共查询到20条相似文献,搜索用时 562 毫秒
1.
Mei-Chun Lu 《Plant Cell, Tissue and Organ Culture》2004,78(1):93-96
High frequency plant regeneration was induced from protocorm-derived callus cultured on half-strength of Murashige—Skoog medium
with 2,4-dichlorophenoxyacetic acid (2,4-D, 0–5 mg l−1) and 1-phenyl-3-(1,2,3-thiadiazol-5-yl, 0–1 mg l−1) urea (TDZ) in the dark. Twelve totipotent callus lines were selected within 76 callus lines regenerated on half-strength
of Murashige—Skoog (MS) medium with 0.5 mg l−1 TDZ. The proliferation rate was 4–5-fold in fresh weight after 30 days of culture on half-strength MS medium containing 5
mg l−1 2,4-D and 0.5 mg l−1 TDZ in the dark. The maximum number of shoot buds generated by 0.01 g callus explant was 134 after 4 months of culture. These
calli were regenerated to plantlets via protocorm-like bodies (PLBs) after 75–150 days of culture. The shoots, with two true
leaves, were transferred to hormone-free medium, rooting and eventually formed plantlets. Totipotent callus lines of Pleione formosana Hayata have been successfully established in this study. 相似文献
2.
Summary Callus of Phalaenopsis Nebula was induced from seed-derived protocorms on 1/2 Murashige and Skoog (MS) basal medium plus 0–1.0 mg l−1 (0–4.52 μM) N-phenyl-N′-1,2,3,-thiadiazol-5-yl urea (TDZ) and/or 0–10 mg l−1 (0–45.24 μ M) 2,4-dichlorophenoxyacetic acid (2,4-D). Protocorms 2 mo. old performed better than 1-mo.-old protocorms for callus induction.
More calluses formed on 1/2 MS basal medium supplemented with 0.1–1.0 mg l−1 (0.45–4.52 μM) TDZ. These calluses could be maintained by subculturing every month with basal medium supplemented with 0.5 mg l−1 (2.27 μM) TDZ and 0.5 mg l−1 (2.26 μM) 2,4-D. Protocorm-like bodies were formed, and plants regenerated from these calluses on 1/2 MS basal medium alone or supplemented
with 0.1–1.0 mg l−1 (0.45–4.52 μM) TDZ. Plantlets were then potted on sphagnum moss in the greenhouse and grew well. No chromosomal abnormalities were found
among the root-tip samples of 21 of the regenerated plantlets that were successfully acclimatized. 相似文献
3.
An in vitro protocol for efficient plant regeneration has been developed from mature embryo explants of highland barley (Hordeum vulgare L. var. nudum Hk. f.) under endosperm-supported culture. Embryos with (endosperm-supported culture, ES) or without endosperm
(non-endosperm-supported culture, NES) were excised from mature seeds and cultured on MS medium supplemented with various
concentrations of 2,4-D (1–5 mg l−1) for callus induction. The percentage of callus induction from ES explants was significantly (P < 0.05) lower than that from NES. The highest frequency (97.6%) of callus induction was obtained from NES explants on MS
medium containing 3 mg l−1 2,4-D. When the primary calli were maintained at a reduced concentration of 2,4-D (0.5 mg l−1) for 3 weeks, embryogenic calli were formed. The embryogenic calli were then transferred to MS medium supplemented with different
concentrations of BA (1–5 mg l−1) and 500 mg l−1 casein hydrolysate (CH) for shoot regeneration. However, the capacity of plant regeneration from ES explant-derived calli
was significantly (P < 0.05) higher than that from NES. The best response (81.3%) was observed from ES explant-derived calli on MS medium containing
2 mg l−1 BA. Regenerated plantlets with well-developed root systems were transferred to pots where they grew well, attained maturity
and produced fertile seeds. This method could be employed for genetic manipulation studies. 相似文献
4.
Qian Zhang Jianjun Chen Richard J. Henny 《Plant Cell, Tissue and Organ Culture》2006,84(2):100161-100168
A simple and effective method of regenerating Syngonium podophyllum ‘Variegatum’ via direct somatic embryogenesis has been established. Leaf and petiole explants were cultured on Murashige
and Skoog (MS) medium supplemented with N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU) or N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ) with either α-naphthalene acetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D). Somatic embryos directly formed at one or two sides of petiole explants on MS medium supplemented 2.5 mg l−1 TDZ with 0.5 mg l−1 NAA or 2.0 mg l−1 TDZ with 0.2 mg l−1 NAA or with 0.2 and 0.5 mg l−1 2,4-D, respectively. The frequency of petiole explants with somatic embryos produced was as high as 86% when cultured on medium
containing 2.5 mg l−1 TDZ with 0.5 mg l−1 NAA. Up to 85% of somatic embryos were able to germinate after transferring onto medium containing 2.0 mg l−1 6-benzylaminopurine (BA) and 0.2 mg l−1 NAA. Approximately 50–150 plantlets were regenerated from a single petiole explant. However, there was no somatic embryo
formation from leaf explants regardless of growth regulator combinations used. Regenerated plantlets from petiole explants
were stable and grew vigorously after transplanting to a soilless container substrate in a shaded greenhouse. 相似文献
5.
Hippolyte Kodja Isabelle Robene-Soustrade Jacques Figier 《Acta Physiologiae Plantarum》1997,19(3):359-366
Callus cultures of Tabernaemontana persicariaefolia, (Apocynaceae), an endangered species endemic to the Mascarene Islands, were established from leaf explants on MS medium containing either
5 mg·l−1 2,4-D and 0.5 mg·l−1 BA or 5 mg·l−1 2,4-D, 0.5 mg·l−1 BA and 200 mg·l−1 DFMO. Histological studies showed regenerating nodules resembling globular embryos in calli after 4 weeks on the DFMO medium.
Green shoot formation was achieved by sequential subculture of the induced calli on media with gradually decreasing 2,4-D
concentrations (5→1→0 mg·l−1). Regeneration was greatly stimulated in the presence of DFMO. The first emergence of shoots occured 3 weeks earlier than
in untreated callus cultures. 相似文献
6.
Wan-Jun Zhang Jiang-Li Dong Ben-Guo Liang Yong-Sheng Jin Tao Wang 《In vitro cellular & developmental biology. Plant》2006,42(2):114-118
Summary We report a protocol for efficient plant regeneration of four tall fescue (Festuca arundinacea Schreb.) cultivars (‘Surpro’, ‘Coronado’, ‘Summer Lawn’, and ‘Fawn’) via somatic embryogenesis. Calli were initiated from
mature seeds grown on modified Murashige and Skoog (MMS) medium supplemented with 7.0mgl−1 (31.7μM) 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.05 mgl−1 (0.23 μM) kinetin (Kin). Calli were maintained and proliferated by subculture at monthly intervals on MMS medium containing 4.5 mgl−1 (20.4 μM) 2,4-D and 0.2mgl−1 (0.9 μM) Kin. Somatic embryos (SE) were induced from seed-derived calli on SE-induction medium (MMS supplemented with 2.0 mgl−1 2,4-D and 0.2mgl−1 Kin). Plantlets were regenerated from somatic embryogenic calli grown on modified SH medium supplemented with 2 mgl−1 Kin. Using this optimized protocol, 78.6–82.3% of mature seeds of all four cultivars produced SE clusters, of which 93.5–95.3%
regenerated into plants within 10 wk. The regenerants showed no phenotypic abnormalities. 相似文献
7.
Plant regeneration from protoplast culture of Crocus cancellatus was investigated using regenerable embryogenic calli obtained from shoot meristem culture on LS (Linsmaier and Skoog, 1965)
medium containing 4 mg l−1 kinetin and 1 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Protoplasts were isolated directly from embryogenic calli. The best protoplast growth
was found on those embedded in Ca-alginate beads and cultured with nurse cells in MS (Murashige and Skoog, 1962) medium supplemented
with 2 mg l−1 kinetin, 1 mg l−1 2,4-D and 100 mg l−1 ascorbic acid at 25 °C in darkness. After 4–5 weeks of culture, microcalli appeared on the surface of the Ca-alginate beads,
but the protoplasts without immobilization in Ca-alginate beads showed very low cell division. Growth of the microcalli in
the medium with nurse cells was much better than in the medium without nurse cells. Transferring beads onto half strength
MS medium supplemented with 0.2 mg l−1 kinetin and 0.1 mg l−1 2,4-D, increased the growth of embryogenic calli. Somatic embryo development was observed either on half strength MS medium
growth regulator free or with 1 mg l−1 abscisic acid. Matured embryos germinated on half strength MS medium containing 25 mg l−1 of gibberelic acid. Plantlet formation was obtained on half strength MS medium containing 1 mg l−1 6-benzyladenine and 1 mg l−1 α-naphthaleneacetic acid at 20 °C in a 16/8 h light/dark cycle.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
8.
In an attempt to optimize somatic embryo formation in Oncidium ‘Gower Ramsey’, the effects of five auxins (2,4-D, IAA, IBA, NAA and picloram) and five cytokinins (2iP, BA, kinetin, TDZ
and zeatin), used alone, was tested in vitro using root-derived callus. In general, kinetin (0.5 and 2 mg l−1) and zeatin (0.5 mg l−1) were found to be more effective than other auxin and cytokinin treatments to induce somatic embryogenesis from root-derived
callus. 相似文献
9.
Calli were induced from mature caryopses of timothy grass (Phleum pratense L.) on MS medium (Murashige and Skoog 1962) supplemented with 500 mg·dm−3 casein hydrolysate and 5 mg·dm−3 2,4-D (2,4-dicholorophenoxyacetic acid) or 2 mg·dm−3 dicamba (3,6-dichloro-o-anisic acid). Twelve-week-old calli were passaged on media with reduced levels of auxins (2 mg·dm−3 2,4-D or 1 mg·dm−3 dicamba). Tissues induced on medium with 2,4-D were transferred on medium with 2,4-D and on medium with dicamba; parallely
calli initiated on medium with dicamba were passaged on medium with 2,4-D or dicamba. Calli from various media sequences were
used to establish cell suspension cultures in media containing 2 mg·dm−3 2,4-D or 1 mg·dm−3 dicamba. An assessment of regeneration ability of calli was made on MS medium containing 0.2 mg·dm−3 kinetin. Callus tissue induced and/or subcultured on any of the media with 2,4-D did not regenerate plants while dicamba
added to the media was the effective stimulator of regenerability. In the presence of 2,4-D calli and suspensions produced
a jelly-like extracellular matrix. In cell suspension this phenomenon was observed 4–5 days after each passage. The measurements
of electric potential of calli, growing on MS medium with kinetin were performed. Non-regenerating callus areas had an electric
potential close to 0 mV while parts of tissue with meristematic centres were characterized by lower values of electric potential. 相似文献
10.
Wenyan Wang Chenggang Wang Bang-Lian Huang Bangquan Huang 《Plant Cell, Tissue and Organ Culture》2008,92(2):165-171
A protocol was developed for regeneration and Agrobacterium-mediated genetic transformation of Lesquerella fendleri. Calli were first induced from hypocotyls and cotyledons on MS plus 0.5 mg l−1 BA, 1 mg l−1 NAA and 1 mg l−1 2,4-D, then co-cultivated for 2–3 days in darkness on MS supplemented with 0.5 mg l−1 BA, 0.2 mg l−1 NAA and 100 μmol l−1As together with Agrobacterium tumefaciens strain EHA105/pCAMBIA1301 that harbored genes for uidA (GUS) and hygromycin resistance. Following co-cultivation, calli transfected by A. tumefaciens were transferred to MS with 0.5 mg l−1 BA, 0.2 mg l−1NAA, 500 mg l−1 Cef and 10 mg l−1 hygromycin and cultured for 10 days, then the hygromycin was increased to 20 mg l−1 on the same medium. After 4 weeks the resistant regenerants were transferred to MS with 0.5 mg l−1BA, 0.2 mg l−1 NAA, 500 mg l−1 Cef and 25 mg l−1 hygromycin for further selections. Transgenic plants were confirmed by polymerase chain reaction analysis, GUS histochemical
assay and genomic Southern blot hybridization. With this approach, the average regeneration frequency from transfected calli
was 22.70%, and the number of regenerated shoots per callus was 6–13. Overall results described in this study demonstrate
that Agrobacterium-mediated transformation is a promising approach for improvement of this Lesquerella species. 相似文献
11.
Organogenic cultures were induced from zygotic embryo and megagametophyte explants of the Central American cycad species,
Dioon edule. Plant growth medium consisted of B5 major salts, Murashige and Skoog minor salts and organics, 400 mg l−1 glutamine, 100 mg l−1 arginine, 100 mg l−1 asparagine, 60 g l−1 sucrose, 8 g l−1 Difco Bacto agar and was supplemented with kinetin (0 – 13.94 μM) and 2,4-dichlorophenoxyacetic acid (2,4-D) (0 – 9.05 μM)
arranged as a 5×4 factorial in a randomized block design. Callus initiation occurred on a wide range of medium formulations
from megagametophyte explants; however, shoot formation occurred only on medium supplemented with 2.26 μM 2,4-D. In comparison,
callus initiation from explanted zygotic embryos occurred on fewer medium formulations, and adventitious shoot induction occurred
from callus on formulations with 9.29–13.94 μM kinetin + 0.45–9.05 μM 2,4-D. Rooted shoots, derived from megagametophyte and
zygotic embryo cultures, have been regenerated.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
12.
Callus of Orthosiphon stamineus could be induced successfully from petiole, leaf and stem tissues but not roots when cultured on MS medium containing different concentration of NAA (0–4.0 mg l–1) and 2,4-D (0–2.0 mg l–1). Highest fresh weight callus production was obtained from leaf explants and those with best friability were obtained on MS medium plus 1.0 mg l–1 2,4-D plus 1.0 mg l–1 NAA. Cell suspension cultures were established from these cultures. The appropriate cell inoculum size for the best cell growth was 0.75 g of cells in 20 ml culture medium. Cell suspension culture using MS medium supplemented with 1.0 mg l–1 2,4-D promoted the best cell growth with maximum biomass of 8.609 g fresh weight and 0.309 g dry weight 24 days after inoculation. Cells that grew in MS medium supplemented with 1.0 mg l–1 2,4-D reached the stationary growth phase in 15 days as compared to the cells that grew in MS medium supplemented with 1.0 mg l–1 2,4-D + 1.0 mg l–1 NAA reached the stationary phase in 24 days. MS medium supplemented with 1.0 mg l–1 2,4-D was considered as the maintenance medium for maintaining the optimum cell growth of O. stamineus in the cell suspension cultures with 2-week interval subculture. 相似文献
13.
Manjula S. Thomas Anita Daniel Benny Nair G.M. 《Plant Cell, Tissue and Organ Culture》1997,51(2):145-148
Protocols for in vitro plant regeneration via axillary and adventitious shoot regeneration were established in an important
medicinal plant, Aristolochia indica L. (Aristolochiaceae). Basal Murashige and Skoog's (MS) medium supplemented with 0.54
μM α-naphthaleneacetic acid (NAA) and 13.31 μM benzyladenine (BA) induced the maximum number of shoots (45-50) from shoot
tip and nodal segment cultures. Phenolic accumulation in leaf and internodal stem derived callus cultured in MS medium containing
NAA or 2,4-dichlorophenoxyacetic acid and BA or kinetin was controlled by the addition of 1.0 mg l-1 phloroglucinol (PG) to the callus induction medium. Basal medium supplemented with 2.69 μM NAA, 13.31 μM BA and 1.0 mg l-1 PG induced the best results in terms of shoot bud regeneration from leaf derived callus. Direct de novo development of shoots
from leaf segments was achieved using 13.31 μM BA along with 50 mg l-1 activated charcoal. The microshoots were rooted in White's medium supplemented with 2.46 μM indolebutyric acid. More than
85% of rooted plants survived in the soil.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
14.
Tissue culture and plant regeneration of blue grama grass, Bouteloua gracilis (H.B.K.) Lag. Ex Steud
Gerardo Armando Aguado-Santacruz José Luis Cabrera-Ponce Víctor Olalde-Portugal M. A. Rosario Sánchez-González Judith Márouez-Guzmán Luis Herrera-Estrella 《In vitro cellular & developmental biology. Plant》2001,37(2):182-189
Summary As a first step towards applying biotechnology to blue grama, Bouteloua gracilis (H. B. K.) Lag. ex Steud., we have developed a regenerable tissue culture system for this grass. Shoot apices were isolated
from 3-d-old seedlings and cultured in 15 different growth regulator formulations combining 2,4-dichlorophenoxyacetic acid
(2,4-D), Picloram (4-amino-3, 5,6-trichloropicolinic acid), N6-benzyladenine (BA) or adenine (6-aminopurine). The highest induction of organogenic callus was obtained with formulations
containing 1 mg l−1 (4.52 μM) 2,4-D plus 0.5 mg l−1 (2.22 μM) BA. and 2 mg l−1 (8.88 μM) BA plus 1 mg l−1 (4.14 μM) Picloram with or without 40 mg l−1 (296.08 μM) adenine. Lower frequencies of induction were obtained for embryogenic as compared to organogenic callus. The most efficient
treatments for induction of embryogenic callus contained 2 mg l−1 (9.05 μM) 2,4-D combined with 0.25 (1.11 μM) or 0.50 mg l−1 (2.22 μM) BA, or 1 mg l−1 (4.52 μM) 2,4-D with 0.50 mg l−1 (2.22 μM) BA. Regeneration was achieved in hormonefree Murashige anmd Skoog (MS) medium, half-strength MS medium or MS medium plus
1 mg l−1 (1.44 μM) gibberellic acid. The number of plantlets regenerated per 500 mg callus fresh weight on MS medium ranged from 9 for 2 mg
l−1 (9.05 μM) 2,4-D to 62.2 for induction medium containing 2 mg l−1 (8,28 μM) Picloram, 1 mg l−1 (4.44 μM) BA and 40 mg l−1 (296.08 μM) adenine. Regnerated plants grown in soil under greenhouse conditions reached maturity and produced seeds. 相似文献
15.
Summary Callus induction was observed from hypocotyl, root, and cotyledonary leaf segments, grown on Murashige and Skoog (MS) medium
supplemented with various concentrations and combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (KN). Maximum
callusing (100%) was obtained from root and cotyledonary leaf segments grown on MS medium supplemented with a combination
of 2 mg l−1 (9.1 μM) 2,4-D and 0.2 mg l−1 (0.9 μM) KN. The calluses, when subcultured in the same medium, showed profuse callusing. However, these calluses remained recalcitrant
to regenerate regardless of the quality and combinations of plant growth regulators in the nutrient pool. When hypocotyl segments
were used as explants, callus induction was noticed in 91% of cultures which showed shoot regeneration on MS medium supplemented
with 2 mg l−1 2,4-D and 0.2 mg l−1 KN. These shoots were transferred to fresh medium containing various concentrations and combinations of 6-benzyladenine (BA)
and N6-(2-isopentenyl)adenosine (2-iP). Maximum shoot multiplication was observed after 60 d of the second subculture on MS medium
containing 2 mg l−1 (8.9 μM) BA. These shoots were rooted best (87%) on MS medium containing 2 mg l−1 (9.9 μM) indole-3-butyric acid (IBA). The plantlets were transferred to the field after acclimatization and showed 60% survival. 相似文献
16.
In Iris germanica L., 'G1', 'Adorn' and 'Rococo', induction and proliferation of embryogenic calli were achieved by culture
of leaf-base explants on Murashige and Skoog (MS) medium supplemented with 1 mg l−1 2,4-D, 1 mg l−1 kinetin, 200 mg l−1 casein hydrolysate, 250 mg l−1 proline, 30 g l−1 sucrose and 2.5 g l−1 gellan gum. Among these cultivars, however, only in 'G1' could a suspension culture be established using a liquid N6 medium
with 1 mg l−1 2,4-D, 1 mg l−1 kinetin, 200 mg l−1 casein hydrolysate, 250 mg l−1 proline and 30 g l−1 sucrose. Murashige and Skoog medium with 1 mg l−1 gibberellic acid (GA3), 30 g l−1 sucrose and 2.5 g l−1 gellan gum was suitable for somatic embryo formation from suspension cells. When the somatic embryos were transferred to
solid, growth regulator-free MS medium and subcultured monthly, 36 shoots were obtained from 20 mg suspension cells.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
17.
A. Muthusamy K. Vasanth D. Sivasankari B. R. Chandrasekar N. Jayabalan 《Biologia Plantarum》2007,51(3):430-435
The embryogenic calli (EC) were obtained from hypocotyl explants of groundnut (Arachis hypogaea L.) cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid
(2,4-D) in combination with 0.5 mg dm−3 6-benzylaminopurine (BAP). The EC were exposed to γ-radiation (10–50 Gy) or treated with 1–5 mM of ethyl methane sulphonate
(EMS) or sodium azide (SA). The mutated EC were subcultured on embryo induction medium containing 20 mg dm−3 2,4-D. Somatic embryos (SE) developed from these calli were transferred to MS medium supplemented with BAP (2.0 mg dm−3) and 0.5 mg dm−3 2,4-D for maturation. The well-developed embryos were cultured on germination medium consisting of MS salts with 2.0 mg dm−3 BAP and 0.25 mg dm−3 naphthaleneacetic acid (NAA). Well-developed plantlets were transferred for hardening and hardened plants produced normal
flowers and set viable seeds. The fresh mass of the EC, mean number of SE per explant and regeneration percentage were higher
at lower concentrations of mutagens (up to 30 Gy/3 mM). Some abnormalities in regenerated plants were observed, especially
variations in leaf shape. 相似文献
18.
A system for rapid plant regeneration through somatic embryogenesis from shoot tip explants of sorghum [Sorghum bicolor (L.) Moench] is described. Somatic embryogenesis was observed after incubation of explants in dark for 6–7 weeks through
a friable embryogenic callus phase. Linsmaier and Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid (2 mg l−1) and kinetin (0.1 mg l −1) was used for induction of friable embryogenic calli and somatic embryos. Germination of somatic embryos was achieved about
5 weeks after transfer onto Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (2 mg l−1) and indole-3-acetic acid (0.5 mg l −1) under light. Seeds from in vitro-regenerated plants produced a normal crop in a field trial, and were comparable to the crop grown with the seeds of the mother
plant used to initiate tissue culture. The simplicity of the protocol and possible advantages of the system for transformation
over other protocols using different explants are discussed.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
19.
Deeks Shannon J. Shamoun Simon F. Punja Zamir K. 《Plant Cell, Tissue and Organ Culture》2001,66(2):97-105
A procedure for in vitro culture of the parasitic flowering plant western hemlock dwarf mistletoe, Arceuthobium tsugense (Rosend.) G.N. Jones subsp. tsugense, is described. A factorial experiment evaluated the effects of media (Harvey's medium (HM) and modified White's medium (WM)),
temperatures (15 °C and 20 °C), presence or absence of light, and plant growth regulators (the auxin 2,4-dichlorophenoxyacetic
acid (2,4-D) and the cytokinin 6-benzylaminopurine (BAP) at varying concentrations (0.001 mg l−1 to 1 mg l−1)). Seed explants germinated in less than one week in culture and produced radicles. Optimal conditions for radicle elongation
were WM at 20 °C in the presence of light and without plant growth regulators. Some of the radicles split at the tip to yield
callus while others swelled to become spherical holdfasts. Holdfasts were also produced at the tips of radicles, and callus
arose from split holdfasts. Factors that promoted holdfast production were Harvey's medium, light, and 2,4-D at 1 mg l−1. Callus development from split radicles and split holdfasts was optimal on WM with 0.5 mg l−1 2,4-D and 1 mg l−1 BAP at 20 °C in the dark.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
20.
Embryogenic cultures were induced from leaflets from new vegetative flushes of mature ‘Brewster’ litchi trees on B5 medium
containing 400 mg l−1 glutamine, 200 mg l−1 casein hydrolysate, 30 g l−1 sucrose, 4.52 μM 2,4-D, 9.30 μM kinetin and 3 g l−1 gellan gum in darkness. Embryogenic cultures consisting of proembryonic cells and masses were maintained either on semi-solid
MS medium supplemented with 4.52 μM 2,4-D and 0.91 μM zeatin or as embryogenic suspension cultures in liquid medium of the
same composition. Maturation of somatic embryos occurred on semi-solid MS medium with 5–20% (v/v) filter-sterilized coconut
water in darkness. Recovery of plants from somatic embryos was improved with 14.4 μM GA3 on half-strength MS medium with 0.2 g l−1 activated charcoal under a 16 h photoperiod provided by cool white fluorescent lights (60–80 μmol s−1 m−2). Plants have been successfully acclimatized in the greenhouse. 相似文献