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1.
Identification of a novel elite genotype for in vitro culture and genetic transformation of cotton 总被引:4,自引:0,他引:4
Hypocotyls of cotton (Gossypium hirsutum L.) cultivars cv. YZ-1, Coker 312 and Coker 201 were inoculated on Murashige and Skoog callus induction medium. YZ-1 exhibited
a very high regeneration potential, with 81.9 % of the explants inoculated differentiated into embryogenic callus within 8–10
weeks. During the process of callus maintenance (subculture for 1 to 3 years), the total embryos number in Coker 312 and Coker
201 calli dropped sharply, and the percentage of embryo germination decreased. On the contrary, the callus of YZ-1 consistently
maintains a high frequency of plant regeneration after long-time subculture. Transgenic kanamycin-resistant calli of Coker
201 partially lost the ability of somatic embryogenesis and plant regeneration. The stress produced by the transformation
procedure slightly affected somatic embryogenesis and plant regeneration of YZ-1, which showed minimum loss of plant regeneration
ability. 相似文献
2.
Y. Y. Wu Q. J. Chen X. H. Cui H. Chen J. Chen X. C. Wang 《Russian Journal of Plant Physiology》2007,54(4):524-529
We utilized gene transfer technology for genetic perennial ryegrass improvement, efficient regeneration, and Agrobacterium-mediated transformation of phosphinothricin acetyltransferase gene (bar). Four growth regulator combinations were compared and intact seeds of six turf-type cultivars as mature embryo sources were
tested to optimize the regeneration conditions. Callus formation and regeneration were observed in all seeds. The highest
callus formation frequency was observed in the seeds cultured on MS medium supplemented with 9 mg/l 2,4-D, without benzyladenine.
Cv. TopGun revealed the highest callus induction and regeneration frequencies of 96 and 48.9%, respectively. By using an optimized
regeneration system, embryogenic calli were transformed by an Agrobacterium strain LBA4404 containing the plasmid pCAMBIA3301. After the selection of the potentially transgenic calli with phosphinothricin,
a herbicide, 22 transgenic resistant plants were regenerated. With PCR, Southern-blot hybridizations, and GUS expression techniques,
we confirmed that some regenerants were transgenic. Two of the tested transgenic plants showed herbicide resistance. Our results
indicated that embryogenic calli from mature seeds can be directly used for perennial ryegrass efficient regeneration and
transformation and this protocol is applicable for genetic engineering of herbicide-resistant plants.
Published in Russian in Fiziologiya Rastenii, 2007, Vol. 54, No. 4, pp. 590–596.
The text was submitted by the authors in English. 相似文献
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5.
Harsh Chauhan Srinivas A. Desai Paramjit Khurana 《Plant Cell, Tissue and Organ Culture》2007,91(3):191-199
An efficient genotype independent, in vitro regeneration system was developed for nine popular Indian wheat cultivars, three
each of Triticum aestivum L. viz., CPAN1676, HD2329 and PBW343, Triticum durum Desf. viz., PDW215, PDW233 and WH896, and Triticum dicoccum Schrank. Schubl. viz., DDK1001, DDK1025 and DDK1029, by manipulating the concentration and time of exposure to the growth
regulator, thidiazuron (TDZ). A total of 18 (for immature inflorescence and embryo explant) and six (for mature embryo explant)
different combinations of growth regulators were tried for callusing and regeneration, respectively. Media combination with
low concentration of TDZ (2.2 μM) in combination to auxin and/or cytokinin (depending upon culture stage), was found to be
effective for immature and mature explants. Compact, nodular and highly embryogenic calli were obtained by using immature
embryo, immature inflorescence and mature embryo explants, and regeneration frequency up to 25 shoots/explant with an overall
80% regeneration was achieved. Comparable regeneration frequency was achieved for mature embryo explants. No separate hormone
combination for rooting was required and plantlets ready to transfer to soil could be obtained in a short period of 8–10 weeks.
This protocol can be used for raising transgenic plants for functional genomics analysis of agronomically important traits
in the three species of wheat. 相似文献
6.
A good culture system provides considerable quantities of highly regenerable target tissues. Embryogenic callus cultures are ideal for micro-projectile-mediated transformation, because regenerable cells are not very stable. Effective exploitation of genetic transformation requires good regeneration systems. We selected three sugarcane genotypes for the establishment and optimization of good in vitro regeneration systems, viz., S-2003-us-359, S-2006-sp-30, and S-2003-us-165. Three callus induction media were investigated. These media were composed of Murashige and Skoog (MS) medium salt plus 1, 2, and 3 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). Medium with 3 mg/L 2,4-D gave the greatest mass of embryogenic calli. The calli produced on the three callus induction media were transferred to 18 types of regeneration media (RM1-RM18). They varied with respect to plant growth regulators and sucrose levels but the basal medium was MS. Two levels of sucrose (30 and 40 g/L), three levels of 2,4-D (0.1, 0.25, 0.5 mg/L) and three levels of 6-benzylaminopurine (0, 0.25 and 0.5 mg/L) were studied in the regeneration media. The effects of callus age on regeneration were evaluated by transferring the calli to regeneration media after 15, 21, 28, and 35 days of culture. The 21-day-old callus of the genotype S-2003-us-359 on RM3 yielded the largest number of plants and was selected as the best for transformation. Six RAPD DNA primers were used to check genetic stability; this medium did not affect the sugarcane genomes. 相似文献
7.
Somatic embryogenesis and plant regeneration of turf-type bermudagrass: Effect of 6-benzyladenine in callus induction medium 总被引:17,自引:0,他引:17
In order to optimize tissue culture conditions for bermudagrass, an important warm-season turfgrass species, tissue culture
responses of young inflorescences of a hybrid bermudagrass cultivar `Tifgreen' (Cynodon dactylon×Cynodon transvaalensis) and a common bermudagrass cultivar `Savannah' (Cynodon dactylon) were investigated. When cultured on Murashige and Skoog medium with 4.52 to 13.57 μM (1–3 mg l-1) 2,4-D, young inflorescence segments yielded non-embryogenic calli which were unorganized and had loosely associated, long
tubular cells on the surface. However, inclusion of 6-benzyladenine (BA) in callus induction medium at a level of 0.044 μM
(0.01 mg l-1) induced formation of a compact, nodular embryogenic structure on approximately 20% of the calli. Calli with such a compact
embryogenic structure were highly regenerable. When young inflorescences smaller than 0.75 cm were cultured, the embryogenic
structure yielded green plantlets with regeneration rates of 79.5% and 83.3%, respectively for the two cultivars. All 96 plants
regenerated from calli induced in the BA-containing medium were green and morphologically normal. The embryogenic nature of
the compact structure was confirmed by scanning electron microscopy.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
8.
Optimization of in vitro plant regeneration and genetic transformation of apomictic species such as Dichanthium annulatum would enable transfer of desirable genes. Seven genotypes of this grass species were screened through mature seed explant for embryogenic callus induction, callus growth and quality (color and texture), and shoot induction. Genotype IG-1999, which produced highly embryogenic, rapidly growing good-quality callus capable of regenerating at a high frequency, was selected for transformation experiments. Using a binary vector (pCAMBIA1305), frequency of GUS expression was compared between two methods of transformation. Bombardment of embryogenic calli with gold particles coated with pCAMBIA1305 at a distance of 11 cm, pressure of 4 bars, and vacuum of 27 Hg passing through 100 muM mesh produced maximum GUS expression (23%). Agrobacterium infection was maximum at an optical density of 2.0 when cocultured under vacuum for 15 min and cocultivated for 3 days at 28 degrees C in constant dark on MS medium of pH 5.8 with 3 mg/l 2,4-D, and 400 muM acetosyringone. Among two binary vectors used for Agrobacterium-mediated transformation, pCAMBIA1301 showed higher frequency of GUS expression while pCAMBIA1305 recorded more of the GUS spots per callus. Supplementation of acetosyringone in the cocultivation medium was found indispensable for Agrobacterium-mediated transformation. Injuring the calli through gold particle bombardment before their cocultivation with Agrobacterium improved the transformation efficiency. Several transgenic plants were developed using the PIG method, while stable GUS-expressing calli were multiplied during selection on MS medium containing 250 mg/l cefotaxime and 50 mg/l hygromycin, incubated in constant dark. A highly significant difference was observed between two methods of transformation for both frequency of GUS expression and GUS spots per callus. PIG-mediated transformation resulted in higher GUS expression compared to the Agrobacterium method. These results demonstrate that Dichanthium annulatum is amenable to Agrobacterium-mediated genetic transformation using a binary vector. 相似文献
9.
A procedure was established for the induction of regenerable calli from immature inflorescence segments of high-tannin cultivars of sorghum (Sorghum bicolor (L.) Moench). Murashige & Skoog's medium with several components altered was utilized for inducing, maintaining, and regenerating the cultures. Embryogenic calli formed at a frequency of 8–70% depending on the genotype. During a ten-month period, 3600 plants were regenerated from eight genotypes tested. Among the developmental stages of immature inflorescence tested (from differentiation of secondary branch primordia to floret formation) no critical differences were found in potential for callusing, embryogenesis or regeneration. Genotypic differences were observed in pigment production, embryogenic callus formation, shoot differentiation, and in maintenance of regeneration capacity.Abbreviations 2,4-D
dichlorophenoxyacetic acid
This is Journal Paper Number 11972 from the Purdue University Agricultural Experiment Station 相似文献
10.
Manal M. Abdel-Rahman Jack M. Widholm 《In vitro cellular & developmental biology. Plant》2010,46(6):509-515
Type II maize callus of the HiII genotype can be separated into regenerable and non-regenerable types based on the visible
morphology of the callus. When the non-regenerable morphotype of callus initiated 6 mo or a yr earlier was treated with from
2% to 20% polyethylene glycol (PEG; 3,350 molecular weight) for three subculture periods of 21 d each, the morphotype changed
to regenerable, and the callus did become highly regenerable. The PEG treatments did not improve the plant regeneration ability
of the regenerable morphotype or of an old culture initiated 4 yr earlier. 相似文献
11.
Optimization of the conditions for an efficient induction of somatic embryogenic calli and regeneration of plants from mature seeds of japonica rice cultivars was attempted. The number, color, size, shape, and appearance time of the induced embryogenic calli varied among the rice cultivars depending on the type of basal medium (LS, MS, N6). Presence of adequate amount of sucrose in the medium was an absolute requirement for embryogenic callus formation and shoot induction. Induction of the embryogenic calli, whose overall rates ranged from 30 to 56%, was most efficient in N6 medium supplemented with 3.0 mg l–1 of 2,4-D and 30 g l–1 of sucrose. Agar concentration in the regeneration medium was also critical for the shoot induction. Kinetin was found to be more effective for shoot regeneration compared with BA, while the highest shoot regeneration frequencies were observed when either cytokinin was combined with high concentration (2.0 mg l–1) of NAA. The optimal concentration of kinetin for the highest shoot regeneration frequency (6777%) was different among the cultivars tested. The embryogenic calli-derived shoots rooted on a plant growth regulator-free MS medium were successfully established in soil, producing fertile seeds. 相似文献
12.
Araceli Rodríguez-Sahagún Gustavo Acevedo-Hernández José M. Rodríguez-Domínguez Benjamín Rodríguez-Garay Jesús Cervantes-Martínez Osvaldo A. Castellanos-Hernández 《Plant Cell, Tissue and Organ Culture》2011,104(2):271-275
Somatic embryogenesis in Agave tequilana Weber var. Azul was affected by the interaction between the light regimes applied during the induction phase and the expression
phase. When embryogenic calli was exposed to white or red light during the expression phase, an average of two germinated
embryos per explant was obtained regardless of the light treatment used for callus induction. Conversely, the highest number
of germinated embryos, an average of 18 per explant, was obtained when applying either white or red light during the induction
phase and then wide-spectrum light during the expression phase. Culture medium had also a great influence in this process,
with embryo germination being reduced by up to 70%, depending on the light treatment, when using Schenk and Hildebrandt (SH)
medium instead of Murashige and Skoog (MS) medium. 相似文献
13.
The Stimulatory Effect of the Antibiotic Cefotaxime on Plant Regeneration in Maize Tissue Culture 总被引:1,自引:0,他引:1
In order to elucidate the effects of the antibiotic cefotaxime on callus growth and morphogenesis, we incubated embryogenic maize calli (Zea mays L.) of A188 and R91 lines and of their F1 hybrid with 50–500 mg/l cefotaxime throughout several subcultures. Cefotaxime did not affect the induction frequency and growth of the embryogenic callus but enhanced its morphogenesis. In both tested lines and a hybrid, the highest increase in the number of regenerated plants was observed at the antibiotic concentration of 150 mg/l. The degree of morphogenesis stimulation and the range of cefotaxime concentrations effective in stimulation of plant regeneration depended on the properties of calli obtained from tested genotypes. 相似文献
14.
Russian wildrye [Psathyrostachys juncea (Fisch.) Nevski] is a cool-season forage grass with a broad adaptation to semi-arid regions of North America. In order to explore the potential of biotechnology for genetic improvement of this important forage species, we developed an efficient tissue culture system. Embryogenic calli were induced from mature embryos with an induction frequency in the range of 2-7%. The selected highly embryogenic calli allowed the regeneration of dozens of plants from a single callus. Individual embryogenic calli were then used to establish single genotype-derived suspension cultures. Eighteen embryogenic cell suspension lines were established from three cultivars (Bozoisky-Select, Sawki and Tetracan). A relatively high green plant regeneration frequency, up to 70%, was achieved from plated cell clusters of the established suspension cultures. The regenerated plants were fertile after two winters of vernalization in the field. This efficient plant regeneration system provides a solid basis for generating transgenic plants. 相似文献
15.
Bernadine D. Metzinger Charles M. Taliaferro Becky B. Johnson Earl D. Mitchell Jr. 《Plant Cell, Tissue and Organ Culture》1987,10(1):31-38
Explants from immature inflorescences of four genotypes of Old World bluestem grasses, (Bothriochloa spp.), produced callus tissue on Linsmaier and Skoog (RM) and 1/2 Murashige and Skoog (1/2 MS) media containing high levels of growth regulators. Callus masses were composed of two distinct tissue types, one a compact, white, embryogenic portion (E calli), the other soft, translucent, gelatinous and nonembryogenic (NE calli). When transferred to medium with a reduced level of 2,4-D, and/or supplemented with zeatin, E callus underwent further organization culminating in shoot production. Light and scanning electron microscopy confirmed the embryogenic pathway of differentiation. Genotype significantly affected callus induction frequency and the number of plants regenerated. The RM medium induced more explants to initiate callus compared to the 1/2 MS medium. Age of the inflorescence explant, as indicated by size, was critical for callus induction. Inflorescences with racemes 8 mm in length were superior to older ones. Five-hundred-twenty-two plantlets were regenerated and grown to maturity. 相似文献
16.
In this paper, we describe the transformation of regenerable maize tissues by electroporation. In many maize lines, immature zygotic embryos can give rise to embryogenic callus cultures from which plants can be regenerated. Immature zygotic embryos or embryogenic type I calli were wounded either enzymatically or mechanically and subsequently electroporated with a chimeric gene encoding neomycin phosphotransferase (neo). Transformed embryogenic calli were selected from electroporated tissues on kanamycin-containing media and fertile transgenic maize plants were regenerated. The neo gene was transmitted to the progeny of kanamycin-resistant transformants in a Mendelian fashion. This showed that all transformants were nonchimeric, suggesting that transformation and regeneration are a single-cell event. The maize transformation procedure presented here does not require the establishment of genotype-dependent embryogenic type II callus or cell suspension cultures and facilitates the engineering of new traits into agronomically relevant maize inbred lines. 相似文献
17.
Regeneration potential of different wheat, rye and barley species in leaf explant culture. Comparative analysis of the induction ability of morphogenetic processes in vitro has been carried out in 16 wheat genotypes, 4 barley species and 6 rye genotypes. It has been shown that tetra- and hexaploid wheat species as well as wild barley species exhibited the highest embryogenic potential in the leaf explant culture while diploid wheat species and rye genotypes showed the lowest one. Genotypic dependence of processes of callus formation, induction of embryogenic calli and regeneration was revealed in the studied species. 相似文献
18.
Response of twenty eight cultivars of durum wheat (Triticum turgidum var. durum) to immature embryo culture, callus production and in vitro salt tolerance was evaluated. For assessment of cultivars to salt tolerance, growing morphogenic calli were exposed to different
concentrations of NaCl (0, 0.3, 0.6, 0.9, 1.2, 1.5, 1.8 and 2.1% w/v) added to the culture medium during two subsequent subcultures
(4 weeks each). Comparison of cultivars for callus induction from immature embryo was based on callus induction frequency
and fresh weight growth of callus (FWG). While, for salt tolerance, the relative fresh weight growth (RFWG) and necrosis percent
of callus were used. There were significant differences among cultivars for potential of regeneration from immature embryo,
and ‘Shahivandi’ a native durum wheat cultivar originating from western Iran was superior among the cultivars tested. The
FWG distinguished cultivars more than callus induction frequency did for callus induction evaluation. Hence, a range of FWG
from 1.23 to 14.65 g was observed in ‘Mexical-75’ and ‘Omrabi-5’ cultivars, respectively. Growing calli derived from cultivars
‘PI 40100’ and ‘Dipper-6’ showed superiority for tolerating salinity under in vitro conditions.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
19.
The origin and development of somatic embryos in calli initiated from immature zygotic embryos of Picea abies (L.) Karst. (Norway spruce) and P. glauca (Moench) Voss (white spruce) was studied. Immature zygotic embryos cultured on callus induction medium produced two types of white calli that were phenotypically different from one another. The callus that proliferated from the hypocotyl region was white to translucent, glossy, mucilaginous and embryogenic. The callus mass which originated from the radicle end was reddish-white, nonmucilaginous and nonembryogenic. Whole mount preparations of the entire explant with two different types of calli showed the presence of embryogenic cells in the mucilaginous callus mass derived from the hypocotyl region of the zygotic embryo. The origin of somatic embryos in both Norway and white spruce could be traced to single cells of the hypocotyl callus.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
6-benzylaminopurine 相似文献
20.
Lambé Pascal Mutambel Hity S.N. Deltour Roger Dinant Monique 《Plant Cell, Tissue and Organ Culture》1998,55(1):23-29
Three genotypes of Pearl millet were screened in vitro for induction of embryogenic callus, somatic embryogenesis and regeneration.
Shoot apices excised from in vitro germinated seedlings or immature embryos isolated from green house established plants were
used as primary explants. The frequency of embryogenic callus initiation was significantly higher in shoot apices in comparison
with immature zygotic embryos. Moreover, differences between genotypes were minimal when using shoot apices. Friable embryogenic
calli (type II) developed on the initial nodular calli after 1 to 3 months of culture. The frequency of type II callus is
related to the composition of the maintenance medium and they were more often found in ageing cultures. The transfer of embryogenic
calli onto auxin-free medium was sufficient for inducing somatic embryo development in short-term culture (3 months) while
a progressive loss in regeneration potential was observed with increasing time of subcultures. Maturation of embryogenic calli
on medium supplemented with activated charcoal, followed by germination of somatic embryos on medium supplemented with gibberellic
acid, restored regeneration in long-term cultures.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献