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Agrobacterium tumefaciens-mediated transformation of Vigna mungo (L.) Hepper

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Abstract

Transformed Vigna mungo (blackgram) calli were obtained by cocultivating segments of primary leaves with Agrobacterium tumefaciens vir helper strains harbouring the binary vector pGA472 having kanamycin resistance gene as plant transformation marker. Transformed calli were selected on Murashige and Skoog medium supplemented with 50 mg/l kanamycin and 500 mg/l carbenicillin. Transformed calli were found to be resistant to kanamycin up to 900 mg/l concentration. Expression of kanamycin resistance gene in transformed calli was demonstrated by neomycin phosphotransferase assay. Stable integration of transferred DNA into V. mungo genome was confirmed by Southern blot analysis.

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Abbreviations

BAP:

6-benzylaminopurine

2,4-D:

2,4-dichlorophenoxyacetic acid

2iP:

6-(γ,γ-dimethylallylamino)purine

Kn:

kinetin

nptII:

neomycin phosphotransferaseII

MS:

Murashige and Skoog (1962) medium

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Communicated by G. C. Phillips

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Karthikeyan, A.S., Sarma, K.S. & Veluthambi, K. Agrobacterium tumefaciens-mediated transformation of Vigna mungo (L.) Hepper. Plant Cell Reports 15, 328–331 (1996). https://doi.org/10.1007/BF00232365

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  • DOI: https://doi.org/10.1007/BF00232365

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