p5cr基因在金龟子绿僵菌合成苦马豆素中相关性分析

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中图分类号：S859.87 文献标志码：A 文章编号：0366-6964（2025）09-4718-12

Correlation Analysis ofthe p 5cr Gene in Swainsonine Biosynthesis in Metarhizium anisopliae

SUN Pinzhi，QU Yingying， ZHANG Qin，YANG Liwen，LI Yange， ZHANG Yiqingqing，ZHANG Yu，LU Hao*（College of Veterinary Medicine，Northwest A&F University， Yangling 7121oo， China）

Abstract： This study aims to utilize the CRISPR/Cas9 gene editing technology to investigate the role of ϕ5cr （pyrroline-5-carboxylate reductase） in the synthesis of swainsonine by Metarhizium anisopliae，providing a theoretical basis for the biosynthesis and industrial production of swainsonine. In this study，Metarhizium anisopliae was used as the object of study，qRT-PCR method was used to detect the gene expression levels of ϕ5cr ，sdh，ρks，and ϕ450 in the swainsonine biosynthetic pathway. The correlation analysis was conducted on the production of swainsonine and the expresson levels of related genes of Metarhizium anisopliae after 1 to 7 days of fermentation. The ϕ5cr gene was selected as the target gene. CRISPR/Cas9 gene editing technology was applied to knock out and overexpress the catalytic gene ϕ5cr ，in order to validate the correlation between the ϕ5cr gene and swainsonine synthesis in Metarhizium anisopliae. The results demonstrated a strong positive correlation between swainsonine production and the expression level of the pks gene，while extremely strong correlations （ P<0.001 ）wereobserved with the expression levels of the ϕ5cr ， sdh ，and ϕ450 genes. Knockout and overexpression of the ϕ5cr genein Metarhizium anisopliae led to a significant decrease （ P<0.01 ）and increase （ P<0.001 ） in swainsonine yield，respectively. This study reveals that the ϕ5cr gene exhibits an extremely strong association with swainsonine biosynthesis in Metarhizium anisopliae and plays a crucial regulatory role in its biosynthetic pathway. These findings provide a foundational framework for further investigations into the function and regulatory role of the ϕ5cr gene.

Keywords： Metarhizium anisopliae ; ϕ5cr gene; swainsonine; locoweed; CRISPR/Cas9 gene editing technology

∗ Corresponding author：LU Hao，E-mail：luhao@nwsuaf.edu.cn

疯草（locoweed）是危害国内外畜牧业的主要毒草，动物采食疯草后引起以慢性神经机能紊乱为特征的中毒病。（剩余18513字）