TRANSFORMATION AND CHARACTERIZATION OF ANTIFUNGAL GENES AGAINST TURCICUM LEAF BLIGHT DISEASE IN MAIZE (Zea mays) AND TOBACCO
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Date
2023-03-30
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University of Agricultural Sciences, Bangalore
Abstract
Maize (Zea mays L.) is a vital food crop of world importance is markedly affected by disease leaf blight generally called as turcicum leaf blight of maize caused by fungal pathogen Exserohilum turcicum. The existing available management practices including chemicals are having major limitation, hence transgenic approach is one of the alternative approaches in providing resistance against turcicum leaf blight of maize. The present investigation aims at transformation of antifungal genes into maize and tobacco using different methods of transformation which will be useful in providing resistance against turcicum leaf blight infection. The plasmid from transformed pB4NU vector + RACK1 gene and pB4NU vector + Glucanase gene were amplified using gene specific primers. The amplification of genes was confirmed by PCR using gene specific primers with amplification of 1005 bp (RACK1) and 1051bp (Glucanase). The transformation was carried out using in-planta mediated transformation in maize and agroinfiltration in tobacco respectively. The transformed maize and tobacco plants was confirmed by using gene specific primers and expression of the antifungal proteins was confirmed by SDSPAGE analysis. The characterization of expressed antifungal proteins including bioassay was done using transformed tobacco plants. Bioassay of antifungal proteins was carried out using poison food technique under invitro against turcicum leaf blight fungus using antifungal proteins from agroinfiltrated tobacco plants