Cloning And Expression Analysis Of Glutathione Synthesis Related Genes For The Huipizhi Heidou Resistant To Heterodera Glycines

The soybean cyst nematode (SCN) is one of the most important disease of soybean in the world.This disease was wildly distributed and seriously disserved in China.The researches were systematically conducted the number of the second stage juveniles, the contents of active oxygen expression of Glutathion synthetase, homoglutathione synthetase,and y-glutamylcystein syntherase under H.glycines infection in Huipizhi Heidou and Liaodou 15,To study Huipizhi Heidou gene profiles under soybean cyst nematode infection to explore the gene involved in soybean-nematode incompatible interaction and resistance mechanism at molecular level.The main results were as followed:1. The resistant Huipizhi Heidou had no effection on the invading of SCN race 3 but had an effection on the development of SCN race 3.Histopathological responce in roots of the resistant Huipizhi Heidou and the susceptible Liaodou 15 infected with SCN race 3 was studied by cryo-sectioning technique.2. The varieties of confrontation feeling within the root active oxygen detection, the soybean after receiving SCN3 infect, resistance, feeling both varieties within the root H2O2, begin with Huipizhi Heidou in the early 5d grow bigger, then though growth but smaller; In Liaodou 15,10,15 d dramatically increased, peak appeared late compared with grey leather black beans, speculation within the root of the resistant varieties of defense enzymes activity is relatively high susceptible cultivars, keep clear of H2O2 in the root. Showed that root H2O2 in the SCN resistant varieties different performance is associated with resistance.3. The expression of the hGSHS、GSHS、γ-ECS genes were detected in the resistant Huipizhi Heidou and the susceptible Liaodou 15 infected by SCN race 3 with method of Real-Time Quantitative PCR,the results showed that the expression of gene y-ECS was higher in roots after infection;The expression of gene GSHS was higher in Liaodou 15 than Huipizhi Heidou in 15 day after infection; The expression of gens hGSHS was lower in resistance soybean than control. Lower GSH related genes with resistance to SCN3 in beige with Huipizhi Heidou were positively correlated.4. Gene of hGSHS、GSHS、γ-ECS were cloned and sequenced from soybean roots infected H.glycines by Reverse Transcription PCR,which were crucial enzyme in plant cells regulating Glutahione.The cDNA length of cloned genes were 1781bp、1654bp、1559bp,the open reading frame of cDNA which encoded polypeptide of 498、544、503 amino acid residues. Using Clustal, DNAMAN analysis software and the analysis of the commonly used database and online tools such as, to clone the gene encoding protein has carried on the physical properties, the transmembrane domain, signal peptide, feeling of water-borne and other characteristics.