Scholarlink

<p>Plant chitinase, a member of the pathogenesis-related proteins and glycoside hydrolase family 19, plays a crucial role in plant resistance to pathogenic fungi. In this study, we isolated a chitinase gene encoding PaChi from <i>Picea asperata</i>, containing an open reading frame of 768&#xa0;bp that encodes 255 amino acid residues. PaChi includes the chitin-binding domain, catalytic domain, and active site characteristic of class IV chitinases. Homologous modeling and phylogenetic analysis indicated that PaChi exhibits the highest structural similarity and closest genetic relationship with the chitinase of Norway spruce, suggesting potential functional similarities. RT-qPCR analysis demonstrated that <i>PaChi</i> expression is relatively high in the roots of <i>P. asperata</i>. Additionally, <i>PaChi</i> transcription levels were significantly upregulated during the early stages of <i>Lophodermium piceae</i> infection, indicating active involvement in the defense response of <i>P. asperata</i> against <i>L. piceae</i> infection. SDS-PAGE analysis revealed that the recombinant PaChi has a molecular weight of approximately 27&#xa0;kDa. The purified recombinant PaChi exhibited a chitinase activity of 4.61 U/mg, with optimal pH and temperature at 7.0 and 40&#xa0;°C, respectively. Notably, purified recombinant PaChi significantly affected the mycelial morphology of <i>L. piceae</i>, inclusion body concentration and cell swelling. These findings lay the groundwork for further understanding the molecular mechanisms by which <i>PaChi</i> is involved in biological stress.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Molecular identification and functional characterization of a chitinase gene in Picea asperata

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年5月13日

查看全文
  • Yufeng Liu,
  • Xumei Zhang,
  • Siqi Yuan,
  • Jun Liu

摘要

Plant chitinase, a member of the pathogenesis-related proteins and glycoside hydrolase family 19, plays a crucial role in plant resistance to pathogenic fungi. In this study, we isolated a chitinase gene encoding PaChi from Picea asperata, containing an open reading frame of 768 bp that encodes 255 amino acid residues. PaChi includes the chitin-binding domain, catalytic domain, and active site characteristic of class IV chitinases. Homologous modeling and phylogenetic analysis indicated that PaChi exhibits the highest structural similarity and closest genetic relationship with the chitinase of Norway spruce, suggesting potential functional similarities. RT-qPCR analysis demonstrated that PaChi expression is relatively high in the roots of P. asperata. Additionally, PaChi transcription levels were significantly upregulated during the early stages of Lophodermium piceae infection, indicating active involvement in the defense response of P. asperata against L. piceae infection. SDS-PAGE analysis revealed that the recombinant PaChi has a molecular weight of approximately 27 kDa. The purified recombinant PaChi exhibited a chitinase activity of 4.61 U/mg, with optimal pH and temperature at 7.0 and 40 °C, respectively. Notably, purified recombinant PaChi significantly affected the mycelial morphology of L. piceae, inclusion body concentration and cell swelling. These findings lay the groundwork for further understanding the molecular mechanisms by which PaChi is involved in biological stress.