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Agrobacterium tumefaciens Mediated Genetic Transformation of Pigeonpea [Cajanus cajan (L.) Millsp.]  

Kumar, S.Manoj (AP-NL Biotechnology Project, Department of Botany, Osmania University)
Syamala, D. (AP-NL Biotechnology Project, Department of Botany, Osmania University)
Sharma, Kiran K. (AP-NL Biotechnology Project, Department of Botany, Osmania University)
Devi, Prathibha (AP-NL Biotechnology Project, Department of Botany, Osmania University)
Publication Information
Journal of Plant Biotechnology / v.6, no.2, 2004 , pp. 69-75 More about this Journal
Abstract
Optimal protocol for efficient genetic transformation has been defined to aid future strategies of genetic engineering in pigeon pea with agronomically important genes. Transgenic pigeonpea plants were successfully produced through Agrobacterium tumefaciens-mediated genetic transformation method using cotyledonary node explants by employing defined culture media. The explants were co-cultivated with A. tumefaciens strain C-58 harboring the binary plasmid, pCAMBIA-1301 [con-ferring $\beta$-glucuronidase(GUS) activity and resistance to hygromycin] and cultured on selection medium (regeneration medium supplemented with hygromycin) to select putatively transformed shoots. The shoots were then rooted on root induction medium and transferred to pots containing sand and soil mixture in the ratio of 1:1. About 22 putative TO transgenic plants have been produced. Stable expression and integration of the transgenes in the putative transgenics were confirmed by GUS assay, PCR and Southern blot hybridization with a transformation efficiency of over 45%. Stable integration and expression of the marker gene has been confirmed in the TO and T1 transgenics through PCR, and Southern hybridization.
Keywords
Agrobacterium tumefaciens; genetic transformation; pigeonpea;
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