Isolation and Identification of a Bacillus thuringiensis Strian

doi:10.7668/hbnxb.2007.04.043

Abstract

Abstract: In this paper a Bacillus thuringiensis strain was isolated from the soil sample of Diaoling Mountain in Yunnan province. The cry gene-types of Bt isolate was identified by use of PCR-RFLP and SDS-PAGE methods. This strain which had a cry-type includes cry1Aa, cry1Ac, cry1Db, cry2Ab, cry1Ia produced Bipyramidal shape crystal, the protein composed of the crystal were 130, 70 and 56 kDa, and exhibited insecticidal activity against Laphygma exigua (first-instar), LC₅₀ was 1.43 ng/mg. It can be used in composing the bacteria of gene engineering.

Key words: Bacillus thuringiensis, Isolation, Insecticidal gene, Identification

CLC Number:

S432

SU Xu-dong, MA Wen, MA Xiao-yan, WANG Xue-jing, LI Ying-jun, TAN Jian-xin. Isolation and Identification of a Bacillus thuringiensis Strian[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2007, 22(4): 184-188. doi: 10.7668/hbnxb.2007.04.043.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2007.04.043

http://www.hbnxb.net/EN/Y2007/V22/I4/184

References

[1] Ishiwata S. On a kind of severe flacherie(sotto disease)[J]. Dainihon Sanshi Kaiho, 1901, 114: 1-5.
[2] Pinto L M, Azambuja A O, Diehl E. Pathogenicity of Bacillus thuringieusis isolated from two species of Acromyrmex (Hymenoptera, Formicidae)[J]. Braz J Biol, 2003, 63 (2): 301-306.
[3] Schnepf E, Crickmore N, Rie J V, et al. Bacillus thuringiensis and its pesticidal crystal proteins[J]. Microbiol Mol Biol Rev, 1998, 62: 775-806.
[4] Sayyed A H, Raymond B, Ibiza-Pallcios M, et al. Genetic and biochemical characterization of field-evolved resistance to Bacillus thuringiensis toxin Cry1Ac in the diamondback moth, Plutella xylostella[J]. Appl Environ Microbiol, 2004, 70(12): 7010-7017.
[5] Prabagaran S R, Nimal S J, Jayachandran S. Phenotypic and genetic diversity of Bacillus thuringiensis strains isolated in India active against Spodoptera litura[J]. Appl Biochem Biotechnol, 2002, 102-103(1-6): 213-226.
[6] Uribe D, Martinez W, Ceron J. Distribution and diversity of cry genes in native strains of Bacillus thuringiensis obtained from different ecosystems from Colombia[J]. J Invertebr Pathol, 2003, 82 (2): 119-127.
[7] Vilas-Boas G T, Lemos M V. Diversity of cry genes and genetic characterization of Bacillus thuringiensis isolated from Brazil[J]. Can J Microbiol, 2004, 50(8): 605-613.
[8] Wang J, Boets A, Van R J, et al. Characterization of cry1, cry2, and cry9 genes in Bacillus thuringiensis isolates from China[J]. J Invertebr Pathol, 2003, 82 (1): 63-71.
[9] Chen F C, Tsai M C, Peng C H, et al. Dissection of cry gene profiles of Bacillus thuringiensis isolates in Taiwan[J]. Curt Microbiol, 2004, 48(4): 270-275.
[10] Kuo W S, Chak K F. Identification of novel cry-type genes from Bacillus thuringiensis strains on the basis of restriction fragment length polymorphism of the PCR-amplified DNA[J]. Appl Envir Microbiol, 1996, 62: 1369-1377.
[11] Song F P, Zhang J, Gu A, et al. Identification of cry1I-type genes from Bacillus thuringiensis strains and characterization of a novel cry1I-type gene[J]. Appl Envir Microbiol, 2003, 69: 5207-5211.
[12] Song F P, Zhang J, Huang D F, et al. Establishment of PCRRFLP identification system of cry genes from Bacillus thuringiensis[J]. Scientia Agricultura Sinica, 1998, 31 (3): 13-18.
[13] 翟兴礼, 杨霞. Bt与无机物添加剂混合对菜青虫杀虫效果的研究[J]. 河南农业科学, 2005, 7: 20-22.
[14] 王容燕, 冯书亮, 范秀华. 对金龟子幼虫有杀虫活性的苏云金杆菌HBF-1菌株发酵培养基优选[J]. 华北农学报, 2003, 18(院庆专辑): 96-98.
[15] 郭亚平, 马恩波, 任竹梅. 苏云金杆菌制剂防治细胸金针虫的初步研究[J]. 华北农学报, 2001, 16(2): 108-112.
[16] 尹新明, 郭线茹, 贾新成. 提高苏云金杆菌对棉铃虫毒力的研究[J]. 华北农学报, 1999, 14(2): 1-5.
[17] 张永安, 宋福平, 戴莲韵, 等. 我国不同森林立地带土壤中苏云金芽孢杆菌cry基因资源研究[J]. 林业科学研究, 2003, 16(1): 19-25.
[18] 李长友, 袁强. 我国不同森林立地带Bt分离株杀虫晶体蛋白及基因分析[J]. 林业科学, 2002, 38(3): 102-105.
[19] 黄志鹏, 关雄. 苏云金杆菌新菌株WB9 cryl基因型的PCR-RFLP分析[J]. 福建农业大学学报, 2001, 30(3): 304-308.
[20] 张杰, 宋福平, 左雅慧, 等. 31株苏云金芽孢杆菌杀虫晶体蛋白基因型鉴定及表达产物研究[J]. 微生物学报, 2000, 40(4), 372-378.
[21] Yu H, Zhang J, Huang D, et al. Characterization of Bacillus thuringiensis strain Bt185 toxic to the asian cockchafer: Holotrichia parallela[J]. Curr Microbiol, 2006, 53 (1): 13-17.
[22] 冯书亮, 范秀华, 王容燕, 等. 苏云金杆菌在华北果园土壤中消长动态的研究[J]. 中国病毒学, 2000, 25: 107-111.
[23] Bravo A, Sarabia S, Lopez L, et al. Characterization of cry genes in a Mexican Bacillus thuringiensis strain collection[J]. Appl Envir Microbiol, 1998, 64: 4965-4972.
[24] Van Rie J. Bacillus thuringiensis and its use in transgenic insect control technologies[J]. Int J Med Microbiol, 2000, 290: 463-469.

Please choose a citation manager

Content to export