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在大肠杆菌中表达的苏云金芽孢杆菌Cry1Ia36的杀线虫活性。

The nematicidal activity of Bacillus thuringiensis Cry1Ia36 expressing in Escherichia coli.

作者信息

Cheng Feixue, Wang Dongwei, Wang Jian, Wang Xin, Long Minzhi, Sun Shu-E, Zhu Chunhui, Cheng Ju-E, Tan Xinqiu, Zhang Deyong, Liu Yong

机构信息

Key Laboratory of Integrated Management of the Pests and Diseases on Horticultural Crops in Hunan Province, Institute of Plant Protection, Hunan Academy of Agricultural Sciences, Changsha 410125, China.

Hunan Institute of Agricultural Economics and Information, Hunan Academy of Agricultural Sciences, Changsha 410125, China.

出版信息

Pestic Biochem Physiol. 2025 Jun;211:106419. doi: 10.1016/j.pestbp.2025.106419. Epub 2025 Apr 21.

Abstract

Plant-parasitic nematodes (PPNs) are a significant threat to numerous agricultural crops. Biocontrol is an effective and safe method of the managing PPNs. Bacillus thuringiensis (Bt) and its parasporal crystal proteins (ICPs) are important biocontrol resources for PPNs. In this study, the 2160-bp cry1Ia36 gene from the Bt YC-10 strain was expressed in Escherichia coli and the 81-kDa protein was purified. The nematicidal activity test yielded an LC of 12.79 mg/L at 96 h for the second stage juveniles (J2s) of Meloidogyne incognita, and the Cry1Ia36 protein exhibited a pronounced inhibitory effect on the infection of M. incognita to cucumber roots with the pot experiment, which the control effect was 76.95 % treated with 40 mg/L Cry1Ia36. Transgenic tomatoes that expressed the cry1Ia36 gene exhibited efficient control of M. incognita, showing a notable reduction in the number of galls and eggmasses compared to the control. Subsequent transcriptome analysis revealed substantial alterations in the expression patterns of numerous genes in M. incognita J2s treated with Cry1Ia36 protein. Furthermore, the yeast two-hybrid analysis and pull-down assays demonstrated an interaction protein in M. incognita and the interaction protein is associated with the nematicidal activity of Cry1Ia36. Consequently, these findings suggest that the Cry1Ia36 protein could be a promising nematicidal agent for the control of PPN diseases, which offers a viable alternative to chemical pesticides because of its association with environmental concerns, although no previous reports have been found that demonstrate its activity against PPNs. Concurrently, this study has also expanded the spectrum of Bt ICPs.

摘要

植物寄生线虫(PPNs)对众多农作物构成重大威胁。生物防治是管理PPNs的一种有效且安全的方法。苏云金芽孢杆菌(Bt)及其伴孢晶体蛋白(ICPs)是防治PPNs的重要生物防治资源。在本研究中,来自Bt YC - 10菌株的2160 bp cry1Ia36基因在大肠杆菌中表达,并纯化了81 kDa的蛋白。杀线虫活性测试表明,对于南方根结线虫的二龄幼虫(J2s),在96小时时的致死浓度(LC)为12.79 mg/L,并且Cry1Ia36蛋白在盆栽试验中对南方根结线虫侵染黄瓜根表现出显著的抑制作用,用40 mg/L Cry1Ia36处理时防治效果为76.95%。表达cry1Ia36基因的转基因番茄对南方根结线虫表现出有效的防治效果,与对照相比,根结和卵块数量显著减少。随后的转录组分析揭示了用Cry1Ia36蛋白处理的南方根结线虫J2s中许多基因表达模式的显著变化。此外,酵母双杂交分析和下拉试验证明了南方根结线虫中有一个相互作用蛋白,并且该相互作用蛋白与Cry1Ia36的杀线虫活性相关。因此,这些发现表明Cry1Ia36蛋白可能是一种有前景的防治PPN病害的杀线虫剂,由于其与环境问题相关,它为化学农药提供了一种可行的替代方案,尽管此前尚未发现有报道证明其对PPNs的活性。同时,本研究也拓宽了Bt ICPs的作用谱。

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