CRISPR/Cas9-Mediated Knockout of the PxJHBP Gene Resulted in Increased Susceptibility to Bt Cry1Ac Protoxin and Reduced Lifespan and Spawning Rates in Plutella xylostella

Juvenile hormone binding protein (JHBP) is a key regulator of JH signaling, and crosstalk between JH and 20-hydroxyecdysone (20E) can activate and fine-tune the mitogen-activated protein kinase cascade, leading to resistance to insecticidal proteins from Bacillis thuringiensis (Bt). However, the inv...

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Published in:Journal of agricultural and food chemistry 2024-04, Vol.72 (14), p.8180-8188
Main Authors: You, Shijun, Yao, Shuyuan, Chen, Xuanhao, Hou, Qing, Liu, Zhaoxia, Lei, Gaoke, Xie, Xuefeng, Liang, Zhengjiao, Yuchi, Zhiguang, You, Minsheng, Liu, Yuanyuan, Xiong, Lei
Format: Article
Language:English
Subjects:
Animals
Bacillus thuringiensis - genetics
Bacillus thuringiensis - metabolism
Bacillus thuringiensis Toxins - metabolism
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
CRISPR-Cas Systems
Endotoxins - genetics
Endotoxins - metabolism
Female
Food Safety and Toxicology
Hemolysin Proteins - genetics
Hemolysin Proteins - metabolism
Insecticide Resistance - genetics
Larva - metabolism
Longevity
Moths - genetics
Moths - metabolism
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Summary:Juvenile hormone binding protein (JHBP) is a key regulator of JH signaling, and crosstalk between JH and 20-hydroxyecdysone (20E) can activate and fine-tune the mitogen-activated protein kinase cascade, leading to resistance to insecticidal proteins from Bacillis thuringiensis (Bt). However, the involvement of JHBP in the Bt Cry1Ac resistance of Plutella xylostella remains unclear. Here, we cloned a full-length cDNA encoding JHBP, and quantitative real-time PCR (qPCR) analysis showed that the expression of the PxJHBP gene in the midgut of the Cry1Ac-susceptible strain was significantly higher than that of the Cry1Ac-resistant strain. Furthermore, CRISPR/Cas9-mediated knockout of the PxJHBP gene significantly increased Cry1Ac susceptibility, resulting in a significantly shorter lifespan and reduced fertility. These results demonstrate that PxJHBP plays a critical role in the resistance to Cry1Ac protoxin and in the regulation of physiological metabolic processes associated with reproduction in adult females, providing valuable insights to improve management strategies of P. xylostella.
ISSN:0021-8561
1520-5118
DOI:10.1021/acs.jafc.3c08721