Transformation of Finger millet (Eleusine coracana L. Gaertn.); variety A404, through Agrobacterium tumefaciens
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Date
2013
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Birsa Agricultural University, Kanke, Ranchi, Jharkhand
Abstract
The present research work has been carried out with a view to standardize the
protocol for Agrobacterium tumefaciens mediated transformation on Eleusine
coracana L. a valuable crop plant, using Agrobacterium strains GV2260 having
pCAMBIA2300 plasmid. Aseptic explants for transformation were obtained by
inoculating seeds of Eleusine coracana L. on MS medium supplemented with 2, 4-D
4.0 mg/L, callus was induced. Callus was inoculated on MS medium supplemented
with BAP 2.0 mg/L for multiplication of shoots. For kanamycin sensitivity test of
shoots showing maximum etiolation (98%), blackening (84%) and death (82%) have
occurred on MS medium supplemented with 100.0 mg/L of kanamycin after 21 days
of inoculation. Agrobacterium strain GV2260 was maintained at 26˚C for 20 hrs at
150 rpm in dark condition in LB medium supplemented with 50.0 mg/L rifampicin,
50.0 mg/L kanamycin and 100.0 mg/L acetocyringone. The explants were treated for
different co-culture periods after which those were inoculated in solid MS medium
supplemented with BAP 2.0 mg/L and kept at 25±2˚C and 16 hr light/8 hr dark. After
10 days of infection the explants were subculture on MS medium supplemented with
100.0 mg/L of kanamycin for the confirmation of transformation. After 21 days it
was observed that only one explant survived out of nine explants in 30 min of coculture
with Agrobacterium tumefaciens.
Description
Transformation of Finger millet
(Eleusine coracana L. Gaertn.); variety
A404, through Agrobacterium tumefaciens
Keywords
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