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1.
Fertile regenerated plants were obtained from protoplasts via somatic embryogenesis in Coker 201 (Gossypium hirsutum L.). Protoplasts were isolated from six different explantsleaves, hypocotyls, young roots, embryogenic callus, immature somatic embryos and suspension cultures and cultured in liquid thin layer KM8P medium. Callus-forming percentage of 20–50% was obtained in protoplast cultures from embryogenic callus, immature embryos and suspension cultures, and visible callus formed within 2 months. Callus-forming percentage of 5–20% in protoplast cultures from young roots, hypocotyls and leaves, and visible callus formed in 3 months. NAA 5.371 μM/kinetin 0.929 μM was effective to stimulate protoplast division and callus formation from six explants. Percentage of callus formation in the medium with 2,4-D 0.452 μM/kinetin 0.465 μM was over 40% from suspension cultures and immature embryos, 25% from embryogenic callus and 10% from hypocotyls. Callus from protoplasts developed into plantlets via somatic embryogenesis. Over 100 plantlets were obtained from protoplasts derived from 6 explants. Ten plants have been transferred to the soil, where they all have set seeds. 相似文献
2.
A highly reproducible system for efficient plant regeneration from protoplast via somatic embryogenesis was developed in cotton (Gossypium hirsutum L.) cultivar ZDM-3. Embryogenic callus, somatic embryos and suspension culture cells were used as explants. Callus-forming
frequency (82.86 %) was obtained in protoplast cultures from suspension culture cells in KM8P medium with 0.45 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 0.93 μM kinetin (KIN), 1.5 % glucose and 1.5 % maltose. Protocolonies
formed in two months with plating efficiency of 14 %. However, the callus-forming efficiencies from other two explants were
low. The calli from protoplast culture were transferred to somatic embryo induction medium and 12.7 % of normal plantlets
were obtained on medium contained 3 % maltose or 1 % of each sucrose + maltose + glucose, 2.46 μM indole-3-butyric acid (IBA)
and 0.93 μM KIN. Over 100 plantlets were obtained from protoplasts derived from three explants. The regenerated plants were
transferred to the soil and the highest survival rate (95 %) was observed in transplanting via a new method. 相似文献
3.
Sood Anil Ahuja P.S. Sharma Madhu Sharma O.P. Godbole Savita 《Plant Cell, Tissue and Organ Culture》2002,71(1):55-63
Seedling raised elites of Dendrocalamus hamiltoniiNees et Arn. Ex Munro were chosen as the source of nodal explants from precocious branches. While axillary bud break was accomplished in hormone free 1/2MS medium containing sucrose (3%, w/v), BA supplementation was required for shoot proliferation. A variety of hormonal combinations induced rooting in clumps of shoots. Somatic embryogenesis was also obtained in callus cultures raised in 2,4-D supplemented MS medium and plantlets derived from somatic embryos were hardened for field transfer. Comparative growth performances of plants raised from nodal cuttings of field-grown plants, those from single node cuttings of precocious branches and from somatic embryos indicated that growth performance of the tissue culture raised plants was relatively better than those from nodal cuttings. Improved protocols for efficient micropropagation are visualized to provide an impetus to raising of bamboo nurseries of elite genotypes in bamboo growing areas of western Himalayas. 相似文献
4.
Embryogenic callus (translucent callus) was produced from immature zygotic embryos of Picea wilsonii Mast. Subsequently somatic embryogenesis occurred on the brown callus. The somatic embryos could be stimulated to developinto plantlets on the medium without hormone. Young somatic embryos were produced from embryogenic callus in liquid suspension culture, in which suspensor was several or more than ten times the size of the somatic embryo. The somatic embryo showed very similar to zygotic embryos in micro-section and living material. 相似文献
5.
Immature and mature zygotic embryos of Paspalum scrobiculatum L. cv. PSC 1 cultured on MS or N6 nutrient medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), formed embryogenic callus. Induction of embryogenic callus and subsequent somatic embryogenesis was possible at a lower concentration of 2,4-D on N6 than MS medium. Immature embryos were highly totipotent, forming somatic embryos at a higher frequency than mature embryos. Addition of amino acids (L-proline or L-tryptophan) to 2,4-D medium resulted in significant enhancement of embryogenesis on culture of mature embryos. Silver nitrate also supported an increased frequency of embryogenesis. Thus it is possible to have high frequency of somatic embryogenesis on culture of mature embryos, which are available in abundance and with ease than immature embryos. The somatic embryos readily germinated and formed plantlets on hormone-free regeneration medium. The regenerated plantlets were successful on transfer to soil and set seed. 相似文献
6.
Jingli Yang Chenguang Zhou Lihui Liu Du Jia Zhiyang Yin Myongjo Kim Changyeon Yu Chenghao Li 《Plant Cell, Tissue and Organ Culture》2012,110(2):289-298
Embryogenic callus was obtained from young petiole, stem, and root explants of 4-year-old Siberian ginseng plants on medium supplemented with 2,4-dichlorophenoxyacetic acid. Embryogenic callus differentiated into somatic embryos (SEs), most of which could germinate but developed abnormally. Friable embryogenic callus was induced mainly from the root regions of germinated primary SEs or regenerated plantlets on plant growth regulator-free medium. Histological studies showed that the embryogenic callus initiated from the subepidermal cells of young roots. The bioreactor system was more efficient than suspension culture regarding the number and growth of SEs, although a similar amount of embryonic tissue was used. An average of 115,370 germinated SEs developed from an initial 400?mg of embryogenic callus, and 64.7?% of germinated SEs converted into plantlets after a 4-weeks culture on agar medium. During the bioreactor culture process, secondary SEs were induced directly from SEs at various stages, a phenomenon that rarely occurred in suspension culture. These secondary SEs developed quickly and germinated during the bioreactor culture process. Proline content and peroxidase and catalase activities of SEs cultured in bioreactors were higher than in SEs cultured in suspension culture. 相似文献
7.
探索华山松(Pinus armandii)体细胞胚胎发生技术对其实施规模化无性繁殖和开展遗传转化具有重要意义。本文以1/2LM为基本培养基, 通过激素调节等措施对华山松的胚性愈伤组织诱导和幼胚的离体培养技术进行了初步研究。研究结果: 胚性愈伤组织诱导率最高可达52.71%, 但愈伤组织继代培养后没有体细胞胚胎的分化; 首次从其子叶期的幼胚中直接诱导出具有根和茎的完整植株, 诱导率达92%以上。文章确认了采集的幼胚发育状态对胚性愈伤组织的诱导有重要影响, 并对诱导的培养条件等进行了探讨。 相似文献
8.
M. Cristina Pedroso M. Salom Pais 《In vitro cellular & developmental biology. Plant》1995,31(1):31-35
Summary Two methods (I and II) for somatic embryo production from embryogenic suspension cultures ofCamellia japonica are presented. Method I, embryogenic suspension cultures, was established from suspension cultures initiated from leaf-derived
callus. These cultures were maintained by reducing agitation and increasing subculture interval. Induction of somatic embryogenesis
was achieved in MS28 medium, 6, 12, 24, and 36 mo. after culture establishment. Embryo production decreased after 1 yr of
culture. Method II, suspensions of single embryogenic cells and proembryos, was obtained from leaves cultured in liquid MS13
medium 6 wk after culture initiation. Embryo production was 23 embryos/ml. Germination of cell suspension-derived embryos
on MS56 medium was 16.7 % (±4.2%) for method I, and 35.4% (±5.1%) for method II. The embryos germinated into
plantlets with 0 to 7 axillary shoots. 相似文献
9.
V. Vasil I. K. Vasil 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1987,73(6):793-798
Summary Protoplasts isolated from a totipotent embryogenic cell suspension culture of Zea mays L. (cultivar Dekalb XL82) underwent sustained cell divisions when cultured in liquid as well as agarose media. Optimal colony formation (5%) occurred in a liquid medium containing 0.5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). A soft and unorganized callus was formed when the protocolonies were transferred to agar solidified suspension maintenance medium. Compact, organized and yellow to pale green folded structures and somatic embryos were formed upon subsequent transfer of this callus to a low 2,4-D medium. Clusters of somatic embryos germinated precociously but no plants were recovered. 相似文献
10.
Summary Methods of plant regeneration from callus and protoplasts of Helianthus giganteus L. are described. Embryogenic callus was obtained from leaf explants and plants were regenerated from these calli on MS media with different combinations of benzyladenine and naphtaleneacetic acid. Leaf protoplasts isolated from in vitro grown plants formed somatic embryos when cultured in agarose solidified droplets of V-KM medium containing benzyladenine and naphtaleneacetic acid. Embryos developed into plantlets on media with reduced auxin contents. Regenerated plants were successfully planted in soil.Abbreviations BA
benzyladenine
- IAA
indoleacetic acid
- MS
Murashige and Skoog medium
- NAA
naphtaleneacetic acid
- V-KM
protoplast culture medium of Binding and Nehls 相似文献
11.
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
12.
C. R. K. Reddy G. Raja Krishna Kumar A. K. Siddhanta A. Tewari K. Eswaran 《Journal of phycology》2003,39(3):610-616
In vitro somatic embryogenesis and regeneration of somatic embryos to whole plants through micropropagules was successfully demonstrated from pigmented uniseriate filamentous callus of Kappaphycus alvarezii (Doty) Doty in axenic cultures. More than 80% of the explants cultured on 1.5% (w/v) agar‐solidified Provasoli enriched seawater (PES) medium showed callus development. The callus induction rate was consistently higher for laboratory‐adapted plants. The excised callus grew well in subcultures and maintained its growth for prolonged periods if transferred to fresh medium in regular intervals. Some subcultured calli (<10%) did undergo transformation and produced densely pigmented spherical or oval‐shaped micropropagules (1–5 mm in diameter) that subsequently developed into young plantlets in liquid PES medium. The micropropagule production was further improved through somatic embryogenesis by a novel method of culturing thin slices of pigmented callus with naphthaleneacetic acid (NAA) or a mixture of NAA and 6‐benzylaminopurine. Transfer of embryogenic callus along with tiny somatic embryos to liquid medium and swirling on orbital shaker facilitated rapid growth and morphogenesis of somatic embryos into micropropagules that grew into whole plants in subsequent cultivation in the sea. The daily growth rate of one tissue cultured plant was monitored for seven generations in field and found to be as high as 1.5–1.8 times over farmed plants. The prolific somatic embryogenesis together with high germination potential of somatic embryos observed in this study offers a promising tool for rapid and mass clonal production of seed stock of Kappaphycus for commercial farming. 相似文献
13.
Efficient methods in totipotent callus formation, cell suspension culture establishment and whole-plant regeneration have been developed for the goosegrass [ Eleusine indica (L.) Gaertn.] and its dinitroaniline-resistant biotypes. The optimum medium for inducing morphogenic calli consisted of N6 basal salts and B5 vitamins supplemented with 1-2 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D), 2 mg l(-1) glycine, 100 mg l(-1) asparagine, 100 mg l(-1) casein hydrolysate, 30 g l(-1) sucrose and 0.6% agar, pH 5.7. The presence of organogenic and embryogenic structures in these calli was histologically documented. Cell suspension cultures derived from young calli were established in a liquid medium with the same composition. Morphogenic structures of direct shoots and somatic embryos were grown into rooted plantlets on medium containing MS basal salts, B5 vitamins, 1 mg l(-1) kinetin (Kn) and 0.1 mg l(-1) indole-3-acetic acid (IAA), 3% sucrose, 0.6% agar, pH 5.7. Calli derived from the R-biotype of E. indica possessed a high resistance to trifluralin (dinitroaniline herbicide) and cross-resistance to a structurally non-related herbicide, amiprophosmethyl (phosphorothioamidate herbicide), as did the original resistant plants. Embryogenic cell suspension culture was a better source of E. indica protoplasts than callus or mesophyll tissue. The enzyme solution containing 1.5% cellulase Onozuka R-10, 0.5% driselase, 1% pectolyase Y-23, 0.5% hemicellulase and N(6) mineral salts with an additional 0.2 M KCl and 0.1 M CaCl(2) (pH 5.4-5.5) was used for protoplast isolation. The purified protoplasts were cultivated in KM8p liquid medium supplemented with 2 mg l(-1) 2,4-D and 0.2 mg l(-1) Kn. 相似文献
14.
Chellappan Soundar Raju Krishnan Kathiravan Abubakker Aslam Appakan Shajahan 《Plant Cell, Tissue and Organ Culture》2013,112(3):387-393
A reproducible protocol for somatic embryogenesis was established for mango ginger (Curcuma amada Roxb.)—an important horticultural aromatic rhizomatous plant. Embryogenic callus induction was obtained from leaf sheath explants of in vitro raised plants on Murashige and Skoog (MS) agar medium containing 2.0 mg/L 2,4-dichlorophenoxyacetic acid and 0.5 mg/L 6-benzyladenine (BA). Embryogenic callus proliferation, somatic embryo (SE) formation and subsequent plantlet conversion occurred under optimal culture conditions. The effects of MS medium strength, sucrose and BA on SE formation were also evaluated. Half strength MS liquid medium necessary for SE formation and optimal sucrose concentration was found to be 3.0 %. BA at 0.3 mg/L produced the highest number (84.71 %) of SEs from leaf sheath explants. Secondary somatic embryos originated from primary somatic embryos on the same medium supplemented with 0.4–0.6 mg/L BA. Stereo microscopic and scanning electron microscopic observation revealed that the globular and torpedo shaped somatic embryos resulted in suspension culture during development. Mature somatic embryos germinated readily and developed into normal plantlets after 3 weeks on half strength MS basal agar medium under dark condition. Well rooted plantlets were successfully acclimatized at the survival rate of 70 %. 相似文献
15.
Embryogenic callus was induced from leaf explants of Eleutherococcus sessiliflorus cultured on Murashige and Skoog (MS) basal medium supplemented with 1 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D), while no plant growth regulators were needed for embryo maturation. The addition of 1 mg l(-1) 2,4-D was needed to maintain the embryogenic culture by preventing embryo maturation. Optimal embryo germination and plantlet development was achieved on MS medium with 4 mg l(-1) gibberellic acid (GA(3)). Low-strength MS medium (1/2 and 1/3 strength) was more effective than full-strength MS for the production of normal plantlets with well-developed shoots and roots. The plants were successfully transferred to soil. Embryogenic callus was used to establish a suspension culture for subsequent production of somatic embryos in bioreactor. By inoculating 10 g of embryogenic cells (fresh weight) into a 3l balloon type bubble bioreactor (BTBB) containing 2l MS medium without plant growth regulators, 121.8 g mature somatic embryos at different developmental stages were harvested and could be separated by filtration. Cotyledonary somatic embryos were germinated, and these converted into plantlets following transfer to a 3l BTBB containing 2l MS medium with 4 mg l(-1) GA3. HPLC analysis revealed that the total eleutherosides were significantly higher in leaves of field grown plants as compared to different stages of somatic embryo. However, the content of eleutheroside B was highest in germinated embryos. Germinated embryos also had higher contents of eleutheroside E and eleutheroside E1 as compared to other developmental stages. This result indicates that an efficient protocol for the mass production of E. sessiliflorus biomass can be achieved by bioreactor culture of somatic embryos and can be used as a source of medicinal raw materials. 相似文献
16.
Embryogenic callus derived from zygotic embryos of black pepper (Piper nigrum Linn.) were induced to form somatic embryos on solid and liquid Schenk and Hildebrandt basal medium. Callus proliferation,
somatic embryo-genesis and germination of embryos were achieved in about 8 months in static cultures while it took only 8
weeks in liquid suspension cultures. The highest number of embryos and plantlets was produced from cells grown as suspension
cultures raised in half-strength medium without growth regulators and sucrose level reduced from 3% to 1.5%. Regenerated plants
were established in soil. 相似文献
17.
Embryogenic callus derived from zygotic embryos of black pepper (Piper nigrum Linn.) were induced to form somatic embryos on solid and liquid Schenk and Hildebrandt basal medium. Callus proliferation, somatic embryo-genesis and germination of embryos were achieved in about 8 months in static cultures while it took only 8 weeks in liquid suspension cultures. The highest number of embryos and plantlets was produced from cells grown as suspension cultures raised in half-strength medium without growth regulators and sucrose level reduced from 3% to 1.5%. Regenerated plants were established in soil. 相似文献
18.
A. C. Bonfils S. C. Gleddie J. A. Webb W. A. Keller 《In vitro cellular & developmental biology. Plant》1992,28(3):137-142
Summary Rapidly growing cell suspension cultures of shepherd’s purse (Capsella bursa-pastoris L. Medic.) were established from leaf-derived calli. These suspensions remained unorganized in the presence of 2,4-D, but
underwent extensive root organogenesis in a growth regulator-free liquid medium. Attempts to induce direct embryogenesis in
liquid cultures were unsuccessful, but numerous embryos were obtained from cells plated onto growth-regulator-free solid medium.
These embryos were frequently abnormal, and secondary embryogenesis was problematic for plant recovery but fertile plants
were recovered. Viable protoplasts could readily be isolated from these cell suspensions. After 1 wk of culture, protoplast
viability was 62%, and 7% of the cells had divided. Embryogenesis was observed from protoplast-derived microcolonies, plated
on growth-regulator-free medium. Although these somatic embryos were difficult to root, plants were recovered. New cell suspensions
were more recently established, which were only 4 to 6 mo. old when plant regeneration was attempted. Numerous shoots were
obtained when these cells were plated onto growth-regulator-free solid media. However, these shoots differed from the embryos
previously obtained in that they readily rooted and rapidly developed into plantlets. This system may allow the use of shepherd’s
purse as a gene source for introgression of agronomically interesting traits intoBrassica crop species through protoplast manipulation and somatic hybridization. 相似文献
19.
A procedure is described for the regeneration of plants from protoplasts of tetraploid leek (Allium ampeloprasum L.), 2n = 4x =32. Regeneration-competent protoplasts could only be obtained from an embryogenic suspension culture that was initiated with friable, embryogenic callus derived from immature embryos. The generally low plating efficiency could be increased by embedding the protoplasts in Ca-alginate, compared to culturing the protoplasts in liquid or agarose-solidified medium. A minimum plating density of 2 × 105 pps/ml was required to obtain microcalli. Upon transfer of the protoplast-derived calli on agarose-solidified BDS medium, morphologically different callus types proliferated. After transfer to regeneration medium, compact or friable calli with an embryogenic appearance produced somatic embryos and plantlets at a frequency of up to 80%. Calli that had been classified as heterogeneous also regenerated shoots, but mainly via organogenesis, at a frequency of 46%. After transfer of shoots to half strength MS medium, healthy, well-rooted plants were obtained, that were successfully transferred to soil. All plants contained the tetraploid DNA level. 相似文献
20.
Efficient plant regeneration through somatic embryogenesis was achieved from callus cultures derived from semi-mature cotyledon explants of Dalbergia sissoo Roxb., a timber-yielding leguminous tree. Somatic embryos developed over the surface of embryogenic callus and occasionally, directly from cotyledon explants without intervening callus phase. Callus cultures were initiated from cotyledon pieces of D. sissoo on Murashige and Skoog (1962) medium supplemented with 4.52, 9.04, 13.57, and 18.09 mumol/L 2,4-dichlorophenoxyacetic acid and 0.46 mumol/L Kinetin. Maximum percentage response for callus formation was 89% on MS medium supplemented with 9.04 mumol/L 2,4-D' and 0.46 mumol/L Kn. Somatic embryogenesis was achieved after transfer of embryogenic callus clumps to 1/2-MS medium without plant growth regulators (1/2-MSO). Average numbers of somatic embryos per callus clump was 26.5 on 1/2-MSO medium after 15 weeks of culture. Addition of 0.68 mmol/L L-glutamine to 1/2-MSO medium enhanced somatic embryogenesis frequency from 55% to 66% and the number of somatic embryos per callus clump from 26.5 to 31.1. Histological studies were carried out to observe various developmental stages of somatic embryos. About 50% of somatic embryos converted into plantlets on 1/2-MSO medium containing 2% sucrose, after 20 days of culture. Transfer of somatic embryos to 1/29-MSO medium containing 10% sucrose for 15 days prior to transfer on 1/2-MS medium with 2% sucrose enhanced the conversion of somatic embryos into plantlets from 50 to 75%. The plantlets with shoots and roots were transferred to 1/2 and 1/4-liquid MS medium, each for 10 days, and then to plastic pots containing autoclaved peat moss and compost mixture (1:1). 70% of the plantiets survived after 10 weeks of transfer to pots. 120 regenerated plantlets out of 150 were successfully acclimatised. After successful acclimatisation, plants were transferred to earthen pots. 相似文献