Sanchis V, Gohar M, Chaufaux J, Arantes O, Meier A, Agaisse H, Cayley J, Lereclus D
Unité de Biochimie Microbienne, Institut Pasteur, Centre National de Recherche Scientifique, Paris, France.
Appl Environ Microbiol. 1999 Sep;65(9):4032-9. doi: 10.1128/AEM.65.9.4032-4039.1999.
The main problems with Bacillus thuringiensis products for pest control are their often narrow activity spectrum, high sensitivity to UV degradation, and low cost effectiveness (high potency required). We constructed a sporulation-deficient SigK(-) B. thuringiensis strain that expressed a chimeric cry1C/Ab gene, the product of which had high activity against various lepidopteran pests, including Spodoptera littoralis (Egyptian cotton leaf worm) and Spodoptera exigua (lesser [beet] armyworm), which are not readily controlled by other Cry delta-endotoxins. The SigK(-) host strain carried the cry1Ac gene, the product of which is highly active against the larvae of the major pests Ostrinia nubilalis (European corn borer) and Heliothis virescens (tobacco budworm). This new strain had greater potency and a broader activity spectrum than the parent strain. The crystals produced by the asporogenic strain remained encapsulated within the cells, which protected them from UV degradation. The cry1C/Ab gene was introduced into the B. thuringiensis host via a site-specific recombination vector so that unwanted DNA was eliminated. Therefore, the final construct contained no sequences of non-B. thuringiensis origin. As the recombinant strain is a mutant blocked at late sporulation, it does not produce viable spores and therefore cannot compete with wild-type B. thuringiensis strains in the environment. It is thus a very safe biopesticide. In field trials, this new recombinant strain protected cabbage and broccoli against a pest complex under natural infestation conditions.
苏云金芽孢杆菌产品用于害虫防治的主要问题在于其活性谱往往较窄、对紫外线降解高度敏感以及成本效益较低(需要高效能)。我们构建了一种缺失芽孢形成能力的SigK(-)苏云金芽孢杆菌菌株,该菌株表达嵌合的cry1C/Ab基因,其产物对多种鳞翅目害虫具有高活性,包括埃及棉叶虫和甜菜夜蛾,而其他Cryδ-内毒素难以有效防治这些害虫。SigK(-)宿主菌株携带cry1Ac基因,其产物对主要害虫欧洲玉米螟和烟草天蛾的幼虫具有高活性。这种新菌株比亲本菌株具有更高的效能和更宽的活性谱。无芽孢形成能力的菌株产生的晶体保留在细胞内,从而保护它们免受紫外线降解。通过位点特异性重组载体将cry1C/Ab基因导入苏云金芽孢杆菌宿主,从而消除不需要的DNA。因此,最终构建体不包含非苏云金芽孢杆菌来源的序列。由于重组菌株是在芽孢形成后期受阻的突变体,它不产生有活力的孢子,因此在环境中无法与野生型苏云金芽孢杆菌菌株竞争。因此,它是一种非常安全的生物农药。在田间试验中,这种新的重组菌株在自然侵染条件下保护甘蓝和西兰花免受害虫复合体的侵害。