GENETIC TRANSFORMATION OF SORGHUM
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Date
2004
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Professor Jayashankar Telangana State Agricultural University
Abstract
Sorghum is an important dryland crop used as food, feed and fodder. Due to increasing demand towards its usage in diverse applications, it is imperative to deploy modern bioteclinological tools to improve its productivity and quality. Two principal methods viz., Agr-obacteriunz infection and particle bombardment were used for genetic transformation. In vitro response of sorghuni is low, monocalcitrant and is higlily genotype dependent. Tissue culture and regeneration response was low when compared to other cereals. During the course of study, efficiency of different expl;tnts were studied in two inbred lines AKMS 14B and 296B through transient g~ls ex, ession profile and regeneration percent. Particle bon~bardn~ent though showed high percent of regeneration in all the explants, Agrobacteritrr?~ method of transfor11i:ltion was preferred because of its cost efficacy and low copy number and transient gin expression was more through Agrobacten'ur~z infection than tl~rough pa~ticle bombardment. Shoot meristems isolated from germinated seedlings were no st suited explants in terms of irz vitr-o handling and transformation efficiency. 3d and 7d old seedlings were studied through electron microscopy to nleasure the nleristen~atic zone that correlate with the transformation parameters. Infection at two OD60O values, 1.0 and 2.0 absorbiincc were studied. Absorbance at ODboo 1.0 gave low transient grr.r; expression but controllecl Agrobacterizlnz growth after cocultivation; whereas at OD6oo 2.0 absorbancc ga1.e high transient gtls expression with overgrowth of Agr-obncteriunl. Different antibiotic treatments and culture conditions were tested that can control Agrobactc~-izlrn growth and facilitate growth of plant tissues. Host specific response towards antibiotics was evident. Addition of L-Cysteine at 400mg~-' improved the growth of shoot tips during cocultivation. The study suggests that, better transformability from direct explants than from the callus derived explants in two sorghum inbred lines using Agr.ohc[c.tc/-i~rn~ as the method of gene transfer.
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D7080