Synchronized Somatic Embryogenesis Of Gossypium Barbadense L. Induced By Various Treatments And The Cloning Of GbAIL5?GbAGL15

The asynchronization development of somatic embryos causes a lot of difficulties in the tissue culture and plantlets reproduction of Gossypium barbadense L.and brings about adverse impact on the transgenic research in Gossypium barbadense L.This study,with the purpose of improving the efficiency of somatic embryogenesis in Gossypium barbadense L.,chose embryonic callus of Xinhai 16,a breed of Gossypium barbadense L.,as the material to cultivate under different conditions for synchronization induction.Then,the coding sequences of GbAIL5 gene and GbAGL15 gene in Gossypium barbadense L.were cloned and their expression trends after injured by low temperature for 3 d were analyzed by qRT-PCR technology.The main results of this study were as follows:(1)Three kinds of abiotic stress,low temperature,starvation and osmotic stress were used for testing their effects of inducing the synchronization of somatic embryogenesis.The induced results was analysed by statistic software,which showed that the synchronization could be significantly induced by conditions of 4?-3 d,phosphate starvation-5 d and nitrogen starvation-5 d,and the most remarkable effect was induced by 4?-3 d.(2)The coding sequences of GbAIL5 and GbAGL15 in Xinhai 16 were cloned.The length of the coding sequence of GbAIL5 gene was 1626 bp,encoding 541 amino acids,and the length of the coding sequence of GbAGL15 gene was 753 bp,encoding 250 amino acids.AIL5 had two specific conserved AP2 domains in its structure.AGL15 contained a MADS-box domain and a K-domain.(3)Expression trends of these two genes cultivated at 28? for 0-5 d were studied by qRT-PCR after low temperature treatment for 3 d.The results showed that the expression quantities of the two genes were both changed after induced at 4? for 3 d.The relative expression quantity of GbAIL5 was observably higher than control in the first day moving back to 28?;but its expression was then declined during 2 d and 3 d,and then went up again in 4 d and 5 d.The expression of GbAGL15 induced by low temperature was increased during the first 4 d and the difference was up to significant level.