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Development of herbicide-resistant transgenic tomato plants with Agrobacterium-mediated genetic transformation using bar gene

Vasudha Marapaka,Kranthikumar Gande,Raghu Banoth,Dulam Sandhya,Venkataiah Peddaboina

Published 2025 in Discover Plants

ABSTRACT

Tomato (Solanum lycopersicum L.) is an essential vegetable crop cultivated worldwide under various agro-climatic conditions, its productivity inflicted by several biotic and abiotic stresses. The Agrobacterium tumefaciens strain LBA4404 harbours pCAMBIA3301 with the bar and gusA genes as selectable, and reporter genes were applied for developing an efficient transformation procedure using cotyledon explants of tomato cv Arka Samrat. The evaluation of many factors, like explant pre-cultured for two d, acetosyringone at 100 μM, co-cultivated for two d, bacterial cell density at 0.6 (OD = 600), and 10 min infection duration showed an approximately five-fold increase in the transformation frequency. The wild-type (control) and infected with Agrobacterium suspension explants were placed on shoot regeneration medium (SRM) augmented with Phosphinothricin (PPT) (0.5 mg/l) and cefotaxime (400 mg/l). The putatively transgenic shoots were inoculated onto the rooting medium comprising IAA (1.0 mg/l), PPT (0.5 mg/l), and cefotaxime (100 mg/l). The putatively transgenic tomato plants were phenotypically the same as their mother plants and showed normal flowering. The stable integration of the transgenic gene was confirmed, and the presence of 450 bp size in putative transgenic (T0) plants was validated using PCR analysis. The highest transformation efficiency was observed in Arka Samrat (40.78%), followed by Pusa Ruby (37.21%), Arka Rakshak (30.95%), Arka Abha (30.55%), and Arka Meghali (29.27%). The present study demonstrates the significant effect of genotypes on varying transformation efficiency in different genotypes using cotyledon explants.

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