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1.
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
2.
Summary Direct somatic embryo formation and plantlet regeneration was achieved from immature leaflets of chickpea (Cicer arietinum L.). Optimal somatic embryogenesis was obtained when immature leaflets were exposed to media supplemented with 15 μM 2,4-dichlorophenoxyacetic acid (2,4-D) for 7 d, to 2000 μM 2,4-D for 3 d, and to 50 μM 2,4-D for 10 d, followed by transfer onto Murashige and Skoog (MS) basal medium. Exposure of explants to high 2,4-D levels
(200–2000 μM) for 3 d produced bottle-shaped embryos, while exposure to low 2,4-D levels (<50 μM) and 50–2000 μM for 10 d produced spherical-shaped embryos. Two percent of embryos converted into plants upon culture on MS medium containing
15 μM gibberellic acid and 1 μM 3-indolebutyric acid. All regenerated plants were phenotypically normal. 相似文献
3.
Summary Several factors that may affect induction of somatic embryogenesis in loblolly pine (Pinus taeda L.) were investigated in 1994 and 1995. Megagametophytes containing immature zygotic embryos were excised from seeds as explants.
Potassium chloride, silver nitrate, myo-inositol, coconut water, or polyamine was added to the control media (U.S. patent
no. 5,036,007) to determine the effects of each single ingredient or their combinations on the initiation of embryogenic tissue.
Supplements of myo-inositol at 22.2 mM resulted in increases in frequencies of cell mass extrusion and proliferation compared with the control media in consecutive
years. Addition of silver nitrate showed the potential to promote initiation of embryogenic culture. The combination of 10
mM potassium with 29.4 μM silver nitrate achieved the highest frequencies in both extrusion and proliferation of embryogenic tissue. The combination
of silver nitrate at 29.4 μM with addition of myo-inositol at 11.1 or 22.2 mM achieved a higher conversion rate from extrusion to proliferation. Polyamine did not significantly affect the induction of
somatic embryogenesis, but coconut water was inhibitory.
Published with approval of the Director of Arkansas Agricultural Experimental Station. 相似文献
4.
Agrobacterium-mediated transformation of American chestnut (Castanea dentata (Marsh.) Borkh.) somatic embryos 总被引:1,自引:0,他引:1
Linda D. Polin Haiying Liang Ronald E. Rothrock Mutsumi Nishii Deborah L. Diehl Andrew E. Newhouse C. Joseph Nairn William A. Powell Charles A. Maynard 《Plant Cell, Tissue and Organ Culture》2006,84(1):69-79
These studies were designed to test if a binary vector containing the gfp, bar and oxalate oxidase genes could transform American chestnut somatic embryos; to see if a desiccation treatment during co-cultivation would affect the transformation frequency of different American chestnut somatic embryo clones; to explore the effects of more rapid desiccation; and to see if the antibiotics used to kill the Agrobacterium were interfering with the regeneration of the somatic embryos. Two days of gradual desiccation was found to significantly enhance transient GFP expression frequency. When this treatment was tested on six American chestnut clones, five were transformed and four of these remained embryogenic. Transformation was confirmed by Southern hybridization. Phenotypically normal transgenic shoots were regenerated and rooted. Vascular tissue specific expression of the oxalate oxidase gene was detected in one transgenic line. Carbenicillin, cefotaxime, and tricarcillin were found to not interfere with the regeneration of transformed embryos. 相似文献
5.
6.
Tran Thanh Thu Tran Thi Xuan Mai Eric Dewaele Slama Farsi Yohannes Tadesse Geert Angenon Michel Jacobs 《Molecular breeding : new strategies in plant improvement》2003,11(2):159-168
A requirement for generating transgenic pigeonpea [Cajanuscajan (L.) Millsp] plants is the development of a highly efficientin vitro regeneration procedure. This goal was achieved byusing germinated seedlings grown on B5 medium supplemented with 10 mgl–1 6-benzylaminopurine, which induced differentiatingcallus formation in the cotyledonary node region. The calli were transferred onB5 medium with 0.2 mg l–1 6-benzylaminopurine toobtain shoot induction. Elongated shoots were then further cultured on a B5hormone-free medium for rooting. Using this regeneration system transgenicpigeonpea plants were obtained both by particle bombardment andAgrobacterium tumefaciens-mediated gene transfer. Thepresence of the transgenes in the pigeonpea genome was confirmed by GUS assays,PCR and Southern hybridisation. The transgenic rooted plants were successfullytransferred to soil in the greenhouse. GUS and PCR assays of T1 progeniesconfirmed that the transgenes were stably transmitted to the next generation.This is the first report of successful use ofAgrobacteriumas well as particle bombardment for production of transgenic pigeonpea plants. 相似文献
7.
Kui Shin Voo Clayton L. Rugh Joe C. Kamalay 《In vitro cellular & developmental biology. Plant》1991,27(3):117-124
Summary We describe a tissue culture procedure for somatic embryogenesis and plantlet regeneration in cotton (Gossypium hirsutum L. cv. Coker 312). Callused explants or individual globular embryos were transferred to basal media to induce somatic embryogenesis.
To determine characteristic early indicators of successful germination and conversion, we identified six types of embryos
that developed on basal media. Two of the six embryo types, designated as tulip-shaped and trumpet-shaped, could undergo conversion
in preliminary tests, whereas the others had little or no developmental potential. Several media treatments designed to enhance
the maturation of globular somatic embryos failed to increase the fraction of embryos which matured to form recoverable types.
In efforts to improve plantlet recovery, tulip-shaped embryos were used in limited trials to contrast the effects of chemical
and physical desiccation treatments on germination and conversion. The selective use of tulip-shaped somatic embryos, coupled
with partial desiccation, seems to have augmented plant recovery. Growth habit, flowering, seed set, and lint production of
most of the regenerated plants were comparable to seed-derived plants grown under the same conditions.
Partial research support was provided by state and federal funds appropriated to the Ohio Agricultural Research and Development
Center, The Ohio State University. 相似文献
8.
Summary Immature zygotic embryos from open-pollinated and selfed Carica papaya L. fruits, 90 to 114 days post-anthesis, produced 2 to 20 somatic embryos on apical domes, cotyledonary nodes, and radicle meristems after culture for three weeks on half-strength Murashige and Skoog (MS) medium supplemented with 0.1 to 25 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D), 400 mg l–1 glutamine, and 6% sucrose. After six weeks of culture, about 40 to 50% of the zygotic embryos had become embryogenic, and each embryogenic embryo yielded hundreds of somatic embryos within five months of culture on media supplemented with 2,4-D. Somatic embryos matured on half-strength MS medium, germinated on MS medium containing 5 mg l–1 kinetin, and grew large enough for greenhouse culture on MS medium. Shoots were rooted in vermiculite and grown in the greenhouse.Journal Series no. 3449 of the Hawaii Institute of Tropical Agriculture and Human Resources 相似文献
9.
B. N. S. Murthy Jerrin Victor Rana P. Singh R. A. Fletcher Praveen K. Saxena 《Plant Growth Regulation》1996,19(3):233-240
In vitro regeneration in chickpea (Cicer arietinum L.) was achieved by direct culture of mature seeds on Murashige and Skoog (MS) medium supplemented with either N-phenyl-N(-1,2,3-thidiazol-5-yl) urea (thidiazuron, TDZ) or N6-benzylaminopurine (BAP). Multiple shoots formed de novo without an intermediary callus phase at the cotyledonary notch region of the seedlings within 2 to 3 weeks of culture initiation. TDZ was found to be more effective compared to BAP as an inductive signal of regeneration. The former induced multiple shoot formation at all the concentrations tested (1 M to 100 M), although, maximum morphogenic response was observed at 10 M concentration. Addition of naphthaleneacetic acid (NAA) alone or in combination with BAP to the MS medium failed to invoke a similar response. When the TDZ supplemented medium was amended with L-proline, the resultant regenerants were mostly somatic embryos. Histological investigations confirmed the switch in the regeneration pathway from directly formed adventitious shoots to embryogenesis. For obtaining plantlets, adventitious shoots were rooted on MS medium supplemented with 2.5 M NAA; somatic embryos were germinated and established on MS medium. Normal plants were regenerated from both adventitious shoots and somatic embryos and transferred to soil.Abbreviations BAP
6-benzylaminopurine
- MS
Murashige and Skoog [14] basal medium
- NAA
naphthaleneacetic acid
- TDZ
thidiazuron [N-phenyl-N(-1,2,3,-thidiazol-5-yl)-urea] 相似文献
10.
Rubén álvarez José M. álvarez Jaime M. Humara ángeles Revilla Ricardo J. Ordás 《Biotechnology letters》2009,31(9):1477-1483
The bar gene was introduced into the cork oak genome. Cork oak embryogenic masses were transformed using the Agrobacterium strain AGL1 which carried the plasmid pBINUbiBar. This vector harbours the genes, nptII and bar, the latter under control of the maize ubiquitin promoter. The transgenic embryogenic lines were cryopreserved. Varying activities
of phosphinothricin acetyl transferase were detected among the lines, which carried 1–4 copies of the insert. Molecular and
biochemical assays confirmed the stability and expression of the transgenes 3 months after thawing the cultures. These results
demonstrate genetic engineering of herbicide tolerance in Quercus spp.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
Rubén álvarez, Ricardo J. Ordás are contributed equally. 相似文献
11.
Various leaf sections of Gasteria verrucosa Haw. and Haworthia fasciata Haw. were cultured on media to examine the effect of picloram (4-amino 3, 5, 6-trichloropicolinic acid) and 2, 4-D (2, 4-dichlorophenoxy acetic acid) on somatic embryogenesis. Picloram (0.5, 1.0, 2.0, 3.0 mgl-1) outperformed 2, 4-D (0, 1.0, 2.0, 3.0 mgl-1) as the auxin source of both earliness of callus and embryo induction and final yield of embryos produced at both kinetin levels examined (0.25, 1.0 mgl-1). Embryos arose initially as a yellow, compact globular masses from the area just beneath the epidermis in linear pattern parallel with the main axis of the leaf and then developed a heartshaped appearance. Embryo formation was preceded by growth of callus almost crystalline in appearance on the cut surface. Subsequent shoot formation developed from green pigmented loci in crystalline callus derived from embryos. Shoot and root development in Gasteria was induced on a defined medium containing quarter strength MS or B5 salts with no hormonal supplementation. 相似文献
12.
Auxin pulse treatment holds the potential to enhance efficiency and practicability of somatic embryogenesis in potato 总被引:1,自引:1,他引:0
The objective of the current study was to simplify existing somatic embryogenesis systems in potato (Solanum tuberosum L.) cv. Desiree. The project targeted the agar-based induction phase of the potato somatic embryogenesis process as the key
area for improvement. Experiments were established to ascertain the effect of a 2,4-D (2,4 dichlorophenoxyacetic acid) pulse,
applied to the primary internodal section explant source and its subsequent effect on embryo induction. Parameters tested
were the duration of the auxin pulse in a range from 0 to 300 min, and the concentrations of 2,4-D applied, in a range from
0 to 5,120 μM. The mean number of somatic embryos formed per explant was recorded after 4 and 8 weeks culture. Our findings
indicated that the somatic embryogenesis in potato internodal segments could be evoked by an auxin (2,4-D) pulse treatment
over a wide concentration and duration range. The results further suggested that a simple 20 μM 2,4-D pulse treatment could
replace a lengthy 2 week induction phase in potato somatic embryogenesis and thus improve the system’s practicability for
wider uptake. 相似文献
13.
Thidiazuron stimulates shoot organogenesis and somatic embryogenesis in white ash (Fraxinus americana L.) 总被引:3,自引:0,他引:3
Sharon Bates John E. Preece Nadia E. Navarrete J. W. Van Sambeek Gerald R. Gaffney 《Plant Cell, Tissue and Organ Culture》1992,31(1):21-29
Immature and mature nonstratified seeds of white ash (Fraxinus americana L.) were dissected transversely and 2/3 of each seed was placed onto agar-solidified Murashige and Skoog medium. Adventitious buds, shoots, and somatic embryos formed on callus, cotyledons, and hypocotyls of the resulting seedlings. Shoot organogenesis was induced on explants cultured on medium with 10 M thidiazuron but not on explants on media with benzyladenine (BA) or isopentenyladenine. Not all seed sources were equally capable of shoot organogenesis and embryogenesis. Atypical of adventitious regeneration of other woody plants, mature seed explants of white ash were more organogenic with shoots that elongated better than explants from immature seeds. Somatic embryogenesis was observed in cultures where mature seeds were first cultured for 4 weeks on a medium containing 10 M adenine 2,4-dichlorophenoxyacetic acid in combination with 0.1 and 1.0 M thidiazuron, followed by transfer to a medium containing 0.05 M 6-benzyladenine and 0.5 M naphthaleneacetic acid. Adventitious shoots and epicotyls from both seedlings and germinated somatic embryos were rooted under intermittent mist and acclimatized to the greenhouse.Abbreviations BA
6-benzyladenine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- IBA
indolebutyric acid
- 2iP
isopentenyladenine
- NAA
naphthaleneacetic acid
- TDZ
thidiazuron-N-phenyl-N-1,2,3-thiadiazol-5-ylurea
- WPM
woody plant medium 相似文献
14.
A direct somatic embryogenesis protocol was developed for four cultivars of Nicotiana species, by using leaf disc as an explant. Direct somatic embryogenesis of Nicotiana by using BAP and IAA has not been investigated so far. This method does not require formation of callus tissues which leads to somaclonal variations. The frequency of somatic embryogenesis was strongly influenced by the plant growth hormones. The somatic embryos developing directly from explant tissue were noticed after 6 d of culture. Somatic embryogenesis of a high frequency (87–96%) was observed in cultures of the all four genotypes (Nicotiana tabacum, N. benthamiyana, N. xanthi, N. t cv petihavana). The results showed that the best medium for direct somatic embryogenesis was MS supplemented with 2.5 mg/l, 0.2 mg/l IAA and 2% sucrose. Subculture of somatic embryos onto hormone free MS medium resulted in their conversion into plants for all genotypes. About 95% of the regenerated somatic embryos germinated into complete plantlets. The plants showed morphological and growth characteristics similar to those of seed-derived plants. Explants were transformed using Agrobacterium tumifacious LBA4404 plasmid pCAMBIA1301 harboring the GUS gene. The regenerated transgenic plants were confirmed by PCR analysis and histochemical GUS assay. The transformation efficiency obtained by using the Agrobacterium- mediated transformation was more than 95%. This method takes 6 wk to accomplish complete transgenic plants through direct somatic embryogenesis. The transgenic plantlets were acclimatized successfully with 98% survival in greenhouse and they showed normal morphological characteristics and were fertile. The regeneration and transformation method described herein is very simple, highly efficient and fast for the introduction of any foreign gene directly in tobacco through direct somatic embryogenesis. 相似文献
15.
A rapid and effective system of somatic embryogenesis and organogenesis from the in vitro needles of redwood (Sequoia sempervirens (D.Don.) Endl.) had been established. The influences of plant growth regulators (PGRs) and days of seedlings in vitro on adventitious bud regeneration and somatic embryogenesis were studied. The process of somatic embryo formation was also observed. The results showed that embryogenic callus was induced and proliferated on Schenk and Hildebrandt (SH) medium with BA (0.5 mg/l), KT (0.5 mg/l) and IBA (1.0 mg/l). SH medium containing BA (0.5 mg/l), KT (0.2 mg/l) and IBA (0.2 mg/l) effectively promoted adventitious bud regeneration. The highest frequency (66.3%) of direct somatic embryogenesis was obtained in the combination of BA (0.5 mg/l) and IBA (0.5 mg/l). The optimal days of seedling in vitro for adventitious bud and somatic embryogenesis were 30 days and 30–40 days, respectively. The developments of somatic embryos were similar to that of zygotic embryogenesis. The result of histocytological studies indicated that proteins were gradually accumulated in the process of somatic embryo formation and there were two peaks of starch grains accumulation that one was in the embryogenic callus and the other was in the globular embryos. These results indicated that starch and protein were closely related with the energy supply and the molecular base of somatic embryogenesis, respectively. 相似文献
16.
Plant regeneration via somatic embryogenesis in many cultivars of cotton (Gossypium hirsutum L.) 总被引:4,自引:0,他引:4
Summary Embryogenic callus was formed from several cultivars of cotton (Gossypium hirsutum L.) when sections of hypocotyl and cotyledon were cultured on medium supplemented with 5 mg/liter 6-(γ, γ-dimethylallyl-amino)-purine
(2iP) and 0.1 mg/liter α-naphthaleneacetic acid (NAA) for callus initiation and proliferation, and subcultured on medium supplemented
with 5 mg/liter NAA and 0.1 to 1 mg/liter 2iP for embryogenic callus induction. It seems that a high 2iP:auxin ratio is preferred
for callus initiation and proliferation, but should be exchanged with a higher NAA:cytokinin ratio before differentiation
will occur. Embryogenic calluses were recovered at a frequency of 2 to 85% depending on the cultivar used. Coker cultivars
produced embryogenic callus faster and at higher frequencies than other cultivars. Embryogenic callus produced somatic embryos
on phytohormone-free medium. This medium was used to maintain and proliferate embryogenic callus for a perid of 18 to 24 mo.
Somatic embryos were converted to plants on a lower ionic strength medium supplemented with 0.1 mg/liter gibberellic acid
(GA3) and 0.01 mg/liter NAA. Glucose was the only carbohydrate used through all phases of tissue culture and was much better than
sucrose, on which phenolic production was very high. High temperature (30° C) and low light intensity (9 μE · m−2 · s−1) were optimal conditions for callus initiation, embryogenic callus induction, and maintenance, whereas lower temperature
(25° C) and high light intensity (90 μE · m−2 s−1) were the optimal conditions for somatic embryo maturation, germination, and plantlet development. Plants could be regenerated
within 10 to 12 wk in Cokers or 7 to 8 mo. in others. 相似文献
17.
Nutritional effect of nitrate salts of potassium and ammonium, together with different concentrations of sulphate salts of aluminium, potassium, magnesium, and ammonium on secondary somatic embryogenesis, wereinvestigated. Nitrate salts of potassium (9.39 mmol/L) and ammonium (10.31 mmol/L) with only 1.5 mmol/L potassium sulphate produced maximum number of synchronous secondary embryos (i.e. 20-25 secondary embryos per primary embryo in 91.6 percnt; responsive explants).Of the different factorial combinations of glutamine, BAP, and IBA tested, maximum number of synchronous secondary embryos developed on a medium supplemented with 8.88 μmol/L BAP, 0.98 μmol/L IBA and 10 mmol/L glutamine.Synchronous and normal development of secondary embryos could thus be obtained when optimal concentrations of PGRs, glutamine, nitrates, and salts of potassium sulphate were combined together.Germination of the embryos (up to 52 percnt;) was acheived only when sulphate salts of potassium were removed from the medium. 相似文献
18.
Barbara Stefaniak 《Plant cell reports》1994,13(7):386-389
Summary Friable embryogenic callus and somatic embryos of 4 Gladiolus cultivars were obtained on Murashige and Skoog (MS) medium with various concentration of auxins from the following explants: corm slices, young leaf bases and whole, intact plantlets. Somatic embryos transferred on MS hormone-free medium regenerated into plantlets. All plantlets obtained through embryogenesis did not differ phenotypically from the parental clones. The embryogenic friable callus has been maintained for over 2 years in culture and has retained a very high regeneration capacity.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- KIN
kinetin
- NAA
naphthaleneacetic acid
- MS
Murashige and Skoog Medium (1962)
- E
embryogenic callus
- NE
non-embryogenic callus 相似文献
19.
Trabelsi El Bahri Bouzid Sadok Bouzid Monji Elloumi Nedra Belfeleh Zina Benabdallah Abdallah Ghezel Rachida 《Journal of Plant Biology》2003,46(3):173-180
We induced somatic embryogenesis from the cotyledon segments ofOlea europaea (L) cvs. ‘Chetoui’, ‘Chemleli’, and ‘Arbequina’. Calli were established from all three cultvars on OMc media supplemented
with IBA and 2i-R The greatest success was obtained with media that contained zero or low concentrations of growth regulators.
High levels of hormones (i.e.,>0.5 mgL-1 IBA and 2i-P) inhibited embryogenesis. Embryos at different maturation stages were observed with continuously proliferating
secondary embryogenesis. Abnormally shaped embryos and teratoma were also noted. Four weeks was the optimal incubation period
for inducing embryogenesis on the auxin-containing medium. In addition, 30 to 40 gL-1 sucrose was more effective than glucose in stimulating the growth and maturation of somatic embryos. Embryogeic efficiency
was also higher when multivariate combinations of nitrogen sources (inorganic and organic nitrogen forms) were used. The plantlets
that were derived from our germinating somatic embryos were similar to those obtained from axillary buds. 相似文献
20.
We established an efficient plant regeneration system for Catharanthus roseus L. (G.) Don through somatic embryogenesis. Embryogenic callus was induced from hypocotyl of seed germinated in vitro. Somatic
embryogenesis in Catharanthus has been categorized into three distinct stages: (1) initiation and proliferation of embryo; (2) maturation, and; (3) germination
or plantlet conversion. Beside plant growth regulators, various stages of embryogenesis were screened for their response to
a wide variety of factors (pH, gelrite, light, sugar alcohols, polyethyleneglycol and amino acids), which affect embryogenesis.
All of the tested factors had a small to marked influence on embryogeny and eventual conversion to plantlets. The plantlets
were acclimatized successfully in a greenhouse. To our knowledge, this is the first report describing a detailed study of
various cultural factors which regulate embryogenesis in C. roseus. The results discussed in this paper may be used in mass propagation to produce medicinal raw material, and the embryo precursor
cells could be used in genetic modification programmes that aim to improve the alkaloid yield as well. 相似文献