Identification And Main Trait Analysis Of GmWRI1a Gene High Oil Soybean

Soybean [Glycine max(L.)Merrill] is an important oil crop in the world,and chinise people are big consumers of soybean oil.However,soybean,in China,depents on imports to meet the needs because of the lower oil content and lower yield of soybean seeds.Therefore,an increase in the oil content of soybean seeds would dramatically contribute to reducing the shortage of vegetable oil.The high demand for soybean oil has sparked an interest in increasing the relative oil content per seed.Genetic engineering not only can complement conventional plant breeding methods to break long cycle but can also precisely regulate trait expression in short periods of time without any lineage drag in crop plants.With the rapid development of genetic engineering recent years,breeding high oil soybean cultivars using transgenic technology has become the most important research method.WRI1 transcription factor belongs to AP2/EREBP transcription factor family,which is a key regulator to regulate fatty acid biosynthesis in plants.Soybean Gm WRI1 a gene was isolated from soybean and had beentransformed into soybean cultivar DN50 in our lab.The results showed that the oil content of transgenic soybean seeds was significantly increased.In order to further screen high oil transgenic soybean lines,the six T3 generation transgenic soybean lines obtain ed in our laboratory were used as experimental materials in this research.At the DNA level and protein level,the transgenic soybean lines were detected using molecular biology method.The quality traits such as seed oil content,protein content,agronomic traits,resistance to non-target organisms and gene drift of transgenic soybean plants were analyzed.The main results are as follows:(1)The transgenic soybean linesoverexpressing Gm WRI1 a gene were screened and identified using 130mg/L phosphine glyphosate smear,RCR amplification and Southern blot.The results showed that the Gm WRI1 a and the Bar gene were suceedly inserted into transgenic soybean genome in the form of low copy and multiple copies.(2)The expression production of the Bar protein in transgenic soybean was detected by Western blot method.The results showed that the Bar protein was overexpressed in transgenic soybean plants.(3)The oil content and protein content of soybean seeds overexpressing Gm WRI1 a gene were determined.The results showed that the overexpression of Gm WRI1 a gene increased the seed oil contents,and decreased the protein content in soybean seeds.(4)The agronomic characters of Gm WRI1a-overexpressing soybean plants were investigated.The results showed that there were no differences in germinating rate,growth period and other agronomic traitsof Gm WRI1a-overexpressing soybean lines compared with wild type DN50.However,the plant height,the number of nodes and branches of main stem changed significantly in some transgenic lines compared with DN50,which indicated that the overexpression of Gm WRI1 a gene might have the effect on the plant height,node numbers and branch numbers.(5)The gene drift of transgenic soybean plants were identified.The results showed that no gene drift was found within 50 m from the planting area.(6)The resistances of Gm WRI1a-overexpressing soybean plants to non-target diseases and insect pests were identified.The results showed that the resistances of soybean plants overexpressing Gm WRI1 a gene to Sclerotinia sclerotiorum,Plutella xylostella and Soybean moth were not changed compred with wild type.(7)The quality characters of Gm WRI1a-overexpressing soybean seeds were determined.The results showed that there were no significant differences in soluble reducing sugar,starch,cellulose,moisture,ash content of transgenic soybean plants compared with wild tpye.However,the fatty acid composition is significantly changed.