华北农学报 ›› 2026, Vol. 41 ›› Issue (1): 184-193. doi: 10.7668/hbnxb.20195733

所属专题: 植物保护 生物技术 热点论文

• 资源环境·植物保护 • 上一篇    下一篇

苏云金芽孢杆菌杀虫蛋白基因型鉴定及对鳞翅目害虫高毒力菌株筛选

丰硕, 黄国强, 程佳旭, 曹伟平, 贾海民, 宋健   

  1. 河北省农林科学院 植物保护研究所,河北省农业有害生物综合防治技术创新中心, 河北省作物有害生物综合防治国际科技联合研究中心,河北 保定 071000
  • 收稿日期:2024-06-02 出版日期:2026-02-28
  • 通讯作者:
    宋健(1980—),女,河北涿州人,研究员,硕士,主要从事害虫综合防治研究。
  • 作者简介:

    丰硕(1994—),男,河北沧州人,助理研究员,硕士,主要从事农业害虫生物防治研究。

  • 基金资助:
    河北省农林科学院基本科研业务费项目(2023120101); 河北省自然科学基金项目(C2024301038); 河北省农林科学院现代农业科技创新专项课题(2022KJCXZX-ZBS-6)

Identification of Insecticidal Protein Genotypes of Bacillus thuringiensis and Screening of Highly Virulent Strains Against Lepidopteran Pests

FENG Shuo, HUANG Guoqiang, CHENG Jiaxu, CAO Weiping, JIA Haimin, SONG Jian   

  1. Plant Protection Institute,Hebei Academy of Agriculture and Forestry Sciences,IPM Innovation Center of Hebei Province,International Science and Technology Joint Research Center on IPM of Hebei Province,Baoding 071000,China
  • Received:2024-06-02 Published:2026-02-28

摘要:

苏云金芽孢杆菌(Bt)能够产生多种杀虫蛋白,是重要的生防资源。为了发掘对鳞翅目害虫高毒力的Bt菌株,缓解害虫抗性压力并扩大资源储备,以河北地区土壤样品中分离的109株Bt菌株为研究对象,通过油镜观察伴孢晶体形状,利用40对通用引物(包括cry、cyt、vip 3种基因型)PCR鉴定所有菌株的基因型。以小菜蛾、斜纹夜蛾等5种鳞翅目害虫为靶标,筛选高杀虫活性菌株,并利用SDS-PAGE分析其晶体蛋白。结果表明,伴孢晶体形状主要是圆球形(90株),少部分为菱形(19株)。杀虫蛋白基因型检出率为93.6%,主要包括29种不同的cry基因型、1种cyt基因型以及3种vip基因型,91株具有至少2种杀虫蛋白基因型的组合,其中26株含有6种以上,多为cry+vip型。室内生测发现19株对小菜蛾等多种鳞翅目害虫的高毒力菌株,主要表达130,65 ku的蛋白,包含杀鳞翅目的cry1类、cry2类、cry15类等基因型,与PCR鉴定结果一致。获得了对鳞翅目害虫广谱、高毒力的Bt菌株,并初步明确了其杀虫基因,对新型Bt产品的开发具有重要意义。

关键词: 苏云金芽孢杆菌, 基因型, 鳞翅目害虫, 杀虫活性

Abstract:

Bacillus thuringiensis(Bt)produces multiple insecticidal proteins and serves as a vital biological control agent.This study aims to identify Bt strains with insecticidal activity against lepidopteran pests,thereby alleviating pest resistance pressure and expanding resource reserves.A total of 109 Bt strains were selected from soil of Hebei Province.The shape of parasporal crystals was observed under an oil microscope,and the insecticidal protein genotypes of the Bt strains were identified by PCR using 40 pairs of primers(including cry,cyt,and vip genotypes).The insecticidal activity bioassay was performed using five lepidopteran pests,including Plutella xylostella and Spodoptera litura.The molecular weight of crystal proteins was detected via SDS-PAGE.The shape of parasporal crystals from 90 strains was spherical,and the crystals from the remaining 19 strains were diamond-shaped.The detection rate of insecticidal protein genotypes reached 93.6%,with 29 distinct cry genotypes,1 cyt genotype,and 3 vip genotypes identified.A total of 91 strains possessed combinations of at least two insecticidal protein genotypes,among which 26 strains contained six or more,predominantly exhibiting cry+vip patterns.We found that 19 Bt strains showed high insecticidal activity against various lepidopteran pests such as P.xylostella. These strains primarily expressed proteins at 130,65 ku,encompassing lepidopteran-killing genotypes such as cry1,cry2,and cry15,consistent with PCR identification results.Here,we identified broad-spectrum insecticidal and highly toxic Bt strains against lepidopteran pests,and revealed its insecticidal genes.Our work provides an important candidate for the development of bioinsecticide.

Key words: Bacillus thuringiensis, Genotype, Lepidopteran pests, Insecticidal activity

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引用本文

丰硕, 黄国强, 程佳旭, 曹伟平, 贾海民, 宋健. 苏云金芽孢杆菌杀虫蛋白基因型鉴定及对鳞翅目害虫高毒力菌株筛选[J]. 华北农学报, 2026, 41(1): 184-193. doi: 10.7668/hbnxb.20195733.

FENG Shuo, HUANG Guoqiang, CHENG Jiaxu, CAO Weiping, JIA Haimin, SONG Jian. Identification of Insecticidal Protein Genotypes of Bacillus thuringiensis and Screening of Highly Virulent Strains Against Lepidopteran Pests[J]. Acta Agriculturae Boreali-Sinica, 2026, 41(1): 184-193. doi: 10.7668/hbnxb.20195733.