Isolation of BT and analysis of ICP gene in Indonesia.

Research Project

Project/Area Number	07044173
Research Category	Grant-in-Aid for international Scientific Research
Allocation Type	Single-year Grants
Section	Joint Research
Research Institution	Hokkaido University
Principal Investigator	IIZUKA Toshihiko Hokkaido University, Fac. of Agriculture, Prof., 農学部, 教授 (50001441)
Co-Investigator(Kenkyū-buntansha)	HASTOWO Sugyo Institut Pertanian, Bogor, 準教授 LAY Bibiana W. Institut Pertanian, Bogor, 準教授 SAHARA Ken Hokkaido University, Fac. of Agriculture, Instructor, 農学部, 助手 (30241368) ASANO Shinichiro Hokkaido University, Fac. of Agriculture, Instructor, 農学部, 助手 (60222585) BANDO Hisanori Hokkaido University, Fac. of Agriculture, Asso. Prof., 農学部, 助教授 (20189731)
Project Period (FY)	1995
Project Status	Completed (Fiscal Year 1995)
Budget Amount *help	¥2,600,000 (Direct Cost: ¥2,600,000) Fiscal Year 1995: ¥2,600,000 (Direct Cost: ¥2,600,000)
Keywords	Insecticidal crystal protein(ICP) / Bacillus thuringiensis / Crystal protein gene(cry) / Microbiological control of insect / Soil bacterium
Research Abstract	Bacillus thuringiensis(Bt)is a gram-positive soil bacterium distinguished from other bacilli by its production of insecticidal protein. Because of their strong insecticidal activity and highly restricted host specificity, Bt has been used for microbiological control of plant-destroying insect. Recently, new Bts with unique characteristic traits have been isolated from various area in the world. Aim of our project is to isolate Bts with unique characteristics from Indonesia where is the habitant of huge number of insect species. In the last two years, we isolated many Bts from the soil collected from more than 50 areas in Indonesia. This year, we tried to obtain further Bts with unique properties from the areas where we have been able to obtain Bts before and characterized them. We newly obtained 17 Bt strains from 6 areas including forests and sericultural farms, resulting in obtaining more than 80 Bt isolates in the three years-collaboration. In the H-serotyping, the isolates were classified into three subspecies, alesti, kenyae and indiana. It was demonstrated by the following researches such as analysis of ICP by SDS-PAGE,ICP gene-typing by PCR method established in our Lab, and bio-assay of the host specificity that our Bt collection contained three unique isolates which had never been reported. The results from the studies on the properties of these three unique Bt isolates lead us to the tentative conclusions that the minor mutation in the ICP gene results in the change of host specificity and that there is new ICP gene which has quite different nucleotide sequence in nature.