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Bacillus Thurungiensis isolates from indonesia toxic to mosquito insects

Research Paper | March 1, 2018

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Akhmad Rizali

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Int. J. Biosci.12( 3), 154-161, March 2018

DOI: http://dx.doi.org/10.12692/ijb/12.3.154-161


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B. thuringiensis has been studied worldwide over the past decades, mainly because this gram-positive bacterium produce significant amount of crystal proteins with toxic activity against economically important insect larvae. Most strains of B. thuringiensis produce delta-endotoxin crystals toxic to lepidopteran insects such moth. But some strains of B. thuringiensis produce delta-endotoxin crystals toxic to dipteran insects such as mosquitoes and blackflies. One gram of soil samples was suspended in 9ml of sterile distilled water and shaken for 5min. the upper layer of the soil suspension was transferred to a test tube and heated at 80oC for 5min in water bath to kill non-spore-forming organism and vegetative cells.to prepare the sporulated culture, bacteria were grown on nutrient agar pH 7.2, at 30oC for 4 days., it was observed and photographed with (SEM). The results that serovar entomocidus (original starin) produced bipyramidal-formed and entomocidus INA288 produced large cuboidal-form crystals. The PCR screening showed that cry4 Aa had a novel mosquitocidal cry gene and had 70 kDa peptide, but serovar entomocidus (original strain) encodes only cry1Aa, cry1Ab, cry1B and cryIC, which have not been thought to be dipteracidal activity.they had differences form crystal protein, and unique as insecticidal to control the same orders (mosquitocidal).


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Bacillus Thurungiensis isolates from indonesia toxic to mosquito insects

Aroson A. 2002. Sorulation and delta-endotoxin synthesis by Bacillus thuringiensis. Cell Mol. Life Sci 59, 417-425.

Asano S, Bando H, Iizuka T. 1993. Amplification and identification of cryII genes from Bacilus thuringiensis by PCR procedures. J. Seric. Sci. Jpn 62, 223-227.

Asano S. 1996. Identification of cry gene from Bacillus thuringiensis by PCR and isolation of unique insecticidal bacteria. Mem. Fac. Agric. Hokkaido Univ. 19, 529-563.

Baba F, Asano S, Iizuka T. 1990. Purification of crystals from Bacillus thurigiensisby using Percoll. J. Sci. Jpn 59, 487-489.

Balarman K, Hoti SL, Manonmani LM. 1981. An Idigenous virulent strain of Bacillus thuringiensis, highly pathogenic and specific to mosquitoes. Current Science 50, 199-200.

Boonserm P, Mo M, Angsuthana Sombat C, Lescar J. 2006. Structure of the functional form of the mosquito larvicidal cry 4Aa toxin from Bacillus thuringiensis at 2.8-angstrom resolution. J. Bacteriol 188, 3391-3401.

Bourquet S. 2004. Resistance to Bacillus thuringiensis toxin in the Eruropean corn borer: what chance for Bacillus thuringiensis maize. Physiol. Entomol 29, 251-256.

Bulla L, Jr A, Kramer KJ, Davidson LI. 1977. Characterization of the enmocidalparasporal crystal of Bacillus thuringiensis. J. Bacteriol 130, 375-383.

Bulla LA, faust RM, Andrews R, Goodman N. 1985. Insecticidal bacilli, pp. 185-209. In: the Moleculer Biology of the Bacilli, Volume II. D. A. Dubnau (Ed.). Academic Press, New York.

Burges HD. 2001. Bacillus thuringiensis in pest control: now and the future, Pest outlook 12, 90-97.

Delucca AJ, II, Simonson JG, Larson AD. 1981. Bacillus thuringiensis distribution in soils of the United States. Canadian J. Maicrobiol 27, 865-870.

Dulmage HT. 1992. Insecticidal activity of Bacillus thuringiensis and their potential for pest control in Microbial control for pests and plant diseases and plant diseases 1970-1980 (Ed.H.D Burges). Acad. Press. N.Y PP.

Golberg LJ, Margalit J. 1977. A bacterial spore demonstrating rapid larvicidal activity against Anopheles sergentii, Uranotaenia unguiculata, Culex univitattus, Aedes aegypti and Culexpipiens, Mosq. New 37, 355-358.

Hastowo S, Lay BW, Ohba M. 1992. Naturally occurring Bacillus thuringiensis in Indonesia. J. Appl. Bacteriol 73, 108-113.

Heimpel AM. 1967. A critical review of Bacillus thuringiensis Berl.And other crystalliferous bacteria. Ann. Rev. Entomol 12, 287-322.

Held GA, Kawanishi CY, Huang YS. 1990. Characterization of the parasporal inclusion of Bacillus thuringiensis subsp. Kyushuensis. J. Bacteriol 481-483.

Iizuka T, Ishino M, Nakajima T. 1982. Comparative morphology of Parasporal crystal and characterization of plasmid DNA from various subspecies of entomopathogenic bacteria, Bacillus thuringiensis. J. Fac. Agric. Hokkaido Univ 13, 423-431.

Iizuka T, Sasaki J, Asano S, Bando H. 1995. Comparative studies on isolation and identification of Bacillus thuringiensis. Biotechnology and Enviro. Benefits, Vol. I, 143-153.

Iizuka T, Yamamoto T. 1984. Serological properties of the mosquitocidal protein of Bacillus thuringiensis and the morphology of its parasoral crystal. J. fac. Hokkaido Univ. 62, 98-114.

Ishii T, Ohba M. 2013. Investigation of mosquito-specific larvicidal activity of a soil isolate of Bacillus thuringiens is serovar Canadensis. Curr. Microbiol 35, 40-43.

Kalman S, Kiehne KK, Libs JL, Yamamoto T. 1993. Cloning of novel cryIC-type gene from a strain Bacillus thuringiensis subs. Galleriae. Appl. Enviro. Microbio 59, 1131-1137.

Kawalek MD, Benjamin S, Lee HL, Gill SS. 1995. Isolation and identification of novel toxin from a new mosquitocidal isolate from Malaysia, Bacillus thuringiensis subsp. Jegathesan. Apl. Enviro. Microbiol 2965-2969.

Kim K. Ohba H, aizawa K. 1984. Purification of the toxic protein from Bacillus thuringiensis serotype 10 isolate demonstrating a preferential larvicidal activity to mosquito. J. Invertebr. Pathol 44, 214-219.

Kreig A, Huger A, Langenbruch G, Schentter W. 1983. Bacillus thurigiensis isolate with activity againtColeoptera. In Biotechnology in invertebrate pathology and cell culture. KarlMaramorosch (ed.) p. 101-114.

Laemmli UK. 1970. Cleavage of structural proteins during tha assembly of the head of bacteriophage T4. Nature (London) 277, 680-685.

Lee SG, Eckblad W, Bulla L, Jr A, 1985. Diversity of protein inclusion bodies and identification of mosquitocidal protein in Bacillus thuringiensis subsp. Israelensis. Biochem. Biophys. Res. Commun 126,953-960.

Lopez-Pazos SA, Martinez JM, Castilo AX, Samanca JAC. 2009. Present and significant of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleotera: Curculionidae).

Ohba M, Aizawa K. 1986. Insect toxicity of Bacillus thuringiensis isolated from soils of Japan. J. Invertebr. Pathol 47, 12-20.

Padua LE, Ohba M, Aizawa K. 1984. Isolation of a Bacillus thuringiensis strain (serotype 8a:8b) highly and selectively toxic against mosqouito larvae. J. Invertebr. Pathol 44, 12-17.

Pfannenstiel MA, Ross EJ, Kramer VC, Nickerson KW. 1984. Toxicity and composition of protease-inhibited Bacillus thuringiensis var. israelensiscrystal. FEMSMicrobio.Lett 21, 39-42.

Poopathi S, Abidha S. 2010. Mosquitocidal bacterial toxin (Bacillus spaeharicus and Bacillus thuringiensis serovarisraelensis): mode of action, cytopathological effects and mechanism of resistance.

Rizali A, Shin-ichiro Asano, Ken Sahara, Hisanori Bando, Bibiana W, Lay, Sugyo Hastowo and Toshihiko Iizuka. 1998. Novel Bacillus thuringiensis serovaraizawai strains isolated from mulberry leaves in Indonesia. Appl. Entomol. Zool 33(1), 111-114.

Shorey HH, Hall IM. 1962. Effect of chemical and microbial insecticides on several insect pests of lettuce in southern California. J. Econ. Entomol 56, 169-174.

Siegel JP. 2000. Bacteria in field manual of techniques in Invertebrate pathology (Lacy, L.L. and Kaya, H. K. Eds). Pp. 209-230. Dordrechat, Netherlands: Kluwer Scientific Publishers.

Siegel JP. 2001. The mammalian safety of Basillusthuringiensis. Basedinsecticides. J. inverteb. Pathol 77, 13-21.

Yamamoto T, McLaughlin RE. 1981. Islation of a protein from the parasporal crystal of Bacillus thuringiensis var. kurstaki toxic to the mosquito larva, Aedes taeniorhynchus. Biochem. Biophys. Res. Commun 103, 414-421.

Yu Y–M, Ohba M, Gill SS. 1991. Characterization of mosquitocidal activity of Bacillus thuringiensis subsp. Fukuokaensis crystal proteins. Appl. Enviro. Microbiol 1075-1081.