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1.
Five different genotypes from in vitro as well as greenhouse grown melon plants were shown to be highly responsive for in vitro shoot formation from leaf explants when placed on basic MS medium supplemented with 1 mg/l 6-benzylaminopurine. In addition, a very suitable regeneration system was obtained when cotyledon pieces of mature seeds were incubated on the same culture medium. In this case, the first shoots already appeared after 10 days of incubation, and hundreds of shoots were formed on the cut surface 3 to 4 weeks later. Explants from mature cotyledons derived from seedlings did not lead to any shoot formation.Abbreviations MS
Murashige and Skoog
- IAA
3-indoleacetic acid
- BA
6-benzylaminopurine 相似文献
2.
Gaurav Krishna P. Sairam Reddy Pramod W. Ramteke Pogiri Rambabu Sayed S. Sohrab Debashis Rana Parthasarathi Bhattacharya 《Physiology and Molecular Biology of Plants》2011,17(4):375-385
In vitro regeneration of pigeon pea through organogenesis and somatic embryogenesis was demonstrated with pigeon pea cv. JKR105. Embryonic axes explants of pigeon pea showed greater regeneration of shoot buds on 2.5 mg L−1 6-benzylaminopurine (BAP) in the medium, followed by further elongation at lower concentrations. Rooting of shoots was observed on half-strength Murashige and Skoog (MS) medium with 2 % sucrose and 0.5 mg L−1 3-indolebutyric acid (IBA). On the other hand, the regeneration of globular embryos from cotyledon explant was faster and greater with thidiazuron (TDZ) than BAP with sucrose as carbohydrate source. These globular embryos were maturated on MS medium with abscisic acid (ABA) and finally germinated on half-strength MS medium at lower concentrations of BAP. Comparison of regeneration pathways in pigeon pea cv. JKR105 showed that the turnover of successful establishment of plants achieved through organogenesis was more compared to somatic embryogenesis, despite the production of more embryos than shoot buds. 相似文献
3.
Hassan Rostami Archana Giri Amir Sasan Mozaffari Nejad Amir Moslem 《Saudi Journal of Biological Sciences》2013,20(3):251-255
Barley is the fourth most important crop in the world. Development of a regeneration system using immature embryos is both time consuming and laborious. The present study was initiated with a view to develop a regeneration system in six genotypes of Indian barley (Hordeum vulgare) cultivars as a prerequisite to transformation. The mature embryos were excised from seeds and cultured on MS medium supplemented with high and low concentrations of cytokinins and auxins respectively. The MS medium containing 3 mg/L N6-benzylaminopurine (BA) and 0.5 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) was found to be the most effective for multiple shoot formation in HOR7231 cultivar that could produce 12 shoots per explant. The other cultivars HOR4409 and HOR3844 produced a minimum number of adventitious shoots (1.33 and 1.67 respectively) on MS medium supplemented with 1 mg/L BA and 0.3 mg/L 2,4-D. The elongated shoots were separated and successfully rooted on MS medium containing 1 mg/L indole-3-acetic acid (IAA). The response of different barley cultivars was found to be varying with respect to multiple shoot production. This is the first report of multiple shoot induction and plantlet regeneration in Indian cultivar of barley which would be useful for genetic transformation. 相似文献
4.
A procedure has been developed which allows high frequency adventitious shoot regeneration from immature cotyledons of pea. Prolific shoot regeneration occurred following an initial callus growth on a Murashige and Skoog (MS) medium containing 0.5 mg/l 6-benzylaminopurine (BAP) and 4 mg/l -naphthaleneacetic acid (NAA). Cotyledon expiants proximal to the embryonic axis had the highest regeneration potential, however, the presence of an embryonic axis inhibited adventitious shoot regeneration. Addition of silver nitrate (AgNO3) to the medium did not promote the number of regenerated shoots but resulted in shoots with well developed tendrils and large stipules which had a reduced rooting capacity. Regenerated shoots rooted readily (80–90%) in half strength MS medium containing 1 mg/l indole-butyric acid (IBA) and further established well in compost. 相似文献
5.
Adventitious shoots were successfully regenerated from leaf explants of in vitro cultures of Platanus acerifolia Willd. The leaves of three clones (genotypes), designated as PH1, PH2 and PC, respectively, were wounded by three to four transverse cuts through the midvein and cultured on 26 media based on Murashige and Skoog (MS) basal medium, containing different concentrations of 6-benzylaminopurine (BAP) in combination with different concentrations of indole-3-butyric acid (IBA). The highest regeneration rate (>90%) and the largest number of shoot clumps per regenerating leaf (>4 shoot clumps/explant) were obtained with leaves of genotype PH2 cultured on MS basal medium supplemented with 17.76 microM BAP and 4.92 microM IBA. The other two genotypes, PH1 and PC, showed very low capability of shoot regeneration (<10%) on all the media tested. Shoots on leaf explants originated mainly from callus that developed around the cut end of petioles and along the cuts across the midvein. The regenerated shoots were micropropagated, rooted and transplanted to the soil successfully. 相似文献
6.
In vitro regeneration of plantlets and multiplication of Sesbania bispinosa (Jacq.) W.F. Wight plants from cultured callus tissue were demonstrated. Callus was established from both cotyledons and mature leaflets on Murashige and Skoog (MS) basal medium supplemented with BAP (0.5 mg/l) and 2,4-D (2 mg/l). Callus mediated shoot bud differentiation was studied under defined nutritional, hormonal and cultural conditions. Various concentrations of BAP or kinetin (Kn) with coconut milk (CM) in MS media induced different levels of shoot bud differentiation as well as multiplication. Multiple shoot bud differentiation occurred in most of the primary calli. The best medium for shoot bud differentiation from cotyledon derived callus, contained BAP (2 mg/l) and 15% CM (V/V). More efficient shoot bud organogenesis was recorded with BAP than Kn. Supplementation with CM in MS media accelerated shoot bud organogenesis in differentiating callus tissue. Rooting of differentiated shoots was achieved by a three step culture procedure involving (a) MS solid medium containing IBA (2 mg/l), (b) growth regulator free half strength MS medium with 1% charcoal, and (c) half strength MS liquid medium free of vitamins, growth regulators and charcoal.Abbreviations IAA
indoleacetic acid
- IBA
indole-3-butyric acid
- NAA
naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
6-benzylaminopurine
- Kn
kinetin
- CM
coconut milk
- MS
Murashige and Skoog's medium
- SBI
shoot bud inducing medium 相似文献
7.
Jing Qin Mao Mohsin Abbas Zaidi John Thor Arnason Illimar Altosaar 《Plant Cell, Tissue and Organ Culture》2006,87(2):121-125
An in vitro regeneration system was developed in cowpea [Vigna unguiculata (L.) Walp.] Blackeye. Among several explants studied, shoot initiation response was observed from shoot apices of 3–5-day-old seedlings. The optimal medium for maximum shoot initiation comprised MS salts, B5 vitamins, 8.88 μM N
6-benzylaminopurine, 1 gl-1 casein hydrolysate, 342 μM L-glutamine, 3% sucrose, 0.3% phytagel, adjusted to pH 5.8. A shift in pH from 5.8 to 7.0 had no effect on shoot initiation and on number of shoots per explant. The highest shoot initiation frequency (77%) was obtained using this preferred medium, reaching a maximum of eight shoots per explant. For shoot elongation, 14 μM gibberellic acid was supplemented in the shoot initiation medium. Presence of indolebutyric acid in the rooting medium had no effect on root induction. The regenerated plants were fertile and developed normally. 相似文献
8.
High frequency plant regeneration from immature embryos of an elite barley cultivar (Hordeum vulgare L. cv. Morex) 总被引:1,自引:0,他引:1
An efficient plant regeneration system was developed for Hordeum vulgare L. 'Morex' barley, an important United States malting cultivar. The protocol was based on a series of experiments involving the sizes of immature embryos and the culture media. We found that the embryo size is critical for the establishment of embryogenic callus. Smaller embryos (0.5-1.5 mm) showed a much higher ability to produce embryogenic callus capable of regenerating green plants with fewer albinos than did the larger embryos (1.6-3.0 mm). Either 3 mg/l 2,4-dichlorophenoxyacetic acid or dicamba in modified Murashige and Skoog's (MS) medium was optimum for the induction of embryogenic callus. The embryogenic callus maintained high regeneration during six subcultures in the callus induction medium. Efficient shoot regeneration was obtained on modified MS medium containing 0.5-1.0 mg/l 6-benzylaminopurine (BA). Regenerated shoots were rooted on half-strength MS medium containing 0.2 mg/l IBA. Plants were successfully transferred to soil and grown to maturity in the greenhouse. This efficient plant regeneration system provides a foundation for generating transgenic plants of this important barley cultivar. 相似文献
9.
Summary Development of an efficient transformation method for recalcitrant crops such as sugar beet (Beta vulgaris L.) depends on identification of germplasm with relatively high regeneration potential. Individual plants of seven sugar
beet breeding lines were screened for their ability to form adventitious shoots on leaf disk callus. Disks were excised from
the first pair of true leaves of 3-wk-old seedlings or from partially expanded leaves of 8-mo.-old plants and cultured on
medium with 4.4 μM 6-benzylaminopurine for 10 wk. At 5 wk of culture, friable calluses and adventitious shoots began to develop. Rates of callus
and shoot formation varied between breeding lines and between individual plants of the same line. Line FC607 exhibited the
highest percentage (61%) of plants that regenerated shoots on explants. Among the plants with a positive shoot regeneration
response, line FC607 also had the highest mean number (8.3±1.1) of shoots per explant. Individual plants within each line
exhibited a wide range of percentages of explants that regenerated shoots. A similar variation was observed in the number
of shoots that regenerated per explant of an individual plant. No loss of regeneration potential was observed on selected
plants maintained in the greenhouse for 3 yr. Regenerated plants exhibited normal phenotypes and regeneration abilities comparable
to the respective source plants. Based on our results, it is imperative to screen a large number of individual plants within
sugar beet breeding lines in order to identify the high regenerators for use in molecular breeding and improvement programs. 相似文献
10.
Three genotypes of chickpea ICCV-1, ICCV-6 and a Desi (local) variety were tested for plant regeneration through multiple shoot production. The embryo axis was removed from mature seeds, the root meristem and the shoot apex were discarded. These explants were cultured on medium containing MS macro salts, 4X MS micro salts, I35 vitamins, 3.0 mg/1 BAP, 0.004 mg/1 NAA, 3% (w/v) sucrose and incubated at 260C. The explants were transformed withAgrobacterium tumefaciens strain LBA4404 with binary vector pBI121 containing theuidA andnptIl genes. Multiple shoots were repeatedly selected with kanamycin. The selected kanamycin resistant shoots were rooted on MS medium supplemented with 0.05 mg/1 113A. The presumptive transformants histochemically stained positive for GUS. Additionally, nptll assay confirmed the expression ofnptII in kanamycin resistant plants. Transgenic plants were transferred to soil and grown in the green house.Abbreviations BAP
6-benzylamino purine
- 2,4-D
2,4dichlorophenoxy acetic acid
- IAA
Indole acetic acid
- IBA
Indole butaric acid
- NAA
Naphthalene acetic acid 相似文献
11.
Lining Tian Yan Wen S. Jayasankar Susan Sibbald 《In vitro cellular & developmental biology. Plant》2007,43(4):343-347
Plant regeneration of Prunus
salicina (Japanese plum) using mature seeds was studied and evaluated. Shoots were effectively induced from hypocotyl slices of mature
seeds on media containing cytokinins. Among three plant growth regulators evaluated, thidiazuron (TDZ) was the most effective
for shoot induction overall. Shoots were also induced using 6-benzylaminopurine (BA), but the effectiveness was reduced at
low concentrations. Low regeneration was induced using kinetin. Three plum varieties were evaluated and the regeneration appeared
to be genotype dependent. Induced shoots elongated, roots formed, and plantlets developed upon transfer of the shoots to the
rooting medium. Primary shoots, when sub-cultured on fresh induction medium, produced multiple shoots, and such multiplication
could continue for more cycles. The plantlets were transferred to soil, and the full plants were readily recovered in a greenhouse.
The regeneration process was relatively fast as plants could be recovered in 4 to 5 mo. after the culture initiation. 相似文献
12.
G. Franklin L. Carpenter E. Davis C.S. Reddy D. Al-Abed W. Abou Alaiwi M. Parani B. Smith R.V. Sairam 《Plant Growth Regulation》2004,43(1):73-79
Soybean (Glycine max L.) plantlets were e?ciently regenerated from mature and immature cotyledons of?ve different genotypes by studying various parameters affecting regeneration. Green organogenic noduleswere induced at the proximal end, which subsequently differentiated into shoot buds on modi?ed MS(Murashige T. and Skoog F. 1962. Physiol. Plant. 15: 473-497) medium. The presence of 6-benzylaminopurine(BAP) and thidiazuron (TDZ) in the medium exerted a synergistic effect, in that regenerationeffciency was higher than for either cytokin alone. The regenerated shoot buds elongated and rooted onMS medium containing 0.29 lM gibberellic acid (GA3) and 2.69 lM a-naphthaleneaceticacid (NAA),respectively. Rooted plants were established in the greenhouse with 87% success and produced viable seeds.Preliminary studies with Agrobacterium show great promise for soybean transformation based on theregeneration protocol reported here. 相似文献
13.
Gunvant Patil Ramesh Patel Ranjeet Jaat A. Pattanayak Pradeep Jain R. Srinivasan 《Acta Physiologiae Plantarum》2009,31(5):1077-1084
The use of glutamine has been shown to increase the frequency of organogenesis and regeneration in the in vitro culture of
several plants. The effect of glutamine on hormone-induced multiple shoot formation in desi and kabuli genotypes of chickpea
(C-235 and PUSA-1053) were evaluated. Embryo axes with or without attached cotyledons were cultured in thidiazuron (TDZ) or
6-benzylaminopurine (BAP)-containing medium, respectively, with various concentrations of glutamine. Glutamine improved and
prolonged the multiple shoot regeneration ability of the embryo axis. Chickpea embryo axis with attached cotyledon and cultured
in TDZ-containing medium showed improved and prolonged shoot regeneration with 5 mM glutamine, while embryo axis without cotyledon
and cultured in BAP-containing medium showed prolonged regeneration ability in 10 mM glutamine. Glutamine, however, did not
serve as a substitute for cotyledon. Desi genotype (C-235) showed better response for multiple shoot formation as compared
to the kabuli genotype (PUSA-1053). Glutamine at a concentration of 5 mM also improved root formation in excised in vitro
shoots. 相似文献
14.
In vitro regeneration techniques have been optimized for seven strains and cultivars of sugar beet (Beta vulgaris L.) bred in Russia. The frequency of shoot regeneration from somatic cells and tissues of sugar beet varies from 10 to 97% depending on the explant type, culture-medium composition, and genotype. The in vitro regeneration potential has been estimated in plants with different genotypes. The effect of medium composition (phytohormones and carbohydrates) on the frequency of the formation of a morphogenic callus competent for plant regeneration has been determined. The effect of the types and concentrations of various cytokines (zeatin, kinetin, and 6-benzylaminopurine) on direct shoot regeneration from cotyledon nodes has been estimated. The culture-medium composition has been optimized for direct shoot regeneration from petioles. The effects of different concentrations of abscisic acid on the frequency of shoot regeneration from a morphogenic callus has been studied. Micropropagation has been used to obtain petiole explants and reproduce the shoots obtained by direct regeneration from cotyledon nodes, petioles, and calluses. Improved shoot-regeneration methods can be used for both agrobacterial and bioballistic genetic transformation of the sugar beet genotypes studied. 相似文献
15.
In vitro regeneration techniques have been optimized for seven strains and cultivars of sugar beet (Beta vulgaris L.) bred in Russia. The frequency of shoot regeneration from somatic cells and tissues of sugar beet varies from 10 to 97% depending on the explant type, culture-medium composition, and genotype. The in vitro regeneration potential has been estimated in plants with different genotypes. The effect of medium composition (phytohormones and carbohydrates) on the frequency of the formation of a morphogenic callus competent for plant regeneration has been determined. The effect of the types and concentrations of various cytokines (zeatin, kinetin, and 6-benzylaminopurine) on direct shoot regeneration from cotyledon nodes has been estimated. The culture-medium composition has been optimized for direct shoot regeneration from petioles. The effects of different concentrations of abscisic acid on the frequency of shoot regeneration from a morphogenic callus has been studied. Micropropagation has been used to obtain petiole explants and reproduce the shoots obtained by direct regeneration from cotyledonnodes, petioles, and calluses. Improved shoot-regeneration methods can be used for both agrobacterial and bioballistic genetic transformation of the sugar beet genotypes studied. 相似文献
16.
This study demonstrates the importance of preconditioning ofsource tissue in regeneration of multiple shoot buds from severalspecies of Lathyrus. Preconditioned multiple shoots of Lathyruscicera L., L. ochrus (L.) DC. and L. sativus L. were obtainedby germinating seeds on Murashige and Skoog (MS) medium containing50 µM N5-benzylaminopurine (BAP) for 2 to 3 weeks. Multipleshoot bud formation occurred when epicotyl explants of preconditionedshoots were cultured on MS medium containing 550 µMBAP. No shoot regeneration was observed from epicotyl explantswhich were obtained from non-preconditioned shoots. Shoot budswere formed directly on explants without an intervening callusphase after 2 to 3 weeks of culture. Regenerated shoot budsformed healthy shoots which developed prolific and strong rootswhen transferred to MS medium supplemented with 2.5 µMnaphthaleneacetic acid (NAA). Lathyrus cicera L., L. ochrus (L.) DC., Ochrus Vetch, L. sativus L., Lathyrus pea, de novo differentiation, epicotyl, preconditioning with BAP 相似文献
17.
Genotypic response of cowpea Vigna unguiculata (L.) to in vitro regeneration from cotyledon explants
M. S. Brar J. M. Al-Khayri T. E. Morelock E. J. Anderson 《In vitro cellular & developmental biology. Plant》1999,35(1):8-12
Summary A plant regeneration system applicable to 17 cowpea genotypes was developed. Cotyledons were initiated on 1/3 MS medium containing
15 to 35 mg N6-benzyladenine (BA) per 1 (66.6 to 155.3 μM) for 5 to 15 d. For shoot regeneration, the explants were transferred to a medium containing 1 mg BA per 1 (4.4 μM). Within 1 wk, shoot formation was visible at the proximal end of the cotyledons. Regeneration percentages (1% to 11%) and
the numbers of shoots (4 to 12 per explant) were significantly influenced by genotype. Culture duration and BA concentration
in the initiation stage significantly affected regeneration capacity. Explants initiated on media containing 15 mg BA per
1 for 5 d resulted in the highest percentage of explants capable of regeneration. Conversely, the highest number of shoots
was obtained from explants initiated on media supplemented with 35 mg BA per 1. Whole plants were obtained on a plant growth
regulator-free medium. To our knowledge, this is the first report of plant regeneration from U.S. commercial cowpea cultivars
and breeding lines. This system is adaptable to diverse cowpea genotypes and will facilitate cowpea genetic transformation.
Published with the approval of the Director of the Arkansas Agricultural Experiment Station. 相似文献
18.
Summary Intact seedlings of hybrid seed geranium (Pelargonium x hortorum Bailey) were tested for their ability to produce adventitious shoots and somatic embryos by direct culture of mature seeds on Murashige and Skoog (1962) medium (MS) supplemented with growth regulators BAP, BAP + IAA, or thidiazuron (TDZ). Ten varieties were tested in the presence of different BAP concentrations, four with BAP + IAA, and two with TDZ. Varieties used in this study differed in their response to BAP in the medium. Multiple adventitious shoots were produced by seven of the ten varieties tested. Multiple adventitious shoots were induced at all levels of TDZ in the medium. TDZ also induced callusing from roots and direct embryogenesis from intact hypocotyls. Adventitious shoots were separated, rooted and transferred to soil where they grew as normal healthy plants and flowered.Abbreviations BAP
N6-benzylaminopurine
- IAA
indole-3-acetic acid
- MS
Murashige and Skoog (1962)
- TDZ
thidiazuron 相似文献
19.
Mulpuri Sujatha Sheri Vijay Singareddy Vasavi Natarajan Sivaraj Sankaraneni Chander Rao 《Plant Cell, Tissue and Organ Culture》2012,111(3):359-372
Genetic improvement of sunflower (Helianthus annuus L.) through the use of biotechnological tools requires a reliable in vitro shoot regeneration system. Tissue culture protocols reported to date for sunflower suffer from low efficiency, poor reproducibility, genotype dependence and a tendency for flowering in vitro. The present study describes an efficient protocol system for shoot regeneration via direct adventitious shoot organogenesis from cotyledons of mature seeds of sunflower. About 169 media combinations comprising 12 different growth regulator combinations in various concentrations were assessed for induction of shoots from cotyledons derived from mature seeds and also from seedling tissues of 2?C20-day-old seedlings. Appearance of shoots from seedling tissues was sporadic and the frequency of shoot regeneration was low. Cotyledon explants from mature seeds were consistent with regard to frequency of adventitious shoot regeneration and number of shoots per explant. A high frequency (93.86?%) of adventitious shoot regeneration was obtained within 2?weeks of culture initiation on Murashige and Skoog (MS) medium supplemented with 9.84???M 2-isopentenyladenine (2-iP), 2.85???M indole-3-acetic acid (IAA) and 0.45???M thidiazuron (TDZ). Use of 2-iP in the shoot induction and elongation media prevented precocious flowering. Statistical analysis revealed significant effects of explant orientation, age of seedlings, and genotype on adventitious organogenesis. Maximum shoot regeneration was obtained when cotyledons from 0 and 1-day-old seedlings were placed with their adaxial surface in contact with the medium surface. The protocol developed was tested on 42 genotypes and found to be applicable to a wide range of genotypes. Histological studies indicated that the shoots originated predominantly through adventive organogenesis from the sub-epidermal and cortical regions. 相似文献
20.
Conditions for plant regeneration from excised shoot tips of Vigna radiata were studied. Complete plants were regenerated directly without an intervening callus phase from shoot tips on basal medium (MS salts+B5vitamins). Regeneration frequency varied with genotype, explant size and growth regulator combinations in the medium. Addition of cytokinins induced a variable amount of callus at the base of the shoot tip, followed by multiple shoot formation. Benzyladenine (BA), kinetin and zeatin at 5×10-6 M each induced multiple shoots in 100% of the explants but the highest number of regenerants per explant (9) was produced with BA. The efficacy of BA for shoot multiplication was not improved when it was supplemented with naphthaleneacetic acid (NAA) or indoleacetic acid (IAA). NAA or adenine sulphate, when applied alone, induced complete plantlets. The growth regulator requirement of explants for the induction of multiple shoots varied with explant size. The shoot tip explants maintained proliferation ability on subculture. None of the treatments was effective in inducing shoot bud differentiation from callus. Regenerated shoots were rooted on MS basal medium and MS supplemented with either IAA or indolebutyric acid. The rooted plants were transferred to the field; 60% subsequently survived and grew.Abbreviations BM
basal medium [MS (Murashige & Skoog 1962) salts+B5 (Gamborg et al. 1968) vitamins]
- BA
6-benzyladenine
- AdS
adenine sulphate
- IAA
indole-3-acetic acid
- NAA-1
naphthaleneacetic acid
- IBA
indolebutyric acid 相似文献