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1.
S. Hindson A. R. McElroy C. Portelance 《In vitro cellular & developmental biology. Plant》1998,34(3):181-184
Summary Various preconditioning treatments of alfalfa (Medicago sativa L.) somatic embryos to improve embryo quality and conversion were studied. Four different regenerating genotypes were compared.
Embryogenic cultures were established in liquid culture. Globular embryos were collected and plated on an embryo development
medium until they reached cotyledonary stage. They were then exposed to three treatments: a standard embryo development medium
(control), media supplementation with 1 μM abscisic acid (ABA), 50 mM glutamine and 5% sucrose (T), additional supplementation with 50 μM ABA (TT), and additional supplementation followed by desiccation (TTD). Treatments affected embryo conversion, but not uniformly
for all genotypes. Embryo conversion was increased (P<0.05) by pretreatment (T), while only one exhibited any response to additional ABA (T vs. TT). Desiccation decreased (P<0.05) conversion of pretreated embryos (TT vs. TTD) of all genotypes. The effect of treatments on plantlet weight was less
pronounced and inconsistent across genotypes. 相似文献
2.
Summary The effects of abscisic acid (ABA) (0, 0.09 μM, 0.19 μM, 0.28 μM, and 0.38 μM) or ancymidol (0, 0.98 μM, 1.95 μM, 2.93 μM, 3.90 μM) in embryo germination medium on the conversion of primary embryos to plantlets and secondary embryogenesis were evaluated
for asparagus. ABA and ancymidol each significantly enhanced both responses. ABA was more effective than ancymidol in promoting
the conversion of primary embryos to plantlets, while the converse was true for the production of secondary embryos. The most
effective treatments for embryo conversion were 0.19 and 0.28 μM ABA; 75–77% bipolar and 55–57% globular embryos converted to plantlets. For secondary embryogenesis, the most effective treatments
were 1.95 and 2.93 μM ancymidol; 99–101 and 84–86 somatic embryos were produced from 10 globular and 10 bipolar embryos, respectively. Bipolar
embryos generally converted to plantlets better than globular embryos, but more secondary embryos were produced from globular
embryos than from bipolar embryos in all treatments. ABA and ancymidol also affected the morphology of the plantlets produced.
The plantlets from the embryos incubated on the medium with ancymidol had strong and thick shoots and roots, while those on
the medium with ABA had long, thin shoots and short thin roots. 相似文献
3.
Summary This paper investigates maintenance and proliferation of somatic embryogenesis systems for Ulmus minor and U. glabra. Proliferation occurred with subculture of embryogenic calluses. The calluses were mainly formed by friable nodules composed
of meristematic cells organized into proembryogenic cell masses (PEMs) and thin-walled vacuolated parenchymatic cells. Cotyledonary
embryos, with procambial strands and differentiation of their vascular tissues as well as visible root meristems, were identifiable
after 18d of culture on a proliferation medium with 0.44 μM benzyladenine (BA). The shoot meristem was only occasionally well developed. Somatic embryo multiplication from elm embryogenic
calluses is a clearly asynchronic system, and PEMs as well as embryos at all stages of development are observed simultaneously
at the end of subculture period. Factors affecting the proliferation of elm embryogenic callus, such as culture medium, carbon
source and genotype, were studied. Basal medium (MS) or medium supplemented with 0.44 μM BA produced the highest number of somatic embryos. Somatic embryo production was higher with sucrose or glucose than with
maltose, and significant differences were also found among the four embryogenic lines tested. The use of liquid medium with
filter paper support is an essential step for the survival of isolated somatic embryos during the germination stage. The addition
of 0.22 μM BA′ to liquid MS medium was the best treatment for germination and plantlet conversion of elm somatic embryos. 相似文献
4.
Ana Sofia Duoue Ana Sofia Pires Dulce Metelo Dos Santos Pedro Fevereiro 《In vitro cellular & developmental biology. Plant》2006,42(3):270-273
Summary Plants were suecessfully régenerated via somatic embryos from 3-yr-old cell suspension cultures of Medicago truncatula Gaertin. cv. Jemalong line M9-10a. The cultures were originally initiated from callus induced in well-expanded leaflets of
30 d in vitro-grown plants, Suspension cultures were established in stirred-liquid Murashige and Skoog (MS) basal salts and vitamins supplemented
with 2.3 μM 2.4-dichlorophenoxyacetic acid (2,4-D) and 2.3 μM kinetin (Kin) and subeultured weekly. Somatic embryogenesis induction step was conducted in liquid MS medium containing 0.45
μM 2,4-D and 0.91 μM zeatin (Zea), during 1,2, and 3wk after subculture. Induced and non-induced cultures were transferred to solid embryo proliferation
medium [EPM-MS basal salts and vitamins solidified with 0.2% (w/v) gelrite]. Somatic embryos developed until the late torpedo/dicotyledonary
stages. We found that the best condition for the development of somatic embryos was achieved when suspension cultures were
not subjected to the induction step. Induction of 1 and 2 wk led to a decrease in the recovery of somatic embryos and the
3-wk treatment resulted in no differentiation of somatic embryos. Plant regeneration was obtained in all conditions (except
for 3wk induction) when embryos were transferred to an embryo conversion medium [ECM, similar to EPM but solidified with 0.7%
(w/v) agar]. Embryo conversion rates were 54.5±1.6, 52.5±18.5, and 41.6±8.4% for 0, 1, and 2 wk induction treatments, respectively.
These plants were successfully transferred to the greenhouse where they matured and produced seeds. 相似文献
5.
Blanca Olivia Cañedo Ortiz Martha Evelia Pérez Reyes Eugenio Pérez Molphe Balch 《In vitro cellular & developmental biology. Plant》2000,36(4):268-272
Summary Efficient plant regeneration systems via somatic embryogenesis have been developed for Acacia farnesiana and Acacia schaffneri [Leguminosae (Mimosoideae)]. The protocol used in this study consisted of placing immature, zygotic embryos of these species
in Murashige and Skoog semi-solid basal medium supplemented with 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.65 μM kinetin to induce callus. Some parts of the callus were used for direct embryo differentiation and others for establishment
of cell suspension cultures. In the first case, somatic embryos were produced on semi-solid differentiation media without
growth regulators or with abscisic acid (ABA). The higher number of somatic embryos, 345 and 198 embryos per g callus in A. farnesiana and A. schaffneri, respectively was obtained in media without growth regulators, but adding ABA increased the percentage of embryos that reached
more advanced differentiation stages. The production of somatic embryos was achieved starting from cell suspensions only when
these suspensions were plated into the semi-solid differentiation medium. Somatic embryos germinated on medium containing
217 μM adenine sulfate with efficiencies of 69% in A. farnesiana and 47% in A. schaffneri. Some somatic embryos that developed into plantlets were acclimatized in the greenhouse, and they grew into normal plants. 相似文献
6.
Summary We report a protocol for somatic embryogenesis and plantlet regeneration of Buchanania lanzan Spreng (Family—Anacardiaceae), which is a tropical fruit tree widely distributed in the dry forests of India. Calluses were initiated from immature zygotic
embryos cultured on Murashige and Skoog (MS) medium supplemented with various combinations of 2,4-dichlorophenoxyacetic acid
(2,4-D), 6-benzyladenine (BA) and/or 1-naphthaleneacetic acid (NAA). The highest frequency (60%) of somatic embryo induction
was obtained in cultures grown on MS medium fortified with 4.53 μM 2,4-D, 5.32 μM NAA and 4.48 μM BA. The medium supplemented with 15 μM abscisic acid (ABA) was most effective for maturation and germination of somatic embryos. This is the first report on somatic
embryogenesis in B. lanzan, which may be helpful for in vitro propagation, ex situ conservation and genetic manipulation of this species. 相似文献
7.
H. Brittain-Loucas S. R. Bowley B. D. McKersie 《In vitro cellular & developmental biology. Plant》1998,34(4):281-284
Summary The effects of callus inoculation concentration and culture duration on somatic embryogenesis of orchardgrass,Dactylis glomerata L., were evaluated in suspension cultures of an embryogenic genotype Embryogen-P. Somatic embryo formation was induced in
liquid SH medium containing 30 μM dicamba (SH-30 and 1.5% casein hydrolysate; embryo development was in liquid SH medium without plant growth regulators (SH-0);
and embryo maturation and germination occurred on solid SH-0 medium. Callus proliferation in SH-30 suspension cultures was
greatest when callus was inoculated into the liquid medium at a relatively high concentration of 4% (4 g callus/100 ml medium),
but the induction of somatic embryos was highest in this medium if the callus was inoculated at a lower concentration (<2%).
In a second experiment, somatic embryo yield was highest when SH-0 development medium was inoculated with suspension culture
callus at 0.1% concentration and declined markedly as inoculation concentration increased. Cell concentration is a critical
factor in regulating the somatic embryogenesis response in orchardgrass suspension cultures. 相似文献
8.
Lydia Reidiboym-Talleux Florence Diemer Martine Sourdioux Kathy Chapelain Ghislaine Grenier-De March 《Plant Cell, Tissue and Organ Culture》1998,55(3):199-209
Three different types of morphogenesis were identified in embryogenic cultures of Prunus avium grown on a proliferation medium
containing 0.54 μM NAA, 0.46 μM kinetin and 0.44 μM BA: a friable hyperhydric callus, repetitive embryogenesis and an embryogenic
tissue. Translucent and white somatic embryos were produced from the three types of morphogenesis but mainly from the embryogenic
tissue. These somatic embryos showed histological and cytological teratological features such as highly differentiated cells
with shrunken cytoplasm and destructured nuclei. For the four lines studied, somatic embryo production was improved by transferring
the embryogenic tissue to developmental media without auxin and cytokinin but supplemented with maltose alone or maltose and
10 μM ABA. Three weeks after transfer, the line showing the most embryogenesis produced 1404 somatic embryos per gram of embryogenic
tissue. A concentration of 263 mM maltose significantly increased the number of white somatic embryos for L 10 line, while
translucent somatic embryo production was improved by 88 mM maltose for L 16 line. The combination of maltose and ABA produced
different effects with each line. When used with 88 mM maltose, 10 μM ABA significantly increased white somatic embryo production
for two lines but decreased the production for one line. When combined with 263 mM maltose, ABA had no effect on white somatic
embryo production but significantly decreased the number of translucent somatic embryos. Cells of white somatic embryos contained
protein storage reserves and numerous lipid bodies, while those of translucent embryos did not contain storage reserves or
lipid bodies. After a two-month cold treatment conversion rate of white and translucent somatic embryos reached 8.5% and 35.2%
respectively.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
9.
Summary Regeneration of plants via somatic embryogenesis was achieved from zygotic embryo explants isolated from mature seeds of Schisandra chinensis. Merkle and Sommer's medium, fortified with 2,4-dichlorophenoxyacetic acid (2,4-D; 9.04 μM) and zeatin (0.09 μM), was effective for induction of embryogenic callus. The development of a proembryogenic mass and somatic embryos occurred
on Murashige and Skoog medium (MS) free of plant growth regulators. The embryogenic callus induced on Merkle and Sommer's
medium supplemented with 2,4-D (9.04 μM) and zeatin (0.09 μM) showed development of the maximum number of somatic embryos when transferred to MS medium free of plant growth regulators.
The maximum maturation and germination of cotyledonary somatic embryos (46.3%) occurred on MS medium supplemented with 2,4-D
(0.45 μM) and N6-benzyladenine (1.11 μM). The somatic embryo-derived plants were successfully hardned, with a survival rate of approximately 67%, and established
in the field. 相似文献
10.
S. B. Thencane S. R. Deodhar S. V. Bhosle S. K. Rawal 《In vitro cellular & developmental biology. Plant》2006,42(3):256-261
Summary Immature seeds of Garcinia indica Choiss, were exeised from immature fruits and cultured on Lloyd and McCown (1980), woody plant medium (WPM) with different
combinations of auxins and cytokinins. Somatic embryos were obtained on the media supplemented with 6-benzy laminopurine (BA;
2.2–22.1 μM) alone or in combination with α-naphthalene acetic acid (NAA; 2.6 μM) with 80% frequency within a period of 2–3 wk. Subculture of embryos on medium containing BA (16.0 μM) supplemented with indole-3-acetic acid (IAA: 2.8–5.7 μM) and/or kinetin (4.6 μM) gave rise to clusters of secondary somatic embryos along with maturation of primary embryos. In subsequent subculture on
hormone-free half-strength WPM, the embryo clusters germinated with an increase in the number of secondary somatic embryos.
About 70% of somatic embryos germinated into complete plantlets, which were successfully established under greenhouse conditions. 相似文献
11.
Summary Seedlings from 11 seed sources (lines) of American ginseng from different geographic regions were evaluated on Murashige and
Skoog medium (MS) containing 10 μM α-naphthaleneacetic acid (NAA) and 9 μM 2,4-dichlorophenoxyacetic acid (2,4-D) for callus development and somatic embryo formation. Leaf and stem explants callused
at a frequency of 18.2–100%, while somatic embryos were produced from these calluses at a frequency of 25–87.5% after 5 mo.
Suspension cultures of nine lines were established by transferring embryogenic callus to MS liquid medium with NAA and 2,4-D
at 2.5 and 2.25 μM, respectively, and maintained by subcultures every 8 wk. Globular somatic embryos from these cultures were germinated on
half-strength MS containing 1% activated charcoal, and roots >5 mm in length developed within 3 wk. A 7-d exposure to 3 μM gibberellic acid and 5 μM 6-benzylaminopurine significantly enhanced shoot development and promoted further root development. The chromosome number,
profiles of the common triterpenoid saponins (ginsenosides), and random amplified polymorphic DNA (RAPD) banding patterns
in plantlets derived from suspension culture were compared to those of zygotic seedlings. The chromosome number in root tip
cells and suspension cultured cells was 48. Patterns of the six major ginsenosides, determined by thin-layer chromatography,
in leaves of tissue culture-derived plantlets were identical to those in seedlings. RAPD patterns among plantlets originating
from the same tissue-cultured line were mostly identical; however, altered patterns were observed in some lines that had been
maintained in suspension culture for almost 4 yr. The results from this study indicate that propagation of desired ginseng
genotypes in suspension culture can be achieved, and that biochemical and molecular markers can be used for authentication
of resulting plantlets. 相似文献
12.
Summary The embryogenic potential of different Echinacea purpurea tissues, viz. leaf, cotyledon, and root, was investigated. Maximum embryo-induction was achieved from leaf dises cultured
on Murashige and Skoog medium supplemented with benzylaminopurine (5.0 μM) and indolebutyric acid (2.5 μM) where 95% of the explants responded, yielding an average of 83 embryos per explant within 4 wk of culture. Incubation of
cultures in the dark for an initial period of 14 d significantly increased the frequency of somatic embryogenesis (6–8-fold
in leaf explants). Exposure of the abaxial surface of leaves to the medium significantly increased the number of embryos.
Transfer of somatic embryos to a medium devoid of growth regulators resulted in 80% germination within 7 d. Over 73% of the
somatic embryos developed roots within 28 d of culture on a medium containing naphthaleneacetic acid (10 μM) with a maximum root number of 9.8 per plantlet. Transplanting ex vitro and acclimatization for a period of 7 d were sufficient to promote establishment of plants in the greenhouse, and more than
90% of the regenerated plants survived. 相似文献
13.
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
14.
Summary Efficient in vitro propagation of Ceropegia candelabrum L. (Asclepidaceae) through somatic embryogenesis was established. Somatic embryogenesis depended on the type of plant growth
regulators in the callus-inducing medium. Friable callus, developed from leaf and internode explants grown on Murashige and
Skoog (MS) medium supplemented with 4.52μM2,4-dichlorophenoxyacetic acid (2,4-D), underwent somatic embryogenesis. Compared to solid media, suspension culture was superior
and gave rise to a higher number of somatic embryos. Transfer of the friable callus developed on MS medium containing 4.52μM 2,4-D to suspension cultures of half- or quarter-strength MS medium with lower levels of 2,4-D (0.23 or 0.45 μM) induced the highest number of somatic embryos, which developed up to the torpedo stage. Somatic embryogenesis was asynchronous
with the dominance of globular embryos. About 100 mg of callus induced more than 500 embryos. Upon transfer to quarter-strength
MS agar medium without growth regulators, 50% of the somatic embryos underwent maturation and developed into plantlets. Plantlets
acclimatized under field conditions with 90% survival. 相似文献
15.
G. Vengadesan N. Selvaraj R. Prem Anand V. Gaba A. Ganapathi 《In vitro cellular & developmental biology. Plant》2005,41(6):789-793
Summary Suspension culture of cucumber (Cucumis sativus L.) has been an inefficient method for production of somatic embryos owing to problems with embryo maturation and conversion.
Embryogenic callus of cv. Green Long was induced on semisolid Murashige and Skoog (MS) medium containing 6.8 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.2 μM 6-benzylaminopurine (BA). A large number of globular somatic embryos were obtained on transfer of the callus to MS liquid
medium supplemented with 87.6 mM sucrose, 1.1 μM 2,4-D, and improved by the addition of 342.4 μM
l-glutamine. MS medium supplemented with 87.6 mM sucrose was more effective in somatic embryo production than other sugars. Subsequent development led to the formation of
heart-and torpedo-shaped embryos. Maturation of somatic embryos occurred on plant growth regulator-free MS semi-solid medium
containing 175.2 mM sucrose and 0.5 gl−1 activated charcoal. Conversion of embryos into plants was achieved on half-strength MS semi-solid medium containing 87.6
mM sucrose and 1.4 μM gibberellic acid (GA3) in a 16h photoperiod. Twenty-seven percent of embryos were converted into normal plants. 相似文献
16.
Q. C. Liu H. Zhai Y. Wang D. P. Zhang 《In vitro cellular & developmental biology. Plant》2001,37(5):564-567
Summary Using 15 Chinese and Japanese cultivars of sweetpotato, Ipomoea batatas (L.) Lam., we succeeded in developing an efficient plant regeneration system from embryogenic suspension cultures. The embryogenic
callus derived from shoot apices of the 15 cultivars was used to initiate embryogenic suspension cultures in Murashige and
Skoog (MS) medium containing 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Rapidly proliferating and well-dispersed embryogenic suspension cultures were established.
Cell aggregates 0.7–1.1 mm in size from embryogenic suspension cultures were transferred to solid MS medium supplemented with
9.05 μM of 2,4-D and formed embryogenic callus with somatic embryos. The embryogenic callus with somatic embryos was further transferred
to MS medium supplemented with 3.78 μM of abscisic acid, resulting in the germination of somatic embryos. Within 20 wk after the initiation, the frequencies of
cell aggregates forming plantlets reached approximately 100% for the 15 tested cultivars. These plantlets, when transferred
to soil, showed 100% survival. No morphological variations were observed. 相似文献
17.
Summary
In vitro propagation of Quassia amara L. (Simaroubaceae) was attempted using mature and juvenile explants. Attempts to establish in vitro culture using leaf and internode explants from a plant more than 15yr old were unsuccessful due to severe phenolic exudation.
Plant regeneration through direct and indirect somatic embryogenesis was established from cotyledon explants. Murashige and
Skoog (MS) medium with 8.9 μM N6-benzyladenine (BA) and 11.7 μM silver nitrate induced the highest number (mean of 32.4 embryos per cotyledon) of somatic embryos. Direct somatic embryogenesis
as well as callus formation was observed on medium with BA (8.9–13.3 μM). Semi-mature pale green cotyledons were superior for the induction of somatic embryos. Embryos developed from the adaxial
side as well as from the point of excision of the embryonic axis. More embryos were developed on the proximal end compared
to mid and distal regions of the cotyledons. Subculture of callus (developed along with the somatic embryos on medium with
BA alone) onto medium containing 8.9 μM BA and 11.7 μM silver nitrate produced a mean of 17.1 somatic embryos. Primary somatic embryos cultured on MS medium with 8.9 μM BA and 11.7μM silver nitrate produced a mean of 9.4 secondary somatic embryos. Most of the embryos developed up to early cotyledonary stage.
Reduced concentration of BA (2.2 or 4.4 μM) improved maturation and conversion of embryos to plantlets. Ninety percent of the embryos converted to plantlets. The optimized
protocol facilitated recovery of 30 plantlets per cotyledon explant within 80d. Plantlets transferred to small cups were subsequently
transferred to field conditions with a survival rate of 90%. 相似文献
18.
Summary Indirect somatic embryogenesis, encapsulation, and plant regeneration was achieved with the rare rhoeophytic woody medicinal
plant Rotula aquatica Lour. (Boraginaceae). Friable callus developed from leaf and internode explants on Murashige and Skoog (MS) medium with 0.45 μM 2,4-dichlorophenoxyacetic, acid (2,4-D) was most effective for the induction of somatic embryos. Subculture of the callus
onto half-strength MS medium with the same concentration of 2,4-D resulted in highly embryogenic callus. Suspension culture
was superior to solid medium culture for somatic embryogenesis. Embryogenic callus.during subsequent transfer to suspension
cultures of half-strength MS medium having 0.23 μM 2,4-D induced the highest number of somatic embryos (a mean of 25.6 embryos per 100 mg callus) and the embryos were grown
up to the torpedo stage. Transfer of embryos to half-strength MS basal solid medium allowed development, of 50% of the embryos
to the cotyledonary stage. Of the cotyledonary embryos, 90% underwent conversion to plantlets on the same medium. Encapsulated
cotyledonary embryos exhibited 100% conversion to plantlets. Ninety-five percent of the plantlets established in field conditions
survived, and were morphologically identical to the mother plant. 相似文献
19.
Summary Establishment of fast-growing, highly regenerable callus cultures was examined in Muscari armeniacum Leichtl. ex Bak. in order to develop an efficient genetic transformation system. High-frequency callus formation was obtained
from leaf explants of cv. Blue Pearl on media containing 2,4-dichlorophenoxyacetic acid (2,4-D), α-naphthaleneacetic acid
(NAA) or 4-amino-3,5,6-trichloropicolinic acid (picloram, PIC). Fast-growing, yellowish nodular callus lines and white friable
callus lines containing a few somatic embryos were established on initiation medium supplemented with 4.5 μM 2,4-D and with 54 μM NAA, respectively. The yellowish nodular calluses vigorously produced shoot buds after transfer to media containing 0.44–44
μM 6-benzyladenine (BA), whereas the white friable calluses produced numerous somatic embryos upon transfer to plant growth
regulator-free (PGR-F) medium. Histological observation of shoot buds and somatic embryos indicated that the former consisted
of an apparent shoot meristem and several leaf primordia, and the latter had two distinct meristematic regions, corresponding
to shoot and root meristems. Both shoot buds and somatic embryos developed into complete plantlets on PGR-F medium. Regenerated
plants showed no observable morphological alterations. High proliferation and regeneration ability of these calluses, were
maintained for over 2 yr. 相似文献
20.
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