Development and field performance of a broad-spectrum nonviable asporogenic recombinant strain of Bacillus thuringiensis with greater potency and UV resistance.
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.
Sanchis
V
V
Unité de Biochimie Microbienne, Institut Pasteur, Centre National de Recherche Scientifique, Paris, France. vsanchis@pasteur.fr
Gohar
M
M
Chaufaux
J
J
Arantes
O
O
Meier
A
A
Agaisse
H
H
Cayley
J
J
Lereclus
D
D
D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics / Q000502:physiology* / Q000528:radiation effects*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000633:toxicity; D001427:Bacterial Toxins; D001937:Brassica / Q000382:microbiology; D004269:DNA, Bacterial / Q000235:genetics; D004731:Endotoxins / Q000235:genetics* / Q000378:metabolism / Q000633:toxicity; D006460:Hemolysin Proteins; D007915:Lepidoptera / Q000382:microbiology; D010572:Pest Control, Biological; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism / Q000633:toxicity; D013171:Spores, Bacterial / Q000502:physiology / Q000528:radiation effects; D014170:Transformation, Genetic; D014466:Ultraviolet Rays
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004731:Endotoxins; D006460:Hemolysin Proteins; D011993:Recombinant Fusion Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis