Cloning And Drought Resistance Analysis Of Soybean UDP-glycosyltransferase Gene GmA02G03420

Soybean is an important grain and oil crop in China,and it is an important source of high quality vegetable protein and vegetable oil.It has special strategic position in the development of the national economy.China is the country of origin of soybeans and has a long history of soybean cultivation,which is one of the four largest soybean producers in the world.In recent years,with the improvement of the living standards of our people,the demand for soybeans has increased dramatically.Soybean production is often affected by extreme weather,and drought is the most likely to cause a decline in annual soybean yield.How to overcome the impact of drought on soybean yield has become a major task in current breeding research.Based on the sequencing results of the previous transcriptome,the UDP-glycosyltransferase gene was cloned by the drought-resistant mutant M18,and the overexpression vector and RNA interference expression vector were constructed based on the plant expression vector pCAMBIA3301.The constructed recombinant vector was transferred into the soybean receptor variety 'JN18' by agrobacterium-mediated method.The preliminary detection was carried out by PCR,and the positive seeds obtained were added in the artificial climate chamber.The T2 generation transgenic plants were tested by Southern blotting at the integration level,the detection of fluorescent quantitative PCR at the transcription level and the preliminary identification of indoor drought resistance.In order to provide materials for the study of drought resistance of genetically modified soybeans.The main findings are as follows: 1.The target gene was cloned and the overexpression vector p3301-GmA02G03420 and RNA interference expression vector p3301-GmA02G03420-RNAi were constructed by seamless cloning.2.The soybean receptor variety 'Jinong 18' was transformed by agrobacterium-mediated transformation.By PCR detection,the positive plants of T0 transfer p3301-GmA02g03420 and p3301-GmA02g03420-rnai vector were 4 and 2,respectively.3.The seeds of T0 generation positive plants were germed into soil,we obtained five transformed plants in T1 generation of p3301-GmA02G03420 vector and four of p3301-GmA02G03420-RNAi vector.The seeds of T1 generation positive plants were germed into soil in the artificial climate chamber,we obtained twelve transformed plants in T2 generation of p3301-GmA02G03420 vector and ten of p3301-GmA02G03420-RNAi vector.4.The results of Southern blotting of T2 transgenic plants indicated that the target gene was integrated into the soybean receptor genome in a single copy,and the hybridization signal intensity was different.5.The results of fluorescence quantitative PCR showed that the expression level of the transgenic over-expression vector was significantly higher than that of the control,and the expression level of the trans-interference expression vector was significantly lower than that of the control.6.The results of drought resistance identification showed that the transgenic plants significantly improved the drought resistance of jinong 18.