American-Eurasian J. Agric. & Environ. Sci., 12 (7): 960-966, 2012
ISSN 1818-6769
© IDOSI Publications, 2012
DOI: 10.5829/idosi.aejaes.2012.12.07.1799
Phloroglucinol, BAP and NAA Enhance Axillary Shoot Proliferation and other Growth
Indicators In vitro Culture of Damask Rose (Rosa damascena Mill.)
M. Salekjalali
Young Researchers Club, Ahar Branch, Islamic Azad University, Ahar, Iran
Abstract: Experiments were conducted to study the effect of PG, BAP and NAA on proliferation ability of
damask rose (Rosa damascena Mill.). BAP in four levels (0, 1, 2 and 3 mg l 1), NAA in three levels (0, 0.1 and
1 mg l 1) and PG in three levels (0, 50 and 100 mg l 1) were used and 21 different combinations of them were
tested to investigate the most suitable treatment for multiplication stage as a factorial test on the base of a
completely randomized design. BAP affected on number of green leaves, brown leaves and induced axillary
shoots per explants significantly, whereas 0.1 mg l 1 NAA increased only number of green leaves in compared
with medium without NAA. PG remarkably enhanced the multiplication rates of roses in vitro so that number
of new axillary shoots per explants increased 30% when PG added to medium containing BAP and NAA. Finally
it was shown that the Full-strength MS culture medium containing BAP (2 mg l 1) and NAA (0.1 mg l 1)
supplemented with PG (100 mg l 1) has the best results for shoot proliferation of Rosa damascena Mill.
Microshoots were rooted by ½ strength of MS medium containing IBA at the concentration of 2 mg l 1 with
up to 80% rooting. Plantlets were acclimatized in a soil mixture consisting of mineral perlite and peat moss (2:
1 v/v) and successfully transferred to the greenhouse after 4 weeks.
Key words: Rosa damascena Mill
BAP
NAA
PG
INTRODUCTION
Shoot Proliferation
and Echeverrigary, [9] showed that BAP (6Benzylaminopurine) may cause producing more shoots in
comparison with Kinetin and 2iP (6-[dimethylallyl amino]purine) in rose micropropagation system. Kumar et al.
[10] stated supplementing the media with GA3 (Gibberellic
acid 3) can improve multiplication of Damask rose
explants. They also reported the senescence symptoms of
proliferated shoots and try to cope with the problem by
modifying the media. [11] concluded that the best
hormonal compound for in vitro propagation of
Damask rose is BAP (2 mg l 1) and NAA (1-Naphtalene
acetic acid) (0.1 mg l 1). They also found in vitro rooting
of old roses including Damask rose is much more
difficult than modern roses. Pati et al. [12] found 2 mg l 1
BAP is the most appropriate concentration for in vitro
propagation of Damask rose.
Phloroglucinol (PG) is a phenolic compound
predominantly found in xylem sap of apple and is
known to promote growth and development in a
number of plant species such as Chinchona ledgeriana,
apple cultivars and cocoa [13]. There are some reports
suggest that phloroglucinol enhances growth and rate of
axillary shoot proliferation of some plants in vitro [14, 15].
Rose is the king of flowers and Damask rose (Rosa
damascena Mill.) is classified in old garden roses [1]. The
origin of Damask rose is Iran and the Middle East region
and it is the national flower of Iran.
Damask Rose (Rosa damascena Mill.) is an important
aromatic plant significant from economic point of view in
Iran and some other countries. Besides aromatic and
ornamental values of R. damascena, its medicinal
properties especially anti-depression effects are worth
consideration [2].
Traditionally, rose plants have been propagated
on rootstocks and through cutting method [3]. The
conventional propagation of this species was
associated with various problems such as limitation of
stock plants and prolonged production time [4] and
low adventitious root formation on cuttings. Tissue
culture of Rosa species has become an alternative method
of propagation [5, 6].
The establishment of tissue culture system for
various rose species has been described [5, 7-9] but it is
not much specifically in the case of Damask rose. Carelli
Corresponding Author: M. Salekjalali, Young Researchers Club, Ahar Branch, Islamic Azad University, Ahar, Iran,
Tel: +98 426 2232495, Fax: +98 426 2237718.
960
Am-Euras. J. Agric. & Environ. Sci., 12 (7): 960-966, 2012
Murali et al. [16] reported that somatic embryo
germination of Rosa hybrida cv. Arizona occurred
when explants cultured in ½ MS medium [17]
supplemented with 0.5 mg l 1 NAA (1-Naphtalene
acetic acid) and 100 mg l 1 Phloroglucinol. However, use
of phloroglucinol in the proliferation medium of rose
cultivars has not been reported.
In this paper we describe an optimal protocol for
multiplication of Rosa damascena Mill through high
frequency axillary shoot proliferation from nodal explants.
We examine the influence and interaction of growth
regulators BAP and NAA as well as phloroglucinol in
multiplication of Rosa damascena Mill.
containing full and ½ strength of MS salts. Two
additional treatments (containing or without IBA (3Indolebutyric acid) at the concentration 2 mg l 1) were
tested in the both mediums. Cultures were maintained at
22°C in a culture room with a 16 hrs photoperiod light.
Acclimatization and Transfer to Soil: Plantlets were
removed from the culture flasks and washed with water to
remove all medium attached to the roots. They were then
transplanted to small plastic pots containing mineral
perlite and maintained in a growth chamber at 22°C and
90% Relative Humidity (RH) under a 16 hrs photoperiod.
After 3 days, RH was reduced to 80% and 7 days later
reduced to 70%. After 2 weeks the plants were transferred
to large plastic pots containing mineral perlite and peat
moss (2: 1 v/v) and maintained in growth chamber with
22°C and 60% RH. After 4 weeks the pots were transferred
to the greenhouse for further growth.
MATHERIALS AND METHODS
Plant Materials: Internodal segments (8-10 cm long) of
Rosa damascena Mill.having at least one axillary bud
were collected from plants growing on the botanical
garden of Azerbaijan, Tabriz, Iran. They were cut in 2-3 cm
length segments and placed in sterilized distilled water
containing few drop of Tween 80 for few minutes. Then
surface disinfected using 70% (v/v) ethanol for 60 sec
and then immersed in 10% (v/v) of commercial laundry
bleach (5.25% NaOCl) for 20 min and rinsed three times
in sterilize distilled water. The shoots were trimmed
down to 1 cm long with single node prior to transferring
to establishment medium.
Statistical Analysis: For evaluation of proliferation
stage, two separate experiments as a completely
randomized design with 9 replications were designed.
12 different treatments from composition of BAP and
NAA hormones and 9 treatments from composition of
BAP, NAA and PG were used in the first and second
experiment, respectively. Three characters of the
growth and multiplication rate as dependent factors
(number of green leaves, number of brown leaves and
number of axillary shoots) were measured in the both
experiment. Variance of Data were analyzed using
Multivariate GLM (General Linear Model) and mean of
numbers were compared using Duncan’s Multiple Range
Test at p<0.01.
Establishment Medium: In the establishment stage, all
explants were cultured on a MS medium containing salts,
vitamins and sucrose without any hormones. Explants
were sub cultured to the fresh medium after 3 days
because they were released phenol compounds.
RESULTS AND DISCUSSION
Shoot Proliferation: The basal nutrient medium
containing MS salts and vitamins and supplemented with
BAP, NAA and PG was used in this part of the state. Two
experiments were separately designed. In the first
experiment, BAP at the concentrations of 0, 1, 2 and
3 mg l 1 was combined with NAA at the concentrations
of 0, 0.1 and 1 mg l 1. After identifying the optimum
composition of hormones, in the second experiment,
the effect of PG with 0, 50 and 100 mg l 1 was examined.
Explants were sub cultured to the fresh medium every
3 weeks. Finally, excised single shoot from multiple
shoots were transferred to the fresh medium for root
induction.
In the establishment stage, cultured explants were
grown in the free hormones MS medium with two
subcultures of 3 weeks intervals (Fig. 1-A). Since the
explants released high amount of polyphenol compounds
in the initial days of culturing in this stage, explants sub
cultured to the fresh medium after 3 days. Some explants,
especially those from the shoot apex itself, released a
brown substance from the cut surface into the
establishment medium. These substances noticeably
decreased the bud growth of shoots. The brown
substance is possibly a polyphenol that is toxic to the
explants [4]. This problem resolved with sub culturing
of explants to the fresh medium after 3 days of cultures.
Yan et al. [18] reported that explants exuded less
brown material when transferred to the fresh medium after
3-5 days.
Root Initiation: To establish root proliferation, green and
normal adventitious shoots from shoot proliferation
cultures were excised and cultured on MS medium
961
Am-Euras. J. Agric. & Environ. Sci., 12 (7): 960-966, 2012
Fig. 1: Micropropagation of Rosa damascena Mill. (A) the establishment medium, (B, C) the proliferation stage (D)
Transferred plantlets for acclimatization
Table 1: The main effect of BAP hormone on numbers of green leaves, brown leaves and axillary shoots of Rosa damascena Mill
BAP (mg/l)
Green leaves
Brown leaves
0
18.15±.608**c
10.59±.736**c
Axillary shoots
3.59±.228**c
1
25.04±1.041b
6.02±.412b
4.67±.151b
2
30.41±1.311a
4.19±.362a
5.33±.214a
3
13.33±.842d
14.89±.532d
3.37±.143c
In each column, means followed by the same letters are not significantly different using Multiple Dunkan Test. * and ** are significant at 5% and 1%
respectively and ns is not significant
Table 2: The main effect of NAA hormone on numbers of green leaves, brown leaves and axillary shoots of Rosa damascena Mill
NAA(mg/l)
Green leaves
Brown leaves
Axillary shoots
0
22.00±1.271**b
8.89±.721nsab
4.33±.191nsab
0/1
25.64±1.504a
7.56±.785a
4.53±.205a
1
17.56±.992c
10.32±.930b
3.86±.219b
In each column, means followed by the same letters are not significantly different using Multiple Dunkan Test. * and ** are significant at 5% and 1%
respectively and ns is not significant
In the first experiment of the proliferation stage, BAP
significantly increased number of new green leaves and
axillary shoots and decreased brown leaves at 1% level
(Table 1). Therefore, a significant effect of BAP hormone
was shown in the proliferation stage on growth and
multiplication of Rosa damascena Mill. The optimal
amount of this hormone for Rosa damascena Mill was 2
mg l 1 (Table 1). [10] reported that the best BAP
concentration for micropropagation of Damask rose was
5 µg L 1 (1.25 mg l 1). Inclusion of BAP (1.0-10.0 mg l 1)
in the culture medium was essential for bud break and
shoot multiplication of R. hybrida [19, 20].
It was shown that NAA had a significant effect in the
number of green leaves, whereas it had no effect in
numbers of brown leaves and axillary shoots (Table 2).
The results of our experiment are supported by the
findings of [11], who demonstrated that NAA was
necessary for micropropagation of damask rose. Also, [21]
and [1] reported that NAA had no significant effect in the
proliferation of rose plants.
962
Am-Euras. J. Agric. & Environ. Sci., 12 (7): 960-966, 2012
Table 3: Effect of different concentrations of BAP and NAA on numbers of green leaves, brown leaves and axillary shoots of Rosa damascena Mill
Treatment (BAP + NAA) (mg/l)
Green leaves
Brown leaves
Axillary shoots
T1(0+0)
T2(0+ 0.1)
T3(0+ 1)
T4(1+ 0)
T5(1+ 0.1)
T6(1+ 1)
T7(2+ 0)
T8(2+ 0.1)
T9(2+ 1)
T10(3+ 0)
T11(3 +0.1)
T12(3+ 1)
18.00±.745**fg
20.67±1.027ef
15.78±.703gh
26.11±.754cd
28.67±1.936bc
20.33±1.302ef
30.56±1.773b
36.67±1.667a
24.00±.898de
13.33±1.155hi
16.56±1.501gh
10.11±.857i
10.00±.928**e
8.89±1.379de
12.89±1.207f
7.33±.799cd
5.67±.527bc
5.06±.626bc
4.00±.408ab
2.56±.444a
6.00±.408bc
14.22±.662f
13.11±.873f
17.33±.601g
4.00±.408**cde
4.00±.408cde
2.78±.222f
4.56±.294bcd
4.44±.176bcd
5.00±.289b
5.33±.333ab
6.00±.236a
4.67±.408bc
3.44±.176ef
3.67±.289def
3.00±.236f
In each column, means followed by the same letters are not significantly different using Multiple Duncan Test. * and ** are significant at 5% and 1%
respectively and ns is not significant
Table 4: The main effect of PG on numbers of green leaves, brown leaves and axillary shoots of Rosa damascena Mill
PG Phloroglucinol (mg/l)
--------------------------------------------------------------------------------------------------------------------------------------------------0
50
100
Green leaves
Brown leaves
Axillary shoots
25.26±.776b
4.67±. 325a
4.19±.160c
26.41±.705ab
5.41±.263a
4.74±.137b
27.78 ±.667**a
5.15±.365nsa
5.44±. 180**a
Values indicated with ** are significant at 1% level and ns are not significant.
Table 5: Effect of different concentrations of BAP, NAA and PG on numbers of green leaves, brown leaves, axillary shoots of Rosa damascena Mill
Treatment (BAP+NAA+ PG) (mg/l)
Green leaves
Brown leaves
Axillary shoots
T1(1+0+0)
T2(1+0+50)
T3(1+0+100)
T4(2+0+0)
T5(2+0+50)
T6(2+0+100)
T7(2+0.1+0)
T8(2+0.1+50)
T9(2+0.1+100)
20.22±.465**f
22.11±.611e
26.00±.500cd
26.22±.324cd
27.11±.633c
25.11±.351d
29.33±.441b
30.00±.289b
32.22±.465a
6.22±.324nsef
6.56±.294f
7.11±.261f
4.89±.309cd
5.44±.294de
5.00±.408cd
2.89±.351a
4.22±.401bc
3.33±.408ab
3.67±.289**e
4.56±.294cd
4.89±.309bcd
4.22±.222de
4.44±.176cd
5.44±.176ab
4.67±.236cd
5.22±.147bc
6.00±.333a
In each column, means followed by the same letters are not significantly different using Multiple Dunkan Test. * and ** are significant at 5% and 1%
respectively and ns is not significant
Results also showed that there was significant
difference among of 12 treatments into number of green
leaves, brown leaves and axillary shoots at 1% level
(Table 3). Maximum growth and multiplication rates were
obtained in treatments of 5 (1 mg l 1 BAP and 0.1 mg l 1
NAA), 7 (2 mg l 1 BAP) and 8 (2 mg l 1 BAP and
0.1 mg l 1 NAA) therefore, they were used in the second
experiment (Table 3). However, the highest number of
green leaves and axillary shoots were produced with
2 mg l 1 BAP and 0.1 mg l 1 NAA (Table 3).
In the second experiment, data was shown a
significant effect of PG on number of green leaves and
new axillary shoots per explant at 1% level (Table 4). PG
enhanced the multiplication rate up to 30% compared to
mediums without PG. Improved performance of
963
micropropagation using of PG has already been described
by [22 & 23]. PG is a phloridzin derivative and its main use
is in rooting [24, 25, 26] but sometime it could be used as
a component that reduce vitrification and increase
proliferation [27, 28]. Positive effects of phloglucinol on
the stimulation of growth and shoot proliferation have
been reported for some woody species [29, 30]. PG acted
synergistically with BA in inducing early bud break and
high frequency shoot proliferation of Vitex negundo L.an aromatic medicinal plant [13]. Demiralay et al. [31]
achieved shoot multiplication of Ficus carica var. Bursa
Siyaki on MS medium containing 1 mg l 1 BA and 89 mg
l 1 phloroglucinol. Debabrata and Prakash [32] reported
that phloroglucinol fostered multiple shoot formation,
promoted axillary shoot proliferation in potato.
Am-Euras. J. Agric. & Environ. Sci., 12 (7): 960-966, 2012
CONCLUSION
Phloroglucinol activates ERK, an important component
of intracellular signaling cascades and activated ERK
participates in a wide range of cellular programs including
proliferation, differentiation and movement [33, 34]. In
addition, the ERK pathway is known to influence the
expression of several genes, which are mostly involved in
cell proliferation. Reactive oxygen species (ROS) such as
superoxide anion, hydroxyl radicals and H2O2, are
unwanted and toxic byproducts formed during aerobic
metabolism. ROS can initiate injurious degradative
reactions causing cell death and tissue damage via
apoptosis and/or necrosis in the cells and tissues of
plants. There are many reports that indicate PG exhibits
the antioxidant effect against ROS [35, 36].
Treatments showed a significant effect on number of
green leaves, brown leaves and axillary shoots (Table 5).
On the base of mean comparison, treatment of 9 (2 mg l 1
BAP + 0.1 mg l 1 NAA + 100 mg l 1 PG) was selected as
desirable composition (Table 5). Khosh-Khui and Sink
[11] concluded that the best hormonal compound for
in vitro propagation of Damask rose is BA (2 mg l 1)
and NAA (0.1 mg l 1). Fig. 1-B, C show proliferation
stage and shoot multiplication of Rosa damascena Mill.
In rooting of propagated shoots, ½ strength of
MS medium containing IBA at the concentration 2 mg l 1
with up to 80% rooting were more suitable than
other treatments (Data are not presented). Although
it appears to be relatively easy to proliferate rose
shoots in vitro, rooting is frequently difficult. There
are some reports which state in vitro rooting of old
roses (including Damask rose) are more difficult
than modern roses [37]. Khosh-Khui and and Jabbarzadeh
[38] suggested that rooting is affected by genotype;
MS medium salts concentration, cold dark treatment and
auxin type. In this research, reduced salt concentration
increased rooting in MS medium, in accordance with [4].
In most of the earlier reports, varying concentrations of
different auxins have been used for root induction
[39, 37]. In this report, the most roots and the high
survival percentage of plantlets (up 80 %) were initiated
per explants on half-strength MS medium containing IBA
at the concentration 2 mg l 1. This result is similar with the
reported results of [37, 40, 41].
After 2 weeks, in order to acclimatization plantlets
were transferred to the small pots containing perlite in a
growth chamber (Fig. 1-D) and then, the plants were
transferred to large plastic pots containing perlite and
peat moss and maintained in the growth chamber for
further growth after 4 weeks.
The investigation showed that the best shoot
proliferation was obtained at 2 mg l 1 BAP + 0.1 mg l 1
NAA + 100 mg l 1 PG in full-strength MS medium, while
rooting of shoots improved with half-strength MS medium
containing of IBA at the concentration of 2 mg l 1. To the
best of our knowledge the literature offers no reports of
usefulness of PG in the shoot proliferation of rose, though
there are descriptions of it in other some plants. In this
work, PG strongly enhanced the adventitious shoot
proliferation rate of rose plants. Our present work
provides a practical protocol for efficient axillary bud
multiplication from Rosa damascena Mill.explants using
BAP, NAA and PG.
ACKNOWLEDGEMENTS
This work was supported by a grant from
young researchers club, Ahar Branch, Islamic Azad
University. The author thanks Mr. A. Absalan, master of
young researchers club, Ahar Branch, Islamic Azad
University.
REFERENCES
1.
2.
3.
4.
5.
964
Nikbakht, A., M. Kafi, M. Mirmasoumi and
M. Babalar, 2005. Micropropagation of Damask
Rose (Rosa damascena Mill.) cvs Azaran and
Ghamsar. Int. J. Agr. Biol., 7: 535-538.
Allahverdi Mamaghani, B., M. Ghorbanli, M.H.
Assareh and A. Ghamari Zare, 2010. In vitro
propagation of three Damask Roses accessions.
Iranian J. Plant Physiol., 1: 85-94.
Roberts, A.V. and A. Schum, 2003. Cell, Tissue
and Organ culture (Micropropagation) In:
A.V. Roberts, T. Debener and S. Gudin (eds.),
Encyclopedia of Rose Science, Elsevier Academic
Press.
Skirvin, R.M., M.C. Chu and H.J. Young, 1990. Rose
In: P.V. Ammirato, D.R. Evans, W.R. Sharp and Y.P.S.
Bajaj (eds.), Handbook of plant cell culture, McGrawHill, New York.
Arnold, N.P., M.R. Binns, N.N. Barthakur and D.C.
Cloutier, 1992. A study of the effect of growth
regulators and time of plantlet harvest on in vitro
multiplication rate of hardy and hybrid tea roses. J.
Hort. Sci., 67: 727-35.
Am-Euras. J. Agric. & Environ. Sci., 12 (7): 960-966, 2012
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
Tabaei-Aghdaei, S.R., M.B. Rezaee and K. Jaimand,
2005. Study of genetic variation in essential oil yield
of Rosa damascena Mill. Genotypes from central
regions of Iran. Iranian. Medicinal. Aromatic. Plants.
Res., 2: 35-51.
Telgan, H., V. Elagoz, A. van Mill, A. Paffen and
G. de Klerk, 1992. Role of plant hormones in lateral
bud growth of rose and apple in vitro. Acta Hort.,
319: 137-42.
Rout, G.R., S. Samoantaray, J. Mottley and P. Das,
1999. Biotechnology of the rose: a review of recent
progress. Scientia Hortic., 81: 207-28.
Carelli, B.P. and S. Echeverrigary, 2002. An improved
system for the in vitro propagation of rose cultivars.
Scientia Hortic., 92: 64-74.
Kumar, A., A. Sood, L.M.S. Palni, U.T. Palni and
A.K. Gupta, 2000. In vitro propagation of Bulgarian
rose from selected mature bushes. J. Med. Aromat
Plant Sci., 22: 593-602.
Khosh-Khui, M. and K.C. Sink, 1982. Rooting
enhancement of Rosa hybrida for tissue culture
propagation. Scientia Hortic., 17: 371-6.
Pati, P.K., M. Sharma and P.S. Ahuja, 2001.
Micropropagation, protoplast culture and its
implications in the improvement of scented rose.
Acta Hort., 547: 147-58.
Steephen, M., S. Nagarjan and D. Ganesh, 2010.
Phloroglucinol and silver nitrate enhances axillary
shoot proliferation in nodal explants of Vitex
negundo L.-an aromatic medicinal plant. Iran J.
Biotechnol., 8: 82-89.
Oliveria, A.J.B., V.M. Carvalho, A. Ferreira, F.Y. Sato
and M.F.P. Machado, 2003. In vitro multiplication of
Tabernaemontana Fuchsiaefolia L. (Apocynaceae).
Rev. Arvore, 27: 421-425.
Bhot, M., S. Naphade, J. Varghese and N. Chandra,
2010. In vitro culture studies in three varieties of
Codiaeum Variegatum (L.) blume using node
explants from field Growth plants. J. Cell Tiss. Res.,
10: 2439-2444.
Murali, S., D. Sreedhar and T.S. Lokeswari, 1996.
Regeneration through somatic embryogenesis in
Rosa hybrida L cv Arizona I hybrid tea. Euphytica,
91: 271-5.
Murashige, T. and A. Skoog, 1962. A revised medium
for rapid growth and bioassays with tobacco tissue
culture. Physiol. Plantarum, 15: 473-497.
Yan, M., D.H. Byrne and C. Jing, 1996. Propagation
of rose speciese in vitro. Vitro Cell Dev. Biol. Plant,
32: 103-108.
19. Hasegawa, P.M., 1980. Factors affecting shoot and
root initiation from cultured rose shoot tips. J. Amer.
Soc. Hort. Sci., 105: 216-20.
20. Wulster, G. and J. Sacalis, 1980. Effects of
auxins and cytokinins on ethylene evolution and
growth of rose callus tissue in sealed vessels. Hort.
Sci., 15: 736-7.
21. Cai, J.M.Y. and D.L. Qian, 1984. Induction of
multiple shoots and rapid propagation of clones
of China rose [Rosa Chinensis]. Plant Physiol.
Communications Zhiwu Shenglixue Tongxun, 5: 37-8.
22. Gaspar, T.A., C. Kevers, C. Panel, H. Greppin,
D.M. Reid and T.A. Thorpe, 1996. Plant hormones
and plant growth regulators in plant tissue
culture. In vitro Cell Dev. Biol. Plant, 32: 272-289.
23. Sharma, M., M. Modgil and D.R. Sharma, 2000.
Successful propagation in vitro of apple rootstock
MM 106 and influence of Phloroglucinol. Indian J.
Exp. Biol., 38: 1236-1240.
24. Bassuk, N.L., L.D. Hunter and B.H. Howard, 1981.
The apparent involvement of polyphenol oxidase
and phloridzin in the production of apple rooting cofactors. J. Hortic. Sci., 56: 313-322.
25. Modgil, M., D.R. Sharma and S.V. Bhardwaj, 1999.
Micropropagtion of apple cv. Tydeman's Early
Worcester. Scientia Hortic., 81: 179-188.
26. Wang, Q., 1991. Factors affecting rooting of
microcuttings of the pear rootstock BP10030. Scientia
Hortic., 45: 209-213.
27. Gaspar, T., 1991. Vitrification in micropropagation. In:
Bajaj Y.P.S. (ed.), Springer, Berlin.
28. Aklan, K., S. Cettner, Y. Aka-Kacar and Y. YalcinMendi, 1997. In vitro multiplication of clonal apple
rootstocks M.9 and M.26 and MM.106 by meristem
culture. Acta Hort., 441: 325-327.
29. Jones, O.P., 1976.
Effect
of phloridzin
and phloroglucinol on apple shoots. Nature,
262: 392-393.
30. Jamesa, D.J., V.H. Knighta and I.J. Thurbona, 1980.
Micropropagation of red raspberry and the influence
of phloroglucinol. Scientia Hortic., 12: 313-319.
31. Demiralay, A., Y. Yalacin-Mendi, Y. Aka-kacar and S.
Cettner, 1998. In vitro propagation of Ficus carica L.
var. Bursa siyahi through meristem culture. Acta
Hort., 480: 165-7.
32. Debabrata, S. and S.N. Prakash, 2000. Phloroglucinol
enhances growth and rate of axillary shoot
proliferation in potato shoot tip cultures in vitro.
Plant Cell Tiss. Org., 60: 139-149.
965
Am-Euras. J. Agric. & Environ. Sci., 12 (7): 960-966, 2012
33. Pages, G., P. Lenomand, G. L’Allemania,
J.C. Chambard, S. Meloche and J. Pouyssegur, 1991.
Mitogen activated protein kinases p42mapk and
p44mapk are required for fibroblast proliferation.
Proc. Natl. Acad. Sci., 90: 8319-8323.
34. Robinson, M.J. and M.H. Cobb, 1997. Mitogen
activated protein kinase pathways. Curr. Opin. Cell
Biol., 9: 180-186.
35. Nakamure, T., K. Nagayama, K. Uchida and
R. Tanaka, 1996. Antioxidant activity of
phlorotannins isolated from the brown alga Eisenia
bicyclis. Fisher Sci., 62: 923-926.
36. Kim, J.A., J.M. Lee, D.B. Shin and N.H. Lee, 2004.
The antioxidant activity and tyrosinase inhibitory
activity of phlorotannins in Ecklonia cava. Food
Sci. Biotechnol., 13: 476-480.
37. Jabbarzadeh, Z. and M. Khush-Khui, 2005. Factor
affecting tissue culture of damask Rose (Rosa
damascena Mill.), Scientia Hortic., 105: 475-482.
38. Kim, C.K., J.D. Chung, S.K. Jee and J.Y. Oh, 2003.
Somatic embryogenesis from in vitro grown leaf
explants of Rosa hybrida L. J. Plant Biotechnol.,
5: 161-4.
39. Khosh-Khui, M. and Z. Jabbarzadeh, 2007. Effects of
several variables on in vitro culture of Damask Rose
(Rosa damascena Mill.) Acta Hortic., 751: 389-393.
40. Saffari, V.R., G.R. Sharifi-Sirchi and M.H. TorabiSirchi, 2011. Enhancing rooting consistency in
Rosa damascena scions. Afri. J. Biotechnol.,
10: 16495-16500.
41. Mirza, M.Q.B., A.H. Ishfaq, H. Azhar, A. Touqeer
and A.A. Nadeem, 2011. An efficient protocol for in
vitro propagation of Rosa gruss an teplitz and Rosa
centifolia. Afri. J. Biotechnol., 10: 4564-4573.
966