Effects Of Glutathione Metabolism To Resistant Against Heterodera Glycines In Different Soybean Cultivars

Soybean cyst nematode disease is a plant root disease caused by the infection of soybean cyst nematode(Heterodera glycines,SCN),which seriously harms the quality and yield of soybeans worldwide in production.Therefore,it is extremely important to take appropriate precautions.At present,the widely used measures in the main soybean producing areas mainly include:agricultural control,physical,chemical control and microbial control,etc.,which have played a certain role in the outbreak and spread of soybean cyst nematode disease;The most economical and effective measures for prevention and treatment should be the selection and application of multiple types of disease-resistant varieties.Its main advantages are non-toxic,pollution-free and environmentally friendly.In this study,susceptible varieties and susceptible variety were analyzed for glutathione metabolism and reactive oxygen species.To further analyze the role of glutathione in participating in resistance to soybean cyst nematodes,and help provide an important theoretical basis for the rational use of high-quality soybean resources.The main results were carried out:1.The expressions of glutathione synthesis-related genes y-ECS,GSHS and hGSHS were examined both in resistant(Peking and PI88788)and susceptible(Williams 82)varieties by qRT-PCR.After SCN second-stage juveniles(J2)inoculated at 5d,10d and 15d,it was found that the expression level of both yECS and GSHS genes in Peking were significantly lower than in uninfected roots at 5 dpi.While a significant difference between the expression of y-ECS and GSHS was observed between 10d and 15d,in which they were up-regulated after SCN infection.The expression level of both yECS and hGSH genes in PI88788 were markedly down-regulated at 15 dpi,and the expression of the GSHS gene was significantly lower than that of the controls at 10 dpi.In contrast,the expression of the yECS and GSHS gene expression levels in Williams 82 was always induced during SCN infection at each time point according to qRT-PCR relative to that of the control.The above results showed that the expression of glutathione synthesis-related genes in the roots of resistant soybean varieties Peking and PI88788 were down-regulated in varying degree.And the trenducy were similar to their resistance characteristics.2.Detection of ?-EC,GSH,and hGSH content in roots of different soybean varieties by HPLC.In this study,the results showed that quantitative trends were roughly similar to that of gene expression.At 5 dpi,the contents of ?-EC and GSH in the root system of Peking were decreased.While there was a significant reduction in ?-EC and GSH level in resistant PI88788 roots relative to that in the uninfected control roots at 10 and 15dpi.And the results of nematode development showed that mostly J2 individuals in the Peking root were suppressed at 5d.At 10 and 15d SCN infection,the number of J4 in root of PI88788 were greatly lower than that in Williams 82.The above results indicated that resistant cultivars could inhibit the development of nematodes by regulating the metabolism of glutathione.3.The results of reactive oxygen species(ROS)revealed that H202 accumulation was induced by SCN in the roots of susceptible and resistant soybeans.Importantly,the most significant increase in levels of H2O2 accumulation was recorded in resistant Peking roots infected with SCN at 5 dpi.A high increase in H2O2 accumulation at 10 dpi was observed in the root tissue of resistant PI88788 roots.Interestingly,at 15 dpi,the oxidative burst was detected in the roots of susceptible Williams 82,whereas no significant difference in resistant Peking and PI88788 roots was observed in response to SCN infection.4.The expression of y-ECS,hGSHS and GSHS genes in susceptible Williams 82 had been significantly inhibited after the nematodes by BSO-treated.The development of nematodes in the roots revealed that nematode development in soybean roots treated with BSO had a significant suppression compared with untreated roots and delayed the formation of syncytia.The detection of ROS showed that the production of H2O2 was significantly induced by SCN at 5 dpi in(h)GSH-depleted roots,as evidenced by histochemical staining with 2'.7'-dichlorofluorescein diacetate(DCF)and 3,3-diaminobenzidine(DAB).These results indicated that(h)GSH deficiency could inhibit the development of SCN via modulation of H2O2 accumulation.