Chitinolytic Bacteria Isolated from Chili Rhizosphere: Chitinase Characterization and Application as Biocontrol for Aphis gossypii

*Chitin, a common constituent of insect exoskeleton, could be hydrolyzed by chitinase. This research was conducted to select rhizobacteria isolated from the rhizosphere of chili pepper that produced chitinase and to examine their chitinase activity in degrading chitin of the . A total of 25 rhizobacteria isolates formed a clear zone when grown on chitin agar. Three of them had the highest chitinolytic index and were identified as sp. strain I.5, I.21, and II.14. The II.14 was chosen for characterization of chitinase activity. The isolate showed maximum chitinase activity at 48-h-incubation. Maximum temperature and pH of the chitinase activity were 55°C and 7.0, respectively. The cell culture and the enzyme crude extract of the above three isolates were tested against and the result was compared to the control through microscopic observation. Hydrolytic analysis showed that the enzyme crude extract of these isolates were able to degrade chitin of insect exoskeleton since the first 3-h-incubation. Meanwhile, the cell culture treatment on the chitin showed degrading activity after 12 h ( sp. strain I.21 and II.14), and 9 h ( sp. strain I.5). Chitin degradation of exoskeleton by enzyme crude extract was better than the cell culture treatment. Chitinases produced by sp. strains I.5, I.21, and II.14 are potential as biocontrol agents for .

[1]  Iman Rusmana,et al.  Chitinolytic Bacteria Isolated from Chili Rhizosphere: Chitinase Characterization and Its Application as Biocontrol for Whitefly (Bemisia tabaci Genn.) , 2010 .

[2]  N. Balasubramanian,et al.  Optimization of cultural conditions for production of chitinase by Bacillus laterosporous MML2270 isolated from rice rhizosphere soil , 2008 .

[3]  N. R. Mubarik,et al.  Utilization of Root-Colonizing Bacteria to Protect Hot-Pepper Against Tobacco Mosaic Tobamovirus , 2007 .

[4]  H. Schwarz,et al.  Electron-microscopic and genetic dissection of arthropod cuticle differentiation , 2007 .

[5]  J. D. Funkhouser,et al.  Chitinase family GH18: evolutionary insights from the genomic history of a diverse protein family , 2007, BMC Evolutionary Biology.

[6]  H. Liao,et al.  Bioconversion of shellfish chitin wastes for the production of Bacillus subtilis W-118 chitinase. , 2006, Carbohydrate research.

[7]  F. Meinhardt,et al.  Isolation and Molecular Characterization of Chitinase-Deficient Bacillus licheniformis Strains Capable of Deproteinization of Shrimp Shell Waste To Obtain Highly Viscous Chitin , 2006, Applied and Environmental Microbiology.

[8]  M. G. Aguilar-Uscanga,et al.  Purification and characterization of an exochitinase from Bacillus thuringiensis subsp. aizawai and its action against phytopathogenic fungi. , 2006, Canadian journal of microbiology.

[9]  P. Moore,et al.  Improved Carica papaya Tolerance to Carmine Spider Mite by the Expression of Manduca sexta chitinase Transgene , 2006, Transgenic Research.

[10]  N. Novak,et al.  Expression of Poplar Chitinase in Tomato Leads to Inhibition of Development in Colorado Potato Beetle , 2006, Biotechnology Letters.

[11]  S. Jaoua,et al.  Molecular characterization of a novel chitinase from Bacillus thuringiensis subsp. kurstaki , 2005, Journal of applied microbiology.

[12]  Shu-Chun Chung,et al.  Identification of an antifungal chitinase from a potential biocontrol agent, Bacillus cereus 28-9. , 2005, Journal of biochemistry and molecular biology.

[13]  Rakesh Kumar,et al.  Detection of pathogenesis-related proteins-chitinase and β-1,3-glucanase in induced chickpea , 2005 .

[14]  Y. Pyun,et al.  Characteristics of thermostable chitinase enzymes from the indonesian Bacillus sp.13.26 , 2004 .

[15]  Chien-Jui Huang,et al.  Enhancement of the antifungal activity of Bacillus subtilis F29-3 by the chitinase encoded by Bacillus circulans chiA gene. , 2004, Canadian journal of microbiology.

[16]  H. Merzendorfer,et al.  Chitin metabolism in insects: structure, function and regulation of chitin synthases and chitinases , 2003, Journal of Experimental Biology.

[17]  A. Fereres,et al.  Settlement rate of Aphis gossypii (Hemiptera, Aphididae) and transmission efficiency of Cucumber mosaic virus in melons protected with kaolin-particle films , 2003 .

[18]  J. Prosser,et al.  Molecular Analysis of a Bacterial Chitinolytic Community in an Upland Pasture , 2002, Applied and Environmental Microbiology.

[19]  Y. Pyun,et al.  Biochemical characteristics of chitinase enzyme from Bacillus sp. of Kamojang Crater, Indonesia. , 2002, Journal of Biochemistry, Molecular Biology and Biophysics.

[20]  B. Lugtenberg,et al.  Molecular basis of plant growth promotion and biocontrol by rhizobacteria. , 2001, Current opinion in plant biology.

[21]  K. Mavromatis,et al.  Cloning, Sequences, and Characterization of Two Chitinase Genes from the Antarctic Arthrobacter sp. Strain TAD20: Isolation and Partial Characterization of the Enzymes , 2001, Journal of bacteriology.

[22]  Patil,et al.  Chitinolytic enzymes: an exploration. , 2000, Enzyme and microbial technology.

[23]  San-Lang Wang,et al.  Purification and characterization of two bifunctional chitinases/lysozymes extracellularly produced by Pseudomonas aeruginosa K-187 in a shrimp and crab shell powder medium , 1997, Applied and environmental microbiology.

[24]  M. Wakayama,et al.  Purification and characterization of thermostable beta-N-acetylhexosaminidase of Bacillus stearothermophilus CH-4 isolated from chitin-containing compost , 1994, Applied and environmental microbiology.

[25]  S. Sabry.  Microbial degradation of shrimp‐shell waste , 1992, Journal of basic microbiology.

[26]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[27]  R. E. Buchanan,et al.  Bergey's Manual of Determinative Bacteriology. , 1975 .

[28]  S. T. Cowan.  Bergey's Manual of Determinative Bacteriology , 1948, Nature.

[29]  J. Brown.  Bergey's Manual of Determinative Bacteriology (5th ed.) , 1939 .