Plant regeneration and genetic transformation studies in tomato (Lycopersicon escullentum.Mill.)
An investigation on Lycopersicon esculentus cv. Solan vajara (Tomato) was conducted to standardize the method of high frequency regeneration and to standardize the technique of Agrobacterium mediated gene transfer in tomato tissue. Cotyledon, hypocotyl and leaf explants from 10 days and 16 days old seedlings were used. While using leaf explants the highest shoot regeneration was obtained on MS medium supplemented with 2mg/l BAP and 1 mg/l IAA and highest shoot regeneration from cotyledon explants was obtained on MS medium supplemented with 2 mg/l BAP and 0.5 mg/l IAA. High percentage root regeneration in in vitro raised shoot were obtained on the MS medium supplemented with 0.10 mg/l and 0.05 mg/l NAA. The experiment described on regeneration of whole plants from the explant indicate that seedling age conditions during the culture are as important as the correct plant growth regulator balance for high frequency plant regeneration. Tomato plants are able to regenerate within 2 months and the young plantlets can be transferred to pots for hardening. Hardening procedure for tissue culture raised plantlets was standardized using sterile mixture of sand and soil in plastic pots. Agrobacterium timefaciens strain LBA 4404 was infectious to tomato cells. After cultivation the explants were able to grow on the selective medium, whereas control explants were unable to grow on the selective medium. Transformed calli were selected on the medium containing 50 mg/l kanamycin. Transformation experiment could be scored as early as 4 weeks after selection. Most of the calli obtained on the selective medium were GUS positive.
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This article is written by Shri. Anil Kant of Dr. Y.S. Parmar University of Horticulture and Forestry, Solan on behalf of ASPEE Agriculture Research and Development Foundation. ASPEE Group of Companies is the leading manufacturer and exporter of Agriculture Sprayers and Dusters
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