Research Of Mapping The Powdery Mildew Resistance Gene And Related Proteins In Melon

Melon is a worldwide horticultural crops, Xinjiang is the main producing areas of meloncultivaton in our country.But Xinjiang melon are poor to resistance to powderymildew.Powdery mildew lead to a large area of melon leaves dead at later growth stage, andthe yield and quality increasingly decline, thus caused big economic losses to Farmers.In this experiment, we used Xinjiang melon as the research object, choosed molecularmarkers closely linked to powdery mildew resistance genes, and mapping powdery mildewgenes in genetic.By researching melon powdery mildew resistance gene molecular markerswill be applied to melon powdery mildew resistance gene polymerization breeding.It is verysignificant to cultivate melon innovative germplasm resources and disease resistant breeding.Then extracted the protein of melon Leaf,got Melon protein of higher concentration andpurity.Established and optimized two-dimensional electrophoresis system of melon leaf to digdeeply for melon proteomics by using proteomic technology.The results showed that:(1)Compared with the method of Tirs-phenol extraction,theimproved method Mg/NP-40/PEG3350/TCA acetone extracting leaves protein, and afterSDS-PAGE analysised. The protein content was higher, Rubisco enzyme compared removedgood, bands were clear. When using pH3-10,18cm IPG strips of two-dimensionalelectrophoresis, increasing the low voltage focusing time, get a clearer2-D map. The proteinsample volume800μg, the maximum number of points was obtained in protein, low proteinkurtosis clearer.(2)Using116pairs of SSR molecular marker analysis of mixed DNA separation group, inmelon II linkage group screened12primer combination between disease resistant pool andinfected pool amplified specificity band. Showed that chain with resistance genes.Withpolymorphism, the polymorphism rate was37.5%.Using the above12SSR molecular markeramplified JS♂×H70♀×JS♂DNA isolated populations plant and tested, indicated that theresistance gene is located on LGII in JS♂×H70♀×JS♂isolated populations,and basicallycompletely linkage.