Optimization Of Rapid Transformation System Of Rice

Rice is an important food crop and model plant in China,although the genetic transformation system mediated by Agrobacterium in some japonica rice varieties is relatively mature,most rice varieties have the problem of difficult genetic transformation and low transgenic efficiency.In addition,the traditional genetic transformation process has a long cycle and cumbersome steps,which not only increases somatic variation,but also increases the operation cost.The results of this study are as follows:1)Based on N₆ medium,through two-factor experiments,it was found that the contents of 2,4-D and NAA in induction medium were 3.5 mg/L and 0.2 mg/L to effectively improve callus growth quality,thereby significantly increasing the induction efficiency of ZH11 to more than 90.0%;2)Infection could be carried out on the 7th day after 32℃light induced callus.After 2 days of co-culture in 20℃incubator,washed clean bacteria were transferred to screening medium and incubated under 32℃dark culture.New resistant callus could be grown about 25 days later;3)After adjusting the content of sucrose and sorbitol in differentiation medium,when the content of sucrose was 30 g/L and the content of sorbitol was 0 g/L,the seedlings could be differentiated quickly in about 40 days.In this study,the optimized rapid transformation system could obtain transgenic seedlings within three months,shortening the transformation period by 30-45 days compared with the traditional genetic transformation system.4)Four BBM family gene single-gene knockout mutants were constructed,and the differentiation stage phenotype of single-gene knockout homozygous mutant materials was identified;5)The BBM family gene multigene knockout and overexpression materials were obtained,which could further perform phenotypic observation and functional verification of the BBM gene family.In this study,using the commonly used japonica rice variety ZH11 as the material,an optimized rapid rice transformation system was established by adjusting the medium formula and cultivation conditions in the induction and differentiation stages on the basis of the predecessors.In order to explore the function of Baby Boom（BBM）family genes in plant embryonic growth and development,cell proliferation and differentiation,the genetic mutant resources of this family gene were created.