Expression Analysis Of GmMIOX Gene,Subcellular Localization And Myo-inositol Oxygenase Activity Research Response To SCN Stress

Soybean cyst nematode(SCN)is one of the important diseases that endanger soybean yield.In recent years,the study of SCN disease has gradually become the hotspot in various countries.The protection measures mainly include banker rotation,the use of anti-nematode agents and resistant cultivars cultivation,however resistant cultivar is the most effective and practical measure to control SCN disease.Therefore,cultivating high yield and disease-resistant soybean cultivar has become an important measure to increase soybean yield and farmers' incomes.In this study,the susceptible cultivar Liaodou 15 and the resistant cultivars Huipizhi Heidou,Harbin Xiao heidou and Xiaoli Heidou were infected by SCN3,RNA was extracted at the 0d,5d,10 d,15d,20 d and 25 d respectively.The inositol oxygenase gene was detected by semi-quantitative PCR before and after nematode infection.The main research results include:1.We detected the difference of GmMIOX1,GmMIOX2 and GmMIOX4 gene expression in the resistant cultivars Huipizhi Heidou,Xiaoli Heidou and Harbin Xiao Heidou respectively,The results showed that the GmMIOX1 gene was specifically amplified in the resistant cultivars Huipizhi Heidou,Xiaoli Heidou and Harbin Xiao Heidou,but there was no significant difference between the control and treatment groups.GmMIOX2 gene and GmMIOX4 gene were partially amplified,and the amplification product of the treatment group was obviously.The PCR products of GmMIOX2 and Gm MIOX4 genes were significantly different by nematode infection.However,the GmMIOX1 gene did not change significantly.It showed that the GmMIOX2 and the GmMIOX4 genes are resistant to SCN disease.2.A susceptible cultivar Liaodou 15 and the resistant cultivars were Huipizhi heidou,Harbin Xiao heidou and Xiaoli Heidou were used to research the gene expression of inositol oxygenae family gene from soybeans roots after SCN3 infection.The results showed that in both resistant cultivars the expression level of GmMIOX2 gene reached the maximum on the fifth day,but the changes of susceptible cultivar was not obvious.The expression levels of GmMIOX4 gene in Huipizhi Heidou,Harbin Xiao heidou and Xiaoli Heidou were significantly higher than susceptible cultivar Liaodou 15 on fifth day,twentieth day and tenth day.Indicating that GmMIOX2 and GmMIOX4 genes were related to prevent nematode development.the development of SCN.3.The complete GmMIOX2 and GmMIOX4 genes were amplified intobinary expression vetor to construct pCAMBIA1303-MIOX2 and pCAMBIA1303-MIOX4.4.The complete GmMIOX2 and GmMIOX4 gene were amplified intobinary expression vetor to construct pCAMBIA1303-MIOX2 and pCAMBIA1303-MIOX4.The result of subcellular lacalization showed that GmMIOX2 protein was located on the cytomembrane,the GmMIOX4 protein was located cytoplasm and cytomembrane.5.Through comparing MIOX enzyme activity differences of different resistance and susceptible cultivars.the results show that MIOX enzyme activity in resistant varieties were particularly expressed,embodied in enzymatic activity after inoculation of soybean cyst nematode compared with control group.It proved that MIOX enzyme activity was specificity expressed and played different role in the defense of soybean against soybean cyst nematode.