Advancement of in-vitro Regeneration and Biolistc Genetic Transforamtion Techniques in Tomato cv. Anand Roma (Solanum lycopersicum L.)
Hardiksinh R. Chavda
*
Department of Agricultural Biotechnology, Anand Agricultural University, Anand, Gujarat, India.
Amar A. Sakure
Department of Agricultural Biotechnology, Anand Agricultural University, Anand, Gujarat, India.
Jigar J. Mistry
Department of Genetics and Plant Breeding, B. A. College of Agriculture, Anand Agricultural University, Anand, Gujarat, India.
Ghanshyam. B. Patil
Department of Agricultural Biotechnology, Anand Agricultural University, Anand, Gujarat, India.
Mahesh. B. Vaja
Department of Agricultural Biotechnology, Anand Agricultural University, Anand, Gujarat, India.
*Author to whom correspondence should be addressed.
Abstract
Tomato (Solanum lycopersicum L.) an important Solanaceae crop which was genetically transformed via particle bombardment using cotyledon explants of cv. Anand Roma. An efficient regeneration protocol was optimized through sequential media combinations and resulting in enhanced callus, shoot, and root induction. The MS9 (0.5 mg/L IAA and 1.5 mg/L Zeatin), MSE6 (0.5 mg/L IAA and 1.0 mg/L BAP), RM8 (2 mg/L IBA with half-strength MS media) media combination was found most effective and supporting for early callus induction, maximum shoot regeneration (4.90 shoots/plants), and highest rooting efficiency (84.51%). Among 248 bombarded explants, 127 survived selection on 10 mg/mL hygromycin, of which 107 (84.20%) formed callus, 65 (60.74%) regenerated shoots and 37 (56.92%) developed roots. Ultimately, 27 plantlets survived acclimatization and were confirmed as putative transgenic through PCR amplification of the hptII gene representing a total transformation efficiency of 10.88%. These results highlight the robustness of particle bombardment for stable gene transfer in tomato providing a reliable platform for functional genomics, trait improvement and crop biotechnology particularly in cultivars with limited Agrobacterium susceptibility.
Keywords: Growth regulators, Anand Roma, regeneration, genetic transformation, particle bombardment