Study On The Transformation Of Citrus By Gene-Deletor Technology And With Seedless Genes

Citrus is the most widely grown fruit crops in the world.The development of genetic transformation has provided new opportunities fur plant improvement and made it possible to modify just specific traits maintaining the original characteristics of the cultivar. With the presence of antibiotic or herbicide resistant selective marker genes in transgenic plants causing the concerns of the potential hazards on ecological environment and human health.The Gene-Deletor technology offers a powerful tool to make genetically modified crop safer to grow and to consume. Breeding the seedless exccllent citrus that can challengewith the market is require.The "Gene Deletor" gene, "CK" gene and seedless gene was transferred into the genome of kumquat with explants of epicotyl and mature tissues of Bingtang' orange mediated by AgrobacteHum.Adventitious buds were micro-grafted. Transformed plants were detected by GUS and PCR.The results of this study are as follows:1. Using epicotyls of kumquat seedlings grown in vitro as explants, an efficient regeneration system was established. The results showed that use the 35-day-old epicotyl segments of kumquat, and the optimizing regeneration system would be based on the culture media of MS+6-BA2.0mg/L+NAA1.0mg/L+1.0mg/LKT+3% sucrose+0.8% agar (pH5.7) with the procedures of 26℃for a 16/8 h light/dark cycle.2. Sensitivity analysis to Kanamycin with epicotyl segments of kumquat in vitro. The results showed that the best growth quality of adventitious shoots were obtained when the explants placed in medium supplemented with 50 mg/L kanamycin.3. Genetic transformation with explants epicotyl of kumquat mature tissues of: Bingtang orange, PCR and GUS analysis of different varieties of resistant plants, the plasmid for the Gene-deletor Y-A and its control vector C-F, seedless C-D.The results are as follows:The Y-A gene was integrated into one transgenic plants of kumquat,C-D gene was integrated into three transgenic plants of kumquat, C-F gene was integrated into three transgenic plants of kumquat.The Y-A gene was transferred into five transgenic plants of Bingtang orange.