Hostname: page-component-cd9895bd7-mkpzs Total loading time: 0 Render date: 2024-12-28T13:54:36.930Z Has data issue: false hasContentIssue false

Identification of the cry gene in Bacillus thuringiensis strain WZ-9 and its toxicity against Henosepilachna vigintioctomaculata

Published online by Cambridge University Press:  03 March 2009

Song Ping
Affiliation:
College of Plant Protection, Biocontrol Center of Plant Diseases and Plant Pests of Hebei Province, Agricultural University of Hebei, Baoding 071001, China
Wang Qin-Ying*
Affiliation:
College of Plant Protection, Biocontrol Center of Plant Diseases and Plant Pests of Hebei Province, Agricultural University of Hebei, Baoding 071001, China
Wu Hui-Xian
Affiliation:
College of Plant Protection, Biocontrol Center of Plant Diseases and Plant Pests of Hebei Province, Agricultural University of Hebei, Baoding 071001, China
Lu Xiu-Jun
Affiliation:
College of Plant Protection, Biocontrol Center of Plant Diseases and Plant Pests of Hebei Province, Agricultural University of Hebei, Baoding 071001, China
Wang Yong
Affiliation:
Central Laboratory of Tianjin Academy of Agricultural Sciences, Tianjin 300192, China
*
*Corresponding author. E-mail: wqinying@hebau.edu.cn

Abstract

Bacillus thuringiensis strain WZ-9, isolated from soil in Hebei province, China, was effective against Henosepilachna vigintioctomaculata larvae. The strain presented bipyramidal crystals with a protein band of 130 kDa in SDS–PAGE. The pH changes of the culture media showed important fluctuations during the 24 h growth cycle. The pH varied less in log and stationary phases than it did in the exponential phase. Bioassay results showed that the WZ-9 strain was only harmful to larvae of H. vigintioctomaculata and not to either adults of H. vigintioctomaculata or other several lepidopteran and coleopteran insects. LC50 to second-instar larvae of H. vigintioctomaculata was 2.95×107 cells/ml after 72 h. Genotypic investigations showed that this strain possessed the cry7 gene. Sequence analysis demonstrated that the encoding gene contained an open reading frame (ORF) of 3414 bp and encoded 1138 amino acid residues. The deduced amino acid sequence was 99.65% identical to that of the reported Cry7Ab2 sequences. This gene was designated by the Bt δ-endotoxin nomenclature committee as Cry7Ab3 with accession number BI 1015188 in the GenBank database.

Type
Research Papers
Copyright
Copyright © China Agricultural University 2008

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

First published in Journal of Agricultural Biotechnology 2008, 16(3): 515–520

References

Chen, J, Wang, XP, Lu, MZ, Du, MF and Yin, XM (2004) Selection of Bacillus thuringiensis strain against the longhorned beetles and preliminary characterization of its insecticidal gene. Scientia Silvae Sinicae 40(5): 138142 (in Chinese with English abstract).Google Scholar
Chen, J, Dai, LY, Wang, XP, Tian, YC and Lu, MZ (2005) The cry3Aa gene of Bacillus thuringiensis Bt886 encodes a toxin against long-horned beetles. Applied Microbiology and Biotechnology 67(3): 351356.CrossRefGoogle ScholarPubMed
Crowson, RA (1981) The Biology of the Coleoptera. London: Academic Press.Google Scholar
Dou, LM, Han, LL, Zhang, J, et al. (2007) Cloning, expression and activity of cry1Ia gene from Bacillus thuringiensis isolate. Journal of Agricultural Biotechnology 15(6): 10531057 (in Chinese with English abstract).Google Scholar
Feng, SL, Wang, RY, Fan, XH and Hu, MJ (2000) A new Bacillus thuringiensis strain with high insecticidal activity against scarabaeid species. Chinese Journal of Biological Control 16(2): 7477 (in Chinese).Google Scholar
Gao, MY, Li, RS, Dai, SY, Li, XG and Fu, JH (1999) New coleopterancidal strains of Bacillus thuringiensis and production of coleopterancide. Acta Microbiologica Sinica 39(6): 515520 (in Chinese with English abstract).Google ScholarPubMed
Krieg, A, Huger, AM and Langenbruch, GA (1983) Bacillus thuringiensis subsp. tenebrionsis, a new pathotype effective against larvae of coleoptera. Journal of Applied Entomology 96: 500508.Google Scholar
Laemmli, UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680685.CrossRefGoogle ScholarPubMed
Lambert, B, Höfte, H, Annys, K, Jansens, S, Soetaert, P and Peferoen, M (1992) Novel Bacillus thuringiensis insecticidal crystal protein with a silent activity against coleopteran larvae. Applied and Environmental Microbiology 58(8): 25362542.CrossRefGoogle ScholarPubMed
López-Meza, JE and Ibarra, JE (1996) Characterization of a novel strain of Bacillus thuringiensis. Applied and Environmental Microbiology 62(4): 13061310.CrossRefGoogle ScholarPubMed
Narva, KE, Payne, JM, Schwab, GE, Hickle, LA, Galasan, T and Sick, AJ (1991) Novel Bacillus thuringiensis microbes active against nematodes, and genes encoding novel nematodes – active toxins cloned from Bacillus thuringiensis isolates. European Patent Office, EP0462721.Google Scholar
Ruud, A, Maagd, DE, Bravo, A and Crickmore, N (2001) How Bacillus thuringiensis has evolved specific toxins to colonize the insect world. Trends in Genetics 17(4): 193199.Google Scholar
Sato, R, Takeuchi, K, Ogiwara, K, et al. (1994) Cloning, heterologous expression, and localization of a novel crystal protein gene from Bacillus thuringiensis serovar Japonensis strain buibui toxic to scarabaeid insects. Current Microbiology 28(1): 1519.CrossRefGoogle ScholarPubMed
Schnepf, E, Crickmore, N, Vanrie, J, et al. (1998) Bacillus thuringiensis and its pesticidal crystal proteins. Microbiology and Molecular Biology Reviews 62(3): 775806.CrossRefGoogle ScholarPubMed
Schnepf, HE, Lee, S, Dojillo, J, et al. (2005) Characterization of Cry34/Cry35 binary insecticidal proteins from diverse Bacillus thuringiensis strain collections. Applied and Environmental Microbiology 71(4): 17651774.CrossRefGoogle ScholarPubMed
Storer, NP, Babcock, JM and Edwards, JM (2006) Field measures of western corn rootworm (Coleoptera: Chrysomelidae) mortality caused by Cry34/35Ab1 proteins expressed in maize event 59122 and implications for trait durability. Journal of Economic Entomology 99(4): 13811387.CrossRefGoogle ScholarPubMed
Su, XD (2005) Isolation of Bacillus thuringiensis strains and identification of their cry genes. Masters Degree, thesis of Agricultural University of Hebei, Baoding, China.Google Scholar
Yu, H, Zhang, J, Huang, D and Song, FP (2006) Characterization of Bacillus thuringiensis strain Bt185 toxic to the Asian cockchafer: Holotrichia parallela. Current Microbiology 53(1): 1317.CrossRefGoogle Scholar
Zhang, J, Song, FP, Li, CY, Chen, ZY, Tan, JX and Huang, DF (2002) Cloning and expression of cry3Aa7 gene from Bacillus thuringiensis strain toxic to coleopteran pests. Scientia Agricultura Sinica 35(6): 650653 (in Chinese with English abstract).Google Scholar