Agrobacterium tumefaciens mediated transformation in finger millet (variety RAU-8)
Loading...
Date
2013
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
Birsa Agricultural University, Kanke, Ranchi, Jharkhand
Abstract
Finger millet (Eleusine coracana (L.) Gaertn.) is a cereal grass mostly grown in dry land.
It is a robust, tufted, tillering annual grass, up to 170 cm high. The inflorescence is a
panicle with 4-19 finger-like spikes that resembles a fist when mature, hence the name
finger millet. Its grain is readily digestible, highly nutritious and versatile: it can be
cooked like rice, ground to make porridge or flour or used to make cakes. Finger millet is
a popular food among diabetic patients in the country. Its slow digestion indicates low
blood sugar levels after a finger millet diet thereby reacting as a safer food for diabetics.
Consumption of finger millet prevents constipation and cholesterol. The present study
was undertaken to standardize protocol for in vitro Agrobacterium tumefaciens mediated
transformation of finger millet (variety RAU-8). This would enable application of
biotechnological tools to this crop in future.
Surface sterilized seeds were inoculated on Murashige and Skoog’s medium
supplemented with 2.0, 4.0 mg/l 2, 4-D for callus. For shoot multiplication after 15 days
callus in a MS medium supplemented with combination of 2.0 mg/l IAA and 2.0 mg/l
IBA gave best shoot-root regeneration than 2.0 mg/l BAP and 1.0 mg/l NAA.
Out of different concentrations (25.0 mg/l, 50.0 mg/l, 75.0 mg/l and 100.0 mg/l)
100.0 mg/l kanamycin containing medium was found to be the best for transformant
selection. Kanamycin tolerance test was done considering 3 stages (1) etiolation, (2)
blackening and (3) death.
For transformation, the explants were wounded and dipped in the Agro
suspension. Co-cultivation was done for different time periods (5 min, 10 min, 15 min, 20
min, 25 min, 30 min, 1 hrs, 2 hrs and 3 hrs) of which 30 min was found to be the best
transformation frequency in finger millet. About 25% of explants give rise to entire
kanamycin resistant plantlets upon 21 days.
Description
Agrobacterium tumefaciens mediated transformation in finger millet (variety RAU-8)
Keywords
null