Identification of cry1I-Type Genes from Bacillus thuringiensis Strains and Characterization of a Novel cry1I-Type Gene

ABSTRACT A PCR-restriction fragment length polymorphism method for identification of cry1I-type genes from Bacillus thuringiensis was established by designing a pair of universal primers based on the conserved regions of the genes to amplify 1,548-bp cry1I-type gene fragments. Amplification products were digested with the Bsp119I and BanI enzymes, and four kinds of known cry1I-type genes were successfully identified. The results showed that cry1I-type genes appeared in 95 of 115 B. thuringiensis isolates and 7 of 13 standard strains. A novel cry1I-type gene was found in one standard strain and six isolates. The novel cry1I gene was cloned from B. thuringiensis isolate Btc007 and subcloned into vector pET-21b. Then it was overexpressed in Escherichia coli BL21(DE3). The expressed product was shown to be toxic to the diamondback moth (Plutella xylostella), Asian corn borer (Ostrinia furnacalis), and soybean pod borer (Leguminivora glycinivorella). However, it was not toxic to the cotton bollworm (Helicoverpa armigera), beet armyworm (Spodoptera exigua), or elm leaf beetle (Pyrrhalta aenescens) in bioassays. Subsequently, the Cry protein encoded by this novel cry gene was designated Cry1Ie1 by the B. thuringiensis δ-endotoxin nomenclature committee.

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Genetic Basis of Cry1F-Resistance in a Laboratory Selected Asian Corn Borer Strain and Its Cross-Resistance to Other Bacillus thuringiensis Toxins

PLoS ONE ◽

10.1371/journal.pone.0161189 ◽

2016 ◽

Vol 11 (8) ◽

pp. e0161189 ◽

Cited By ~ 10

Author(s):

Yueqin Wang ◽

Yidong Wang ◽

Zhenying Wang ◽

Alejandra Bravo ◽

Mario Soberón ◽

...

Keyword(s):

Bacillus Thuringiensis ◽

Genetic Basis ◽

Cross Resistance ◽

Corn Borer ◽

Asian Corn Borer

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Characterization of the Cry1Ah resistance in Asian corn Borer and its cross-resistance to other Bacillus thuringiensis toxins

Scientific Reports ◽

10.1038/s41598-017-18586-2 ◽

2018 ◽

Vol 8 (1) ◽

Cited By ~ 16

Author(s):

Muhammad Zeeshan Shabbir ◽

Yudong Quan ◽

Zhenying Wang ◽

Alejandra Bravo ◽

Mario Soberón ◽

...

Keyword(s):

Bacillus Thuringiensis ◽

Cross Resistance ◽

Corn Borer ◽

Asian Corn Borer

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Enhancement of Activity of Bacillus thuringiensis Berliner Against Four Lepidopterous Insect Pests by Nutrient-Based Phagostimulants1

Journal of Entomological Science ◽

10.18474/0749-8004-30.1.29 ◽

1995 ◽

Vol 30 (1) ◽

pp. 29-42 ◽

Cited By ~ 13

Author(s):

Robert R. Farrar ◽

Richard L. Ridgway

Keyword(s):

Bacillus Thuringiensis ◽

Ostrinia Nubilalis ◽

Helicoverpa Zea ◽

European Corn Borer ◽

Insect Pests ◽

Diamondback Moth ◽

Lethal Dose ◽

Corn Earworm ◽

Corn Borer ◽

Whole Plants

To help improve control of insect pests with microbial insecticides, we investigated the interactions of four commercial, nutrient-based phagostimulants (Pheast [AgriSense], Coax [CCT Corp.], Gusto [Atochem North America, Inc.], and Entice [Custom Chemicides] with Bacillus thuringiensis Berliner and four lepidopterous insect pests (gypsy moth, Lymantria dispar [L.] [Lymantriidae]; corn earworm, Helicoverpa zea [Boddie] [Noctuidae]; European corn borer, Ostrinia nubilalis [Hübner] [Pyralidae]; and diamondback moth, Plutella xylostella [L.] [Plutellidae]). Comparisons were made of treated foliage in Petri dishes in the laboratory and of sprayed whole plants in a greenhouse. In general, phagostimulants increased mortality of all species tested, but no consistent differences among phagostimulants were found for any species. Food consumption was generally lower on the treatments that contained phagostimulants causing the highest rates of mortality, possibly as a result of more rapid ingestion of a lethal dose on these treatments. Reduced rates of feeding by insects on treatments with B. thuringiensis alone were seen, probably due in part to intoxication and, possibly, to behavioral effects as well. Indications of potentially significant interactions between host plants and both B. thuringiensis and phagostimulants also were seen.

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Development and Field Performance of a Broad-Spectrum Nonviable Asporogenic Recombinant Strain of Bacillus thuringiensis with Greater Potency and UV Resistance

Applied and Environmental Microbiology ◽

10.1128/aem.65.9.4032-4039.1999 ◽

1999 ◽

Vol 65 (9) ◽

pp. 4032-4039 ◽

Cited By ~ 27

Author(s):

Vincent Sanchis ◽

Michel Gohar ◽

Josette Chaufaux ◽

Olivia Arantes ◽

Alain Meier ◽

...

Keyword(s):

Bacillus Thuringiensis ◽

Recombinant Strain ◽

European Corn Borer ◽

Low Cost ◽

Tobacco Budworm ◽

Beet Armyworm ◽

Cotton Leaf ◽

Uv Degradation ◽

Corn Borer ◽

Activity Spectrum

ABSTRACT 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 chimericcry1C/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 δ-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. Thecry1C/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-typeB. 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.

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Domain III of Bacillus thuringiensis Cry1Ie Toxin Plays an Important Role in Binding to Peritrophic Membrane of Asian Corn Borer

PLoS ONE ◽

10.1371/journal.pone.0136430 ◽

2015 ◽

Vol 10 (8) ◽

pp. e0136430 ◽

Cited By ~ 5

Author(s):

Dongmei Feng ◽

Zhen Chen ◽

Zhiwen Wang ◽

Chunlu Zhang ◽

Kanglai He ◽

...

Keyword(s):

Bacillus Thuringiensis ◽

Peritrophic Membrane ◽

Corn Borer ◽

Asian Corn Borer ◽

Domain Iii

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Bacillus Thuringiensis for Control of Asian Corn Borer, 1993

Arthropod Management Tests ◽

10.1093/amt/20.1.89 ◽

1995 ◽

Vol 20 (1) ◽

pp. 89-89

Author(s):

Use Schreiner ◽

Liborio Dumaliang

Keyword(s):

Bacillus Thuringiensis ◽

Corn Borer ◽

Asian Corn Borer

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Toxicity and Mode of Action of Bacillus thuringiensis Cry Proteins in the Mediterranean Corn Borer, Sesamia nonagrioides (Lefebvre)

Applied and Environmental Microbiology ◽

10.1128/aem.72.4.2594-2600.2006 ◽

2006 ◽

Vol 72 (4) ◽

pp. 2594-2600 ◽

Cited By ~ 33

Author(s):

Joel González-Cabrera ◽

Gema P. Farinós ◽

Silvia Caccia ◽

Mercedes Díaz-Mendoza ◽

Pedro Castañera ◽

...

Keyword(s):

Bacillus Thuringiensis ◽

Binding Site ◽

Mode Of Action ◽

Membrane Vesicles ◽

Bt Corn ◽

Sesamia Nonagrioides ◽

Cry Protein ◽

Cry Proteins ◽

Corn Borer ◽

Mediterranean Countries

ABSTRACT Sesamia nonagrioides is one of the most damaging pests of corn in Spain and other Mediterranean countries. Bt corn expressing the Bacillus thuringiensis Cry1Ab toxin is being grown on about 58,000 ha in Spain. Here we studied the mode of action of this Cry protein on S. nonagrioides (binding to specific receptors, stability of binding, and pore formation) and the modes of action of other Cry proteins that were found to be active in this work (Cry1Ac, Cry1Ca, and Cry1Fa). Binding assays were performed with 125I- or biotin-labeled toxins and larval brush border membrane vesicles (BBMV). Competition experiments indicated that these toxins bind specifically and that Cry1Aa, Cry1Ab, and Cry1Ac share a binding site. Cry1Ca and Cry1Fa bind to different sites. In addition, Cry1Fa binds to Cry1A's binding site with very low affinity and vice versa. Binding of Cry1Ab and Cry1Ac was found to be stable over time, which indicates that the observed binding is irreversible. The pore-forming activity of Cry proteins on BBMV was determined using the voltage-sensitive fluorescent dye DiSC3(5). Membrane permeability increased in the presence of the active toxins Cry1Ab and Cry1Fa but not in the presence of the nonactive toxin Cry1Da. In terms of resistance management, based on our results and the fact that Cry1Ca is not toxic to Ostrinia nubilalis, we recommend pyramiding of Cry1Ab with Cry1Fa in the same Bt corn plant for better long-term control of corn borers.

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