Establishment Of Agrobacterium Tumefaciens-mediated Genetic Transformation System Of Eggplant

Eggplant?Solanum melongena L.?is an important economy vegetable of China.Over the years,eggplant planting patterns and consumer habits had changed a lot,which lead to different requirements of eggplants variety characteristics.Due to conventional breeding moved along at a slow pace,it was extremely urgent to promote the application of transgenic technology in eggplant.Establishing an efficient eggplant transgenic system was the key to Genetic Engineering Breeding of eggplant.Trials were based on the existing efficient eggplant regeneration system.Seven eggplants with different genetic backgrounds preserved in the laboratory were selected as materials.The explants were infected with Agrobacterium strain GV3101 with pHB expression vector containing GUS gene and HPT gene.The culture medium containing Hyg B was used for screen the positive plants.Finally,obtaining the suitable eggplant transformation conditions,which laid the foundation for improving the character and quality of eggplant by genetic engineering technology.The main research results were as follows:1.Established Agrobacterium-mediated eggplant transgenic system?1?50 mg·L^-1 Hyg B was able to distinguish the transformed cells and non-transformed cells.300 mg·L^-1Cb can effectively inhibit the growth and reproduction of Agrobacterium tumefaciens,and didn't affect the normal growth and differentiation of explants.?2?The transformation efficiency of eggplant cotyledon was obviously better than that of hypocotyl,which was more suitable for the eggplant transgenic system.?3?The explants pre-cultured from 1 to 3 d promoted the transformation of eggplant significantly.When explants pre-cultured for 2days,the percentage of resistance callus was highest,and the transformation was the best.?4?The explants grew well if they were infected by Agrobacterium liquid(OD₆₀₀?0.6)for 15 min and co-cultured for 4 days under dark conditions and resistant callus rate was the highest.At the same time,adding100?mol·L^-1 AS to Agrobacterium liquid and co-culture medium had a certain promoting effect on the genetic transformation of eggplant.?5?In the process of delayed screening,explant callus and adventitious bud differentiation were better than direct screening,which was a more suitable screening method.2.Obtaining the best receptor of eggplant transgenic systemSeven hygromycin resistant plants were obtained in this experiment,using PCR and GUS histochemic staining to detect these resistant plants.The results showed that GUS gene have been successfully integrated into their genomes stably.Furthermore,no false positive plants were obtained.The percentage of resistant callus in eggplant was positively correlated with the number of plants obtained.No.140 obtained 3 transgenic plants and the conversion rate was up to 10%.It was the best receptor in this experiment.