Indian Society of Genetics & Plant Breeding

An efficient in vitro regeneration protocol to generate stable transgenic lines of black gram (Vigna mungo L.)

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An efficient and quick in vitro regeneration protocol was
developed for black gram (Vigna mungo L.) using wounded
embryonic axis with cotyledon as explant. Murashige and
Skoog (MS) medium supplemented with 4.44 µM BAP and
2.32 µM Kinetin was found to be effective in producing
maximum number (mean 7.80) of multiple shoots. The
individual shoots elongated to 4.5 cm when MS medium
was supplemented with 2.89 µM GA3 along with 0.44 µM
BAP and 0.46 µM KIN. A novel in vitro rooting technique
was also optimized for black gram using half-strength liquid
MS medium supplemented with 1.34 µM NAA. The shoots
in this medium produced the highest number (mean 7.50)
of roots with root length of 6.02 cm. The plantlets were
transferred to soil mixture and placed in greenhouse where
more than 80% successfully grew to maturity. The same
protocol was successfully used to generate transgenic black
gram lines carrying Bt-Cry2Aa gene through Agrobacteriummediated transformation with a transformation efficiency
of 0.42%. The rooted T0 plants grew to maturity and
produced T1 seeds with the presence and expression of
transgene in T1 plants. Thus, we have standardized an in
vitro regeneration protocol suitable for generation of stable
transgenic black gram plants.

Keywords: Black gram, Vigna mungo, in vitro regeneration, Bacillus thuringiensis, Cry2Aa


Year: 2018
Volume: 78
Issue: 3
Article DOI: 10.31742/IJGPB.78.3.10
Print ISSN: 0019-5200
Online ISSN: 0975-6906



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