Screening And Related Functional Prediction Of Molecular Markers For Resistance Gene In Wheat

Wheat(Triticum aestivum L.)is one of the most important food crops in the world,and its cultivation history can be traced back to more than 8,000 years ago.The demand for wheat yield and quality has gradually increased with the growth of the population.Wheat production is affected by various pathogens.In recent years,wheat disease has been serious.Breeding resistant varieties is through the discovery of new disease resistance sites and cloning,and the design of hybrid combinations or molecular marker-assisted breeding methods is the most effective.With the completion of genome sequencing of wheat and its ancestors,a large amount of analyzable data has been generated.Bioinformatics is a comprehensive interdisciplinary discipline that integrates multiple disciplines such as mathematics,statistics,biological sciences,and computer science,and aims to make use of research in genomics and proteomics.Agricultural bioinformatics arises from bioinformatics,and it plays a significant role in identifying the organizational structure and distribution of important genes,predicting new genes,studying the evolution of resistance genes,analyzing the functional relationship between resistance genes and proteins,and regulating mechanisms.In this paper,specific molecular markers related to wheat powdery mildew resistance and rust resistance were screened,and electronic cloning was performed.Disease resistance related genes were obtained through GO function prediction and functional annotation.Biological information and molecular mechanism of relevant functional genes during the interaction among wheat,powdery mildew and rust were revealed through KEGG analysis.The experimental research content of this paper is as followed:(1)On the Linux platform,the disease resistance markers of wheat were electronically located in Chinese Spring and its ancestors Triticum urartu,Aegilops tauschii to determine the actual location of disease resistance genes on the chromosome.The mapping results were mapped through Map Chart2.1.According to the electronic positioning results,chromosome 2B of hexaploid wheat,chromosome 2 of Triticum urartu,and chromosome 2 of Aogilops tauschii were most.(2)For the sites that can be successfully electronically located,a sequence of 2kb on each side of the two wings was extracted and bioinformatics analysis was performed.The prediction of functional genes on the two wings of the relevant location was conducted using Fgenesh and a total of 96 functional genes were predicted.(3)With Blast2 go,functional annotation and classification of the obtained nucleic acid sequences of functional genes were conducted.Among them,60 functional genes can be successfully annotated,and 36 functional genes cannot be successfully annotated.As a result,we got 58 GO terminology information: 24 were biological pathways(P),accounting for41.38% of the total;10 cell components(C),accounting for 17.24% of the total;24 were molecular functions(F),accounting for 41.38%.The KEGG metabolic pathway analysis obtained the corresponding KO metabolic pathway,EC code and other related information.(4)In order to test the reliability of the predicted functional genes in actual research,based on the sequences of resistance gene,the primers were designed by Primer3.0 software,and the products were analyzed by polyacrylamide gel electrophoresis.