Development of transgenic pearl millet (Pennisetum glaucum (L.) R. Br.) plants resistant to downy mildew

Transgenic pearl millet lines expressing pin gene--exhibiting high resistance to downy mildew pathogen, Sclerospora graminicola--were produced using particle-inflow-gun (PIG) method. Shoot-tip-derived embryogenic calli were co-bombarded with plasmids containing pin and bar genes driven by CaMV 35S promoter. Bombarded calli were cultured on MS medium with phosphinothricin as a selection agent. Primary transformants 1T(0), 2T(0), and 3T(0) showed the presence of both bar and pin coding sequences as evidenced by PCR and Southern blot analysis, respectively. T(1) progenies of three primary transformants, when evaluated for downy mildew resistance, segregated into resistant and susceptible phenotypes. T(1) plants resistant to downy mildew invariably exhibited tolerance to Basta suggesting co-segregation of pin and bar genes. Further, the downy mildew resistant 1T(1) plants were found positive for pin gene in Southern and Northern analyses thereby confirming stable integration, expression, and transmission of pin gene. 1T(2) progenies of 1T(0) conformed to dihybrid segregation of 15 resistant:1 susceptible plants.