgium
1
The creation of new rose varieties is a process that takes at least five years. A general prerequisite for
a successful breeding program is to obtain wide offspring on which selection of seedlings with the
desired traits can be performed. However, modern rose cultivars are characterized by low seed set
and poor seed germination which hinders the achievement of a high number of seedlings. Premature
death of developing embryos due to incompatibility and delayed germination, caused by seed
dormancy, are the most important causes.
The main objective of this study was to improve hybrid tea seed production efficiency. Experiments
were initially conducted to gain insight into the fertility of pollen donors and their effect on a
successful hybridization. To this end, we studied the crossing results of the Nirp International
database. High correlations were found between the number of seeds produced per hybridization
and either the pollen diameter or the percentage of normal pollen, thus allowing the development
and validation of a prediction model for seed set based on pollen quality (Pipino et all., 2011).
Subsequently, to deepen the knowledge about seed development, we studied the embryo
morphogenesis of two floribunda roses at seven time points, from 3 to 60 days after pollination
(DAP). Embryos resulted fully developed at 15 DAP while seeds at 30 DAP. In parallel, the abscissic
acid (ABA) concentration determined in the developing embryos showed basal levels from 30 DAP on.
Based on these results, we focused on in vitro techniques which may be used to rescue abortive or
dormant embryos, thus increasing the numbers of plants obtained from crosses resulting in very few
viable seeds, or to shorten the time needed to obtain seedlings. Our results suggest that the optimal
time point for performing embryo rescue in hybrid tetraploid roses is between 15 and 30 DAP.
49
Poster Session 2: Genetics and Breeding
Poster
Association mapping of phenotypic traits
in roses
Schulz, Dietmar Frank1, Rena Schott1, Ana Priscilla Montenegro1, Dorothee Klostermann1, Marcus
Linde1, Burkhard Spellerberg2 and Thomas Debener1
Gottfried Wilhelm Leibniz Universität Hannover, Department of Molecular Plant Breeding, Herrenhäuser Str. 2, 30419 Hannover, Germany
2
Bundessortenamt, Osterfelddamm 80, 30627 Hannover, Germany
1
In this work we focus on the association of resistance to several important fungal rose pathogens
and different phenotypic traits like scent, anthocyanin and carotinoid content and plant architecture
to molecular markers. A panel of 96 rose cultivars was planted in five different field locations in the
north, middle and western part of Germany. Additionally, one trial was conducted under controlled
conditions under glass in Hannover (Bundessortenamt). The studies included the generation of AFLPand SSR- markers for genome wide association and markers for candidate genes. To achieve this
molecular markers were also useful to estimate population structure (Structure 2.3.4) and genetic
diversity of the association panel, the majority of it tetraploid. Associations between markers and
phenotypic traits were calculated with Tassel 4.1.10. A testcross progeny was developed to verify the
identified marker-trait associations.
50
Poster
Poster Session 2: Genetics and Breeding
THE UTLIZATION OF THE POLYPLOID NATURE OF
ROSES
Jake Ueckert1 David Stelly2, Kevin Crosby1, David H. Byrne1 and George Hodnett2
Department of Horticultural Sciences, Texas A&M University, College Station, Texas, USA
Department of Soil and Crop Sciences, Texas A&M University, College Station, Texas, USA
1
2
Rose chromosome number ranges from 14 to 56 and understanding the dynamics of rose chromosome
numbers can help improve the progress made in breeding programs and enhance the understanding
of certain rose populations. A survey of the pollen size of 80 roses in a breeding collection suggested
that 78% were tetraploid, 7% were triploid, and 14 % were diploid. These results are currently being
verified with chromosome counts. Of these roses, the presence of large pollen grains indicating non
reduction during meiosis occurred in 10% of the roses. Triploid roses most commonly exhibited non
reduction, but there were some diploid plants that had 2n gametes. A common occurrence among
the triploids was the presence of irregular and shrunken pollen grains that were likely aneuploids
leading to pollen abortion. In certain triploids, such as ‘Home Run’ and ‘Tangerine Jewel’, pollen size
suggests that as much as 70% of the viable pollen is 2x while another 25% appears to be 1x, with
the remaining 5% consisting of 3x grains. The progeny of crosses with a triploid rose as the pollen
parent with several diploid and tetraploid female parents are currently being evaluated for ploidy
level to determine if the ratio of progeny ploidy is similar to the ratio of pollen ploidy in the pollen
parent.
51
Poster Session 2: Genetics and Breeding
Poster
DETERMINING THE ASSOCIATION OF SCAR
MARKER ND5E AND RESISTANCE TO RACE 8
OF DIPLOCARPON ROSAE WOLF IN A DIVERSE
GROUP OF LANDSCAPE ROSES
Zlesak, David C.1, Andrea Clark2, James M. Bradeen3, and Stan C. Hokanson2
Univeristy of Wisconsin-River Falls, Department of Plant and Earth Science, River Falls, WI USA
University of Minnesota, Department of Horticultural Science, St. Paul, MN USA
3
University of Minnesota, Department of Plant Pathology, St. Paul, MN USA
1
2
Roses are among the most popular flowering landscape shrubs in the world. Rose black spot disease,
caused by the pathogen Diplocarpon rosae Wolf, is the most serious disease of outdoor-grown
roses worldwide. Both race-specific and horizontal resistances to black spot have been documented
in Rosa spp. and both resistance forms are valuable to pursue in the development of new resistant
cultivars. Having robust markers linked to race-specific resistance genes is of great value to breeders
for use in screening germplasm and working towards stacking multiple race-specific resistances in
new cultivars. Currently the SCAR marker ND5E is the only marker linked to Rdr3, a gene conferring
resistance to race 8 of rose black spot. ND5E was identified in ‘George Vancouver’ and is estimated
to reside 9.1 cM from the gene. Roses of diverse backgrounds (n=89) with known resistance to race
8 (28 resistant and 61 susceptible) were screened for ND5E (n>3 reactions per cultivar). The marker
(~80bp) was definitively recovered in 11 roses (9 resistant and 2 susceptible), and for many of the
other roses faint bands of similar size were inconsistently produced in some of the amplifications.
Although the low false positive rate for susceptible roses clearly possessing the marker is promising
(3.2%), the low detection rate of ND5E among resistant roses (32.1%) suggests lack of universality
of this marker. Relatives of resistant roses possessing ND5E were subsequently screened for ND5E
and presence of the marker was found to be common in family lines. Efforts are underway to identify
new and more tightly linked markers to Rdr3.
52
Poster
Poster Session 3: Rose Hips
Genetic Resources of the Wild Rose
(Rosa spp.) in Latvia
Antra Balode
Institute of Agrobiotechnology, Faculty of Agriculture, Latvia University of Agriculture
Liela 2, Jelgava, LV 3001, Latvia
Latvia is situated between the vegetation zones of Northern and Central Europe; therefore, most rose
species cannot survive the winter, and it is essential to develop freeze-resistant cultivars. Working
with wild plants, breeders try to bring the excellent properties of wild materials into cultivars.
Evaluation of wild species and cultivars is in progress. The latest research indicates many wild species
are scarce - endangered. Altogether – so far – eleven (11) wild species have been found. Evaluation
of wild roses and rose cultivars that originate in Latvia is the main goal of the research. To evaluate
the survival potential of Rosa spp., natural habitats have been surveyed in 2012. Currently, the
Red Data Book has played a significant role in the protection of endangered species. In both open
habitats and among shrubs, in a small area of Kurzeme and Zemgale (west and central Latvia), the
following species were found: R. caesia, R. coriifolia, R. mollis, R. pomifera (subsp. glandulosa) and
R. rubiginosa. In eastern and central parts of Latvia, in the open as well as in shrub-covered areas, two
rare species – R. sherardii and R. ciesielskii were found either in small groups or as solitary specimens.
Common all over Latvia, growing in shrubs on edges of woods and forests, are: R. canina (subsp.
virens), R. majalis, R. subcanina and R. vosagiaca. Identification has been based on morphological
characteristics – growth habit, prickles, floral morphology (flower, colour, diameter, flagrance) and
foliage (size, colour, and type). Research results indicate the diversity in genotypes varies according
to location and distribution. The species R. rugosa has been used, for more than 50 years, in breeding
programs at the National Botanical Garden of Latvia. Sixteen cultivars have been registered and
distributed. In 2003, a large collection of R. rugosa cultivars was planted in Rundale Castle Park
based on Baroque garden designs. The cultivars were winter hardy, as well as black-spot resistant.
53
Poster Session 3: Rose Hips
Poster
STRATEGIES FOR IMPROVING SEED
GERMINATION IN HYBRID TEA ROSES
Luca Pipino1,2, Valentina Scariot2, Laura Gaggero3, Andrea Mansuino4, Marie-Christine Van Labeke1,
and Annalisa Giovannini3
Department of Plant Production, Ghent University, Coupure links 653, B-9000, Gent, Belgium,*Fax:
+ 32 09 2646225, *E-mail: [email protected], [email protected]
2
Department of Agriculture, Forest and Food Sciences, University of Turin, Via Leonardo da Vinci 44,
I-10095, Grugliasco (TO), Italy, Fax: + 39 011 6708798, E-mail: [email protected]
3
Consiglio per la Ricerca e la Sperimentazione in Agricoltura-Unità di Ricerca per la Floricoltura e le
Specie Ornamentali, Corso Degli Inglesi 508, 18038 Sanremo (Imperia), Italy;
Fax: + 39 0184-694856; E-mail: [email protected]; [email protected]
4
NIRP International, Az. Agricola di Ghione L. & Figli, Via San Rocco 1, Fraz. Bevera, I-18039, Ventimiglia (IM), Italy, Fax: + 39 0184 210259, E-mail: [email protected]
1
Rose breeders need reliable and efficient germination protocols to exploit all vital embryos in their
breeding programme. A combined physical and physiological dormancy is present in the achenes
of hybrid tea roses resulting in variable and not uniform seed germination. In this study, different
treatment combinations were assessed to overcome physical dormancy and to enhance germination.
Both chemical and microbial seed scarification were carried out. A mild scarification with sodium
hypochlorite (NaOCl) before stratification promoted a more uniform germination and enhanced the
percentage of germination from 49.2% to 65.9%. Epiphytic bacteria, EmercalTM (a product composed
by bacteria and co-metabolites produced by bacterial fermentation), or Remedier® (a commercial
formulation of Trichoderma harzianum and Trichoderma viridae), added to the stratification sand,
enhanced percentage of germinated seeds but had no effect on the uniformity of germination.
Addition of a compost activator to the stratification sand did not improve seed germination.
After stratification, pre-sowing treatments with gibberellic acid (GA) were carried out. Immersion
of stratified seeds in 1 g l-1 GA3 highly enhanced percentage of seed germination (77.6%), mean
germination time (26.0 days) and uniformity of germination (2.5%) compared to immersion in water
(respectively 64.8%, 32.8 days and 7.9%). Therefore, this study proposes a combination of treatments
as a reliable optimisation of seed germination protocols for hybrid tea roses.
54
Poster
Poster Session 3: Rose Hips
WILD SPECIES OF ROSA MAINTAINED IN A FIELD
GENE BANK AT THE RESEARCH INSTITUTE OF
HORTICULTURE IN SKIERNIEWICE, POLAND
Dariusz Sochacki
Research Institute of Horticulture, Konstytucji 3 Maja 1/3, 96-100 Skierniewice, Poland
In a field collection of the genus Rosa established at the Research Institute of Horticulture in
Skierniewice, Poland, 21 wild species and their hybrids from different regions of the world have been
collected. Only two of them are native to Poland. However, at least 15 species of roses occur in wild
habitats in Poland. Most of them belong to the section Caninae, including the most common species
– the dog rose (Rosa canina). To make the collection richer in native species, a search in other regions
of Poland was started in 2009.
The first botanical expedition was organized to very old Cambrian mountains called the Pepper
Mountains (Góry Pieprzowe) on the west bank of the Vistula (Wisła) river, close to the town of
Sandomierz. These low mountains occupy a small area but are the biggest natural rosarium in Poland
and one of the biggest in Europe, with several thousands of rose shrubs of several species. In the
autumn of 2009, rose hips from 16 shrubs, initially identified as 6 different species, were collected.
In the autumn of 2010, during the next expedition to the Pepper Mountains, rose hips from 15
shrubs, including 12 located in the previous year as well 3 new shrubs, were collected. The obtained
seeds were germinated and in 2011 a total of 400 seedlings were produced from the seeds of 15
different shrubs discovered in 2009; the seedlings were transplanted to pots. In November 2011, an
expedition to the Sandomierz region was organized. During the expedition, 15 rose shrubs in their
natural habitats and in home gardens in the villages around Sandomierz, Opatów and Staszów were
located and initially identified as 8 different species. From the rose hips collected from 14 shrubs
more than 13 thousand seeds were obtained and germinated. In the spring of 2012, an expedition to
the Sandomierz region was organized during the flowering period of some species.
The last expedition took place in December 2012 to the surroundings of the town of Przemyśl in the
south-eastern part of Poland. Nine rose shrubs were found in home gardens in the local villages as
well as along field borders.
In total, forty-four rose shrubs were located in wild habitats and home gardens in the villages in the
Sandomierz and Przemyśl regions. Current work aims at the proper identification of the species and
taxon nomenclature.
55
Poster Session 4: Rose Genetics
Poster
A genetic, genomic and molecular frame
to study floral development in roses
Annick Dubois1, Sébastien Carrère2, Philippe Vergne1, Olivier Raymond1, Ludovic Cottret2, Shu-Hua
Yang1, Aurélie Vialette1, Jeremy Just1, Jérôme Gouzy2, Mohammed Bendahmane1
Laboratoire Reproduction et Développement des Plantes, UMR CNRS-INRA-Lyon1-ENSL, Ecole
Normale Supérieure, Lyon, France.
2
Laboratoire des Interactions Plantes-Microorganismes, UMR INRA-CNRS, Castanet-Tolosan France.
1
Wild and cultivated roses harbor a huge diversity of flower morphologies, colors and scents, making
them a particularly intriguing model for the study of natural and artificial evolution of these floral
traits. Our group focuses on floral architecture, especially on the genetic control of petal shape and
petal number. While wild roses all have five petals, most cultivated roses have double flower ranging
from 10 petals to as many as 200. We will describe our current approach to study the molecular
framework of double flower formation as well as the establishment of the necessary tools allowing
genetic, genomic and molecular studies in roses. We chose as a model a diploid rose cultivar, Rosa
chinensis Old Blush. This cultivar has a double colored and scented corolla and is a common ancestor
of many modern rose cultivars. We believe that in the near future these tools will be helpful for
genetics, genomics and functional genetics studies in roses and other related Rosaceae.
56
Poster
Poster Session 4: Rose Genetics
PRODUCTION OF SOMATIC EMBRYOS AND
TRANSGENIC PLANTS DERIVED FROM BREEDING
LINES OF ROSA HYBRIDA
Su Young Lee1, Jeong-Ho Kim2, Jin Yong Ko3, Seung Tae Kim4, Eun Kyung Lee1, Won Hee Kim1, O
Hyeon Kwon1, Hye JinLee1, and Kyeong-Seong Cheon1
National Institute of Horticultural & Herbal Science, Floriculture Research Division, Suwon 441440, Korea
2
National Institute of Horticultural & Herbal Science, Vegetable Research Division, Suwon 441-440,
Korea
3
Gumi Floriculture Experimental Farm, Gyeongsangbuk-do Agricultural Research & Extension Services, Gumi 730-831, Korea
4
National Institute of Horticultural & Herbal Science, Planning and Coordination Division, Suwon
441-440, Korea
1
This study was conducted to obtain transgenic plants carrying the GUS-intron gene by using somatic
embryos (including embryogenic calluses) derived from 2 in vitro root explants of breeding lines
(KR056002 and KR056006) bred by crossing between Rosa hybrida ‘Tineke’ and ‘Mirinae Gold’.
Calluses were induced from root explants cultured in Schenk and Hildebrandt (SH) medium, which
was supplemented with 5 or 11 mg·L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D). Somatic embryos
were generated from the calluses, which were cultured in SH medium supplemented with 3 mg·L-1 of
2,4-D. The ratio of callus formation from in vitro root explants was dependent on the concentration
of 2,4-D supplement in the SH medium. Somatic embryos were generated from both KR056002
and KR056006, and embryogenesis was observed from calluses around the somatic embryos. The
regenerative capacity of the embryos was maintained longer in calluses derived from in vitro root
explants cultured on SH medium supplemented with 11 mg·L-1 of 2,4-D than with 5mg·L-1 of 2,4-D.
Six transgenic lines carrying GUS-intron constructs were obtained. The expression rate of the GUSintron gene in multi-shoots was 100%. After the formation of healthy roots, 3 transgenic lines were
transferred to the greenhouse. All 3 lines were confirmed as GUS-intron transgenic plants by PCR
and Southern analyses.
57
Poster Session 4: Rose Genetics
Poster
ISOLATION OF ROSA LEAFY (RLFY) AND ITS
EFFECTS ON RECURRENT FLOWERING IN ROSES
LiJuan Lian1, YunFeng Liu1, YuMan Zhang2, RongXiang Fang2 and QingLin Liu1
Department of Ornamental Horticulture and Landscape Architecture, China Agricultural University,
Beijing 100193, China
2
State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences,
Beijing 100101, China
1
Recurrent flowering is an essential economic character in roses. To investigate its relationship with
the expression patterns of LEAFY (LFY) homologues, Rosa Leafy (RLFY) genes were amplified from Rosa
chinensis, R. multiflora and R. rugosa using a combination of degenerate and gene-specific primers by
thermal asymmetric interlaced-PCR (TAIL-PCR) and normal PCR. The cDNAs of RLFY were amplified
by RT-PCR and their sequences were compared. The phylogenetic relationships and tertiary structure
of the RLFY proteins were analyzed; their subcellular localization was also determined. The expression
levels of RLFY in roses with different flowering habits were determined by semi-quantitative RT-PCR
and real-time RT-PCR. There was high homology among RmLFY from R. multiflora, RcLFY from R.
chinensis ‘Slater’s Crimson China’, and RrLFY from R. rugosa, while four amino acids were deleted
from the protein encoded by RpLFY from R. chinensis ‘Old Blush’. The RLFY protein is located in the
nucleus as with other transcription factors, and its predicted tertiary structure is nearly the same
as FaLFY from strawberry. There were large differences in expression levels at the early stage, but
essentially no difference at other stages between the non-recurrent species, R. multiflora and R.
rugosa, and the recurrent flowering Rosa chinensis. These results suggest that once floral initiation
is complete, recurrent flowering roses can continuously flower without repeated accumulation of
RLFY mRNA.
58
Poster
Poster Session 4: Rose Genetics
Investigation of malformed flower types
of Rosa hybrida L by using direct in situ
RT-PCR
Heiko Mibus1*, Agata Jedrzejuk2 and Margrethe Serek1
Faculty of Natural Sciences, Institute for Ornamental and Woody Plant Science, Leibniz University
of Hannover, Herrenhaeuser Str. 2, 30419 Hannover, Germany
2
On leave from: Department of Ornamental Plants, Faculty of Horticulture and Landscape Architecture, Warsaw University of Life Sciences, Nowoursynowska 166, 02-787 Warszawa, Poland
*Corresponding author: [email protected], phone: +49-51176219265; fax: +495117622654
1
Different MADS-box-genes were investigated in normally developed flowers and in malformed flowers
(similar C-function mutation) to clarify the molecular mechanism of flower development in Rosa
hybrida L. Expression patterns of MADS-box genes in early organ development were cytologically
investigated by in-situ RT-PCR. Expression levels of flower organs of full developed flowers and
different buds stages were examined by qRT-PCR. Exclusive expression of RhAP1-1 and RhAP1-2
in whorl 1 and 2 of rose flowers demonstrated that the expression patterns were similar to genes
homologous to APETALA1 in other plant species. In contrast, RhFUL showed a unique expression
pattern and was expressed only in sepals. Previous qRT-PCR analysis of full develop flower organs
and total buds showed that all A class genes were up regulated and the two C class genes RhC1
and RhC2 were down regulated in all organs of malformed flower types. Expression patterns of the
two different investigated B class genes were similar in normal and malformed flowers. Cytological
investigation by in-situ RT-PCR demonstrated that only one C class gene RhC2 was suppressed and
the A class gene RhFUL was expressed in whorls 3 and 4 of malformed flowers. The suppression of
RhC2 may be the reason for expression of RhFUL in whorls 3 and 4 that lead to the malformed flower
phenotype.
59
Poster Session 4: Rose Genetics
Poster
The nature of Rdr1 locus sequence
diversity along the Rosaceae
Terefe-Ayana, Diro1 and Thomas Debener1,
Gottfried Wilhelm Leibniz Universität Hannover, Department of Molecular Plant Breeding, Herrenhäuser Str. 2, 30419 Hannover, Germany
1
Rdr1 stands for resistance to Diplocarpon rosae 1. It is the first fully characterized and completely
sequenced resistance gene in roses. The Rdr1 locus is completely sequenced from the resistant Rosa
multiflora hybrid 88/124-46 and a susceptible Rosa rugosa genotype. The related orthologous locus
was obtained from the completely sequenced genome of Prunus persica (peach), Fragaria vesca
(strawberry) and Malus x domestica (apple). Comparative analyses of the Rdr1 orthologous locus
of these five species revealed that R. multiflora and R. rugosa share a high degree of synteny. A
less pronounced synteny was observed in strawberry but was missing in peach and apple plants.
Instead, in peach and apple genes from the Rdr1 locus are distributed on two different chromosomes.
Furthermore, phylogenetic analysis of 53 complete TIR-NBS-LRR genes from the five Rosaceae species
revealed that with the exception of some genes from apple and peach, most of the genes occur in
species-specific clusters, indicating that recent TIR-NBS-LRR gene diversification began prior to the
split of Rosa from Fragaria in the Rosoideae and peach from apple in the Spiraeoideae and continued
after the split in individual species.
60
Poster
Poster Session 4: Rose Genetics
A TALEN based approach to obtain mlo
based broad spectrum resistance to
powdery mildew in roses
Wehmeyer, Juliane, Kaufmann, Helgard and Debener, Thomas
Gottfried Wilhelm Leibniz Universität Hannover, Department of Molecular Plant Breeding, Herrenhäuser Str. 2, 30419 Hannover, Germany
Powdery mildew caused by Podosphaera pannosa (Wallr.: Fr.) de Bary is one of the most severe diseases
of rose. Especially for the production in the greenhouse it depicts a great problem. Biology and race
structure of P. pannosa limit the effectiveness of race-specific, monogenic resistances making the
application of fungicides indispensable. However, this application is not very desirable because of
high costs, increasing legal restrictions and public concerns. An alternative to overcome this problem
can be the mildew resistance locus o (mlo)-mediated resistance. The mlo-based resistance has been
characterized in barley, A. thaliana, tomato and pea where the loss-of-function of specific members
of the MLO gene-family leads to a recessive broad-spectrum resistance. This resistance displays
a high durability since it has been used in barley for over thirty years without being broken. The
MLO proteins are heptahelical transmembrane proteins which probably manipulate or suppress the
SNARE protein-dependent and vesicle-associated defense mechanisms of the cell and by that confer
susceptibility towards the pathogen.
In rose four MLO homologs (RhMLO1, RhMLO2, RhMLO3 and RhMLO4) closely related to the
functional orthologues in A. thaliana, pea and tomato have been sequenced and mapped. The aim
of this work is the verification of functionality of these four rose MLOs regarding the mediation of
susceptibility towards P. pannosa. An RNAi approach aimed to knock out single genes as well as
producing double and triple mutants represents the initial analysis where first steps have already
been taken. In a second approach knock out of the four rose MLO genes shall be achieved using
transcription activator-like effector nucleases (TALENs). TALENs consist of amino acid repeats which
target single nucleotides of the DNA and in fusion with a nuclease specifically create double-strand
breaks in the desired DNA region.
61
Poster Session 5: Postharvest Physiology
Poster
IMPROVED METHOD OF CALCULATING THE
ORNAMENTAL VALUE OF FLOWERS BASED ON
PETAL DISCOLOURING MEASURED IN CIEDE2000
Boronkay, Gábor1
Research Institute for Fruit Growing and Ornamentals, Park utca 2., H-1223 Budapest, Hungary
1
A method for quantifying the ornamental value of a flower of garden rose (Rosa hybrida Hort.) was
created at the Rose Garden Budatétény, based strictly on measurements. Here attractiveness means
the capacity to make a uniform colour effect. The blooming time of a flower was distributed into 23
phenological sub-stages from the immature bud to the falling of petals. CIEDE2000 standard chromatic
differences (∆E00) were calculated between the petal-colours of each stage and stage 6 (the optimal
state of the flower). The Ornamental Value of a colour is OVc=15-∆E00. The Ornamental Value of a
stage is OVs=Av%*OVc, where Av% is the relative visible surface of the flower as a percentage of the
surface of stage 6. The total Ornamental Value of a section of stages is OVt=Σ(Av%*(15-∆E00)*ls),
where ls is the length of each stage measured in days. The OVt should be calculated for three different
sections of subsequent flowering stages: a) the whole life of the flower (2-9 stages), b) the life of
the flower before dead-heading (2-7.5 stages), c) the middle (5.5-7) stages. In 2012 the method was
further improved, by the interpolation of the colour parameters at the non-measured stages.
This method is presented with an example of 9 Hungarian red floribundas and polyanthas and ‘La
Sevillana’ floribunda. The data were assessed from 2003-2008 in Budapest, Hungary. According to
the scores, 9 of the examined varieties are highly attractive to stage 7.5 and the best Hungarian was
the variety ‘Petőfi Sándor’ (score: 155). If the whole life of the flower is taken into consideration,
another two varieties have relatively low total OVt-s. It means that these roses need dead-heading,
while the others don’t need this process. According to the results, this Ornamental Value calculation
is suitable for determining the extent of maintenance the roses require in outdoor conditions.
62
Poster
Poster Session 5: Postharvest Physiology
STUDY ON THE IMPROVEMENT OF THE
ENVIRONMENT FOR SELECTED VARIETIES OF
ROSE GRAY MOLD DISEASE RESISTANCE
Seung Tae Kim1, Won Hee Kim1, Eun Kyung Lee1, Su Young Lee1, O Hyeon Kwon1, Hye Jin Lee1, and
Kyeong-SukHan2
National Institute of Horticultural & Herbal Science, Floriculture Research Division, Suwon 441440, Korea
2
National Institute of Horticultural & Herbal Science, Horticultural & Herbal Crop Environment
Division, Suwon 441-440, Korea
1
The purpose of this study was to increase the reliability of conventional screening methods for
rose gray mold (Botrytis cinerea). Two screening systems for instance the screening and the tray
screening method have been used, but the efficiency of the methods is too unreliable because of
low repeatability and uniformity and furthermore because of variable environmental factors. To
overcome these problems, one petal per the petri-dish was inoculated with the plant pathogens
Botrytis. Thus the shortcomings of the existing systems could be improved. The most stable infection
symptoms occurred at a concentration of 105/ml rose gray mold conidia. Lower as well as higher
concentrations lead to less uniform symptom development. In addition, the results of comparing the
gray mold development after infection through plant wounds, we found that approximately 24hr
after inoculation symptoms rapidly appeared. Therefore, a conidial concentration of 105/ml in petal
infection assays seem to be optimal for reproducible results.
63
Poster Session 5: Postharvest Physiology
Poster
Antimicrobial effect of chlorine dioxide
on vase life extension of cut rose flowers
Young Boon Lee1, Wan Soon Kim1*, Seul Ki Lee1, and Won Hee Kim2
Department of Environmental Horticulture, University of Seoul, Seoul 130-743, Korea
Floriculture Research Division, National Institute of Horticultural & Herbal Science, Suwon 441440, Korea
1
2
This study was conducted to investigate the antimicrobial effect of chlorine dioxide (ClO2) on
the vase life of cut rose ‘Beast’ (Rosa hybrida L.). The postharvest measures to extend the vase
life of cut rose flowers were divided into two groups: holding solution and pulsing treatments.
In holding solution treatment, cut flowers were treated with the preservative solutions containing
tap water (TW, control), distilled water (DW), and ClO2 solutions of 2 to 8 µL·L-1, comparing with
an commercialized antimicrobial compound of 8-hydroxyquinoline sulfate (8-HQS) solution of 200
µL·L-1. In pulsing solution treatment, cut flowers were dipped into the ClO2 solutions of 50 to 250
µL·L-1 for 60 seconds and then were placed in DW. The longest vase life was observed in the holding
solution with ClO2 4 µL·L-1 as 13.8 days, which were about four days longer than the control, followed
by ClO2 pulsing treatment. The concentration of ClO2 8 µL·L-1 was toxic to cut flowers causing dried
leaves and bleached stems. Bacteria were fully inhibited by ClO2 of even 2 µL·L-1in holding solution,
while detected in TW and DW with 3.7x105 and 6.3x105 CFU∙L-1 respectively. Considering that ClO2
treated in holding solution was completely exhausted in only four days, the vase life extension of cut
rose flowers by ClO2 treatment might be due to its initial antibacterial ability.
64
Poster
Poster Session 6: Secondary Metabolites and Biochemistry
EVALUATE AND COMPARE THE CONTENTS OF THE
ESSENTIAL OILS Rosa×damascena Miller and
Rosa×damascena Mill. var. trigintipetala
(Dieck) Koehne
Alp Şevket1 Erdem Aslan Sinem2 and Karagöz Suzan3
Yüzüncü Yıl University, Faculty of Agriculture, Department of Landscape Architecture Van
Ankara University, Faculty of Pharmacy, Department of Pharmacognosy Ankara Turkey
3
Yüzüncü Yıl University, Environmental Studies Research Center, 65080 Van, Turkey
1
2
Geographic and climatic conditions of the province of Van have revealed a unique garden culture.
Roses have been one of the indispensable elements in Van garden culture. Rosa x damascena Miller
grown in Van gardens has an important place in the traditional Van cuisine. The aim of this study
is to evaluate and compare the contents of the essential oils obtained from Isparta rose oil (Rosa x
damascene var trigintipetala) which is traded and Gül-i Muhammed (Rosa x damascena) using GC
and GC/MS. High contents of β-citronellol (35.19%), nonadecane (14.20%), geraniol (9.35%) and
nerol (8.65%) were found in Rosa x damascene var. trigintipetala rose oil; spathulenol (10.12%),
α-amorphene (6.89%), salvial-4(14)-en-1-one (4.78%) and 1,5-epoxysalvial-4(14)-ene (4.72%)
were also found to a high amount in Rosa X damascene rose oil
65
Poster Session 6: Secondary Metabolites and Biochemistry
Poster
Multiple uses of Rugosa hybrids
A. Sparinska1 and N.Rostoks2
University of Latvia Botanical Garden (2 Kandavas Str., Riga, LV-1083, Latvia)
Faculty of Biology, University of Latvia (4 Kronvalda Blvd., Riga, LV-1586, Latvia)
2
1
In the climatic conditions of the Baltic countries and North Europe Rosa rugosa hybrids are the
most winter hardy group of roses, therefore bred for use in landscape architecture for public places.
Unfortunately it has spread out of gardens and made huge populations on the Baltic coast and
roadsides. Understanding uses of this plant could provide sustainable management solutions instead
of costly and complicated destruction of these rose communities. This study is aiming at identifying
new qualities of Latvia bred R. rugosa Hybrids by focusing on testing, if R. rugosa hybrids flowers are
suitable for production of rose water and, furthermore, rose oil, or their hips as a source of vitamin C.
From 10 cultivars volatiles of rose petals and their rose water were extracted using solid phase
microextraction (SPME) with subsequent separation by gas chromatography and identification by
comparison of their mass spectra with mass spectral libraries (Nist98). Content of ß-carotene and
ascorbic acid was detected spectrophotometrically in R.rugosa and six Rugosa Hybrid rose hips.
Fifty volatile aroma compounds were identified in petals. Among those, phenylethylalcohol,
ß-citronelol and nerol were predominant, but the composition of compounds varied among varieties.
Species Rosa rugosa and ‘Plena’ showed the highest total level of volatiles and contained 26%
and 31% ß-citronelol, respectively, and 17-19% of phenylethyl alcohol. ‘Raita’ produced a smaller
total amount of volatiles, but a higher proportion of ß-citronelol (ca. 44%) and phenylethyl alcohol
(ca. 30%). Main volatile compound were detected in rose water in the same proportions, but
their concentration was twice as high as in petals. The best source of ascorbic acid were cultivars
‘Schneezwerg’ and ‘Raita’ containing ca. 15 mg/g fruits.
In summary, cultivar ‘Raita’ is recommendable for use as a decorative plant, as a source of volatile
oil or rose water and as a fruit plant - source of ascorbic acid. Thus, it could be a good garden rose
plant for allotment gardeners. Wild populations of R.rugosa are suitable for production of rose water
and nutrients.
66
Poster
Poster Session 7: Growth Physiology and Production/Propagation
The Recent Status of Rose Research and
Production in Iran
M. Mirzaei and N. Ahmadi*
Department of Horticultural sciences, Tarbiat Modares University, P. O. Box 14115-336, Tehran,
Iran
*E-mail address: [email protected]
The value of rose (Rosa spp.) plants as an important ornamental and fragrant plant is demonstrated
by the large amount of scientific research reports on virtually all aspects of these plants reported
worldwide. Having a variety of favorable climate conditions and four distinguished seasons, the area
under ornamental plant production in Iran is considerable and rose accounts for the largest volume
of flower production consumed as cut flowers or by the rose oil extraction industry in this country.
Iran is famous for its organic rose essential oil obtained exclusively from fresh gathered Damask rose
flowers (Rosa damascena Mill.). Iran is also famous for producing rose water which is known for its
healing properties and used in folk medicines. In the present paper, we represent recent findings on
rose researches in Iran in different fields of investigation including rose propagation and cultivation,
post-harvest management, genotyping, pest and disease management. This paper includes some
trade information relating to market of fresh and processed products of rose plants over the most
recent years. This review would serve as reference material for interested groups such as producers,
consumers, investors and researchers who are concerned and interested about the issue of rose
research and production in Iran.
67
Poster Session 7: Growth Physiology and Production/Propagation
Poster
Evaluation of Rose Flower Production in
Taif, Saudi Arabia
Fahed A. Al-Mana and Yahia A. Ahmad
Plant Production Department, College of Food & Agricultural Sciences, King Saud University, P.O.
Box 2460 Riyadh 11451, Saudi Ar
[email protected]
The city of Taif in Saudi Arabia is popular for its great cultivation of roses the most common of which
is known as the Taif Rose (Rosa damascena trigintipetala), a Damask type rose. Taif Rose is mainly
produced for its aromatic oil (attar) which is considered the best quality perfume in the Middle East
such that it has important economic value in Saudi Arabia. Taif roses are mostly cultivated on about
2000 farms located in the suburbs and uplands of Taif city and its associated valleys. This study
is aimed to evaluate the flower production of Taif roses grown on farms at various locations and
mainly cultivated and harvested for rose oil and rose water. Five farms were selected for the study
and various grown rose species and varieties were investigated. Plant and flower characters such as
shrub height and diameter, number of branches, flower stem length and diameter, number of flowers
and petals, and essential oil content were evaluated. The study showed that there is a significant
difference in flower production among the studied farms. Flower production also varied among
the various rose species and varieties. It was observed that the main Taif Rose is Rosa damascena
trigintipetala. However, there are other Damask type rose varieties such as Autumn Damask Rose
(R.X damascena semperflorens) and Summer Damask Rose (R.X damascena damascena). Taif Rose
grows as a small (1.5 m) thorny bush with whitish hairy leaves and pink 30-petal flowers which
have a strong fragrance. This main Taif rose had the most intensely fragrant flowers which produced
precious oil. The price of one tolah (about 1/3 oz) of attar is usually sold for $ 530- $ 800.
68
Poster
Poster Session 7: Growth Physiology and Production/Propagation
EFFECT OF SEED COAT SCARIFICATION,
MACERATION AND STRATIFICATION ON SEED
GERMINATION IN GARDEN ROSES
Conev, Rumen, and Parminderjit Sandhu,
Vineland Research and Innovation Centre, Department of Applied Genomics, 4890 Victoria Ave. N.,
Box 4000, Vineland Station, ON, L0R 2E0, Canada
Email: [email protected]
Many landscape roses have poor and erratic seed germination, which poses a significant challenge
to hybridizers. A series of multi-factorial replicated experiments were conducted in Vineland, Canada
since 2010 to study the effect of seed coat scarification, maceration, and warm and cold stratification
on seed germination in garden roses.
The response of 11 cultivars of garden roses to different seed treatments (T) was studied in a pilot
experiment in 2010-2011. Seeds from ‘Champlain’, ‘Morden Fireglow’, ‘Morden Blush’, ‘Morden Snow
Beauty’, ‘Morden Sunrise’, ‘Winnipeg Parks’, ‘Parkwood Scarlet’, ‘Carefree Spirit’, ‘Cape Diamond’,
‘Lupo’ and ‘Night Owl’ were subjected to the following 6 treatments (3 replications/ treatment, 50
seeds/replication): T1-6 weeks cold stratification (WCS); T2-scarification with 50% H2SO4 (SSA)
for 30 sec followed by 6WCS; T3-SSA for 60 sec followed by 6WCS; T4-3 weeks warm stratification
(WWS) followed by SSA for 60 sec followed by 6WCS; T5-3WWS followed by seed coat maceration
with 0.5% cellulase for 36 hours followed by 6WCS; T6-9WWS followed by 12WCS. After the
treatments, seeds were sown in potting mix and germination was recorded for 17 weeks. Morden
Snow Beauty, Morden Sunrise and Carefree Spirit did not respond to any treatment. 6WCS (T1)
did not induce germination in any genotype. Scarification followed by 6WCS (T2 and T3) slightly
increased germination in Parkwood Scarlet and Lupo. Adding 3WWS (T4) provided modest benefit
only in Morden Blush, which along with Cape Diamond had max germination response (31% and
16% respectively) in T5. The combination of extended exposure to warm and cold stratification (T6)
secured the best germination in 6 out of 11 genotypes (up to 52% in Lupo).
In order to further investigate the effect of the duration of the exposure to, and the concentration of
sulfuric acid and cellulase, seeds from the cultivar Morden Blush were subjected to 9 treatments in
2011-2012, as follows: T1-SSA for 1 min; T2-SSA for 30 min; T3-SSA for 60 min; T4-0.5% cellulase
for 30 h; T5-1% cellulase for 30 h; T6-0.5% cellulase for 40 hours; T7-1% cellulase for 40 hours;
T8-0.5% cellulase for 50 hours; and T9-1% cellulase for 50 hours. All treatments (3 replications
per treatment, 50 seeds each) were followed by 48 hours leaching in water followed by 5 weeks
warm and 12 weeks cold stratification. After the treatment, seeds were sown in potting mix and
data were recorded for 12 weeks. Seed coat maceration significantly increased germination (32 %49%) compared to seed scarification (15%-17%). Macerating the seed coat with 0.5% cellulase for
50 hours produced the best germination response among the studied pre-stratification treatments
Detailed experiments on various regimes of warm and cold stratifications are currently underway.
69
Poster Session 7: Growth Physiology and Production/Propagation
Poster
EFFECT OF PRUNING SEVERITY ON LANDSCAPE
ROSES
Downer, James1, Karlik, Jan2, and Anna Howell1
University of California, Cooperative Extension 669 County Square Drive, suite 100, Ventura, Ca
93003
2
University of California Cooperative Extension, 1031 South Mount Vernon Avenue, Bakersfield, CA
93307
1
Roses grown in landscapes typically receive a pruning treatment after bloom or during the dormant
period of winter months. Rosarians, horticulturists and gardeners have adopted a variety of paradigms
for pruning roses. While some advocate pruning close the ground, others leave a large structure each
year. Despite a wealth of anecdotal information, little research has been conducted on pruning roses
grown in landscape settings. In our study we selected eight varieties, four hybrid tea roses, two
own-root roses, and two floribunda rose varieties as pruning subjects. Pruning treatments consisted
of four treatments in increasing severity. These were unpruned, leave 5 canes at 91cm, leave 4
canes at 46cm, and leave 3 canes at 15cm. Each spring, flowers were counted, stem lengths of
selected varieties measured and overall quality rated. While pruning severity decreased quantity of
blooms, it did not have a measurable effect on stem length of hybrid tea roses or overall quality in
most varieties and in most years. There were significant differences between varieties in response to
pruning severity.
70
Poster
Poster Session 7: Growth Physiology and Production/Propagation
EFFECT OF DIFFERENT IRRIGATION REGIMES
AND FREQUENCIES ON YIELD AND QUALITY OF
HYDROPONIC CUT ROSES
Fascella, Giancarlo, Giovanni Gugliuzza, Michele Massimo Mammano and Pierantonio Maggiore
Consiglio per la Ricerca e la Sperimentazione in Agricoltura, Unità di Ricerca per il recupero e la
valorizzazione delle specie floricole mediterranee (Research Unit for Mediterranean Flower Species),
S.S.113 - Km 245.500, 90011 Bagheria (Palermo), Italy.
As irrigation management can influence crop response and water use, studies on the effect of
irrigation regime and frequency on soilless cultivations may help to reduce water consumption and
production costs. With this objective, two greenhouse experiments were carried out on cut roses (cv.
Red France) grown in a closed hydroponic system.
In the first one, three irrigation regimes [high (HIR), medium (MIR) and low (LIR)] were tested: every
regime was characterized by the same duration of deliveries (3 minutes each) which were performed
daily when a prefixed level of accumulated radiation (300, 600 and 900 kJ/m2) was reached to
get to different total water supplies. Results showed that LIR was more effective for hydroponic
cut roses and more suitable for low-input irrigation management as evidenced by no significant
differences on flower yield (average 15.5 stems/plant), quality and biomass production than MIR and
HIR but showed lower water supply (0.5 vs. 0.8 and 1.3 L/plant/day, respectively) and higher water
use efficiency (1.8 vs. 1.1 and 0.7 g/L, respectively).
In the second trial, the same water supply was daily distributed to plants during different time
intervals: three durations of deliveries (1.5, 3 and 6 minutes) were differently performed [4, 2 and 1
time/day, – high (HIF), medium (MIF) and low irrigation frequency (LIF), respectively] in order to reach
an equal water amount for all treatments. Irrigation frequency did not influence cut rose production
(average 15.0 stems/plant) but only quality traits (higher stem length and weight recorded with HIF)
as well as WUE with higher values obtained on HIF plants.
71
Poster Session 7: Growth Physiology and Production/Propagation
Poster
Effect of rates and application times of
gibberellic acid on ‘Osiana’ cut rose
Grossi, José Antonio Saraiva1, Milanez, André Malacarne1, Tufik, Camila Borges Antonio1, Rodrigues,
Ernesto José Resende2, Barbosa, José Geraldo1, Finger, Fernando Luiz1
Plant Science Department, Federal University of Viçosa, 36570-000, Viçosa, Minas Gerais, Brazil
Federal Institute Technologic, 38.400-970, Uberlândia, Minas Gerais, Brazil
1
2
Rose is an important cut flower commercialized in the Brazilian market. With the goal to evaluate the
quality of flowering stems as influenced by the number of gibberellic acid applications, the cultivar
Osiana was grown under greenhouse conditions during the summer season in Viçosa, Brazil. The
experiment was arranged in a random block design with five rates of gibberellic acid (0, 1000, 2000,
3000 and 4000 mg/L) sprayed 1, 2, 3 or 4 times on young rose shoots from 6 to 12cm in length. Five
replicates of each treatment were used. Gibberellic acid treatment decreased the flower diameter
regardless of application time. Flower diameter at 0 and 1000 mg/L rates of gibberellic acid were
110.6 and 94.8 mm, respectively. When treated with 1000 mg/L gibberellic acid, flower peduncle
length was 96% longer than the control treatment regardless of application times. Also the length,
fresh and dry weight of flower stems increased in response to gibberellic acid treatments regardless
of application times. At the 1000 mg/L gibberellic acid rate, flower stems were 48% longer than the
control treatment. Increasing the gibberellic acid rate or application time decreased the leaf number
per stem. The control treatment exhibited 10 leaves per flowering stem; however, the 4000 mg/L
gibberellic acid rate at 4 application times exhibited only 3.4 leaves per flowering stem.
72
Poster
Poster Session 7: Growth Physiology and Production/Propagation
Growth and development of rose
cultivars influenced by gibberellic acid
rates
Grossi, José Antonio Saraiva1, Milanez, André Malacarne1, Tufik, Camila Borges Antonio1, Rodrigues,
Ernesto José Resende2, Barbosa, José Geraldo1, Finger, Fernando Luiz1
Plant Science Department, Federal University of Viçosa, 36570-000, Viçosa, Minas Gerais, Brazil
Federal Institute Technologic, 38.400-970, Uberlândia, Minas Gerais, Brazil
1
2
This experiment was carried out to evaluate the growth and flower development of cut rose cultivars
Grand Gala, Carola and Greta as affected by rates of gibberellic acid. The experiment was established
in random blocks with five rates of gibberellic acid (0, 1000, 2000, 3000 and 4000 mg/L) sprays and
five replicates. Gibberellic acid rates were weekly sprayed when the new shoots initially exhibited
from 6 to 12cm in length. The rate of 1000 mg/L gibberellic acid increased the flowering stem length
by 55% on Carola in comparison with the control treatment. The average flowering stem length of
Carola, Grand Gala and Greta were 108.6, 79.4 and 76.8cm, respectively, at 46 days after the first
spray application. Carola and Grand Gala cultivars exhibited higher stem diameter and greater fresh
and dry leaf weight than Gretaplants. Gibberellic acid increased the flower peduncle length but
decreased the flower diameter of all rose cultivars. Gibberellic acid did not affect the flower peduncle
diameter. The Grand Gala cultivar exhibited the largest flower peduncle diameter.
73
Poster Session 7: Growth Physiology and Production/Propagation
Poster
STRATOSPHERIC OZONE DEPLETION AND
CALIFORNIA’S OUTDOOR PRODUCTION: A
SURPRISING LINKAGE
Karlik, Jan1 and Mark Sabin2
University of California Cooperative Extension, 1031 S. Mt. Vernon Avenue, Bakersfield, California,
93307 USA
2
Agricultural Commissioner’s Office, County of Kern, 1001 S. Mt. Vernon Avenue, Bakersfield, California, 93307 USA
1
Stratospheric ozone depletion is a significant environmental problem. The role of long-lived chlorinecontaining compounds in promoting ozone destruction was recognized beginning in the 1970s,
leading to international policy agreement to limit their use beginning with the Montreal Protocol
in 1987. Quantification of ozone-depleting potential of various compounds and further research
led to recognition of bromine as a halogen with greater ozone depletion potential than chlorine,
and methyl bromide has been the most important anthropogenic source of atomic bromine in the
atmosphere. Since the U.S. was a signatory to the Montreal Protocol and subsequent amendments,
methyl bromide was listed for phaseout in year 2000. However, methyl bromide has been a mainstay
of outdoor rose production in the major growing area of Kern County, California. Research in Kern
County and elsewhere did not identify a ready replacement for that compound. Through California
total pesticide reporting, we present data for the amount of methyl bromide used in Kern County
against the global budget for that compound and its diminished use in roses, and suggest the loss of
methyl bromide has played a key role in the decline of California’s outdoor rose production industry.
In addition, the loss of methyl bromide provides an excellent case study of the interaction of science
and policy, as well as highlighting the risk of technology-forcing when no suitable alternative can
be found.
74
Poster
Poster Session 7: Growth Physiology and Production/Propagation
PROPAGATION OF ROSA BEGGERIANA
‘POLSTJÅRNAN’ WITH LEAF-BUD CUTTINGS
USING BIOSTIMULANTS
Monder, Marta Joanna1, Pacholczak Andrzej2, Kozakiewicz Paweł3, Niedzielski Maciej4, Jankowska
Agnieszka3, Woliński Konrad4
Polish Academy of Science Botanical Garden – Center for Biological Diversity Conservation in
Powsin, Department of Botanical and Garden Collection, Prawdziwka 2, 02-973 Warsaw, Poland,
[email protected]
2
Warsaw University of Life Science, Department of Ornamental Plants, Nowoursynowska 159, 02776 Warsaw, Poland
3
Warsaw University of Life Science, Department of Woody Science and Wood Preservation, Nowoursynowska 166, 02-787 Warsaw, Poland
4
Polish Academy of Science Botanical Garden – Center for Biological Diversity Conservation in
Powsin, Department of the Evaluation and Plant Biodiversity Conservation, Prawdziwka 2, 02-973
Warsaw, Poland.
1
Historical roses are valued for their high resistance to unfavourable growth conditions and frost, as
well as maintaining biodiversity and heritage of garden plants. This research aimed to specify the
possibility of effective propagation of them with the use of an environmentally friendly method.
The developmental phase of shoots and the use of biostimulants from seaweed were studied for
effects on rooting and on the content of biologically active components. Leaf-bud cuttings were
taken from shoots 1) with flower buds closed; 2) flowers open; 3) just after petal fall; and 4) 7-14
days after petal fall. Cuttings were rooted using 14 treatment combinations including an untreated
control, treatment with standard rooting stimulator powders containing 0.4 % IBA and 0.2 % NAA,
or watered with solutions of different concentrationsof BioRhizotonic, RootJuice, or BioRoots. Two
substrates were used: peat-perlite and peat-sand. The evaluation included the percentage of rooted
cuttings, their weight and quality of the root system, total content of proteins, chlorophyll a/b
content, carotenoid content, polyphenolic acid content, reducing and total carbohydrate content,
SDS-page analysis, and analysis of the anatomical structure of shoots in the place of rhizogenesis.
The highest percentage of rooted cuttings was obtained from shoots with flower buds closed (85.63
%; peat-sand 91.79 % - peat-perlite 79.46 %), and the lowest – 7-14 days after flowering (56.07
%). The use of biostimulants had a positive influence on the percentage of rooted cuttings and
their weight and quality of roots. It also slightly inhibited the decrease in the content of proteins,
polyphenolic acids and carbohydrates in leaves which was taking place during the process of rooting;
on the other hand, the decrease in the content of dyes was intensified. A lower percentage of rooted
cuttings and a particularly large callus at the excision site on the cutting appeared after NAA was
used. The callus grew toward the inside and the core tissues were stretched, thus creating empty
spaces, especially in cuttings from the last developmental phase when using RootJuice and BioRoots.
The biostimulants under examination, in appropriate concentrations, can be successfully used to
improve rooting of Rosa ‘Polstjårnan’ cuttings. The research was financed by the National Science
Centre (Poland) project NN 310008240.
75
Poster Session 7: Growth Physiology and Production/Propagation
Poster
GROWTH AND EARLY YIELD OF STENTINGPROPAGATED CUT ROSES AFFECTED BY
ROOTSTOCK
Park, Yoo Gyeong1 and Byoung Ryong Jeong1,2,3
Department of Horticulture, Division of Applied Life Science (BK21 Program), Graduate School of
Gyeongsang National University, Jinju 660-701, Republic of Korea
2
Institute of Agriculture & Life Science, Gyeongsang National University, Jinju 660-701, Republic of
Korea
3
Research Institute of Life Science, Gyeongsang National University, Jinju 660-701, Republic of
Korea
Corresponding autor: T. 055-772-1913; F. 055-757-7542; [email protected]
1
A study was conducted to investigate the effect of rootstock on growth and early yield of stentingpropagated cut roses (Rosa hybrida Hort.) ‘Pink Aurora’ and ‘Yellow King’. The scions, prepared as
single node cuttings, each with a five-leaflet leaf, were grafted on cuttings of the various rootstocks
Rosa indica ‘Major’, Rosa multiflora ‘Wonye No. 1’, Rosa multiflora ‘K-1’, or Rosa ‘Burr’. All leaves
and buds were removed from the rootstock cuttings. Both the base of the scions and the top of the
rootstocks were held together and simultaneously cut at a 45° angle for grafting. Scion-rootstock
unions were planted in rockwool cubes and were placed in a graft-take chamber for five days before
being placed on a misted greenhouse bench. Scion-rootstock unions were rooted in rockwool cubes
before being transplanted into rockwool slabs. In ‘Pink Aurora’, stem length, stem diameter, fiveleaflet leaves per stem, and stem fresh weight were not affected by the rootstock. The greatest total
yield of ‘Pink Aurora’ was obtained in plants grafted on the Rosa indica ‘Major’ rootstock. Overall
growth of ‘Yellow King’ was the greatest in plants grafted on Rosa ‘Burr’ rootstock. Total yield of
‘Yellow King’ was not affected by the rootstock. These results suggest that Rosa indica ‘Major’ is the
most effective rootstock.
This study was carried out with the support of “On-Site Cooperative Agriculture Research Project
(Project No. 006330)”, RDA, Republic of Korea. “Yoo Gyeong Park was supported by a scholarship
from the BK21 Program, the Ministry of Education and Human Resources Development, Republic of
Korea.”
76
Poster
Poster Session 7: Growth Physiology and Production/Propagation
Difficult-to-graft Rosa genotypes from
the Europa Rosarium Sangerhausen –
Comparison of different propagation
methods by cuttings and budding on two
rootstocks
Winkelmann, Traud1, Alesja Busch1, Katrin Kluge1, Simon Richartz1 and Thomas Hawel2
Gottfried Wilhelm Leibniz Universität Hannover, Institute of Floriculture and Woody Plant Science,
Herrenhäuser Str. 2, D-30419 Hannover, Germany
2
Europa Rosarium der Stadt Sangerhausen, Steinberger Weg 3, D-06526 Sangerhausen, Germany
1
The Europa Rosarium Sangerhausen owns the largest collection of roses (8,300 accessions) worldwide
and is part of the German genebank for ornamental plants. Maintaining the species and cultivars is
achieved mainly by budding on Rosa corymbifera ‘Laxa’, but results in high losses in some genotypes.
Moreover, the collection of Rosa species is aspired to be propagated on own roots in order to see the
true species habit. Thus, the objective of this project was to compare bud-grafting on two different
rootstocks (pentaploid Rosa corymbifera ‘Laxa’ and diploid Rosa multiflora seedlings) with cutting
propagation.
62 genotypes, including Rosa species as well as rose cultivars and representing 23 rose classes,
were selected for this experiment, 25 of which showed severe losses in graftings of previous years.
Budding was carried out with 52 genotypes in August 2012 with 2-3 buds per genotype-rootstockcombination, both in Sangerhausen under open field conditions and in Hannover in container-culture
followed by overwintering in a frost-free PE-tunnel. At the same time for 48 of the 62 genotypes,
5-20 cuttings, depending on the amount of available material were harvested. They were 20-50 cm
in length and were set in peat substrate with 2 kg m-3 slow release fertilizer (Osmocote exact 5-6 M)
under high-pressure fog.
A first evaluation of grafting success in March 2013 revealed no difference in the overall numbers
of successful buddings for both rootstocks. In Hannover, for 16 out of the 52 genotypes propagation
by grafting failed, while the data for Sangerhausen will be collected in late April for the first time.
Rooting was observed in all genotypes except one, although in different frequencies and qualities.
Detailed evaluations will follow once in April and once in July 2013 combined with histological
analyses of root formation and the grafting units.
77
Poster Session 8: Abiotic Stress
Poster
EFFECTS OF AIR HUMIDITY AND AIR
MOVEMENT ON GROWTH, VISUAL QUALITY AND
POSTHARVEST STRESS TOLERANCE OF POT ROSE
Dália R.A. Carvalho1, Sissel Torre2, Marta W. Vasconcelos1, Domingos P.F. Almeida3, Ep Heuvelink4,
Susana M.P. Carvalho1,4
CBQF - Centro de Biotecnologia e Química Fina – Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa/Porto, Rua Dr. António Bernardino Almeida, 4200072 Porto, Portugal
2
Department of Plant and Environmental Sciences, Norwegian University of Life Sciences, PO Box
5003, 1432 Aas, Norway
3
Faculty of Sciences, University of Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal
4
Department of Plant Sciences, Horticultural Supply Chains Group, Wageningen University, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands
1
High relative air humidity (RH > 85%) during growth reduces postharvest life of roses due to
unfunctional stomata and water stress. Since several plant species respond to mechanical stimuli
by closing stomata, the objective of this work was to assess if a high level of air movement would
improve stomatal closure, counteracting the negative effect of high RH on postharvest quality.
Additionally, its impact on plant growth, visual quality and postharvest stress tolerance to dark and
drought (simulating shipping and/or retailing) was evaluated.
Pot rose ‘Toril’ were grown in controlled growth chambers under moderate (60%) and high (90%) RH,
combined with no additional air movement or with a continuous additional air velocity of 0.92 ± 0.03
ms-1 throughout plant development. At flowering stage, plants were transferred to darkness under
drought conditions at 20 °C and 40% RH to evaluate postharvest stress tolerance and storability.
High RH during growth significantly increased plant height (9%), whereas flower dry weight, flower
diameter and peduncle diameter were significantly reduced (13%, 4% and 12%, respectively). High
air movement during growth did not affect plant height nor flower size, but it significantly increased
the peduncle diameter by 6%. Total dry weight and time to flowering were not significantly affected
either by RH or air movement. During the postharvest storage period, plants grown at elevated RH
increased their transpiration rate by 4.7-fold, but the air movement during growth had no effect
on this parameter. Air movement enhanced storability from 6 to 8 days in plants grown at high RH
while it decreased from 26 to 23 days in plants grown at moderate RH. In conclusion, high RH has a
minimal effect on plant growth and visual quality parameters but it drastically reduces postharvest
stress tolerance. High air movement during growth improved stress tolerance in high RH-grown
plants and it had a minor effect on visual quality.
78
Poster
Poster Session 8: Abiotic Stress
Screening Methods for Drought
Resistance in Rosa
Katrien De Dauw1, Leen Leus2, Johan Van Huylenbroeck2, Marie-Christine Van Labeke1
Department of Plant Production, Ghent University,
Coupurelinks 653, 9000 Gent, Belgium
2
ILVO (Institute for Agricultural and Fisheries Research),Plant Sciences Unit
Caritasstraat 21, 9090 Melle, Belgium
1
Drought stress severely impacts plant production worldwide and is predicted to increasingly affect
crop production due to climate change. The challenges presented hereby require solutions fitting
in a sustainable and modern horticulture. One of the main strategies to cope with the changing
environment is the development of new cultivars with an improved resistance against drought
stress. This study aims at applying polyploidy in this process, through the comparison of drought
stress resistance in diploid Rosa genotypes, originating from a cross between R. ‘Yesterday’ x R.
wichurana, and their mitotically induced tetraploid counterparts. The first step in this research is
the development of adequate screening methods for the characterisation of the diploid segregating
rose population. After a first broad screening (69 F1-hybrids), based on ecophysiological parameters
including relative water content and leaf water potential, we selected a drought tolerant and a
drought sensitive diploid genotype. These genotypes were then subjected to two different irrigation
regimes, reducing the amount of water received by the drought stress group to less than 10% of the
optimal amount given in the control group. Substrate volumetric water content, stomatal resistance
and leaf water potential were monitored during the experiment. Biochemical analyses on leaf material
included osmotic potential, soluble sugar content, proline content, total antioxidant content, total
protein content and enzyme (CAT, GPX, APX, SOD) activity. Results showed that both the drought
sensitive and tolerant genotypes responded to drought stress with increased stomatal resistance, a
more negative leaf water potential and accumulation of protein, soluble sugars and proline. In the
control group we also noticed higher initial proline levels in the drought sensitive genotype. Sucrose
and proline content and specific catalase activity during drought stress allowed a clear distinction
between both levels of drought tolerance in this experiment. In further experiments, diploids and
tetraploids will be compared.
79
Poster Session 8: Abiotic Stress
Poster
Plant growth and flowering trend
of landscaping roses under low
maintenance and xeriscaping.
M. E. Giorgioni
Dip.to di Scienze Agrarie, Alma Mater Studiorum, University of Bologna, Italy
The 6-year trial is a step of a wider and long research programme focused on the selection of rose
shrubs well suited to a low maintenance regime, without annual winter pruning, disease control
and fertilization and especially, in xeric conditions. The study was carried out in an open field in an
area located in North Italy, marked by hot and dry summers, with temperature up to 45 °C, yearly
rainfall around 550 mm and a xeric 2.5 month long period, from June to mid August. Floriferousness,
flowering trend from May to October, number of flower flushes, canopy height and diameter, covering
ground area of 40 cvs were investigated throughout 5 years after planting and on the 6th year, after
a mechanic, not selective pruning. Before pruning, ‘Cristal Fairy’, ‘Dolce Fairy’, ‘Galatea’, ‘Supernova’
(white flower), ‘Les Quatre Seasons’, ‘Neon’, ‘Siesta’, ‘Sweet Haze’ (pink flower), ‘Andromeda’, ‘Knock
Out’, ‘Palmengarten’ (dark pink flower) and ‘Cubana’ were the cvs with the widest plants, over 1 m2,
whereas ‘Florshow’ cvs and ‘Nipper’ the smallest, with an area under 0.5 m2. ‘Cristal Fairy’, ‘Toscana’,
‘Schneewittchen’, Knock Out’ and ‘Tremenda’ resulted the cvs with the highest flower Cover Index
(over 1) and ornamental value index (35-50), therefore with the most remarkable chromatic impact of
plants at maximum flowering time and the most prolonged flowering period, with several appreciable
peaks. Detailed data related to specific cultivars, 6 year growth and flowering trends are reported.
80
Poster
Poster Session 8: Abiotic Stress
DO PLOIDY LEVELS PLAY A ROLE IN (A)BIOTIC
STRESS RESISTANCE IN ROSES?
Leen Leus1, Katrien De Dauw2, Johan Van Huylenbroeck1 and Marie-Christine Van Labeke2
Institute for Agricultural and Fisheries Research (ILVO), Plant Sciences Unit, Caritasstraat 21, 9090
Melle, Belgium
2
Department of Plant Production, Ghent University,Coupure links 653, 9000 Gent, Belgium
1
Polyploidy is known to arise spontaneously in many plant species in nature. Frequently increased
chromosome copies are associated with changes in plant anatomy and morphology. In addition,
altered stress resistances in polyploids have been reported in ecological studies and in some
agricultural crops. Nevertheless knowledge on the effect of polyploidy on stress resistance is limited.
In our study rose is used as model crop to get better insight in the effects of chromosome doubling
on both biotic(powdery mildew) and abiotic (drought) stress resistance. Mitotic polyploidisation of
diploid roses isused to evaluate the influence of more genomic copies excluding the influence of
extra allelic diversity.
A diploid population (90 genotypes) resulting from the hybridisation of ‘Yesterday’ x Rosa wichurana
is used. In a first step the progeny was characterized for both drought and powdery mildew resistance.
Subsequently, 19 F1-hybrids and both parents were selected for chromosome doubling. Genotypes
were either susceptible or tolerant for drought and/or powdery mildew. Mitotic polyploidisation was
obtained using an oryzalin(5 µM) treatment in liquid medium on nodal sections of tissue cultured
plantlets.
Polyploidisation resulted in the first polyploid roses. Currently experiments to compare stress
resistance at the diploid and tetraploid level are started. Preliminary results on the effect of ploidy
level on plant morphology and on stress resistance in the selected rose genotypes will be given.
81
Poster Session 9: Rose Diseases and Pests
Poster
INSECTICIDAL EFFECT OF EXTRACTED SOLUTIONS
OF SEVERAL CONDIMENT VEGETABLES ON TWOSPOTTED SPIDER MITE OF ROSE
D.C. An1, J.G. Kim1, B.J. Lee1, C.W. Ro2, B.R. Jeong3
Flower Research Institute, Gyeongsangnam-do Agricultual Research & Extension Services, Changwon 641-923, Korea.
2
Research and Development Bureau, Gyeongsangnam-do Agricultual Research & Extension Services, Jinju 660-360, Korea
3
Institute of Agriculture & Life Science, Gyeongsan National University, Jinju 660-701, Korea
1
This study was conducted to develop an environment-friendly control method for the two-spotted
spider mite which is one of the most serious arthropod pests on rose crops in South Korea. Some
condiment vegetables, such as garlic, hot pepper, and onion, that have a strong spicy fragrance,
were blended with the same weight of water and filtered. These solutions were diluted with either
10, 20, 50, or 100 times with water. The solutions were sprayed to the rose crops infested with twospotted spider mites and the condition of the spider mites was observed under a microscope after 30
minutes. All garlic solutions except for the 100 fold dilution showed the strongest insecticidal effect
by melting the skin of all the adults, larvae, and eggs. The undiluted and the 50 fold garlic dilution
had an EC of 8.76 and 0.55dS·m-1 and pH of 6.3 and 7.0, respectively. Onion and pepper solutions
diluted up to 20 times also showed a similar effect. Physiological disorders observed three days after
spraying were some necrotic spots on the new shoots sprayed with the undiluted garlic solution.
It will be necessary to do further investigations on the extraction and safer treatment methods to
improve the control efficiency of the solutions on the spider mite in the future.
82
Poster
Poster Session 9: Rose Diseases and Pests
ROSE ROSETTE DISEASE: A COMPREHENSIVE
RESEARCH AND EDUCATION PLAN
Dobres, Michael1,2, Kelly Kemp1.2 and Christina Quinn1,2
NovaFlora LLC, 25 Lewis Rd, West Grove, PA 19390, USA
The Conard-Pyle Co., 25 Lewis Rd, West Grove, PA 19390, USA
1
2
Rose Rosette Disease (RRD) is an increasing threat to the garden rose industry in the USA. The disease
is characterized by deformed shoots and flowers that form a witches broom like structure. Roses
infected with RRD die within three years.
The disease has only been confirmed in North America, though scientific reports of similar witches
broom like infections in Europe and Asia exist. RRD was first identified in the Rockies in the 1940s
and spread through the Mid-West in the 1980s to the Eastern seaboard by the mid-1990s.
The primary host for RRD is Rosa multiflora. Distribution maps of RRD and R. multiflora indicate that
the geographic boundaries of RRD are largely set by the distribution of R. multiflora, though other
non-native species may also serve as hosts.
The putative causal agent of RRD is a negative strand RNA virus that is transmitted by an eriophyoid
mite (Phyllocoptes fructiphilus).
We have developed a 96-well RT-PCR assay based on published sequences of the RRD virus. We have
used this assay to screen more than 500 rose plants. We confirm a one hundred percent correlation
between symptoms and the presence of the virus. The virus is only detected in symptomatic leaves.
The scientific literature indicates that several native North American species are resistant to the
disease. We have begun a screening program to identify highly resistant lines and with the goal of
eventually breeding varieties with increased resistance to RRD and/or the eriophyoid mite.
We are forming a coalition of growers, breeders, extension agents and university scientists to help
combat the disease. Our initial focus is to educate growers and landscapers on how to recognize and
prevent the disease. Our secondary focus is to learn more about the disease and develop varieties
that are resistant to RRD.
83
Poster Session 9: Rose Diseases and Pests
Poster
CHARACTERIZATION OF ROSA SPP. BREEDING
POPULATIONS TO BLACK SPOT FOR QTL
IDENTIFICATION
Qianni Dong1,2, Dave Byrne2, Kevin Ong3, Xinwang Wang1,2
Texas A&M AgriLife Research and Extension Center, Dallas, TX 75252
Department of Horticultural Sciences, Texas A&M University, College Station, TX 77843
3
Department of Plant Pathology & Microbiology/ Bioenvironmental Sciences, Texas A&M University,
College Station, TX 77845
1
2
Black spot disease, caused by the fungus Diplocarpon rosae Wolf, is the most serious disease of
landscape roses (Rosa hybrid L.) worldwide. Dominant genes for complete resistance to specific races
of the pathogen were identified in roses as Rdrs. From a breeding perspective, a rapid screening of
potential hybrid materials by molecular markers is beneficial for identifying the resistant germplasm
efficiently. Although partial resistance has also been documented, the responsible QTLs remain
unidentified. In this project, responses to D. rosae of 16 genotypes of roses that were used as parents
in hybrids were characterized with two inoculation methods: the detached leaf assay (DLA) and
the whole plant inoculation (WPI) method. The correlation between the relative resistances among
genotypes as determined by each method was analyzed. Although DLA is more sensitive than WPI
in measuring relative resistance, the correlation among the two methods is high (r2 >0.8) which
indicates that either can be utilized to characterize D. rosae. Six diploid hybrid populations which
are segregating for strong partial resistance derived from Rosa wichurana have been planted in the
field for black spot evaluations. These plants were also measured by DLA for resistance to race 8. This
phenotypic data will be combined with the genotyping data for the populations identify QTLs for
partial resistance to black spot.
84
Poster
Poster Session 9: Rose Diseases and Pests
DOES PESTICIDE USE IN CALIFORNIA OUTDOOR
ROSES REFLECT IMPLEMENTATION OF IPM
PRINCIPLES?
Karlik, Jan1 and Mark Sabin2
University of California Cooperative Extension, 1031 S. Mt. Vernon Avenue, Bakersfield, California,
93307 USA
2
Agricultural Commissioner’s Office, County of Kern, 1001 S. Mt. Vernon Avenue, Bakersfield, California, 93307 USA
1
Production of outdoor rose plants in California has been centered in Kern County. Pesticide use patterns
are important for understanding progress and potential targets for integrated pest management
(IPM) or other pesticide reduction strategies, as well as providing background information for worker
safety, and for risk assessment as urban areas expand into agricultural hinterland. The University of
California has a statewide Special Program to foster dissemination of IPM knowledge and application,
including emphasis on non-chemical pest management methods where available, as well as use
of softer and bio-rational pesticides. California’s total reporting system for pesticides provides
specific geolocated pesticide use data for roses and other crops so that pesticide use patterns can
be analyzed. We investigated pesticide application data for outdoor rose fields from 1997-2010 to
see changes in pesticide use per ha and compare applied mass of older broad-spectrum pesticides
with applications of compounds more environmentally friendly. We note kg of active ingredient per
ha for major pesticide groups, with an emphasis on insecticides due to their relatively higher human
toxicities as compared to other pesticide types, and note trends in applications by chemistry type and
toxicity category. We then infer the trend toward IPM use in roses and opportunities for additional
implementation of IPM principles.
85
Poster Session 9: Rose Diseases and Pests
Poster
Reverse genetics strategies to obtain
broad spectrum powdery mildew
resistance in roses
Kaufmann, Helgard1, Juliane Wehmeyer1 and Thomas Debener1
Gottfried Wilhelm Leibniz Universität Hannover, Department of Molecular Plant Breeding, Herrenhäuser Str. 2, 30419 Hannover, Germany
1
Powdery mildew is a major disease of economic importance in cut and pot a rose which is caused
by the obligate biotrophic fungus Podospaera pannosa. As an alternative to conventional resistance
breeding strategies utilizing single-dominant genes or QTLs, mildew resistance locus O (MLO)-based
resistance might offer some advantages. In dicots such as Arabidopsis, pea and tomato, loss-offunction mutations in MLO genes confer high levels of broad-spectrum resistance. This resistance is
characterized by durability and recessive inheritance. We have isolated and characterized four MLO
homologues from a large rose EST collection isolated from leaves. These genes are phylogenetically
closely related to other dicot MLO genes that play a role in powdery mildew susceptibility. Therefore,
they are candidates for MLO genes involved in rose powdery mildew interactions. The aim of our work
is a reverse genetics approach to select rose plants that are homozygous for loss of function in one
or more MLO genes. From a collection of 960 rose cultivars and species MLO genes were amplified
and analyzed by high-throughput sequencing to discover rare mutations.
86
Poster
Poster Session 9: Rose Diseases and Pests
Analysis of potential effector genes in the
genome of Diplocarpon rosae
Klein, Enzo1; Marcus Linde1, Thomas Debener1
Gottfried Wilhelm Leibniz Universität Hannover, Department of Molecular Plant Breeding, Herrenhäuser Str. 2, 30419 Hannover, Germany
1
The immune response of plants consists of two major reactions, the PAMP-triggered immunity (PTI)
and the effector-triggered immunity (ETI). PAMP-triggered immunity is a general mechanism which
uses receptor proteins from the host to recognise conserved molecular patterns (PAMP = pathogenassociated molecular patterns) on the surface of a pathogen. In the case of the effector-triggered
immunity, effector proteins of the pathogen. which have the purpose to suppress the immune defence
of the host, are recognised through specific R-proteins (resistance gene products). This interaction
of R-genes and Avr-genes (effectors) is called the gene-for-gene model. Thus far all the molecular
analyses of the interaction between the pathogen Diplocarpon rosae and the rose has concentrated
on the host, resulting in the mapping of two R-genes, Rdr1 and Rdr3.
The present work focuses on the effector proteins of the pathogen, Diplocarpon rosae, since knowledge
about them can help to detect new R-genes and lead to a better understanding of the interaction
mechanism.
The sequence of the isolate DortE4 was used as a source of candidates for effectors genes. A gene
prediction was made and the resulting gene models were annotated. In combination with other
bioinformatic analyses, several candidate genes were identified. The expression of these genes
was tested by PCR in four different interaction systems, including compatible and incompatible
interactions, and at different developmental stages of Diplocarpon rosae. The present results indicate
that most of the genes are expressed in all investigated situations.
87
Poster Session 9: Rose Diseases and Pests
Poster
Development of Multiplex SSR-marker for
Diplocarpon rosae
Kohlenberg, Max1; Marcus Linde1 and Thomas Debener1
Gottfried Wilhelm Leibniz Universität Hannover, Department of Molecular Plant Breeding, Herrenhäuser Str. 2, 30419 Hannover, Germany
1
Black Spot, caused by the hemibiotrophic ascomycete Diplocarpon rosae, is one of the most
serious fungal diseases on roses worldwide. Due to the great economic impact of the disease the
characterization of the genetic diversity of D. rosae isolates from different locations all over the
world is of high interest.
Because of their relatively high frequency in the genome and the high mutation rates SSR markers
were considered as the method of choice for fingerprinting genotypes. As isolation of genomic DNA
of D. rosae samples is a limiting factor for the marker analysis, we tried to develop a multiplex PCR
assay. By next-generation-sequencing of the D. rosae isolate DortE4 over the last few years we
gained enough data to establish a number of D. rosae specific SSR primers. Three to five primer pairs
were pooled in different multiplex PCR approaches to display their polymorphic character with just
little PCR effort.
According to the PCR conditions, several critical parameters had to be considered. Especially annealing
temperatures and product sizes as well as the DNA template and dNTPs amounts were fundamental
for the success of the multiplex approach.
The application of these multiplex sets to mixed or pooled samples is possible as well as to investigate
the genetic diversity of D. rosae in general. Here, they could complement inoculation experiments,
which are still essential to specify D. rosae races.
88
Poster
Poster Session 9: Rose Diseases and Pests
ASSOCIATION MAPPING OF HORTICULTURAL
TRAITS IN ROSES
Montenegro Alonso, Ana Priscilla1, Dietmar Schulz1, Marcus Linde1 and Thomas Debener1
Gottfried Wilhelm Leibniz Universität Hannover, Department of Molecular Plant Breeding, Herrenhäuser Str. 2, 30419 Hannover, Germany
1
Understanding of the genetic architecture of economically important complex traits have eluded
plant breeders, mainly due to the fact that complex traits present quantitative patterns of
inheritance. One of these complex traits is resistance to black spot disease on roses, which is caused
by Diplocarpon rosae (Wolf) and is considered the most important disease affecting garden roses
worldwide. Currently, association mapping has become the alternative method to explain phenotypic
variation in terms of changes in the DNA sequence of an individual genome. This research was mainly
conducted to explore loci linked to the genes responsible for the variation of important horticultural
traits in roses through association mapping. Specifically, this study was developed with the intention
of (1) determine if the quantitative variation of black spot resistance is related to single genes in
the rose genome; (2) determine if genes of the Rdr1 family are associated with resistance to black
spot; and (3) determine if candidate genes encoding defense signaling proteins are associated with
resistance to black spot. In this study, 96 diverse cultivars of roses were evaluated phenotypically for
resistance to black spot through artificial inoculations and field evaluations in different locations
throughout Germany (North of Hamburg, West of Hannover and Hannover). The data collected was
associated to molecular markers (microsatellites and AFLPs) developed in parallel. Also, candidate
genes from the Rdr1 gene family (MicLRR SSR and RGA1) and the defense signaling pathway (NPR1,
EDS1, AOS, MPK4, EDS5, LSD1, HSP90.1) were developed and used to genetically analyze the cultivars
and correlate them to the disease and morphological data. Until now, differences in the phenotypic
reaction to two races of this pathogen (DoE4 and R6) have been observed between genotypes and
between environments. First association experiments indicate weak associations between candidate
genes encoding defense signaling proteins and disease resistance.
89
Poster Session 9: Rose Diseases and Pests
Poster
Functional Characterization of rose TNLgenes for resistance against Diplocarpon
rosae isolates
Peng, Weiyan1, Diane Prüß1, Diro Terefe_Ayana1 and Thomas Debener1
Gottfried Wilhelm Leibniz Universität Hannover, Department of Molecular Plant Breeding, Herrenhäuser Str. 2, 30419 Hannover, Germany
1
Black spot is a severe disease that leads to considerable yield losses in the cultivation of roses.
The disease is caused by the hemibiotrophic fungus Diplocarpon rosae. In contrast to the use of
fungicides, breeding for resistant cultivars is environmentally friendly and consumer-preferred way
of overcoming the disease. So far, Rdr1 is the most extensively studied resistance gene against black
spot disease in roses. The Rdr1 gene comprises a cluster of nine highly related TIR-NBS-LRR (TNL)
resistant gene analogs (RGAs). A preliminary transient expression study revealed two of the gene
clusters (RGA 1 and RGA8) as possible candidate genes. To verify the function, the two candidate
genes were transformed into the susceptible rose genotypes “Pariser charme” and further analysed.
Inoculations of the transgenic plants with multiple Diplocarpon rosae isolates indicated a universal
resistance against the races Dort E4, F004, D002, DÜA3 and S009 for the RGA1. The RGA8-transgenic
plants are however susceptible to all of the five isolates. Southern blot hybridization analyses revealed
a high copy numbers (more than 7) in the RGA1 transgenic plants and the same banding pattern for
all of the plants. In an additional inoculation assay, the RGA1 transgenic plants with a high gene
dosage (the multiple copy number) failed to confer resistance to the most aggressive Diplocarpon
rosae race 6.
90
Poster
Poster Session 9: Rose Diseases and Pests
ISOLATION AND CHARACTERIZATION OF MLO
HOMOLOGUES IN ROSE
Thal, Beate1; Helgard Kaufmann1 and Thomas Debener1
Gottfried Wilhelm Leibniz Universität Hannover, Department of Molecular Plant Breeding, Herrenhäuser Str. 2, 30419 Hannover, Germany
1
Powdery mildew is one of the most widespread plant diseases. Podosphaera pannosa, the causal
agent of rose powdery mildew, hampers the production of cut roses throughout the world. The
infection reduces the host vigor and lowers the aesthetic value. A major tool to control this disease
is the use of resistant plant material.
The biological functions of Mlo (mildew resistance locus o) proteins are mostly unknown, but it is
well established that the powdery mildew fungus is specialized on MLO proteins as an entry portal
into the host cell. MLO is an integral plasma membrane localized protein with seven transmembrane
helices. It belongs to gene families with 10- 15 members per higher plant species.
Mutants of Arabidopsis thaliana (A.t.), barley, tomato and pea without functional Mlo are resistant
to all tested isolates of powdery mildew. The role of Mlo as a susceptibility factor is not its main
function. A.t. Mlo gene family members AtMlo5 and 9 have been shown to be involved in the process
of pollen germination. AtMlo4 and 11 are associated with root development.
In the present work we characterized leaf expressed members of the rose Mlo family. 27 rose Mlo
homologues were identified in two EST libraries of untreated and “stressed” leaves. The sequences
were grouped in a phylogenetic tree together with AtMlo1- 15. Rh (Rosa hybrida) Mlo 1- 4 were
identified as AtMlo2, 6 and 12 homologues and have already been described. They are candidates
for powdery mildew susceptibility genes. Five additional rose Mlos were characterized for the first
time. They were mapped in the diploid rose population 97/7. In addition the tissue specificity of gene
expression of was analyzed.
Mlo in rose is an interesting but so far uncharted field. The characterization of Mlo genes is a first
prerequisite of the potential final aim to obtain loss-of-function mlo mutants in rose which is so far
unrivaled.
91
Poster Session 9: Rose Diseases and Pests
Poster
POWDERY MILDEW RESISTANCE IDENTIFICATION
IN WILD ROSA GERMPLASM
Xianqin Qiu1,2, Hao Zhang1,2, Ye Hua2, Hongying Jian2, Qigang Wang2, Huijun Yan2, Shubin Li2 and
Kaixue Tang2
College of Horticulture and Forestry, Huazhong Agricultural University/ Key Laboratory of Horticultural Plant Biology, Ministry of Education, 430070 Wuhan, China
2
Yunnan Flower Breeding Key Lab, Flower Research Institute, Yunnan Academy of Agricultural
Sciences, 650205 Kunming, China
1
Powdery mildew caused by Podosphera pannosa is a major disease of roses causing significant losses
especially in greenhouse production. The powdery mildew resistance of wild Rosa species was assessed
to determined their usefulness as sources of resistance for rose disease-resistant breeding. In this
research, 50 accessions from nine botanical sections including nine ancestor species of modern roses
and ten endemic species to China were evaluated by isolated identification and field identification.
The results of both methods were mostly consistent. The accessions of R. laevigata, R. longicuspis,
R. luciae, and R. banksiae that were evaluated were immune to Podosphera pannosa. High levels
of powdery mildew resistance was found among the accessions of the Section Pimpinellifoliae, R.
rubus, and R. rugosa among others. Fifteen others e.g. R. lichiangensis, R. gallica, R. bracteata were
moderately resistant to this pathogen. However, accessions of eleven species including those in the
Section Chinenses, R. praelucens, and R. centifolia, were highly sensitive. The results indicated that
the Section Pimpinellifoliae, Laevigatae and Banksianae contain higher resistance to Podosphera
pannosa than the others. The resistant species were characterized either by thick and shiny young
leaves or by rugose young leaves with prominent veins. Moreover, the Section Chinenses was the
most susceptible section. The study will provide useful information for utilizing these resistant species
in rose disease-resistant gene isolation and new cultivar breeding.
92
Authors index
Ahmadi, N. 67
Al-Mana, Fahed A. 68
Alp, Şevket 41, 42, 43, 65
An, D.C. 82
Balode, Antra 53
Boronkay, Gábor
62
Boyd, Peter2
Bradeen, James M. 17
Byrne, David H. 6, 27, 32, 51, 84
Carvalho, Dália R.A. 31, 78
Chao, Yu44
Conev, Rumen69
De Riek, Jan 19
De Vries, D.P.
13
Debener, Thomas
33
Dobres, Michael
83
Downer, James70
Dubois, Annick 56
Fascella, Giancarlo
Foucher, Fabrice 71, 3
15
Giday, Habtamu Giorgioni, M. E. Giovannini, Annalisa Greyvenstein, Ockert
Grossi, José Antonio Saraiva
Grout, Brian 7
80
46, 54
47
73
36
Hibrand-Saint Oyant, Laurence 8
Hokanson, Stan C. 52
Horibe, Takanori 29
Jeong, Byoung Ryong 28
Kanda, Momoyo 25
Karlik, Jan
74, 85
Kaufmann, Helgard
86
Kawamura, Koji18
Kim, Seung Tae 63
Kim, Wan Soon 23, 64
Klein, Enzo87
Kohlenberg, Max
88
Lee, Su Young Leus, Leen Li, Shubin 57
34, 81
5
Liu, QingLin 20, 58
Mibus, Heiko 59
Mollov, Dimitre37
Monder, Marta Joanna
75
Montenegro Alonso, A.P.
89
Nybom, Hilde 12
Otagaki, Shungo 9
Park, Yoo Gyeong
76
Paudel, Dinesh Babu
48
Pemberton, H. Brent
26
Peng, Weiyan90
Rachid, Boumaza Resende, Ernesto José 45
72
Schmitzer, Valentina Schulz, Dietmar Frank
Smulders, M.J.M. Sochacki, Dariusz Sparinska, A. 14
21,5
16
55
66
Tang, Kaixue 4, 10, 92
Terefe-Ayana, Diro
60
Thal, Beate91
Tzanetakis, Ioannis E. 35, 38
Vainstein, Alexander
24
Van Labeke, Marie-Chiristine 30, 49, 79
Wehmeyer, Juliane
61
Winkelmann, Traud
39, 77
Wissemann, Volker
1
Woltering, Ernst J. 22
Zlesak, David C.
11
Notes
AR 2
S2
Tarifzonen im GVH
Fare zones
Stufenloser Zu-/Abgang
Step-free access
Park+Ride
Paderborn
S5
R13
Bünde
R1
Bielefeld
Rheine
AR 1
Emmerthal
Hessisch Oldendorf
Bad Pyrmont
S1
Minden
Lindhorst
AR 2
Stadthagen
Kirchhorsten
Bückeburg
www.gvh.de
Stand: Dezember 2012
Fare zones for cards
of the GVH-regional fare
S1
S2
Hameln
Weetzen
H-Bornum
Empelde
Ronnenberg
Empelde 9
Ahlem 10
Bad Münder
Springe
Völksen / Eldagsen
Bennigsen
7
H-Linden / Fischerhof
10 17
AR 3
AR 2
AR 1
2
Laatzen
Osterwald
Fasanenkrug
4
Schierholzstraße
Roderbruch
7
Altwarmbüchen
Rethen
5
Anderten
Sarstedt
12
Göttingen
Alfeld
Freden
AR 3
R 13
Nordstemmen
Elze (Han)
Banteln
R12
R
Barnten Emmerke
Lehrte
AR 2
AR 2
AR 1
Sarstedt
1
Messe /Ost (Expo-Plaza)
6 16
Rethen (Leine)
2
Messe / Nord
R 11 R12
1
R12
9
Alte Heide
3
AR 1
H-Anderten / Ahlten
H-Kleefeld H-KarlWiechert-Allee Misburg
8 18
Döhren /Bhf.
Hannover
Messe/Laatzen
2
H-Bismarckstr.
Hannover Hbf
Zoo
8 18
11
R1
S6
S7
S3
Langenhagen
TICKETZONE HANNOVER
Voldagsen
TICKET-/CARDZONE REGION
TICKET-/CARDZONE UMLAND
Laatzen/Eichstraße
(Bahnhof)
CARDZONE HANNOVER 2
Wallensteinstr.
Wettbergen
Coppenbrügge
3
17
H-Nordstadt
1
TICKET-/CARDZONE UMLAND
Isernhagen
Großburgwedel
TICKET-/CARDZONE REGION
Regional- und S-Bahn
CARDZONE HANNOVER 2
CARDZONE HANNOVER 1
H-Ledeburg
H-Vinnhorst
Langenhagen-Mitte
Langenhagen-Pferdemarkt
Langenhagen-Kaltenweide
Bissendorf
Mellendorf
S4 Bennemühlen
Schwarmstedt
Lindwedel
Hodenhagen
Walsrode
Aegidientorplatz
Kröpcke
Steintor
Königsworther Platz 16
Haltenhoffstr. 11
Nordhafen 6
Stöcken
TICKETZONE HANNOVER
Wennigsen Lemmie
(Deister)
Holtensen /Linderte
Egestorf (Deister)
Kirchdorf (Deister)
Barsinghausen
Winninghausen
Bantorf
Bad Nenndorf
Haste
H-Leinhausen
5
Hannover Flughafen S5
Letter
Garbsen
Wunstorf Dedensen/ Seelze
Gümmer
Poggenhagen
Neustadt am Rübenbge.
Eilvese
Hagen (Han)
Linsburg
Nienburg
AR 2
AR 1
Übergang zum Bus
Interchange with bus
AR 1
4
AR 3
R4
R2
AR 3
Soltau
Bremen
Veranstaltungslinie
Special service
Außenringe für
AR 1 Tarifzonen
Cards im GVH-Regionaltarif
16 18
Stadtbahn
Tram line
S-Bahn mit Bahnhof
Suburban train with station
S5
4
Regionalbahn mit Bahnhof
Regional train with station
R2
S7
S4 R13 R14 R15
R14
Wesseln
Bad Salzdetfurth Solebad
Bad Salzdetfurth
Bodenburg
Hoheneggelsen
Hildesheim
Ost
Groß Düngen Derneburg
S3
Hildesheim Hbf
Harsum
Algermissen
Sehnde
AR 1
Hämelerwald
Vöhrum
Peine
R9
Wolfsburg
Calberlah
Gifhorn
AR 3
R11
Bad Harzburg
R15
Braunschweig
R10
Braunschweig
Leiferde
Meinersen
AR 2
Dedenhausen
Dollbergen
AR 1
Celle
Immensen-Arpke
Aligse
Burgdorf
Otze
Ehlershausen
S6
R8
Uelzen
The Faculty of Natural Sciences is the largest faculty at the Leibniz
Universität Hannover and includes the subjects of biology, chemistry,
horticulture, geosciences, geography and food science.
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offering a wide range of subjects including over 30 courses of study. Its
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opportunities for research and PhD programmes.
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