Suppression of Shade- or Heat-induced Leaf Senescence in Creeping Bentgrass through Transformation with the ipt Gene for Cytokinin Synthesis

Cytokinins have been associated with delaying or suppressing leaf senescence in plants. The objectives of this study were to determine whether the expression of the ipt gene that encodes adenine isopentenyltransferase would delay leaf senescence induced by shade or heat stress in a perennial grass species. Creeping bentgrass (Agrostis stolonifera cv. Penncross) was transformed with ipt isolated from agrobacterium (Agrobacterium tumefaciens) using two gene constructs (SAG12-ipt and HSP18-ipt) designed to activate cytokinin synthesis during shade or heat stress. Whole plants of nine SAG12-ipt transgenic lines and the nontransgenic control plants were incubated in darkness at 20 degrees C for 20 days. Chlorophyll content of all transgenic lines and the control line decreased after dark treatment, but the decline was less pronounced in transgenic lines. All transgenic lines had higher isopentenyladenine (iP/iPA) content than the control line after 20 days of treatment. In six of the transgenic lines, iP/iPA content remained the same or higher after dark treatment. Whole plants of nine HSP18-ipt transgenic lines and the control plants were incubated at 35 degrees C for 7 days. Chlorophyll and iP/iPA content declined in the control plants, but the nine transgenic lines had a significantly higher concentration of iP/iPA and were able to maintain chlorophyll content at the prestress level. Our results suggest that expression of SAG12-ipt or HSP18-ipt in creeping bentgrass resulted in increases in cytokinin production, which may have led to the delay and suppression of leaf senescence induced by shade or heat stress.