Agrobacterium mediated transformation of chickpea ( Cicer arietinum L.) embryo axes

Krishnamurthy, K. V.; Suhasini, K.; Sagare, Abhay P.; Meixner, Martin; Kathen, A. de; Pickardt, Thomas; Schieder, O.

doi:10.1007/s002990050005

Cited by 104 publications

(71 citation statements)

References 18 publications

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“…However, too long a co-cultivation period result in overgrowth of Agrobacterium and therefore it is harmful to the plant cells. Role of co-cultivation period correlating with transformation frequencies have been reported by earlier workers (Fontana et al 1993;Krishnamurthy et al 2000;Sarmah et al 2004;Tewari-Singh et al 2004). Previous reports that 48 h of co-cultivation is optimal period for chickpea transformation (Srinivasan et al 1991;Sanyal et al 2003Sanyal et al , 2005 is in agreement with the present study.…”

Section: Discussionsupporting

confidence: 93%

“…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: 72%

“…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%

“…On the basis of maximum GUS positive response, epicotyls and embryonic axes were found to be the explants of choice for transformation of chickpea. Most of the earlier reports on Agrobacterium-mediated transformation in chickpea have used embryonic axes as the explants of choice (Fontana et al 1993;Krishnamurthy et al 2000;Polowick et al 2004;Sarmah et al 2004;Tewari-Singh et al 2004). Leaf and stem explants have also been used (Srinivasan et al 1991).…”

Section: Discussionmentioning

confidence: 99%

“…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: 93%

Section: Discussionsupporting

confidence: 72%

Section: Discussionmentioning

confidence: 47%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

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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Plant Transformation Technology:Agrobacterium‐Mediated Transformation

Komari

Ishida

Hiei

2004

Handbook of Plant Biotechnology

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The soil phytopathogen Agrobacterium tumefaciens induces tumours, known as crown galls, mainly on dicotyledonous plants. Such tumours are generated by a complex, multi‐step transformation process. Another species, A. rhizogenes , causes hairy root disease in higher plants via an identical process. Agrobacterium has been utilised for the transfer of genes to dicotyledonous plants. Now, monocotyledonous plants are routinely transformed by Agrobacterium despite the fact that these plants, including important cereals, were thought until recently to be outside the range of this technology. Most of the common transformation methods heavily depend on tissue culture technology and we refer to such methods as in vitro transformation. The only case of a routine tissue culture free method is the ‘ in planta ’ transformation of Arabidopsis thaliana . Numerous critical factors are involved in both approaches. In transformation in vitro , key factors include choice of vectors and bacterial strains, types of plant tissues to be infected, procedures of preparation the tissues, protocols of infection and co‐cultivation, methods for subsequent culture and selection of transformed cells, antibiotics to remove infecting bacteria, techniques for regeneration of transgenic plants and genotypes of plants. It is our opinion that the type and quality of the starting material is the most important one among them. The capacity to serve as a host plant for crown gall disease is no longer a prerequisite for a host of vector systems based on Agrobacterium . The real prerequisite for in vitro transformation is the availability of technology for dedifferentiation of tissues and regeneration of plants in a given species. On the other hand, the biological processes involved in in planta transformation are yet to be elucidated. Recent evidence suggested that ovules are the primary target, and further basic understanding is likely to help extend the number of species transformed by the method. The plant species that are routinely transformed by Agrobacterium are expanding rapidly. Some gymnosperms, several forest trees and fruit trees, various legumes, and cereal and non‐cereal monocotyledons, which were once considered very recalcitrant, are now in a long list of transformable plants. Arabidopsis , tobacco and rice are the top three species that were transformed during the last two years. Although it is only half a dozen years since the current procedure of rice transformation mediated by Agrobacterium was published, the economic importance and the accumulation of genomic information has made rice the species of focus in plant biotechnology. The advantages of Agrobacterium ‐mediated transformation include the transfer of pieces of DNA with defined ends and minimal rearrangement, the transfer of relatively large segments of DNA, the integration of small numbers of copies of genes into plant chromosomes and the high quality and fertility of transgenic plants. However, transformation does not always produce such ‘clean’ events. Formation of repeats of transgenes, certain rearrangements, integration of non‐target DNA segments and unstable expression of transgenes are among the complications. Although the majority of transgenic plants usually appear ‘good’ in a particular test, accumulation of ‘dropped’ plants is significant after multiple rounds of characterisation and screening. Therefore, further improvement in each of steps is highly desired. Application of the gene transfer mediated by Agrobacterium is further expanding. Transient expression of genes delivered by Agrobacterium is now a useful tool in the study of promoters and gene function. Vectors specifically designed to carry very large segments of DNA have been developed and extensively tested. Insertional mutagenesis by DNA transferred by Agrobacterium is a routine technique in genomics study in Arabidopsis and rice. Methods for targeted integration of transgenes to genomes of higher plants have been drawing considerable attention. Various technologies for the production of ‘selection marker free’ transgenic plants are now in place for better public acceptance of biotechnology products.

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