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1.
Sung R. Min Seung G. Yang Jang R. Liu Pil S. Choi Woong Y. Soh 《Plant cell reports》1992,10(12):621-623
Culture conditions for high frequency somatic embryogenesis and plant regeneration from cotyledonary explants of Codonopsis lanceolata are described. The maximum induction frequency of somatic embryos from cotyledonary explants was 80% on Murashige and Skoog (MS) medium containing 6% sucrose with 1 mg/l 2,4-dichlorophenoxyacetic acid and 10% coconut water. Upon transfer onto MS basal medium containing 3% sucrose, most somatic embryos developed into plantlets.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
gibberellin a3
- MS
Murashige and Skoog 相似文献
2.
Vladimir Chalupa 《Plant cell reports》1990,9(7):398-401
Embryogenic cultures and somatic embryos were obtained from immature zygotic embryos of oak (Quercus robur L.) cultured on a modified MS medium and WPM containing BAP (1 mg·l–1) and GA3 (1 mg·l–1) or BAP and IBA. Germination and conversion of oak somatic embryos into plantlets was achieved on WPM containing a reduced concentration of cytokinin. Linden (Tilia cordata Mill.) somatic embryos developed in embryogenic tissues initiated from immature zygotic embryos cultured on a modified MS medium supplemented with 2,4-D (0.3-2.0 mg·l–1). Germination of linden somatic embryos and plantlet formation occurred on MS medium containing a low concentration of IBA. Oak and linden plantlets produced from somatic embryos were successfully established in soil. Somatic embryos and plantlets were also regenerated from embryogenic cultures of Quercus petraea and Tilia platyphyllos.Abbreviations BAP
6-benzyIaminopurine
- GA3
gibberellic acid
- IBA
indole-3-butyric acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog (1962)
- WPM
woody plant medium 相似文献
3.
Somatic embryogenesis was obtained from mature cassava cotyledons explants. A two-step medium sequence was developed for efficient embryogenesis. Application of 2,4-D (4 mg l-1) yielded proembryogenic masses which developed into somatic embryos after transfer to a medium containing NAA (0.01 mg l-1), BA (0.1 mg l-1) and GA3 (0.1 mg l-1). The 2,4-D concentrations used for embryo initiation strongly influenced embryo development. Among the cultivars tested, TMS 30395 was most responsive. Full strength MS basal medium alone or with 4 x MS micro salts was efficient for the formation of somatic embryos. Casein hydrolysate, adenine sulfate, nicotinic acid, glycine, tryptophan, and serine were ineffective for embryo development. High sucrose concentration (6%, w/v) inhibited the induction of somatic embryos, while 6% sucrose was optimal concentration for the development of somatic embryos after an induction treatment using 2% sucrose. Addition of 0.52 mg l-1 ABA to the induction media resulted in an increase in somatic embryos production. The ploidy levels of the regenerated plantlets were determined by flow cytometry analysis. Fifty regenerants tested were all tetraploids as the source plants and were morphologically normal. The implications of these results are discussed in relation to genetic transformation using the cotyledons as the explant source.Abbreviations ABA
abscisic acid
- BA
6-benzylaminopurine
- DAPI
4,6-diamidino-2-phenylindole
- SR
101 sulforhodamine
- GA3
gibberellic acid
- MCPA
methyl- chlorophenoxyacetic acid
- NAA
naphthalen-acetic acid
- PCPA
P-chlorophenoxyacetic acid
- 2,4-d
2,4-dichlorophenoxyacetic acid
- 2,4,5 T
2,4,5-trichlorophenoxyacetic acid 相似文献
4.
P. I. P. Perera V. R. M. Vidhanaarachchi T. R. Gunathilake D. M. D. Yakandawala V. Hocher J. L. Verdeil L. K. Weerakoon 《Plant Cell, Tissue and Organ Culture》2009,99(1):73-81
Coconut is a cross pollinating palm, propagated only by seeds. Tissue culture is the only vegetative propagation method available
for coconut. Consistent callogenesis was obtained by culturing unfertilised ovaries at -4 stage in CRI 72 medium containing
100 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.1% activated charcoal. Callusing was improved by application of 9 μM thidiazuron
(TDZ). Embryogenic calli were subcultured onto somatic embryogenesis induction medium containing 66 μM 2,4-D. Stunted growth
was observed in the somatic embryos after subculture onto CRI 72 medium containing abscisic acid (ABA). Maturation of somatic
embryos could be achieved in Y3 medium without growth regulators. Conversion of somatic embryos was induced by adding gibberellic acid (GA3) to conversion medium containing 5 μM 6-benzyladenine (BA) while 2-isopentyl adenine (2iP) increased the frequency of plant
regeneration. A total of 83 plantlets was produced from 32 cultured ovaries. 相似文献
5.
Summary Pepper (cv. New Mexico — 6 and Rajur Hirapur) plants were regenerated from immature zygotic embryos via direct somatic embryogenesis. Somatic embryos were formed directly, without any intervening callus, on the zygotic embryo apex, embryo axis and cotyledons on Murashige and Skoog's (MS) medium containing 2,4-D (418 M), thidiazuron (10 M) and a high concentration of sucrose (6–10%). The best response was observed on MS medium containing 2,4-D (9 M), coconut water (10%) and high sucrose (8%). The entire process of induction and maturation of the embryos was completed on the same medium. Histological examination indicated that secondary embryogenesis also occurred directly from the primary somatic embryos. Differentiation of embryos was nonsynchronous, and some embryos were swollen and distorted with fasciation. More than 70% of the mature normal somatic embryos germinated readily on MS medium containing GA3 or TDZ, alone and in combination, and following transfer to pots developed into normal plants.Abbreviations CM
Coconut milk
- 2,4-D
2,4-dichlorophonoxyacetic acid
- GA3
gibberellic acid
- MS
Murashige and Skoog (1962) medium
- NAA
napthaleneacetic acid
- TDZ
thidiazuron 相似文献
6.
An efficient in vitro plant regeneration system characterized by rapid and continuous production of somatic embryos using leaf and petiole expiants has been developed in sweetpotato [Ipomoea batatas L. (Lam.)]. The optimal somatic embryogenic response was obtained in the genotype PI 318846-3 with a two-step protocol: (1) stage I-incubation of expiants in the dark for 2 weeks on Murashige Skoog (MS) medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) (2.5 mg/l) and 6-benzylaminopurine (0.25 mg/l) and, (2) stage II-culture in the light on MS medium with abscisic acid (ABA) (2.5 mg/l). The addition of ABA was critical for enhanced production of somatic embryos. Secondary somatic embryos were produced from the primary embryos cultured on MS medium with 2,4-D at 0.2 mg/l. The somatic embryos were converted into normal plantlets when cultured on basal MS medium. Upon transfer to soil, plants grew well and appeared normal with no mortality. The system of somatic embryogenesis described here will facilitate tissue culture, germplasm conservation and gene transfer research of sweetpotato due to its rapidity (6 to 10 weeks), prolific plant production by direct embryogenesis, ease of secondary somatic embryo production and reproducibility.Abbreviations ABA
abscisic acid
- BAP
6-benzylaminopurine, 2,4-D-2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- KIN
kinetin
- MS
medium of Murashige and Skoog (1962)
- NAA
1-naph-thaleneacetic acid
- PIC
picolinic acid
- TDZ
thidiazuron 相似文献
7.
Ai Hua Chen Jing Li Yang Yu Da Niu Chuan Ping Yang Gui Feng Liu Chang Yeon Yu Cheng Hao Li 《Plant Cell, Tissue and Organ Culture》2010,102(3):357-364
We developed a new protocol for highly efficient somatic embryogenesis and plantlet conversion of Schisandra chinensis. Friable embryogenic callus was induced from cotyledonary leaves and hypocotyls of germinated zygotic embryos on Murashige
and Skoog (MS) agar medium containing 2,4-dichlorophenoxyacetic acid (2,4-D). Preculture of zygotic embryos on 2,4-D-containing
medium increased embryogenic callus induction efficiency. The highest embryogenic callus induction frequency of 56.7% was
obtained from shoot apical meristem-containing hypocotyl explants from 1-week-old germinated embryos on MS medium containing
4.0 mg l−1 2,4-D. Embryogenic callus proliferation, somatic embryo (SE) formation, and subsequent plantlet conversion occurred under
optimal culture conditions. The effects of MS medium strength, sucrose, gibberellic acid (GA3), and 6-benzyladenine (BA) on SE formation and plantlet conversion were evaluated. Low MS medium strength (1/4 to 1/2) was
necessary for SE formation, and the optimal sucrose concentration was 2.0%. Supplementing medium with GA3 negatively impacted SE formation and subsequent development. BA significantly increased the number of SEs and the plantlet
conversion capacity. One-third-strength MS medium with 1.0% sucrose and 0.5 mg l−1 BA produced the highest number of SEs (309 embryos from 9 mg embryogenic callus) and the highest frequency of plantlet conversion
from germinated SEs (52.6%). When transplanted to soil, 90% of the regenerated plants developed into normal plants. 相似文献
8.
Somatic embryogenesis was achieved in callus cultures derived from immature cotyledonary explants ofHardwickia binata Roxb., a multipurpose leguminous tree, on semisolid modified Murashige and Skoog's (mMS) medium containing 2900 mg/l potassium nitrate (KNO3) supplemented with 4.64 µM kinetin (Kn) and 5.37µM a-naphthaleneacetic acid (NAA). Somatic embryos proliferated rapidly after transfer to MS basal medium supplemented with 2052.6 µM L-glutamine and 0.084 µM gibberellic acid (GA3). Maturation of somatic embryos was achieved on half-strength MS basal medium supplemented with 1.23 µM IBA and 2% (w/v) sucrose. Histological studies confirmed different developmental stages of somatic embryogenesis inHardwickia binata.
Abbreviations BA
N6-benzyladenine
- Kn
kinetin
- NAA
a-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IBA
indole-3-butyric acid
- GA3
gibberellic acid
- MS
Murashige and Skoog (1962) medium
- mMS
modified Murashige and Skoog (1962) medium 相似文献
9.
Summary
Kalopanax pictus (Thunb.) Nakai is a tall tree, and its wood has been used in making furniture, while its stem bark is used for medicinal
purposes. Here, we report on the micropropagation of Kalopanax pictus via somatic embryogenesis. Embryogenic callus was induced from immature zygotic embryos. The frequency embryogenic callus
induction is influenced by days of seed harvest. Callus formation was primarily observed along the radicle tips of zygotic
embryos incubated on Murashige and Skoog (MS) medium with 4.4 μM 2,4-dichlorophenoxyacctic acid (2,4-D). Somatic embryogenesis was observed following transfer of embryogenic callus to MS
medium lacking 2,4-D. Somatic embryos at the cotyledonary stage were obtained after 6 wk following culture. Frequency of conversion
of somatic embryos into plantlets was low (35%) on a hormone-free MS basal medium, but it increased to 61% when the medium
was supplemented with 0.05% charcoal. Gibberellic acid (GA3) treatment markedly enhanced the germination frequency of embryos up to 83%. All plantlets obtained showed 98% survival on
moist peat soil (TKS2) artificial soil matrix. About 30 000 Kalopanax pictus plants were propagated via somatic embryogenesis and grown to 3-yr-old plants. These results indicate that production of
woody medicinal Kalopanax pictus plantlets through somatic embryogenesis can be practically applicable for propagation. 相似文献
10.
P. Giridhar Vinod Kumar G. A. Ravishankar 《In vitro cellular & developmental biology. Plant》2004,40(6):567-571
Summary A novel protocol has been developed for inducing somatic embryogenesis from leaf cultures of Decalepis hamiltonii. Callus was obtained from leaf sections in Murashige and Skoog (MS) medium supplemented with α-naphthaleneacetic acid (NAA)+N6-benzyladenine (BA) or 2,4-dichlorophenoxyacetic acid (2,4-D)+BA. Nodular embryogenic callus developed from the cut end of
explants on media containing 2,4-D and BA, whereas compact callus developed on media containing NAA and BA. Upon subsequent
transfer of explants with primary callus onto MS media containing zeatin and/or gibberellic acid (GA3) and BA, treatment with zeatin (13.68μM) and BA (10.65 μM) resulted in the induction of the highest number of somatic embryos directly from nodular tissue. The maturation of embryos
took place along with the induction on the same medium. Embryogenic calluses with somatic embryos were subcultured onto MS
basal medium supplemented with 4.56μM zeatin+10.65 μM BA. After 4wk, more extensive differentiation of somatic embryos was observed. The mature embryos developed into complete
plantlets on growth regulator-free MS medium. A distinct feature of this study is the induction of somatic embryogenesis from
leaf explants of Decalepis hamiltonii, which has not been reported previously. By using this protocol, complete plantlets could be regenerated through indirect
somatic embryogenesis or organogenesis from leaf explants in 12–16 wk. 相似文献
11.
We describe a protocol for somatic embryogenesis of Protea cynaroides, with potential for high frequency production of this important horticultural species. Somatic embryos formed directly on
both P. cynaroides mature zygotic embryos and excised cotyledons cultured on MS medium without growth regulators. The addition of growth regulators
such as naphthalene acetic acid (NAA) (5; 13 and 27 μM) and 2,4-dichlorophenoxyacetic acid (2,4-D) (5; 11 and 23 μM), in combination
with thidiazuron (TDZ) (1 μM), benzylaminopurine (BAP) (1 μM) or kinetin (1 μM) suppressed the formation of somatic embryos.
After eight weeks in culture, formation of somatic embryos was observed. Zygotic explants formed the most embryos when cultured
in a 12-h photoperiod in comparison to explants cultured in the dark. Up to 83% of these embryos germinated after transferal
to the germination medium containing 0.3 μM GA3. Significantly fewer embryos germinated in MS medium with no growth regulators, or supplemented with higher concentrations
of GA3, while low germination percentages were also observed in MS media containing casein hydrolysate and coconut water. The germination
of normal somatic embryos (two separate cotyledons and a single radicle) was observed only in media containing either no growth
regulators, 0.3 μM GA3 or 1 μM GA3. All embryos that germinated in high concentrations of GA3 were malformed. 相似文献
12.
Plant regeneration via somatic embryogenesis was achieved in callus cultures derived from nodal explants of in vitro grown seedlings and excised mature zygotic embryos of three bamboo species on Murashige and Skoog's (MS) basal medium supplemented with 0.5 mg/l kinetin (Kn), 2.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), 10 mg/l adenine sulphate (Ads) and 3% (w/v) sucrose incubated in the light or in the dark. Somatic embryos germinated (95–98%) into normal plants and were transferred to soil with 95% success. In vitro flowering was induced on shoots developed from nodal explants taken from somatic embryo regenerated plants of Bambusa vulgaris, Dendrocalamus giganteus and Dendrocalamus strictus on half-strength MS basal medium supplemented with 0.25 mg/l indole-3-butyric acid (IBA), 0.5 mg/l Ads, 0.5 mg/l gibberellic acid (GA3) and 3% sucrose.Abbreviations BAP
6-benzylaminopurine
- Kn
kinetin
- Ads
adenine sulphate
- IBA
indole-3-butyric acid
- NAA
1-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog (1962) basal medium
- GA3
gibberellic acid 相似文献
13.
Plant regeneration through somatic embryogenesis from mature leaf explants of Eryngium foetidum,a condiment 总被引:1,自引:0,他引:1
Ignacimuthu S. Arockiasamy S. Antonysamy M. Ravichandran P. 《Plant Cell, Tissue and Organ Culture》1999,56(2):131-137
Eryngium foetidum L. is an important plant cultivated as a leafy vegetable and for its essential oil, which are of high economic
value in international trade market. Plants were regenerated through somatic embryogenesis from mature leaf explants of field
grown plants. Leaf explants produced dark brown, compact callus on Linsmaier and Skoog (LS) medium with the combination of
1.0 mg l-1 2,4-dichlorophenoxy acetic acid (2,4-D) and 1.0 mg l-1 benzylaminopurine (BAP). Somatic embryos were induced from embryo-forming callus cultures on Murashige and Skoog (MS) medium
supplemented with 0.1 mg l-1 2,4-D, 2.0 mg l-1 BAP and 1.0 mg l-1 gibberellic acid (GA3). Subsequently, conversion of these somatic embryos into plantlets occurred on MS medium supplemented with 1.0 mg l-1 GA3 and/or 0.1 mg l-1 BAP. The regenerated shoots were rooted and elongated on MS medium supplemented with 0.1 mg l-1 IAA and 1.0 mg l-1 GA3. These plantlets were hardened and transferred to the soil.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
14.
High frequency of plant regeneration in sunflower from cotyledons via somatic embryogenesis 总被引:7,自引:0,他引:7
Summary A plant regeneration methodvia somatic embryogenesis of severalHelianthus annuus L. genotypes was developed. Starting from cotyledonary explants high frequency embryo induction was obtained following several subcultures on defined media. An appropriate cotyledon developmental stage was identified. Etiolated explants and darkness treatment were necessary to obtain somatic embryos in all tested genotypes. After 20–25 days on somatic induction medium containing an auxin:cytokinin ratio of 1:1, the germination of embryos was induced by a reduction of the hormonal ratio (1:2). Shoots were excised from callus and transferred onto a medium containing various vitamins. The range of embryogenesis frequency was 33–72%, depending on the genotype. High frequency of rooting (49–82%) was obtained using a medium supplemented with 0.5 mg/L of ancymidol and by a reduction of photoperiod. A large percentage of somatic embryos developed into normal regenerated plants producing viable seeds.Abbreviations MS
Murashige and Skoog (1962)
- NAA
naphthaleneacetic acid
- BAP
benzylaminopurine
- GA3
gibberellic acid
- EIM
embryo induction medium
- GM
germination medium
- VM
vitamins medium
- RA2
ancymidol rooting medium
- EtOH
ethanol 相似文献
15.
Summary Embryogenic cell suspension cultures and somatic embryos of five genotypes of beech, were obtained from aged cultures derived from immature zygotic embryos cultured on solid medium containing both 2, 4-dichlorophenoxyacetic acid and N6-benzyladenine. The origin of somatic embryos was traced from single cells. Embryos remained arrested at the globular stage on liquid media, further development was achieved after plating embryogenic aggregates on Murashige and Skoog's medium with half strength major salts supplemented with glutamine and low levels of growth regulators. Cultures of different genotypes showed significant differences in maturation frequency which was not affected by the hormone treatments assayed. The frequency of conversion of embryos into plantlets was low. This frequency increased after cold storage of embryos for up to 7 months.Abbreviations BA
N6-benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- EtOH
ethanol
- GA3
giberrellic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- MS
Murashige and Skoog (1962)
- NAA
naphthalene acetic acid
- WPM
woody plant medium (Lloyd and McCown, 1980)
- Z
zeatin 相似文献
16.
R. Lührs H. Lörz 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1987,75(1):16-25
Summary Immature embryos of 41 lines of barley were screened in vitro for callus induction and somatic embryogenesis on different media to establish totipotent cultures. The use of modified MS and CC media, both supplemented with 1 g/l casein hydrolysate, and the substitution of agarose for agar resulted in the highest frequencies of somatic embryo induction. Embryogenic callus was induced and plants regenerated from 23 of the lines tested. The auxins 2,4-D, dicamba, picloram and 2,4,5-T were suitable for embryogenic callus induction. High frequencies of somatic embryo germination occurred on CC medium supplemented with 1 mg/l IAA and 0.05 mg/l zeatin. A strong genotypic effect on the capacity and frequency of embryogenic callus formation was found. Cultivar Golden Promise always gave the best results. Experiments with field grown material in 3 consecutive years showed that environmental factors also strongly influenced the induction of somatic embryogenesis and plant regeneration.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- 2,4,5-T
2,4,5-trichlorophenoxyacetic acid
- dicamba
3,6-dichloro-o-anisic acid
- picloram
4-amino-3,6,6-trichloropicolinic acid
- NAA
naphtaleneacetic acid
- IAA
indole-3-acetic acid
- ABA
abscisic acid
- BAP
6-benzyl amino purine
- 2iP
6-(3-methyl-2 butenyl 1-amino)purine
- GA3
gibberellic acid 相似文献
17.
Summary Somatic embryogenesis was induced in callus cultures derived from nucellar tissue of cashewnut (Anacardium occidentale L.). Callus was obtained from nucellar tissue after 3 wk of culture on semisolid Murashige and Skoog (MS) basal medium supplemented
with 2,4-dichlorophenoxyacetic acid (2,4-D, 5 μM)+gibberellic acid (GA3, 15 μM)+N6-benzyladenine (BA, 5 μM). This callus gave rise to an embryogenic mass after 9 wk on maintenance medium containing 2,4-D (10 μM)+GA3 (15 μM)+4% sucrose +0.5% activated charcoal +10% coconut water (CW) +0.05% casein hydrolysate (CH). The embryogenic mass, after
transfer to medium supplemented with 2,4-D (5 μM)+GA3 (30 μM)+4% sucrose +0.5% activated charcoal +10% CW +0.05% CH, gave rise to somatic embryos. The developmental stages of somatic
embryos were observed using light and stereo microscopes. Histological study of somatic embryo development was also carried
out. The present study would be useful for clonal propagation, and variety improvement in cashewnut, which is essential due
to its increasing demand and export potential. 相似文献
18.
Summary Embryogenic masses were obtained from immature leaves of peanut (Arachis hypogaea L.) cultured on a medium containing 20 mg/l 2,4-D. Somatic embryos developed from these masses following transfer to a medium containing 3 mg/l 2,4-D. The embryo morphology was quite variable. Following transfer to hormone-free medium, these embryos germinated. Shoot elongation was obtained in 25% of the embryos following transfer to a medium supplemented with 0.5 mg/l each of BAP and Kn. The plants grown in vitro by this method survived in sand:soil mixture and were grown to maturity.Abbreviations ABA
abscisic acid
- BAP
6-benzyl amino purine
- 2,4-D
2,4 dichlorophenoxyacetic acid
- GA3
gibberellic acid
- Kn
kinetin
- NAA
1-naphthaleneacetic acid
- 2,4,5-T
2,4,5-trichlorophenoxyacetic acid
- Z
zeatin 相似文献
19.
Heung Kyu Moon Ji Ah Kim So Young Park Yong Wook Kim Ho Duck Kang 《Journal of Plant Biology》2006,49(4):320-325
We tested the possibility of plantlet formation via somatic embryogenesis with leaf segments and mature zygotic embryos from
a rare and endangered tree species,Oplopanax elatus. To induce calli, expiants were cultured under darkness in a solid MS medium containing 3% sucrose, 1g L-1 glutamine, and 0.3% gelrite. Treatment supplements included 2,4-D alone or in combination with thidiazuron. Generally, callus
induction and growth were good from leaf expiants, whereas embryogenic calli could be induced only from zygotic embryos. These
embryogenic calli were white or pale yellow and very friable. ABA and activated charcoal appeared to be important factors
when inducing somatic embryos, with optimum levels being 0.1 mg L-1 and 0.02%, respectively. Many somatic embryos showed abnormalities during their development on the germination medium, but
35% could be converted if placed on a medium containing gibberellic acid (GA3). The germinating embryos sometimes formed secondary embryos at the lower portion of the hypocotyls. Normal or converted
plantlets were acclimatized in an artificial soil mixture; their survival was about 60% after two months. This culturing system
provides a feasible approach for regenerating plants, via somatic embryogenesis, from mature zygotic embryos. 相似文献
20.
Meena K. Cheruvathur G. Krishna Kumar T. Dennis Thomas 《Plant Cell, Tissue and Organ Culture》2013,113(1):63-71
Young healthy cotyledon and leaf explants of Rhinacanthus nasutus (L.) Kurz. were incubated on Murashige and Skoog (MS) medium supplemented with 1.0–5.0 mg/l 2, 4-dichlorophenoxyacetic acid (2,4-D) either alone or in combination with 0.3–1.5 mg/l indole-3-butyric acid (IBA). The optimum callus induction (100 %) was observed from cotyledon explants on MS medium supplemented with 4 mg/l 2, 4-D and 0.5 mg/l IBA. The friable, embryogenic callus when subcultured on half strength MS medium supplemented with IBA (3.0–5.0 mg/l) produced several somatic embryos at various stages of development (globular, heart, torpedo) after 45 days of culture. The highest frequency of callus embryogenesis was observed on ½MS medium supplemented with 4.0 mg/l IBA. Moreover, 47 % of incubated callus responded with a mean number of 16.3 somatic embryos per gram callus. For germination, somatic embryos at the torpedo stage were isolated and subcultured on ½MS medium supplemented with 0.5 mg/l each of 6-benzyladenine and indole-3-acetic acid. After 45 days of culture, plantlets developed with mean lengths of 3.8 cm. Somatic embryos at the torpedo stage were collected and suspended in a matrix of MS medium containing sodium alginate (3 % W/V), dropped into 100 mM calcium chloride (CaCl2·2H2O) solution for the production of synthetic seeds. Optimum growth ability of synthetic seed was obtained on MS medium supplemented with 0.2 mg/l gibberellic acid (GA3). Well developed healthy plantlets derived from somatic embryos and synthetic seeds were hardened and successfully transplanted to soil. 相似文献