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An improved protocol for rapid and efficient agrobacterium mediated transformation of tomato for salt tolerance (solanum lycopersicum l.)

Author: 
Madhu, S. V. and Savithramma, D. L.
Subject Area: 
Life Sciences
Abstract: 

Transformation of tomato with heterologous genes requires rapid and efficient transformation protocols. Agrobacterium mediated transformation protocol of tomato (Solanum lycopersicum L.) cv. 'Pusa Ruby' using Pg NHX1, AVP1 and co-expression of Pg NHX1+AVP1 gene under 35S promoter and poly A signal of CaMV in pBI121 binary vector was optimized by varying parameters such as type of explant, combination of hormones, concentration of antibiotic, bacterial culture and co-cultivation period. Cotyledons were found to be the best explants for shoot regeneration in tomato compared to Hypocotyls with 96.95% and 95.90% shoot regeneration, respectively. The hormonal concentration Zeatin (1mg/l), IAA (0.1mg/l), Adenine sulphate (25 mg/l) produced higher shoot regeneration. The selection of transformants on antibiotic kanamycin was standardized at 100mg/l and the transformation efficiency was optimized at Acetosyringone concentration of 375 µM and bacterial culture of 1 x 108 at 72 h incubation was standardized to obtain the transformation efficiency of 38.81%. With this modified protocol it was possible to obtain transformed plants within a period of 65 to 75 days with a high regeneration and transformation efficiency.

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