İsmail DEMİR, Emine ERYÜZLÜ, Zihni DEMİRBAĞ

A study on the characterization and pathogenicity of bacteria from Lymantria dispar L. (Lepidoptera: Lymantriidae)

Gypsy moth, Lymantria dispar L. (Lepidoptera: Lymantriidae), is one of the most serious pests affecting fruit, forest, industrial, and foliage plants worldwide. The aim of this study was to isolate and characterize pathogenic bacteria from this pest and determine their pathogenicity. L. dispar larvae were collected from various populations in the forest areas of the Black Sea region of Turkey in May and June of 2008 and 2009, and 9 culturable bacterial isolates were obtained from these larvae. Morphological and physiological characteristics of bacterial isolates were determined by conventional and routine techniques, and biochemical properties and metabolic enzyme profiles were revealed using API20E and Phoenix 1000A panel test systems. Additionally, a 16S rRNA gene sequence analysis was performed to identify the isolates at the molecular level and to query against the NCBI genetic database. Consequently, 7 of the bacterial isolates were determined and characterized at the species level while the rest were characterized at the genus level. From the results of all tests, the isolates were identified as Pantotea agglomerans (Lyd1), Klebsiella sp. (Lyd2), Acinetobacter calcoaceticus (Lyd3), Serratia marcescens (Lyd4), Erwinia sp. (Lyd5), Bacillus thuringiensis (Lyd6), B. thuringiensis (Lyd7), B. thuringiensis (Lyd8), and B. thuringiensis (Lyd9). Of all bacterial isolates tested for insecticidal activity, 2 of the B. thuringiensis isolates (Lyd6 and Lyd9) were found to cause the highest insecticidal activity (90%) within 10 days of application. Furthermore, 100% insecticidal activity was observed with a crystal-spore mixture of the same isolates against Lymantria dispar larvae within 10 days.

Anahtar Kelimeler:

Key words: Lymantria dispar, culturable bacterial flora, insecticidal activity, Bacillus thuringiensis

A study on the characterization and pathogenicity of bacteria from Lymantria dispar L. (Lepidoptera: Lymantriidae)

Keywords:

Key words: Lymantria dispar, culturable bacterial flora, insecticidal activity, Bacillus thuringiensis,

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Elkinton JS, Liebhold AM. Population dynamics of gypsy moth in North America. Annu Rev Entomol 35: 571-596, 1990.
Ministry of Agriculture of Turkey. Th e Agricultural Control Technical Recommendations, Vol. 3. Başak Publisher. Ankara; 2008: pp. 182-184.
Wronski, S. Th e Eff ects of Chemical Control Agents Used Against the Gypsy Moth. University of California. Irvine, CA, USA; 1997. Available from http://www.dbc.uci.edu/sustain/ global/sensem/wronski97.html. Accessed 21.03.2011. Ince IA, Demir I, Demirbag Z et al. A cytoplasmic polyhedrosis virus isolated from the pine processionary caterpillar, Th aumetopoea pityocampa. J Microbiol Biotechnol 17: 632- 637, 2007.
Sevim A, Demir I, Höft e M et al. Isolation and characterization of entomopathogenic fungi from hazelnut-growing region of Turkey. Biocontrol 55: 279-297, 2010.
Shimazu M, Soper RS. Pathogenicity and sporulation of Entomophthora maimaiga Humber, Shimazu, Soper and Hajek (Entomophthorales: Entomophthoraceae) on larvae of the gypsy moth, Lymantria dispar L. (Lepidoptera: Lymantriidae). Appl Entomol Zool 21: 589-596, 1986.
Soper RS, Shimazu M, Humber RA et al. Isolation and characterization of Entomophaga maimaiga sp. nov., a fungal pathogen of gypsy moth, Lymantria dispar from Japan. J Invert Pathol 51: 229-241, 1988.
Lau WL, Jumars PA, Armbrust EV. Genetic diversity of attached bacteria in the hindgut of the deposit-feeding shrimp Neotrypaea (formerly Callianassa) californiensis (Decapoda: Th alassinidae). Microb Ecol 43: 455-466, 2002.
Sezen K, Demirbag Z. Isolation and insecticidal activity of some bacteria from the hazelnut beetle (Balaninus nucum L.). Appl Entomol Zool 34: 85-89, 1999.
Demir İ, Sezen K, Demirbağ Z. Th e fi rst study on bacterial fl ora and biological control agent of Anoplus roboris (Sufr., Coleoptera). J Microbiol 40: 104-108, 2002.
Osborn F, Berlioz L, Vitelli-Flores J et al. Pathogenic eff ects of bacteria isolated from larvae of Hylesia metabus Crammer (Lepidoptera: Saturniidae). J Invertebr Pathol 80: 7-12, 2002.
Gokce C, Sevim A, Demirbag Z et al. Isolation, characterization and pathogenicity of bacteria from Rhynchites bacchus (Coleoptera: Rhynchitidae). Biocontrol Sci Techn 20: 973-982, 2010.
Sevim A, Demirbağ Z, Demir İ. A new study on the bacteria of Agrotis segetum Schiff . (Lepidoptera: Noctuidae) and their insecticidal activities. Turk J Agric For 34: 333-342, 2010.
Th iery I, Frachon E. Identifi cation, isolation, culture and preservation of entomopathogenic bacteria. In: Lacey AL. ed. Manual of Techniques in Insect Pathology. Academic Press; 1997: pp. 55-73.
Charles JF, Delecluse A, Nielsen-LeRoux C. Entomopathogenic Bacteria: From Laboratory to Field Application. Kluwer Academic Publisher. Dordrecht; 2000.
Th orpe KW, Podgwaite JD, Slavicek JM et al. Gypsy moth (Lepidoptera: Lymantriidae) control with ground-based hydraulic applications of Gypchek, in vitro-produced virus, and Bacillus thuringiensis. J Econ Entomo1 91: 875-880, 1998.
Hajek AE, Davis CI, Eastburn CC et al. Deposition and germination of conidia of the entomopathogen Entomophaga maimaiga infecting larvae of gypsy moth, Lymantria dispar. J Invertebr Pathol 79: 37-43, 2002.
Krieg NR, Holt JG. Gram-negative aerobic rods and cocci. In: Palleroni NJ. ed. Bergey’s Manual of Systematic Bacteriology. Williams and Wilkins; 1986: pp. 140-218.
Sneath PHA, Mair NS, Sharpe ME et al. Regular, nonsporing gram-positive rods. In: Kandler O, Weiss N. eds. Bergey’s Manual of Systematic Bacteriology. Williams and Wilkins; 1986: pp. 1208-1260.
Claus M. A standardized Gram staining procedure. World J Microbiol Biotechnol 8: 451-452, 1992.
Prescott LM, Harley JP, Klein DA. Microbiology, 3rd ed. William C Brown Publishers. Dubuque, IA, USA; 1996.
Soutourina OA, Semenova EA, Parfenova VV et al. Control of bacterial motility by environmental factors in polarly fl agellated and peritrichous bacteria isolated from Lake Baikal. Appl Environ Microbiol 67: 3852-3859, 2001.
Barry AL, Gavan TL, Badal RE et al. Sensitivity, specifi city, and reproducibility of the automicrobic system (with the Enterobacteriaceae-plus Biochemical Card) for identifying clinical isolates of Gram-negative bacilli. J Clin Microbiol 15: 582-588, 1982.
Weisburg WG, Barns SM, Pelletier DA et al. 16S ribosomal DNA amplifi cation for phylogenetic study. J Bacteriol 173: 697- 703, 1991. Ben-Dov E, Boussiba S, Zaritsky A. Mosquito larvicidal activity of Escherichia coli with combinations of genes from Bacillus thuringiensis subsp. israelensis. J Bacteriol 10: 2581-2587, 1995.
Aptosoglou SG, Sivropoulou A, Koliais SI. Plasmid patterns of Bacillus thuringiensis strains and isolates. Microbios 91: 203- 214, 1997.
Mitchell FL, Smith JW. Pathology and bioassays of the lesser cornstalk borer (Elasmopalpus lignosellus) entomopoxvirus. J Invertebr Pathol 45: 75-80, 1985.
Abbott WS. A method of computing the eff ectiveness of an insecticide. J Econ Entomol 18: 265-267, 1925. Jeyaprakash A, Hoy M, Allsopp M. Bacterial diversity in worker adults of Apis melliferacapensis and Apis mellifera scutellata (Insecta: Hymenoptera) assessed using 16S rRNA sequences. J Invertebr Pathol 84: 96-103, 2003.
Bahar AA, Demirbağ Z. Isolation of pathogenic bacteria from Oberea linearis (Coleoptera: Cerambycidae). Biologia 62: 13- 18, 2007.
Cox CR, Gilmore MS. Native microbial colonization of Drosophila melanogaster and its use as a model of Enterococcus faecalis pathogenesis. Infect Immun 75: 1565-1576, 2007.
İnce İA, Katı H, Yilmaz H et al. Isolation and identifi cation of bacteria from Th aumetopoea pityocampa Den. and Schiff . (Lepidoptera: Th aumetopoeidae) and determination of their biocontrol potential. World J Microbiol Biotechnol 24: 3005- 3015, 2008.
Muratoğlu H, Demirbağ Z, Sezen K. An entomopathogenic bacterium, decemlineata. Turk J Biol 35: 275-282, 2011.
Sezen K, Demir İ, Demirbağ Z. Study of the bacterial fl ora as a biological control agent of Agelastica alni L. (Coleoptera: Chrysomelidae). Biologia 59: 327-331, 2004.
Sezen K, Demir İ, Demirbağ Z. Identifi cation and pathogenicity of entomopathogenic bacteria from common cockchafer, Melolontha melolontha (Coleoptera: Scarabaeidae). New Zeal J Crop Hort 35: 79-85, 2007.
Rausell C, Martinez-Ramirez AC, Garcia-Robles I et al. Th e toxicity and physiological eff ects of Bacillus thuringiensis toxins and formulations on Th aumetopoea pityocampa, the pine processionary caterpillar. Pestic Biochem Phys 65: 44-54, 1999.
Kuzina LV, Peloquin JJ, Vacek D et al. Isolation and identifi cation of bacteria associated with adult laboratory Mexican fruit fl ies, Anastrepha ludens (Diptera: Tephritidae). Curr Microbiol 42: 290-294, 2001.
Reeves WK, Nayduch D. Pathogenic Bacillus from a larva of the Simulium tuberosum species complex (Diptera: Simuliidae). J Invertebr Pathol 79: 126-128, 2002.
Klein M, Kaya HK. Bacillus and Serratia species for scarab control. Memórias do Instituto Oswaldo Cruz Rio de Janeiro 90: 87-95, 1995.
Sezen K, Demir I, Demirbag Z. Investigations on bacteria as a potential biological control agent of summer chafer, Amphimallon solstitiale L. (Coleoptera: Scarabaeidae). J Microbiol 43: 463-468, 2005.
Lopez-Meza JE, Ibarra JE. Characterization of a novel strain of Bacillus thuringiensis. Appl Environ Microbiol 62: 1306-1310, 1996.
Crickmore N, Zeigler DR, Feitelson J et al. Revision of the nomenclature for the Bacillus thuringiensis pesticidal crystal proteins. Microbiol Mol Biol Rev 62: 807-813, 1998.
Feitelson SJ, Payne J, Kim L. Bacillus thuringiensis: insect and beyond. Bio/Technology 10: 271-275, 1992.
Rowe GE, Margaritis A, Dulmage HT. Bioprocess developments in the production of bioinsecticides by Bacillus thuringiensis. Crit Rev Biotechnol 6: 87-127, 1987.