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Chinese Agricultural Science Bulletin ›› 2023, Vol. 39 ›› Issue (27): 103-109.doi: 10.11924/j.issn.1000-6850.casb2022-0814

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Cel61A Gene in Ceratocystis fimbriata: Cloning and Expression Analysis

LUO Qinchuan(), SUN Houjun, TANG Wei, XIE Yiping, YANG Dongjing, MA Jukui, CHEN Jingwei, GAO Fangyuan, ZHANG Chengling()   

  1. Xuzhou Institute of Agricultural Sciences in Jiangsu Xuhuai Area/ Sweet Potato Research Institute of Chinese Academy of Agricultural Sciences/ Key laboratory of Biology and Genetic Improvement of Sweet potato, Ministry of Agriculture and Rural Affairs, Xuzhou, Jiangsu 221131
  • Received:2022-09-26 Revised:2022-12-15 Online:2023-09-25 Published:2023-09-22

Abstract:

The aim was to clarify the expression pattern and the physicochemical characteristics of the encoded protein of polysaccharide monooxygenase gene (CfCel61A), a pathogenic gene of Ceratocystis fimbriata infecting sweet potato. Eleven strains from different regions were used as materials, CfCel61A was cloned, and the physicochemical properties of CfCel61A were determined by bioinformatics analysis software. The gene expression was analyzed by qRT-PCR. The results showed that the coding region of CfCel61A was 1155 bp with a concordance rate of 99.5%-100%, and it encoded 384 amino acids with the concordance rate of 98.4%-100%. The predicted molecular weight was 34.49 kDa, the isoelectric point was 5.59, the hydrophilicity coefficient was -0.195,and it was a stable hydrophilic protein. There was a signal peptide at the N-terminal, and the subcellular localization was in the nucleus without transmembrane structure. There were 58 phosphorylation sites and 2 potential glycosylation sites. The irregular curl of protein secondary structure accounted for 69.01%, which was the highest. The expression of CfCel61A was significantly up-regulated during the infection of C. fimbriata, while the up-regulation ranges were varied at different stages of infection, and it reached the highest level at 12 h and 6 h after the infection in 'Nanjing 92' and 'Xushu 37', respectively. In short, CfCel61A in C. fimbriata may play an important role in the infection of sweet potato and the result will provide a new target for drug development.

Key words: Ceratocystis fimbriata, polysaccharide monooxygenase, bioinformatics, clone, gene expression