Characterization of Cry toxins from autochthonous Bacillus thuringiensis isolates from Mexico

Raquel Camacho-Millán , Elsa Maribel Aguilar-Medina , Héctor Quezada , Óscar Medina-Contreras , Genaro Patiño-López , Héctor Manuel Cárdenas-Cota , Rosalío Ramos-Payán
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Abstract

Background

Chemical pesticides, widely used in agriculture and vector-borne disease control, have shown toxic effects on the environment and the people in contact with them. Bacillus thuringiensis is a widely used bacterium for alternative and safer control of insect pests. Its toxins are specific for insects but innocuous for mammals and may be used as powerful adjuvants when applied with vaccines. The objective of this work was to characterize some autochthonous B. thuringiensis strains, which could be used for the control of a local pest (Diatraea considerata Heinrich) that affects sugar cane crops in Sinaloa, Mexico. Also, to evaluate these strains as a source of Cry toxins, which may be used in the future as adjuvants for some vaccines.

Methods

Eight strains from field-collected dead insects were isolated. These were microbiologically identified as B. thuringiensis and confirmed by amplification and sequencing of 16S rDNA. Bioassays were performed to evaluate their pathogenicity against D. considerata, and Cry toxins were identified by proteomic analyses.

Results

An increased mortality among larvae infected with strain Bt-D was observed, and its toxin was identified as Cry1Ac.

Conclusions

The observed data showed that the selected strain was pathogenic to D. considerata and seemed to produce Cry1Ac protein, which has been reported as an adjuvant in different types of immunization.

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墨西哥本土苏云金芽孢杆菌分离株Cry毒素的鉴定
化学农药广泛应用于农业和病媒传播疾病控制,对环境和接触它们的人具有毒性作用。苏云金芽孢杆菌是一种被广泛应用于替代性和更安全的害虫防治的细菌。它的毒素对昆虫是特异性的,但对哺乳动物无害,当与疫苗一起使用时,可以用作强效佐剂。本研究的目的是鉴定一些本地苏云金杆菌菌株的特征,这些菌株可用于控制影响墨西哥锡那罗亚甘蔗作物的当地害虫(Diatraea考虑海因里希)。此外,为了评估这些菌株作为Cry毒素的来源,这些毒素可能在未来用作某些疫苗的佐剂。方法从野外采集的昆虫尸体中分离8株菌株。经微生物学鉴定为苏云金芽孢杆菌,并经16S rDNA扩增和测序证实。采用生物测定法对其致病性进行了评价,并通过蛋白质组学分析对其进行了鉴定。结果Bt-D感染后,幼虫死亡率升高,毒素为Cry1Ac。结论所选菌株对黄貂鱼具有致病性,并可能产生Cry1Ac蛋白,该蛋白已被报道为不同类型免疫的佐剂。
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