Efficient transgenic plant regenaration through Agrobacterium -mediated transformation of Chickpea ( Cicer arietinum L.)

Kar, Sanchayita; Johnson, Tony M.; Nayak, Pritilata; Sen, S. P.

doi:10.1007/s002990050170

Cited by 23 publications

(47 citation statements)

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“…ICCC37 displayed better response compared to other genotypes, endowing higher transformation efficiency. Similarly, genotypic influence on transformation efficiency in chickpea has been demonstrated by previous workers (Kar et al 1996;Krishnamurthy et al 2000;Senthil et al 2004;Tewari-Singh et al 2004). Preconditioning of epicotyl explants on SR medium played an important role in increasing the transformation frequency, with 48 h of preculture giving the best transformation frequency.…”

Section: Discussionsupporting

confidence: 73%

“…Efficient transformation system for pea was developed based on direct shoot regeneration and meristem proliferation from Agrobacterium-treated seedling explants (Davies et al 1993). To date, a few reports are available on the production of transgenic chickpea plants using Agrobacterium tumefaciens-mediated transformation (Fontana et al 1993;Kar et al 1996;Krishnamurthy et al 2000;Polowick et al 2004;Senthil et al 2004;Tewari-Singh et al 2004;Sanyal et al 2005). In chickpea, earlier work using Agrobacteriummediated gene transfer showed a transformation frequency rate of 0.5-3% (Polowick et al 2004;Senthil et al 2004), while the best transformation frequency in the present study was 14%.…”

Section: Discussionmentioning

confidence: 47%

“…Advances in biotechnology of grain legumes may lead to introduction of novel traits through genetic transformation into chickpea. Although a few reports on Agrobacterium-mediated transformation are available in chickpea (Fontana et al 1993;Kar et al 1996;Altinkut et al 1997;Krishnamurthy et al 2000) the frequency has been low ranging from 0.4 to 4%. Here, we report optimization of conditions for efficient delivery of Agrobacterium T-DNA, harboring cryIAc gene, along with selectable marker nptII and reporter gene uidA into chickpea.…”

Section: Introductionmentioning

confidence: 99%

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Agrobacterium-mediated transformation in chickpea (Cicer arietinum L.) with an insecticidal protein gene: optimisation of different factors

Indurker

Misra

Eapen

2010

Physiol Mol Biol Plants

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Agrobacterium-mediated transformation in chickpea was developed using strain LBA4404 carrying nptII, uidA and cryIAc genes and transformants selected on Murashige and Skoog's basal medium supplemented with benzyladenine, kinetin and kanamycin. Integration of transgenes was demonstrated using polymerase chain reaction and Southern blot hybridization of T 0 plants. The expression of CryIAc delta endotoxin and GUS enzyme was shown by enzyme linked immunosorbent assay and histochemical assay respectively. The transgenic plants (T 0 ) showed more tolerance to infection by Helicoverpa armigera compared to control plants. Various factors such as explant source, cultivar type, different preculture treatment period of explants, co-cultivation period, acetosyringone supplementation, Agrobacterium harboring different plasmids, vacuum infiltration and sonication treatment were tested to study the influence on transformation frequency. The results indicated that use of epicotyl as explant, cultivar ICCC37, Agrobacterium harboring plasmid pHS102 as vector, preculture of explant for 48 h, cocultivation period of 2 days at 25°C and vacuum infiltration for 15 min produced the best transformation results. Sonication treatment of explants with Agrobacteria for 80 s was found to increase the frequency of transformation.

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Section: Discussionsupporting

confidence: 73%

Section: Discussionmentioning

confidence: 47%

Section: Introductionmentioning

confidence: 99%

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Agrobacterium-mediated transformation in chickpea (Cicer arietinum L.) with an insecticidal protein gene: optimisation of different factors

Indurker

Misra

Eapen

2010

Physiol Mol Biol Plants

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“…A few reports are also available on genetic transformation of chickpea (Fontana et al 1993, Kar et al 1996, Khishnamurthy et al 2000, Sarmah et al 2003, Polowick et al 2004and Tewari-Singh et al 2004. These reports clearly demonstrated that Agrobacterium-mediated genetic transformation can be exploited in transforming various types of chickpea explants.…”

Section: Introductionmentioning

confidence: 91%

Agrobacterium-mediated Genetic Transformation of Local Cultivars of Chickpea (Cicer arietinum L.)

Sharmin¹,

Akter

Sarker

et al. 2012

Plant Tissue Cult. & Biotech.

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Agrobacterium-mediated genetic transformation protocol was established for two chickpea (Cicer arietinum L.) varieties, namely Barichhola-4 (Bch-4) and . Transformation ability of different explants such as decapitated embryo with single cotyledon disc (DEC), decapitated embryo (DE) and slice embryo decapitated at shoot end with single cotyledon disc (SEC) were tested using Agrobacterium tumefaciens strain LBA4404 harbouring binary plasmid pBI121, containing the GUS and nptII genes. Maximum transformation ability was exhibited by explants of decapitated embryo (DE) from Barichhola-5 (Bch-5). The optimum regeneration from the transformed tissue was achieved on MS supplemented with 0.5 mg/l BAP, 0.5 mg/l Kn and 0.2 mg/l NAA along with double the amount of CaCl2 and KNO3. Selection of the transformed shoots was carried out by gradually increasing the concentration of kanamycin to 150 mg/l. Stable expression of the GUS gene was detected in various parts of the transformed shoots through GUS histochemical assay. Stable integration of nptII gene within the genomic DNA from these transformed shoots was confirmed through PCR analysis.

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“…This low frequency of transformation was probably due to problems in sampling tissues from chimeric transformants obtained through organogenesis which involves more than one cell in the shoot initiation process. Kar et al (1996) also opined that in an organogenetic system, there is a risk of losing events of transformation due to chimera. Repeated selection in rapidly multiplying shoots is thought to eliminate untransformed tissues.…”

Section: Discussionmentioning

confidence: 99%

Development of Moisture Stress Tolerant Brinjal cv. Utkal Anushree (Solanum melongena L.) Using Agrobacterium Mediated Gene Transformation

Sagare¹,

Mohanty²

2012

JAS

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An efficient and reproducible in vitro regeneration protocol was developed with high frequency regeneration of shoot buds from shoot-tip in MS medium + BAP (2.0 mg/l) in Brinjal cv. Utkal Anushree. Rooting was obtained on MS medium+ NAA (0.01 mg/l). The Agrobacterium tumefaciens strain GV3107 containing a binary vector pCAMBIA2300 carrying rd29A::DREB1A gene has been used for transformation. The shoot-tip from in vitro grown seedling was pre-cultured for 72 hrs and co-cultivated for 24 hrs. Shoot buds were produced on the regeneration medium containing kanamycin (100 mg/l) and cefotaxime (250 mg/l). A transformation frequency of 6.40% was achieved after PCR analysis with gene specific primer. The gene expression for moisture stress tolerance was assessed through morpho-physiological and biochemical analyses.

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Efficient transgenic plant regenaration through Agrobacterium -mediated transformation of Chickpea ( Cicer arietinum L.)

Cited by 23 publications

References 0 publications

Agrobacterium-mediated transformation in chickpea (Cicer arietinum L.) with an insecticidal protein gene: optimisation of different factors

Agrobacterium-mediated transformation in chickpea (Cicer arietinum L.) with an insecticidal protein gene: optimisation of different factors

Agrobacterium-mediated Genetic Transformation of Local Cultivars of Chickpea (Cicer arietinum L.)

Development of Moisture Stress Tolerant Brinjal cv. Utkal Anushree (Solanum melongena L.) Using Agrobacterium Mediated Gene Transformation

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