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Characterization Of A Disulphide Isomerase Gene From Haynaldia Villosa To Reveal The Mechanism Of Resistance Against Powdery Mildew

Characterization Of A Disulphide Isomerase Gene From Haynald...

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江苏省遗传学会2013年学术研讨会

作者： MUHAMMAD Faheem LI Yingbo ZHU Yanfei CHENG Jiangyue wang zongkhuan wang Xiue National Key Laboratory of Crop Genetics and Germplasm Enhancement Nanjing Agricultural University

Haynaldia villosa（or Dasypyrum villosum, 2 n = 2 x = 14, VV）, a wild relative of common wheat, exhibits durable resistance against powdery mildew disease which is one of the key potential threat of wheat production around the globe. In the previous study, an Hv-CMPG gene was identifiedby using GeneChip microarray and c DNA library screening that conferred durable resistance against powdery mildew. A yeast two-hybrid library of H. villosa was constructed to reveal the molecular mechanism of resistance by Hv-CMPG. Screening results identified a disluphide isomerase protein, namely Hv-PDI, to interact with Hv-CMPG. The Hv-PDI c DNA fragment was 1615 bp which included the complete open reading frame of 1323 bp, encoded the protein molecule of 440 amino acids residues. HV-PDI also showed the typical structure of two thioredoxine-like acitive sites（CGHC） and a modified endoplasmic reticulum retention signal（NDEL） at C-terminal. Specific primer pairs were designed according to D-genome sequence to locate this gene on 3 V of H. villosa genome. Subcellular location analysis of Hv-PDI by expression of a fusion to GFP depicted its presence in the compartments of cytoplasm of onion cell. Quantitative RT-PCR analysis demonstrated that transcripts of Hv-PDI substantially increased after the inoculation of Bgt. Transient over-expression of Hv-PDI demonstrated that haustorium index decreased significantly in the epidermal cells of susceptible wheat cultivar "Yangmai-158" as compared to control. Our results reveal the role of Hv-PDI in combating against Bgt and represent a potential novel source to powdery mildew for wheat improvement.

关键词： Haynaldia villosa Protein Disulfide Isomerase(PDI) powdery mildew

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