Effects Of GhWOX11/12 Gene And Red-Blue Light Quality On Cotton Somatic Embryogenesis

Cotton is an important economic crop,fiber crop and oil crop in the world.Many excellent agronomic traits of cotton are obtained through transgenic.Many plant transgenic molecular breeding techniques rely on plant tissue culture technology,but most cotton varieties are difficult to pass somatic cells.The embryogenic pathway is derived from regenerated cotton,which is limited by genotypes,and only a few cotton varieties can.Therefore,how to improve the transgenic efficiency of cotton is particularly important.Cotton somatic embryogenesis mainly undergoes four processes: callus induction stage,embryogenic callus induction stage,somatic embryo induction stage and regeneration stage stage.The quality of callus in early cotton somatic embryogenesis is closely related to the formation of re-differentiation and somatic embryos.WOX11/12 gene activates WOX5 and LBD genes to promote the formation of PSCs with root primordial properties;The ratio of B:R=1:1,B:R=3:1,B:R= 1:3 and DL light quality combination explored the effects of four stages on cotton somatic embryogenesis.Environmental and genetic aspects of its effects on cotton somatic embryogenesis.The main results are as follows:1.The asymmetric distribution of auxin activates the differential expression of Gh WOX11 and Gh WOX12.According to the phenotype of cotton hypocotyl callus initiation,the callus induction was performed at 0 h,24 h,48 h,60 h,72 h,136 h hypocotyl transcriptome sequencing and endogenous auxin content after different treatments.Determination and detection of expression levels.It was found that the asymmetric distribution of auxin caused differential expression of Gh WOX11 and Gh WOX12,thereby promoting callus formation.2.GhWOX11 and GhWOX12 simultaneously regulate callus formation and adventitious root formation.The overexpression vector was constructed to transform the cotton hypocotyl of 7-day-old size without adding exogenous auxin.After 90 days,in the over-expressed 35S::Gh WOX11_Dt and 35S::Gh WOX12_Dt vectors,we found each under On the hypocotyl segment explants,an average of three adventitious roots can be formed,and each hypocotyl explant can only form an adventitious root on average.The number of adventitious roots formed by overexpression of 35S::Gh WOX11_Dt and 35S::Gh WOX12_Dt was significantly different from that of Mock.3.Overexpression genes Gh WOX11 and Gh WOX12 significantly promoted the formation of callus in upland cotton.The CCRI24 cotton hypocotyls were transformed with the constructed 35S::Gh WOX12_Dt and 35S::Gh WOX11_Dt vectors and the empty vector Mock,respectively,through the 35S:: Gh WOX12_Dt and 35 S of the somatic embryogenesis callus stage and embryogenic callus stage of cotton.: Observed analysis and statistics of Gh WOX11_Dt and Mock vector transformants,we found that the over-expressed genes Gh WOX11 and Gh WOX12 significantly promoted the dedifferentiation and re-differentiation of somatic embryogenesis in upland cotton.4.Chimeric Gh WOX11 and Gh WOX12 inhibitors inhibit the formation of embryogenic callus.The Gh WOX11 and Gh WOX12 genes were fused to a suppressor domain SRDX to form a chimeric Gh WOX11 and Gh WOX12 inhibitor and transformed into cotton hypocotyls.It was found that the chimeric genes Gh WOX11 and Gh WOX12 inhibitors significantly inhibited the dedifferentiation of somatic embryogenesis in upland cotton.And during the process of re-differentiation,genes Gh WOX11 and Gh WOX12,by affecting the quality of dedifferentiation callus,and then regulating the re-differentiation into embryogenic callus,have greatly changed the fate of cells.5.The combination of red and blue B?R=1?3 can promote the somatic embryogenesis of cotton.Compared with B:R=1:3 and DL light quality combinations,B?R=1?1 and B:R=3?1 light quality combinations significantly promoted callus proliferation.Compared with B?R=1?1 and B?R=3?1 light quality combinations,B?R=1?3 light quality combination significantly increased weight proliferation of callus without EC differentiation ability.B ? R=1 ? 3 and DL light quality combinations had significantly higher embryonic callus differentiation rate than B?R=1?1 and B?R=3?1 light quality combinations.The number of somatic embryos induced by B?R=1?3 treatment was significantly higher than that of B?R=1?1 and B:R=3?1 light quality combination,but there was no significant difference compared with DL;the number of hypocotyls induced by B?R=1?1 and B?R=3?1 light quality combination was significantly lower than that induced by B?R=1?3 and DL light quality combination,and the number of somatic embryos induced by B?R=1?1 light quality combination and B?R=3?1 light quality combination was significantly lower than that induced by B?R=1 light quality combination and B?R=3?1 light.The plant height of B?R=1?1 and B?R=1?3 combinations was significantly higher than that of B?R=1?3 and DL combinations;the chlorophyll content of B?R=1?3 treatment was higher than that of B?R=1?1 treatment,but the chlorophyll content of B?R=1?3 treatment was significantly higher than that of B?R=3?1 and DL light quality combination.