Genetic Transformation Of Genes Related To Male Sterility For The Thermo-photo-sensitive Male Sterile Line 337S Of Wheat

The thermo-photo-sensitive male sterile line 337 S of wheat is sensitive to the long-day/high temperature and short-day/low temperature,which has the characteristics of sterility stability and wide seeding area.The sterile anthers in short-day and low temperature condition and fertile anthers in normal planting condition are collected for digital gene-expression profiling analysis by RNA sequencing.As a result,many different genes associated with the wheat reproductive development are obtained and the key genes which express the remarkable difference are cloned.At the same time,the over expressing vectors of candidate sterile genes TaMS337S-1 and TaMS337S-2 are also constructed by using the technique of Gateway.Moreover,Gene gun and Agrobacterium transformation are used for transformation of these 2 candidate genes into 6 different varieties to validate the function of regulating the abortive molecular mechanism in the wheat,which can provide the clues fort the abortive molecular mechanism study of 337 S that is sensitive to the short-day in low temperature.The results are as follows: TaMS337S-1 is transformed into Huamai2566 and Huamai2152 by gene gun,but no transgene plants are found.While taking the immature-embryos of Huamai2152,Huamai2566,Zhengmai9023,Longchun23,Kenong199,and Bob White as the acceptor materials,51 regenerated plants are obtained through the TaMS337S-2 gene transformation by gene gun bombardment,including 4 Huamai2566 plants,41 Longchun23 plants,a Bob White plant.PCR results showed that 2 lines of Longchun23 are positive plants while others are negative plants.In addition,7 regenerated plants are transformed with TaMS337S-1 through Agrobacterium infection method,including 3 Huamai2566 plants,4 Longchun23 plants.PCR result showed that 2 lines of Huamai2566 are positive.Meanwhile we found that they had no anthers and the female filaments were exposed.Genetic transformation of TaMS337S-2 was the same as TaMS337S-2.9 regenerated plants were obtained including a Huamai2566 plant,4 Kenong199 plants,4 Longchun23 plants,while no positive plants were checked by PCR analysis.