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1.
Medha Shrikhande S. R. Thengane A. F. Mascarenhas 《In vitro cellular & developmental biology. Plant》1993,29(1):38-42
Summary Media for induction of somatic embryogenesis from immature cotyledonary tissues ofAzadirachta indica (Neem) were determined. Callus was initiated on Murashige and Skoog medium supplemented with 0.5 mg·liter−1 of indol-3 acetic acid, 1.0 mg·liter−1 of 6-benzyl amino purine, and 1000 mg·liter−1 of casein hydrolysate. Effect of kinetin was also studied for embryo induction. Carbohydrate source in the form of sucrose
and glucose alone and in combination was tested for embryogenic efficiency. Seventy percent embryos showed germination. Healthy
plants were potted in sand and soil. Histologic studies confirmed indirect somatic embryogenesis. 相似文献
2.
E. Plata A. Ballester A. M. Vieitez 《In vitro cellular & developmental biology. Plant》1991,27(4):183-189
Summary An anatomical study was carried out during the sequences of events which lead to the differentiation of secondary embryos
ofCamellia reticulata cv ‘Mouchang’. Secondary embryogenesis can be induced by culturing somatic embryos on a modified Murashige and Skoog medium
supplemented with 0.5 mg·liter−1 6-benzylaminopurine and 0.1 mg·liter−1 indole-3-butyric acid. After about 12 days of culture, globular-shaped secondary embryos became apparent, and by 18 to 20
days of culture cotyledonary stages were formed. Embryos developed mainly on the hypocotyl of primary embryos without an intermediate
callus. Histologic monitoring revealed that secondary embryos apparently had a multicellular origin from embryogenic areas
originating in both epidermal and subepidermal layers of the hypocotyl region. This morphogenetic competence is related to
the presence, at the time of culture, of relatively undifferentiated cells in superfical layers of the primary embryo hypocotyl.
Microcomputer image analysis was applied for quantifying cytological events associated with somatic embryogenesis. This method
showed an increasing gradient in the nucleus-to-cell area ratio from differentiated cells passing through preembryogenic cells
to embryogenic cells. The formation of embryogenic areas was preceded by accumulation of starch in the surrounding cortical
cells. The cells underlying globular secondary embryos still contained abundant starch, but it declined as the secondary embryos
developed. 相似文献
3.
Direct somatic embryogenesis from axes of mature peanut embryos 总被引:2,自引:0,他引:2
A. H. McKently 《In vitro cellular & developmental biology. Plant》1991,27(4):197-200
Summary Plant regeneration via somatic embryogenesis was obtained in peanut (Arachis hypogaea L.) from axes of mature zygotic embryos. The area of greatest embryogenic activity was a 2-mm region adjacent to and encircling
the epicotyl. Somatic embryogenesis was evaluated on Murashige and Skoog media supplemented with a variety of auxin treatments.
Maximum production occurred on medium supplemented with 3 mg · liter−1 4-amino-3,5,6-trichloropicolinic acid. Explant cultures were transferred to half-strength medium supplemented with 1 mg ·
liter−1 gibberellic acid for somatic embryo germination and early plantlet growth. Plantlets, transferred to soil, were placed in
a greenhouse and grown to maturity. 相似文献
4.
Summary
Haworthia comptoniana specimens were cultured to determine how benzyladenine (BA) level and in vitro selection for shoot and callus production
affected regeneration capacity and plant phenotype. Leaf explants were cultured on Murashige and Skoog medium containing 0
to 10 mg·liter−1 of BA. The highest number of shoots was obtained with 0.5 mg·liter−1 of BA.H. comptoniana stock cultures (hc) maintained with 0.5 mg·liter−1 of BA produced clumps of small shoots interspersed with friable, white, tan, and green callus. A clump of very large shoots
was isolated and designated cell line Rhc; it differed from the original hc culture in shoot size, the lack of callus growth,
and higher water content. A line of green callus (designated Gc), a line of white callus (Wc), and a line of soft tan callus
(Tc) were also isolated from hc. Optimal BA levels for shoot regeneration from lines Gc and Wc were 2 and 5 mg·liter−1, respectively. No normal shoots could be regenerated from Tc. The phenotypes of these cell lines remained stable for 24 subculture
generations. The hc line that initially required BA for growth became hormone autotrophic whereas the other lines did not.
Culturing using Gelrite and sealing vessels with parafilm promoted vitrification of the hc line. Culturing using GIBCO agar
and unsealed vessels reduced vitrification. The ex-vitro greenhouse survival rates for hc and Rhc plantlets were 10 and 80%,
respectively. The large size of the Rhc shoots apparently resulted in significantly higher survival rates under greenhouse
conditions, but did not result in any phenotypic whole plant changes. 相似文献
5.
Summary Embryogenic callus was initiated from radicles of mature embryos removed from imbibed seeds (24 h). Embryogenic and other
nonembryogenic types of callus proliferated on a modified half-strength Murashige-Skoog medium (MS) basal medium (BM) supplemented
withmyo-inositol, casein hydrolysate (CH), L-glutamine (gln) and growth regulators kinetin (KN), N6-benzyladenine (BAP) each (20×10−6
M), 2,4-dichlorophenoxyacetic acid (2,4-D) (50×10−6
M) Embryogenic callus bearing suspensor-like cells in a mucilaginous gel matrix was isolated and maintained by subculture every
10 to 12 days on BM with KN, BAP each (2×10−6
M) and 2,4-D (5×10−6
M). Somatic embryos developed spontaneously from the callus on this medium at 23±1° C. Closer examination revealed that numerous
polyembryonic clusters, comprised of elongated cells (suspensors) and small dense cells with large nuclei (somatic embryos),
occurred in the viscous gel. When this enriched embryonal-suspensor mass was subcultured to low 2,4-D (1×10−6
M), globular embryos developed by 40 to 60 days. Upon transfer to a liquid medium without growth regulators, the embryos elongated
and developed cotyledons and shoots with needles. Plantlet development was completed by 30 days in a basal medium without
CH, gln and growth regulators. The total culture time was 150 days. Approximately 40±10 embryos were formed from 500 mg of
initial callus. Somatic embryogenesis became aberrant if embryos remained attached to the callus mass and were not subcultured
within 10 to 12 days according to the described protocol. Somatic embryos were encapsulated in an alginate gel and stored
at 4° C for nearly two months without visible adverse effects on viability.
Editor's Statement This paper presents advances in the in vitro regeneration of a commercially useful plant species from stored
seeds. In addition, data is presented on short-term storage of the plantlets, and long-term proliferation of the embryonal
mass in vitro. 相似文献
6.
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
7.
Jameel M. Al-Khayri Christine E. Shamblin Edwin J. Anderson 《In vitro cellular & developmental biology. Plant》1996,32(4):227-232
Summary This study was conducted to establish and optimize a regeneration system for adapted U.S. rice genotypes including three commercial
rice cultivars (LaGrue, Katy, and Alan) and two Arkansas breeding lines. Factors evaluated in the study were genotype, sugar
type, and phytohormone concentration. The system consisted of two phases, callus induction and plant regeneration. In the
callus induction phase, mature caryopses were cultured on MS medium containing either 1% sucrose combined with 3% sorbitol
or 4% sucrose alone, and 0.5 to 4 mg·L−1 (2.26 to 18.10 μM) 2,4-D with or without 0.5mg·L−1) (2.32 μM) kinetin. In the plant regeneration phase, callus was transferred to 2,4-D-free MS medium containing 0 or 2 mg·L−1 (9.29 μM) kinetin combined with 0 or 0.1 mg·L−1 (0.54 μM) NAA. Callus induction commenced within a week, independent of the treatments. Callus growth and plant regeneration, however,
were significantly influenced by interactions among experimental factors. Generally, the greatest callus growth and plant
regeneration were obtained with 0.5 mg·L−1 (2.26 μM) 2,4-D and decreased with increasing 2,4-D concentrations. Kinetin enhanced callus growth only when combined with 0.5 mg·L−1 (2.26 μM) 2,4-D, and 4% sucrose. Inducing callus on kinetin-containing medium generally enhanced regeneration capacity in the presence
of sucrose but not with a sucrose/sorbitol combination. Media containing sucrose alone generally supported more callus proliferation,
but the sucrose/sorbitol combination improved regeneration of some cultivars. NAA and kinetin had little effect on regeneration. 相似文献
8.
Experiments were carried out with three-year-old embryogenic suspension culture of Gentiana pannonica Scop. The initial explant for the suspension determinated both the embryogenic character and embryo production. Cultures
were initiated by culture of hypocotyl, cotyledon and root explants on MS (Murashige and Skoog 1962) medium supplemented with
1.0 mg·l−1 Kinetin and 0.5 mg·l−1 2,4-D, later transferred and maintained in liquid MS medium with 1.0 mg·l−1 Dicamba, 0.1 mg·l−1 NAA, 2.0 mg·l−1 BAP and 80.0 mg·l−1 AS. Regeneration medium included 0.0–1.0 mg·l−1 GA3+0.0−2.0 mg·l−1 Kin.+0.0−160 mg·l−1 AS.
In these culture conditions, the effect of the explant was found to be the most important factor. The curve of growth, growth
coefficient and % of participation of various size aggregates differed in the studied suspensions. Flow cytometry revealed
various DNA content in nuclei from praembryogenic mass depending on the explant origin. To complete embryogenesis the medium
was changed from liquid to solidified in the presence of the same plant growth regulators combination required. The most embryogenic
culture appeared hypocotyl-derived and it yielded the highest number of somatic embryos. The suspension culture originating
from root proliferated the highest number of embryogenic cell clusters but did not produce embryos for fraction 120–450 μm.
One hundred mg of suspension of the fraction that was larger than 450 μm yielded 309, 175, 123 embryos for the following suspensions:
root-, cotyledon-, hypocotyl-derived, respectively. Almost 50 % of non-deformed fully developed embryos from all studied suspensions
passed conversion into germling stage and finally plants were regenerated. 相似文献
9.
Summary A protocol was developed for high frequency somatic embryogenesis and plant regeneration from cotyledon and hypocotyl explants
of Eruca sativa. Explants grown on Murashige and Skoog (MS) medium supplemented with 4.52 μM 2,4-D formed embryogenic callus after 4 wk of culture. Secondary somatic embryos were also produced from primary somatic
embryos on MS medium containing 0.56 μM 2,4-D. Somatic embryos developed into mature embryos on MS medium in the presence of 45 gl−1 polyethylene glycol. After desiccation, somatic embryos developed into plantlets by culturing the mature somatic embryos
on 1/2 x MS medium containing 0.24 μM indole-3-butyric acid. 相似文献
10.
Vishwanath M. Patil 《In vitro cellular & developmental biology. Plant》1998,34(3):240-243
Summary The purpose of this study was to developin vitro techniques for conserving wild and endemic species ofCeropegia by mass multiplication for subsequent reintroduction in their natural habitat. Micropropagation involving a combination of
axillary bud culture, shoot multiplication, somatic embryogenesis andin vitro tuber formation forCeropegia jainii, a rare plant of the Indian sub continent,C. bulbosa var.bulbosa andC. bulbosa var.lushii, common species, was developed. Nodal explants from all species were cultured on 0.5 MS medium with 8.8 μM (2 mg·l−1) N6-benzyl aminopurine (BA) to regenerate the axillary buds. These produced multiple shoots when transferred to multiplication
medium consisting of 0.5 MS medium with 2.2 μM (0.5 mg·l−1) BA, or microtubers when transferred to 0.5 MS medium with 22.2 μM (5 mg·l−1) BA and 23.2 μM (5 mg·l−1) kinetin.In vitro flowering occurred inC. jainii and not in the other two varieties when the plants were cultured on multiplication media with spermine at 0.25 μM (50 μg·l−1) as an additive. Shoot pieces produced callus on MS medium with 9.05 μM (2 mg·l−1) 2,4-dichlorophenoxy acetic acid. Regeneration of the calli by somatic embryogenesis was achieved when they were transferred
to 0.5 MS medium with 2.2 μM (0.5 mg·l−1) BA. Rooting of the shoots was possible both byin vitro andex vitro means. 相似文献
11.
Gyana Ranjan Rout Sanghamitra Samantaray Premananda Das 《Acta Physiologiae Plantarum》1998,20(3):269-275
Nickel tolerant callus lines of Setaria italica L. were developed from callus cultures grown on MS medium supplemented with 0.5 mg·dm−3 kinetin+2.0 mg·dm−3 2,4-D+2.0 mg·dm−3 Ni+2. Standard growth parameters such as callus fresh and dry weight, growth tolerance index were used as indicators of nickel
toxicity. Measurements as early as 2 weeks after the beginning of the treatments did not yield consistent results. However,
growth tolerance index at 4, and 8 weeks after the beginning of treatments yielded significant differences among the non-tolerant
and tolerant calli. The tolerant calli has enhanced growth at 2.0 mg·dm−3 Ni+2 while non-tolerant calli showed a reverse trend in growth in the presence of 2.0–2.5 mg·dm−3 of nickel. The tolerant calli differentiated into mass of embryogenic calli within 4 weeks of culture which could be maintained
for prolonged period without loss of regenerative capacity. 相似文献
12.
Plant regeneration was achieved through direct and indirect somatic embryogenesis in Eucalyptus camaldulensis. Callus was induced from mature zygotic embryos and from cotyledon explants collected from 10, 15, 25, and 30-day-old seedlings
cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of naphthaleneacetic acid (NAA).
Maximum callus induction from mature zygotic embryos was obtained on MS basal medium containing 1 mg l−1 NAA. The frequency of callus development varied based on the age of the cotyledon explants 10-day-old explants giving highest
percentage on MS basal medium supplemented with 1 mg l−1 NAA. Callus obtained from mature zygotic embryos gave highest frequency of somatic embryogenesis on MS basal medium containing
0.5 mg l−1 benzyladenine (BA) and 0.1 mg l−1 NAA. Separate age wise culture of the calli, obtained from cotyledons of different ages cultured separately, revealed high
somatic embryogenic potential on callus from 10-day-old cotyledons. Direct somatic embryogenesis too was obtained from hypocotyl
explants without an intervening callus phase on MS basal medium containing 0.5 mg l−1 BA. The effects of abscisic acid (ABA), sucrose, and different strengths of MS medium on somatic embryo maturation and germination
were also investigated. Number of mature somatic embryos increased with lower concentrations (0–1 mg l−1) of ABA while no significant differences were observed at higher concentrations (2–5 mg l−1) of ABA. Compared to basal medium containing lower concentrations of sucrose (1%), the MS medium supplemented with higher
levels of sucrose (4%) showed significantly lower frequency of mature somatic embryos. Basal medium without any dilution gave
the highest number of immature embryos. However, the number of mature embryos was high at higher medium dilutions. 相似文献
13.
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
14.
A three-stage procedure for embryogenesis in Trachyspermum ammi was developed from cotyledon and cotyledonary node explants cultured in Murashige and Skoog (MS) liquid medium supplemented
with 0.2 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Globular somatic embryos without intervening callus phase developed in 4 wk. The
development of embryos to heart and torpedo stages required second-stage subculture of the explants (along with developing
embryos) in liquid medium with lower concentrations of 2,4-D. Further development of embryos required a third-stage subculture
in hormone-free liquid medium supplemented with 100 mg l−1 casein hydrolysate. Regeneration of complete plantlets occurred after the fully developed somatic embryos were transferred
to solidified half-strength MS medium supplemented with 1 mg l−1 gibberellic acid. 相似文献
15.
A. Śliwińska O. Olszowska M. Furmanowa A. Nosov 《In vitro cellular & developmental biology. Plant》2008,44(2):69-77
Efficient plant regeneration through somatic embryogenesis was achieved in Polyscias filicifolia. Embryogenic calluses were induced on Murashige and Skoog (MS) basal medium supplemented with 0.5 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg l−1 benzylaminopurine (BAP; type I callus) and on MS medium with 2.0 mg l−1 2,4-D and 0.01 mg l−1 kinetin (type II callus) from leaf explants of a 2-yr-old plant. Primary somatic embryos (PSEs) developed after four passages
of suspension culture established from embryogenic callus when cultured in liquid half-strength MS medium (1/2 MS) without
growth regulators. PSEs in the cotyledonary stage were multiplied by adventitious embryogenesis. Single secondary somatic
embryos (SSEs) or their clusters developed at the base of PSE hypocotyls and regenerated into plantlets in a one-step process
on plant growth regulator-free 1/2 MS medium. Low sucrose concentration of 15 g l−1 promoted development of normal SSEs. All SSEs regenerated into single, well-rooted plantlets on a Nitsch and Nitsch medium
supplemented with 0.5 mg l−1 kinetin, 0.1 mg l−1 indole-3-butyric acid, and 10 mg l−1 adenine sulfate. Subsequent two subculture cycles on the same medium were necessary to obtain plantlets sufficiency developed
to allow successful transfer to the soil. Rooted plantlets were established in a peat mixture with 90% survival, with the
plants showing normal morphological characteristics. 相似文献
16.
Ma Dolores Lledó Manuel B. Crespo Juan Bta Amo-Marco 《In vitro cellular & developmental biology. Plant》1995,31(4):199-201
Summary
Vella lucentina M. B. Crespo is a threatened Spanish species that is endemic to a small area in eastern Alicante Province (SE Spain). Micropropagation
techniques were applied forex situ conservation of this plant. Aseptic epicotyls bearing the apical bud were grown in Murashige and Skoog medium supplemented
with 6-furfurylaminopurine (Kin), N6-benzyladenine (BA) or 6-(γ,γ,-dimethilalylamino) purine (2iP). High multiplication rates were obtained with 0.5, 1, or 2
mg·liter−1 BA, or 1 or 2 mg·liter−1 2iP. Indole-3-acetic acid and indole-3-butyric acid were utilized for rooting in half-strength Murashige and Skoog medium.
Regenerated plants were transferred to a potting mix and gradually acclimated to field conditions. No morphological differences
were observed amongin vitro andin vivo plants. 相似文献
17.
R. V. Sairam C. Wilber J. Franklin B. Smith J. Bazil R. Hassel D. Whaling K. Frutiger C. A. Blakey R. Vierling S. L. Goldman 《In vitro cellular & developmental biology. Plant》2002,38(5):435-440
Summary A protocol for high-frequency callus, somatic embryogenesis, and plant regeneration for Tripsacum is described. Plants were regenerated from complete shoot meristems (3–4 mm) via organogenesis and embryogenesis. In organogenesis,
the shoot meristems were cultured directly on a high cytokinin medium comprising 5–10 mgl−1 (22.2–44.4 μM) 6-benzyladenine (BA). The number of multiple shoots varied from six to eight from each meristem. The time required for production
of plants from organogenesis was rapid (4–6 wk). In contrast, callus was induced on an auxin medium and continuously cultured
on an auxin medium for production of somatic embryos. Prolific callus with numerous somatic embryos developed within 3–4 wk
when cultured on an auxin medium containing 5 mgl−1 (22.6μM), 2,4-dichlorophenoxyacetic acid (2,4-D). The number of shoots induced varied from two to five per callus. Regardless of
the cultivars used, the frequency of callus induction and plant regeneration was between 48% and 94%. The seed germination
procedures also were modified and resulted in a maximum of 60–80% seed germination. Finally, the rate of T-DNA transfer to
complete shoot meristems of Tripsacum was high on the auxin medium and was independent of whether super-virulent strains of Agrobacterium were used or not. 相似文献
18.
An efficient somatic embryogenesis system has been established in Catharanthus roseus (L.) G. Don in which primary and secondary embryogenic calluses were developed from hypocotyls and primary cotyledonary somatic
embryos (PCSEs), respectively. Two types of calluses were different in morphology and growth behaviour. Hypocotyl-derived
embryogenic callus (HEC) was friable and fast-growing, while secondary callus derived from PCSE was compact and slow-growing.
HEC differentiated into somatic embryos which proliferated quickly on medium supplemented with NAA (1.0 mg l−1) and BA (1.5 mg l−1). Although differentiation and proliferation of somatic embryos were faster in primary HEC, maturation and germination efficiency
were better in somatic embryos developed from primary cotyledonary somatic embryo-derived secondary embryogenic callus (PCSEC).
At the biochemical level, two somatic embryogenesis systems were different. Both primary and secondary/adventive somatic embryogenesis
and the role of plant growth regulators in two modes of somatic embryo formation have been discussed. 相似文献
19.
Wei Tang Zhongchen Guo Fan Ouyang 《In vitro cellular & developmental biology. Plant》2001,37(5):558-563
Summary Mature zygotic embryos of eight (open-pollinated) families of loblolly pine (Pinus taeda L.) were cultured on eight different basal salt formulations, each supplemented with 36.2 μM 2,4-dichlorophenoxyacetic acid, 17.8 μM 6-benzyladenine, 18.6 μM kinetin, 500 mg l−1 casein hydrolysate, and 500 mg l−1
l-glutamine for 9 wk; embryogenic tissue was formed on cotyledons, hypocotyls, and radieles of mature zygotic embryos. Callus
was subcultured on the callus proliferation medium, the same as the induction medium but with one-fifth concentration of auxin
and cytokinin for 9 wk. On this medium a white to translucent, glossy, mucilaginous embryogenic callus containing embryogenic
suspensor masses (ESMs) was obtained. The highest frequency of explants forming embryogenic tissue, 17%, occurred on a modified
Murashige and Skoog salts basal medium containing the concentration of KNO3, Ca(NO3)2·4H2O, NH4NO3, KCl, ZnSO4·7H2O, and MnSO4·H2O, 720, 1900, 400, 250, 25.8, and 25.35 mg l−1, respectively. Embryogenic suspension cultures were established by culturing embryogenic callus in liquid callus proliferation
medium. Liquid cultures containing ESMs were transferred to medium containing abscisic acid, polyethylene glycols, and activated
charcoal for stimulating the production of cotyledonary somatic embryos. Mature somatic embryos germinated for 4–12 wk on
medium containing indole-butyric acid, gibberellic acid, 6-benzyladenine, activated charcoal, and reduced sucrose concentration
(15 g l−1). Two hundred and ninety-one regenerated plantlets were transferred to a perlite:peatmoss:vermiculite (1∶1∶1) mixture, then
the plants were transplanted to soil in the earth, and 73 plantlets survived in the field. 相似文献
20.
Embryogenic callus in Catharanthus roseus was initiated from hypocotyl on Murashige and Skoog’s (MS) medium supplemented with 1.0–2.0 mg dm−3 of 2,4-dichlorophenoxyacetic acid (2,4-D) or chlorophenoxyacetic acid (CPA). Calli from other sources were non-embryogenic.
Numerous somatic embryos were induced from primary callus on MS medium suplemented with naphthalene acetic acid (NAA) within
two weeks of culture. Embryo proliferation was much faster on medium supplemented with 6-benzylaminopurine (BAP). After transfer
to medium with gibberellic acid (GA3, 1.0 mg dm− 3) mature green embryos were developed and germinated well into plantlets on MS liquid medium supplemented with 0.5 mg dm−3 BAP. Later, embryos with cotyledonary leaves were subjected to different auxins treatments for the development of roots.
Before transfer ex vitro, plantlets were cultivated on half strength MS medium containing 3 % sucrose and 0.5 mg dm−3 BAP for additional 2 weeks. Additionally, the effect of liquid medium has been evaluated at different morphogenetic stages. 相似文献