CLONING AND BIOINFORMATICS ANALYSIS OF WHEAT POWDERY MILDEW RESISTANCE RELATED GENE TAGDSL
Abstract
Powdery mildew disease of wheat is caused by Blumeri agraminis F. sp. tritici. It is one of the primary fungal diseases of wheat. Therefore, it is of great significance to explore and utilize the broad spectrum anti-powdery mildew genes and study their resistance mechanism and molecular mechanism for effectively resisting the harm of powdery mildew. Material is wheat variety Bainong207 supplied by Henan Provincial Research Center of Food Crop Genome Editing Engineering Technology. PmD-19T vector, E. coli 5α, and Agrobacterium GV3101 strain were purchased from Takara Biological Company. PCR apparatus, electrophoresis apparatus, agarose gel electrophoresis imager, autoclaving cooker, water bath cooker, ultra-clean workbench, etc. RT-PCR cloned the entire length of the TaGDSL gene. Bioinformatics analysis of the sequence showed that the total length of ORF was 1269bp, encoding 423 amino acids, with a molecular weight of 38.99 kD and an isoelectric point of 8.19. In addition, the TAGDSL gene has a transmembrane domain, a signal peptide, and the protein is hydrophilic. GDSL lipase is involved in plant physiological metabolism and local and global immunity. It is of great significance for improving disease resistance and yield of wheat. Based on the previous research, this experiment cloned the full-length sequence of a wheat GDSL gene, which enriched the members of the plant GDSL lipase family. She provided a basis for the subsequent exploration of its function and mechanism of action in the resistance to powdery mildew by modern molecular biology methods and the study of its gene function. The TAGDSL lipase gene of wheat was closely related to the durum wheat gene and barley gene by phylogenetic tree analysis. At the same time use bioinformatics method to forecast the gene can be more intuitive and more comprehensive understanding of its structure and properties, for the subsequent use of modern molecular biology methods to explore its function and role in the resistance to powdery mildew mechanism to provide effective basis, and for creating varieties of wheat powdery mildew resistance gene source and theoretical basis.
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