Isolation of BT and analysis of ICP gene in Indonesia.
Grant-in-Aid for international Scientific Research
|Allocation Type||Single-year Grants|
|Research Institution||Hokkaido University|
IIZUKA Toshihiko Hokkaido University, Fac.of Agriculture, Prof., 農学部, 教授 (50001441)
LAY Bibiana w. Institut Pertanian, Bogor, Ass.Prof., 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, Ass.Prof., 農学部, 助教授 (20189731)
|Project Period (FY)
1993 – 1994
Completed(Fiscal Year 1994)
|Budget Amount *help
¥5,500,000 (Direct Cost : ¥5,500,000)
Fiscal Year 1994 : ¥3,000,000 (Direct Cost : ¥3,000,000)
Fiscal Year 1993 : ¥2,500,000 (Direct Cost : ¥2,500,000)
|Keywords||Insecticidal crystal protein (ICP) / Bacillus thuringiensis / Crystal protein gene / Microbiological control of insect / Soil bacteria / Bacillus thuringiensis / ICP遺伝子 / PCR / DNAプローブ|
Bacillus thuringiensis (BT) is known as the bacteria group which produces the insecticidal crystal proteins (ICP) and has been used for microbiological control of plant-destroying insect because of its highly restricted host specificity. Nowadays, new Bts with unique characteristic traits have been isolated from various areas in the world. Aim of our project is to isolate Bts from some areas in Indonesia where is a habitant of huge numbers of insect species. In the last year, we isolated many Bts from the soil under big trees as reported. This year, we tried to obtain further Bts from several areas and studied its biological characteristics.
As a result, we obtained more than 30 BT isolates form about 50 areas containing forest and sericultural field. In the H-serotyping, the isolates were classified into subspp. thuringiensis, sotto, darmstadiensis, aizawai but some were untypable. In the two years-collaboration, we obtained more than 60 BT isolates already and analyzed gene constructi
on especially on crystal protein (cry) gene by PCR method established in our laboratory. Furthermore, the host specificity of the isolates were studied. The results of the experiments showed that the classification by H-serotype is not always reflected in the grouping of BT by its host spectrum. For example, some isolates obtained from Indonesia had the insecticidal activity against Spodoptera litura but it was classified into subsp. kurstaki which was a group of BT without insecticidal activity against the insect. One of those isolates, INA02, had the cryV gene which seemed to have an activity against S.litura. Then we cloned and determined the nucleotide sequence of the cryV gene of INA02.
On the other hand, we found another new type of BT which had the insecticidal activity against Aedes japonicus. We identified a cryII gene in this isolate (SKW isolate) which showed the sequence homology to the cryIIA gene. The function of this gene is under investigation. Another isolate (INA67) which was classified into subspp. tenebrionis showed the insecticidal activity against a coleoptera. However, this isolate contained no cryIII or V gene which were reported as the coleopteracidal crystal protein genes. Less
Research Output (10results)