Study Of ZmAChE Gene Transformation Via Agrobacterium Tumefaciens Tumefaciens-mediated Of Soybean

Soybean plays a significant role in our nation production as traditional crop thatproduce food and oil. Recently, with the rapid increasing emissions of greenhouse gasand release of forest area, high temperature happened with high frequency. Hightemperature had worse influence on soybean yield and quality. Heat stress affected cropyield and quality combined with other stress conditions such as drought stress. Hence, itis necessary and of great importance to produce heat tolerance lines to improve thesoybean yield and fight against adverse environment of global warming.Studies have shown that AChE is a positive regulator of heat tolerance and acontrol factor of stomatal opening and closing in plants. Therefore, this researchintroduced ZmAChE gene into soybean through agrobacterium-mediated transformationto obtain heat tolerant transgenic soybeans. The main research results are as follows:1. Construction of the expression vector PCBM-17.5c-AChE contained Bar,17.5cpromoter and maize AChE gene genetic and transformed soybean. The soybeancotyledonary node was prepared as explants, soaking in the bacteria infection for30minand co-culture for5days. Recovery culture for14days without screening concentration,following2weeks of PPT screening at concentration of5mg/L; after that the survivedexplants were tranferred to elongation medium, PPT screening concentration at1mg/L;when hight of plant is about3-5cm, stained with IBA1min and transfer into rootingmedium. When the root developed, transfered to field.2. About2000cotyledonary node explants was infected, obtained9plants withPPT resistant and2plants with PCR positive bands.3. Transgenic plants and WT were tested by glufosinate and heat experiment, theresult proved that transgenic plants can survive under heat conditions and haveglufosinate resistance.