HIGH-THROUGHPUT AGROBACTERIUM-MEDIATED TRANSFORMATION OF MEDICAGO TRUNCATULA IN COMPARISON TO TWO EXPRESSION VECTORS
Tasawar Sultana*, Farah Deeba* and S.M. Saqlan Naqvi*
ABSTRACT
Legumes have been turbulent to efficient Agrobacteriummediated transformation for a long time. The selection of Medicago truncatula as a model legume plant for molecular analysis resulted in the development of efficient Agrobacterium-mediated transformation protocols. In current study, M. truncatula transformed plants expressing OsRGLP1 were obtained through GATEWAY technology using pGOsRGLP1 (pH7WG2.0::OsRGLP1). The transformation efficiency of this vector was compared with expression vector from pCAMBIA series over-expressing same gene (pCOsRGLP1). A lower number of explants generated hygromycin resistant plantlet for instance, 18.3 with pGOsRGLP1 vector as compared to 35.5% with pCOsRGLP1 vector. Transformation efficiency of PCR positive plants generated was 9.4% for pGOsRGLP1 while 21.6% for pCOsRGLP1. Furthermore 24.4% of explants -1 generated antibiotic resistant plantlet on 20 mgl of hygromycin which was -1 higher than on 15mgl of hygromycin such as 12.2%. T progeny analysis 1 indicated that the transgene was inherited in Mendelian manner. The functionally active status of transgene was monitored by high level of Superoxide dismutase (SOD) activity in transformed progeny.
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