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Chill injury and leaf senescence occur in plants held in prolonged cold, dark storage. To increase tolerance to these conditions, Nicotiana alata and N. tabacum were transformed with either the FAD7 or IPT genes under the control of a cold-inducible promoter (cor15a). FAD7 encodes for omega-3-fatty acid desaturase and was used to resist cold-stress. IPT encodes the cytokinin-pathway enzyme isopentenyl transferase and was used to delay senescence. Independent FAD7 and IPT lines were crossed to produce double transgenic seed. Seedlings from single transgenic (cor15a-IPT or cor15a-FAD7) lines, double transgenic lines, and the wild-type were exposed to prolonged cold, dark conditions. After 3 months in the dark at 2 °C, survival of independent double transgenic N. tabacum lines ranged up to 80% to 90%. However only 40% of FAD7 seedlings survived, 10% of IPT seedlings survived, and no wild-type plants survived. Double transgenic N. alata seedlings average 90% survival under similar conditions and RT-PCR revealed expression of both the IPT and FAD7 genes. Omega-3-FAD enzyme activity increases desaturation in chloroplast membrane fatty acids. When exposed to prolonged cold, the molecular fraction of polyunsaturated fatty acids (18:3 and 16:3) in leaves of wild-type N. alata decreased while monounsaturated (16:1 and 18:1) and saturated fatty acid species (16:0 and 18:0) increased dramatically. In double transgenic N. alata lines exposed to prolonged cold, the molecular fraction of 18:3 and 16:3 increased, while the 16:0 and 18:0 species decreased dramatically compared to nonchilled double transgenic plants.