3/2/2016
About Me
Genetics & New Landscape Plants
Central Gorge Master Gardener Training 2016
Jason Lattier
About Me
Contreras Lab
Oregon State University
Winfield House & Garden
Outline
• What is Plant Breeding?
• Modern Plant Breeding
• Major Goals of Plant Breeding
• Breeder’s Toolbox
• Numbers Game Concept
• Plant Breeding in Landscape Plants
Plant Breeding
+
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3/2/2016
What is Plant Breeding?
What is Plant Breeding?
• Plant breeding is the art and science of improving plant genotypes
(for the needs of human kind)
• Continuation of crop evolution under domestication
• Depends on selection within a genetically variable population
• Agricultural Revolution 10-12K years ago – Mass Selection
• Old practice
• Both natural and artificial selection
What is Plant Breeding?
What is Plant Breeding?
..
• Joseph Kolreuter (1733-1806) – hybrid vigor vs
Essentialism
• Gregor Mendel (1822-1884) – discovered rules of
heredity (genetics)
Outline
• What is Plant Breeding?
• Modern Plant Breeding
• Major Goals of Plant Breeding
• Breeder’s Toolbox
• Numbers Game Concept
• Plant Breeding in Landscape Plants
Modern Plant Breeding
DNA
Genes Turned On
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Modern Plant Breeding
DNA
Modern Plant Breeding
Genes Turned Off
Genes Turned On
DNA
+
Modern Plant Breeding
Modern Plant Breeding
• Modern Plant Breeding – a science that incorporates knowledge from
several related disciplines
1. Command of Mendelian genetics
Eye Color:
Brown
Ocotillo, Nova Scotia Wild Flora Society
Blue
Ocotillo, Nova Scotia Wild Flora Society
KD Talbot,, New Hampshire
Modern Plant Breeding
Modern Plant Breeding
What is the genotype and phenotype?
Mom’s Eyes
• Modern Plant Breeding – a science that incorporates knowledge from
several related disciplines
1. Command of Mendelian genetics
Dad’s Eyes
Dominant Traits
Recessive Traits
½ Brown Eyes
½ Blue Eyes
Dad’s Eyes
Mom’s Eyes
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All children will have brown eyes!
Modern Plant Breeding
Qualitative Traits
vs.
Quantitative Traits
offspring
Modern Plant Breeding
What is the genotype and phenotype?
offspring
Modern Plant Breeding
Modern Plant Breeding
3,000
5,000
• Modern Plant Breeding – a science that incorporates knowledge from
several related disciplines
= 2,000
= 500
2. Knowledge of Statistics for designing experiments and interpreting data
= 0.25
3,500
Modern Plant Breeding
Modern Plant Breeding
• Modern Plant Breeding – a science that incorporates knowledge from
several related disciplines
• Modern Plant Breeding – a science that
incorporates knowledge from several
related disciplines
3. Training in Plant Pathology
4. Familiarity with other plant sciences
(botany, morphology, anatomy, and
taxonomy)
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3/2/2016
Modern Plant Breeding
Modern Plant Breeding
• Cross-Pollinated Crops:
• Self-Pollinated Crops:
Grasses (maize, perennial rygrass), Brassica (broccoli, cabbage, cauliflower)
Vegetables (onion, carrot), composites (sunflower, zinnia, etc)
Modern Plant Breeding
Cereal grains (wheat, barley, oat, rice),
legumes (bean, peas, peanuts, soy),
vegetables (tomato, lettuce)
Modern Plant Breeding
• Wind-pollinated Crops:
Modern Plant Breeding
Modern Plant Breeding
• Modern Plant Breeding – a science that incorporates knowledge from
several related disciplines
4. Familiarity with other plant sciences (botany, morphology, anatomy, and
taxonomy)
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Modern Plant Breeding
Modern Plant Breeding
• Modern Plant Breeding – a science that incorporates knowledge from
several related disciplines
Li et al., 2012
5. Understanding of plant physiology and its relationship to environmental
stresses (cold, drought, salt, soil pH)
Modern Plant Breeding
Modern Plant Breeding
• Modern Plant Breeding – a science that incorporates knowledge from
several related disciplines
• Modern Plant Breeding – a science that incorporates knowledge from
several related disciplines
5. Understanding of plant physiology and its relationship to environmental
stresses (cold, drought, salt, soil pH)
6. Understanding of Cytogenetics when crosses are between different species
or plant with different numbers of chromosomes.
70-20-10 Rule
Environment : Genotype x Environment : Genotype
Modern Plant Breeding
Modern Plant Breeding
• Modern Plant Breeding – a science that incorporates knowledge from
several related disciplines
• Modern Plant Breeding – a science that incorporates knowledge from
several related disciplines
6. Understanding of Cytogenetics when crosses are between different species
or plant with different numbers of chromosomes.
6. Understanding of Cytogenetics when crosses are between different species
or plant with different numbers of chromosomes.
More cross compatible if:
Same genome size (amount of DNA)
Same number of chromosomes
Same ploidy (copy number of each chromosome)
http://data.kew.org/cvalues/
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Modern Plant Breeding
Modern Plant Breeding
• Modern Plant Breeding – a science that incorporates knowledge from
several related disciplines
*
6. Understanding of Cytogenetics when crosses are between different species
or plant with different numbers of chromosomes.
Polyploidy
Normal
Abnormal
*
Modern Plant Breeding
Simplicity
• Modern Plant Breeding – a science that incorporates knowledge from
several related disciplines
0,1
7. Understanding Molecular Markers, Genomics, Bioinformatics (marker assisted
selection)
Complexity
*DNA!
(ACTG)
0,1,2,3,4,5,6,7,8,9
A,B,C,D,E,F,G…Z
Unknown sentence….
• DNA is just a language (ATCG)
• DNA is just a language (ATCG),
but we can only read small chunks…
(4 nucleotides)
Small enough to manage
Large enough to be unique
u just fo
Adenine
Thymine
Cytosine
Guanine
You ju
•
•
•
•
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3/2/2016
• Then, finds the location of the gene (chromosome 6)
• Bioinformatics uses computer programs to assemble the chunks
u just fo
You ju
You ju
u just fo
t found the ge
he gene for res
or resistance to Ea
ance to Eastern Fil
astern Filbe
ilbert Blight!
**
• “You just found the gene for resistance to
Eastern Filbert Blight!”
Modern Plant Breeding
• Build an assay, marker-assisted selection!
EFB Susceptible
-Discard!
EFB Heterozygotes
-Keep!
EFB Resistant
-Keep!
EFB Susceptible Plant
• Modern Plant Breeding – a science that
incorporates knowledge from several related
disciplines
8. Some breeders use Transformation to insert a
discovered gene into other crops
EFB Resistance Gene
Modern Plant Breeding
A. tumefaciens
Modern Plant Breeding
Traits
Crops
Bt
Alfafa
Maize
Soybean
Canola
Cotton
Papaya
Poplar
(Rice)
Roundup-resistant
Increased nutrition
(Disease/Drought Resistant)
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3/2/2016
Outline
Breeding Goals
• What is Plant Breeding?
• Modern Plant Breeding
• Major Goals of Plant Breeding
• Breeder’s Toolbox
• Numbers Game Concept
• Plant Breeding in Landscape Plants
• Resistance to pests and diseases
• Increased yield
• Improved form
• Quality traits
• Sterility
• Physiological Changes
• Beneficial Mutations
Boll Weevil Monument (Enterprise, Alabama)
Breeding Goals
Breeding Goals
• Resistance to pests and diseases
• Increased yield
• Improved form
• Quality traits
• Sterility
• Physiological Changes
• Beneficial Mutations
• Resistance to pests and diseases
• Increased yield
• Improved form
• Quality traits
• Sterility
• Physiological Changes
• Beneficial Mutations
Breeding Goals
Breeding Goals
• Resistance to pests and diseases
• Increased yield
• Improved form
• Quality traits
• Sterility
• Physiological Changes
• Beneficial Mutations
• Resistance to pests and diseases
• Increased yield
• Improved form
• Quality traits
• Sterility
• Physiological Changes
• Beneficial Mutations
Day-neutral Strawberries
Low chill blueberries
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Breeding Goals
Outline
• Resistance to pests and diseases
• Increased yield
• Improved form
• Quality traits
• Sterility
• Physiological Changes
• Beneficial Mutations
• What is Plant Breeding?
• Modern Plant Breeding
• Major Goals of Plant Breeding
• Breeder’s Toolbox
• Numbers Game Concept
• Plant Breeding in Landscape Plants
Thornless Blackberries!
Variegations
Breeder’s Toolbox
Breeder’s Toolbox
• Most important tool:
• Knowledge of Plant Material!
• Knowledge of Plant Material!
• Mutation
How do I grow it?
What is the species diversity?
What are the elite cultivars?
What improvements are needed?
What is the pollination biology?
Breeder’s Toolbox
• Knowledge of Plant Material!
• Mutation
• Irradiation
• Chemical
• Whole Genome Duplication
• Colchicine
• Oryzalin
Gigas Effects
• Irradiation
• Chemical
ATGCCGCAAGATC
ATGCCGTAAGATC
(STOP)
Why?
Cross incompatibility
Plant Breeding: Depends on selection within a genetically variable population
Breeder’s Toolbox
• Knowledge of Plant Material!
• Mutation
• Irradiation
• Chemical
• Whole Genome Duplication
• Colchicine
• Oryzalin
• Tissue Culture
•
•
•
•
Embryo Rescue
Exponential growth of plants
Sterile
Reduced Shipping Costs
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3/2/2016
Outline
Numbers Game Concept
• What is Plant Breeding?
• Modern Plant Breeding
• Major Goals of Plant Breeding
• Breeder’s Toolbox
• Numbers Game Concept
• Plant Breeding in Landscape Plants
• PRI (Perdue, Rutgers, Illinois) – 1940’s apple scab
‘Prima’
• Crossed with Malus floribunda 821
• 100,000 seedlings planted, 60,000 seedlings evaluated by 1992
• 13 cultivar were released
• Only 3 cultivars survived the test of time
• 20,000 planted to produce one stellar cultivar
• A breeder who plants 10,000 apple seedlings/year could release a
truly good cultivar every other year!
Outline
• What is Plant Breeding?
• Modern Plant Breeding?
• Major Goals of Plant Breeding
• Breeder’s Toolbox
• Numbers Game Concept
• Plant Breeding in Landscape Plants
Project idea
All new cultivars begin with an idea…
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3/2/2016
Ornamental Plant Breeding Program
Project idea
Jason Lattier
Dr. Ryan Contreras
Mara Friddle
Hsuan Chen
Project
idea
Project
idea
Seed
Plant
Material
Plant
Material
Potted
plants
Potted
plants
Greenhouse
Disease
screening
Grow outs
Plant Material
Project
idea
Controlled
Crosses
crosses
Seed
Plant
Material
Cuttings
Justin Schulze
Controlled
Crosses
crosses
Seed
Cuttings
Kim Shearer Lattier
Cuttings
Potted
plants
Greenhouse
Disease
screening
Field
evaluation
Grow outs
New cultivar for
licensing and
release
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3/2/2016
Goals and projects
WWpp
Combining Traits
wwPP
• Traditional breeding
• Cercidiphyllum – Combining two traits
• Philadelphus – Combining multiple traits
• Breeding for disease resistance
• Cotoneaster – fireblight resistance
x
• Sterility
• Hibiscus – Seedless cultivars
• Inducing novel mutations
• Ribes – dissected leaves and compact forms
• Galtonia – compact inflorescence
http://jcraulstonarboretum.wordpress.com
http://jcra.ncsu.edu/visit/now-showing/details.php?serial=194
Combining Traits
WWpp
wwPP
W = upright
w = weeping
P = green
p = purple
Combining Traits
WwPp
=WwPp
x
Combining Traits
Combining Traits
WwPp
WwPp
x WwPp
WwPp
Ww x Ww
W w
x
= 1/16 wwpp
W
WW Ww
w
Ww ww
= 1/16 wwpp
¼ ww
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3/2/2016
Combining Traits
WwPp x WwPp
Combining Traits
WwPp
x WwPp
Pp x Pp
P
W w
p
P
PP
Pp
p
Pp
pp
= 1/16 wwpp
W
WW Ww
w
Ww ww
¼ pp
Combining Traits
Albizia julibrissin ‘NCAJ1’
Chocolate Fountain™
¼ ww
P
p
P
PP
Pp
p
Pp
pp
x
= 1/16 wwpp
¼ pp
Goals and projects
• Traditional breeding
• Cercidiphyllum – Combining two traits
• Philadelphus – Combining multiple traits
• Breeding for disease resistance
• Cotoneaster – fireblight resistance
• Sterility
• Hibiscus – Seedless cultivars
• Inducing novel mutations
• Ribes – dissected leaves and compact forms
• Galtonia – compact inflorescence
Katsura Tree
• Weeping: C. japonicum ‘Morioka Weeping’ ♂
• Red foliage: C. japonicum ‘Red Fox’ ♀
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‘Red Fox’ x ‘Morioka Weeping’
‘Red Fox’ inside
GREENhouse
10 November 2015
GREENhouse
‘Red Fox’ outside
Waiting to flower!
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Combining Traits
Goals and projects
• Traditional breeding
• Things get complicated!
• Cercidiphyllum – Combining two traits
• Philadelphus – Combining multiple traits
• Frequency of recessives:
• Breeding for disease resistance
• Cotoneaster – fireblight resistance
• Sterility
• Hibiscus – Seedless cultivars
• Inducing novel mutations
• Single trait:
Purple foliage:
• Two traits:
Weeping:
• Three traits:
Glossy leaves:
• Four traits:
Fragrance:
=¼
¼ × ¼ = 1/16
¼ × ¼ × ¼ = 1/64
‘Snow White’
¼ × ¼ × ¼ × ¼ = 1/256
• Ribes – dissected leaves and compact forms
• Galtonia – compact inflorescence
Combining Traits
‘Miniature Snowflake’
Combining Traits
unknown
unknown
P. mexicanus
P. mexicanus
‘Snow White’
Combining Traits
‘Miniature Snowflake’
‘Snow White’
Combining Traits
‘Miniature Snowflake’
unknown
P. mexicanus
‘Snow White’
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Combining Traits
‘Miniature Snowflake’
• First Generation
unknown
• Grow industry standards
• Cross
all combinations
P. mexicanus
Evaluating fragrance!
• Grow out offspring
• Take phenotype data
• Determine dominant traits
• Make selections
• Rouge poor selection
• Make self-pollinations
• Make new cross combinations
‘Snow White’
Combining Traits
Combining Traits
Combining Traits
Combining Traits
‘Miniature Snowflake’
• Second Generation
unknown
• Evaluate selfs
• Determine recessive traits
P. mexicanus
• Evaluate new hybrids
• Compare traits to industry standards
• Make selections
• Make new cross combinations
• Begin propagating for replicated
field trials
‘Snow White’
• 70-20-10 Rule!
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Goals and projects
• Traditional breeding
• Cercidiphyllum – Combining two traits
• Philadelphus – Combining multiple traits
• Breeding for disease resistance
• Cotoneaster – fireblight resistance
• Sterility
• Hibiscus – Seedless cultivars
• Inducing novel mutations
• Ribes – dissected leaves and compact forms
• Galtonia – compact inflorescence
Breeding for disease resistance
Breeding for Disease Resistance
• 400 species
• Trees, shrubs and
groundcovers
• 52 species
screened over 2
years for sources
of fireblight
resistance
Breeding for Disease Resistance
Breeding for Disease Resistance
How do you screen for fireblight resistance?
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Breeding for Disease Resistance
Fireblight!
Breeding for Disease Resistance
Breeding for Disease Resistance
Measure lesion length after 8 weeks
Breeding for Disease Resistance
Breeding for Disease Resistance
• Cross resistant
species by
industry
standards
• ‘Coral Beauty’
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Breeding for Disease Resistance
Breeding for Disease Resistance
• Self-pollinating
• Must emasculate
• Screen seedlings
using the same
inoculation
method
• Tedious!!!
J. Rothleutner
Selections:
*Ornamental Traits
*Ease of Propagation
Breeding for Disease Resistance
• Many seedlings
resistant to
fireblight
• One seedling
stood out among
the rest
• One gallon
container 
Propagates Well!
Breeding for Disease Resistance
• Seven gallon
container! 
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Breeding for Disease Resistance
OSU Selection
Breeding for Disease Resistance
Selection
‘Coral Beauty’
‘Coral Beauty’
Goals and projects
• Traditional breeding
• Cercidiphyllum – Combining two traits
• Philadelphus – Combining multiple traits
• Breeding for disease resistance
• Cotoneaster – fireblight resistance
• Sterility
• Hibiscus – Seedless cultivars
• Inducing novel mutations
• Ribes – dissected leaves and compact forms
• Galtonia – compact inflorescence
H2011-01-002
Breeding for Sterility
Breeding for Sterility
Hibiscus ‘Diana’
-Sold as sterile
Live mas!
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Breeding for Sterility
Breeding for Sterility
 USNA – Dr. Donald Egolf
• Released sterile (or near sterile)
triploid cultivars
–
–
–
–
Breeding for Sterility
‘Aphrodite’
‘Diana’
‘Helene’
‘Minerva’
Breeding for Sterility
 Egolf Breeding Program:
 Egolf Breeding Program:
(2x)
(4x)
(4x)
X
• H. syriacus selections
H. syriacus 'William R. Smith'
• H. syriacus cultivars
 Assess ploidy of elite cultivars
in the trade
H. syriacus 'William R. Smith'
6x population
3x population
Breeding for Sterility
(8x)
X
Breeding for Sterility
 Flow cytometry
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Breeding for Sterility
Breeding for Sterility
 Root tip squash
Breeding for Sterility
 Root tip squash
‘Diana’ 2n = 4x = 80
‘Diana’ 4.7 pg
Breeding for Sterility
Breeding for Sterility
(6x)
(4x)
• H. syriacus cultivars
X
H. syriacus ‘Pink Giant'
• ~150 cross combinations
• 10,000 plus seedlings
• 200+ progeny to evaluate
5x population
138
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Breeding for Sterility
Breeding for Sterility
Sterile
5x?
139
Breeding for Sterility
140
Breeding for Sterility
• Summer 2016
Evaluate
ornamental traits!
• Summer 2016
Fertility test!
141
142
Goals and projects
Mutation Breeding
• Traditional breeding
• Ribes sanguineum
• Cercidiphyllum – Combining two traits
• Philadelphus – Combining multiple traits
• Breeding for disease resistance
• Cotoneaster – fireblight resistance
• Sterility
• Hibiscus – Seedless cultivars
• Inducing novel mutations
• Native
• Rangy shrub up to 15
feet wide
• Too large for most
home landscapes
• Little genetic variability
in form
• Ribes – dissected leaves and compact forms
• Galtonia – compact inflorescence
144
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Mutation Breeding
EMS
Mutation Breeding
EMS
• Chemical mutagen
• Used to induce variation
• 2011
• 10,000 seeds soaked
• Thousands of seedlings
• Known mode of action, ease of access, and cost
 compared to irradiation
Mutation Breeding
Mutation Breeding
x
‘Oregon Snowflake’
Mutation Breeding
Mutation Breeding
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Goals and projects
• Traditional breeding
• Cercidiphyllum – Combining two traits
• Philadelphus – Combining multiple traits
• Breeding for disease resistance
• Cotoneaster – fireblight resistance
• Sterility
• Hibiscus – Seedless cultivars
• Inducing novel mutations
• Ribes – dissected leaves and compact forms
• Galtonia – compact inflorescence
Galtonia ‘Moonbeam’
Galtonia candicans Decne.
Chemical mutagenesis of Galtonia candicans
Decne. results in altered morphology and
reduced fertility
Tesselaar
• Cold hardy bulb species native
to South Africa
• Self-pollinating
• ~4 – 5’ tall
• drought tolerant
• Summer bloom (until frost)
Kim Shearer Lattier and Ryan Contreras
Galtonia candicans Decne.
Galtonia ‘Moonbeam’
Shipping cart
southernlagniappe.blogspot.com
directgrowerindustries.com
http://mygarden.rhs.org.uk/forums/t/19058.aspx
• Fitting on shipping cart
• Lodging
• potentially weedy
Tesselaar
• Little known cultivar development
• Issues
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Lodging
Potential weediness
http://mygarden.rhs.org.uk/forums/t/19058.aspx
Galtonia ‘Moonbeam’
0%
Ethyl methane sulfonate
• Chemical mutagen
0%
0.2%
0.2%
Ethyl methane sulfonate
• Taken up by embryo during imbibition period
 Results in genome wide point mutations
 replacement of G/C with A/T pairing through genome repair
• Used to induce variation
• Known mode of action, ease of access, and cost
 compared to irradiation
Objective
• Effects: reduced height and fertility, changes in floral
morphology
M1 Germination of seeds
EMS Concentration (%)
0
0.2
0.4
0.6
0.8
1.0
• Produce improved cultivar of Galtonia candicans through the
application of EMS treatments
 reduced height
 decreased lodging
 reduced fertility
Hypothesis: Point mutations resulting from EMS treatment
may result in novel characters such as reduced scape height
Mean germination (%)
39.56 ± 1.09 az
36.78 ± 1.25 b
36.56 ± 0.68 b
11.78 ± 3.90 c
0.89 ± 0.89 d
0.00 ± 0.00 d
zThe
treatment seeds were kept dry at room temperature until EMS treatment application. ANOVA for
completely randomized design performed on germination data. zValues followed by different letters
within a column are significantly different, Fisher’s LSD, P ≤ 0.05.
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M1 Field Evaluation
M1 Germination
0%
0.2%
0.2%
0.4%
0.4%
0.6%
0.8%
0.6%
1.0%
0.8%
1.0%
Germination of G. candicans seedlings 10 weeks after treating seed in 0, 0.2, 0.4, 0.6, 0.8, or
1.0% EMS for 24-h.
Mean scape height (cm)
0%
100
90
80
70
60
50
40
30
20
10
0
Response of plant height (cm) to EMS treatment (%
concentration)
88.51
74.52
60.59
0
0.2
0.4
EMS Concentration (%)
Preliminary M2 Germination Study of OP seed
0%
0%
0.2%
0.2%
Preliminary M2 Germination Study
0%
0%
0.2%
0.2%
Floral Morphology
EMS Concentration
(%)
Average seed per
capsule (M1)
Mean Germination
M2 seed (%)
0
17.15 ± 0.68 az
89.44 ± 2.42 a
0.2
3.06 ± 0.65 b
57.93 ± 4.56 b
0.4
0.00 ± 0.00 c
0 ± 0.00 c
zValues
Preliminary M2 Germination Study of OP seed
followed by different letters within a column are significantly different, Fisher’s LSD, P ≤ 0.05.
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Observed Floral Mutations
Summary
EMS treatment is a viable method to. . .
• reduce plant height which may result in reduced
lodging
• reduce fertility mitigating potential weediness
• induce variation in a population of Galtonia
Summary
EMS treatment is a viable method
to provide an improved cultivar of Galtonia candicans to
the nursery industry
29