A new study on the bacteria of Agrotis segetum Schiff. (Lepidoptera: Noctuidae) and their insecticidal activities

Agrotis segetum Schiff. (Lepidoptera: Noctuidae) is one of the most serious pests of nearly all vegetables in Turkey. In this study, to find a more effective and safe biological control agent, the bacterial flora of Agrotis segetum were investigated in larvae collected from different populations in the vegetable fields of the Black Sea region of Turkey, and 9 bacterial isolates were cultured from these populations. Seven of these bacteria were determined and characterized at the species level and the rest were characterized at the genus level. Morphological, physiological, and biochemical characteristics of the bacterial isolates were determined by conventional and routine techniques. Characterization of the isolates was also performed using the VITEK 32 bacterial identification system. In addition, a 16S rRNA gene sequence was determined for these isolates to query against the NCBI genetic database and to construct a phylogenetic tree using closely related species. Consequently, the isolates were identified as Bacillus cereus (Ags1), Bacillus sp. (Ags2), Bacillus megaterium (Ags3), Enterobacter aerogenes (Ags4), Acinetobacter calcoaceticus (Ags5), Enterobacter sp. (Ags6), Pseudomonas putida (Ags7), Enterococcus gallinarum (Ags8), and Stenotrophomonas maltophilia (Ags9). All isolates were determined the first time from the flora of A. segetum. Furthermore, the insecticidal activities of the bacterial isolates were tested against A. segetum larvae, and Enterococcus gallinarum (Ags8) was found to cause the highest insecticidal activity (60%) 8 days after application.

Anahtar Kelimeler:

Key words: Agrotis segetum, bacterial flora, insecticidal activity

A new study on the bacteria of Agrotis segetum Schiff. (Lepidoptera: Noctuidae) and their insecticidal activities

Keywords:

Key words: Agrotis segetum, bacterial flora, insecticidal activity,

PDF

___

Abbott WS (1925) A method of computing the effectiveness of an insecticide. J Econ Entomol 18: 265-267.
Bahar AA, Demirbag Z (2007) Isolation of pathogenic bacteria from Oberea linearis (Coleptera: Cerambycidae). Biologia 62: 13-18.
Barry AL, Gavan TL, Badal RE, Telenson MJ (1982) Sensitivity, specificity, 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.
Ben-Dov E, Boussiba S, Zaritsky A (1995) Mosquito larvicidal activity of Escherichia coli with combinations of genes from Bacillus thuringiensis subsp. israelensis. J Bacteriol 10: 2581-2587.
Caballero P, Osuna-Vargas E, Santiago-Alvarez C (1991) Efficacy of a Spanish strain of Agrotis segetum granulosis virus (Baculoviridae) against Agrotis segetum Schiff. (Lepidoptera: Noctuidae) on corn. J Appl Entomol 112: 59-64.
Claus M (1992) A standardized Gram staining procedure. World J Microbiol Biotechnol 8: 451-452.
Cox CR, Gilmore MS (2007) Native microbial colonization of Drosophila melanogaster and its use as a model of Enterococcus faecalis pathogenesis. Infect Immun 75: 1565-1576.
Demir I, Sezen K, Demirbag Z (2002) The first study on bacterial flora and biological control agent of Anoplus roboris (Sufr., Coleoptera). J Microbiol 40: 104-108.
Hall TA (1999) Bioedit: A user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucleic Acids Symp 41: 95-98.
Ignoffo CM, Garcia C (1979) Susceptibility of larvae of the black cutworm to species of entomopathogenic bacteria, fungi, protozoa, and viruses. J Econ Entomol 72: 767-769.
Ince IA, Kati H, Yilmaz H, Demir I, Demirbag Z (2008) Isolation and identification of bacteria from Thaumetopoea pityocampa Den. and Schiff. (Lepidoptera: Thaumetopoeidae) and determination of their biocontrol potential. World J Microbiol Biotechnol 24: 3005-3015.
Jakubowska A, Van Oers MM, Ziemnicka J, Lipa JJ, Vlak JM (2005) Molecular characterization of Agrotis segetum nucleopolyhedrovirus from Poland. J Invertebr Pathol 90: 64-68.
Kandler O, Weiss N (1986) Regular, nonsporing gram-positive rods. In: Bergey’s Manual of Systematic Bacteriology (Eds. PHA Sneath, NS Mair, ME Sharpe, JG Holt), Williams and Wilkins, Baltimore, p. 1224.
Kuzina LV, Peloquin JJ, Vacek DC, Miller TA (2001) Isolation and identification of bacteria associated with adult laboratory Mexican fruit flies, Anastrepha ludens (Diptera: Tephritidae). Curr Microbiol 42: 290-294.
Lau WL, Jumars PA, Armbrust EV (2002) Genetic diversity of attached bacteria in the hindgut of the deposit-feeding shrimp Neotrypaea (Formerly Callianassa) Californiensis (Decapoda: Thalassinidae). Microb Ecol 43: 455-466.
Linden PK, Miller CB (1999) Vancomycin-resistant enterococci: the clinical effect of a common nosocomial pathogen. Diagn Microbiol Infect Dis 33: 113-120.
Lipa JJ, Wiland E (1972) Bacteria isolated from cutworms and their infectivity to Agrotis spp. (Lepitoptera: Noctuidae). Acta Microbiol Pol 4: 127-140.
Lossbroek TG, Theunissen J (1985) The entomogenous nematode Neoplectana bibionis as a biological control agent of Agrotis segetum in lettuce. Exp Appl Nematol 39: 261-264.
Lukasova J, Sustackova A (2003) Enterococci and antibiotic resistance. Acta Vet Brno 72: 315-323.
The Ministry of Agriculture of Turkey (2008) The Technical Guide of Agricultural Control, Vol 3, Başak Publisher, Ankara, pp. 262- 265.
Mitchell FL, Smith JW (1985) Pathology and bioassays of the lesser cornstalk borer (Elasmopalpus lignosellus) entomopoxvirus. J Invertebr Pathol 45: 75-80.
Murrell A, Dobson SJ, Yang X, Lacey E, Barker SC (2003) A survey of bacterial diversity in ticks, lice and fleas from Australia. Parasitol Res 89: 326-334.
Ohba M, Aizawa K (1986) Distribution of Bacillus thuringiensis in soil of Japan. J Invertebr Pathol 47: 277-287.
Osborn F, Berlioz L, Vitelli-Flores J, Monsalve W, Dorta B, Lemoine VR (2002) Pathogenic effects of bacteria isolated from larvae of Hylesia metabus Crammer (Lepidoptera: Saturniidae). J Invertebr Pathol 80: 7-12.
Palleroni NJ (1984) Gram-negative aerobic rods and cocci, family I: Pseudomonasdaceae. In: Bergey’s Manual of Systematic Bacteriology (Eds. NR Krieg, JG Holt), Williams and Wilkins, Baltimore, pp. 141-199.
Prescott LM, Harley JP, Klein DA (1996) Microbiology, Wm C Brown Publishers, England.
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, New York.
Sandra WW, Douglas IG (2004) Microorganisms associated with field-collected Chrysoperla rufilabris (Neuroptera: Chrysopidae) adults with emphasis on yeast symbionts. Biol Cont 29: 155-168.
Sezen K, Demir I, Demirbag Z (2005) Investigations on bacteria as a potential biological control agent of summer chafer, Amphimallon solstitiale L. (Coleoptera: Scarabaeidae). J Microbiol 43: 463-468.
Soutourina OA, Semenova EA, Parfenova VV, Danchin A, Bertin P (2001) Control of bacterial motility by environmental factors in polarly elagellated and peritrichous bacteria isolated from Lake Baikal. Appl Environ Microbiol 67: 3852-3859.
Steenberg T, Ogaard L (2000) Mortality in hibernating turnip moth larvae, cylindrosporum. Mycol Res 104: 87-91.
Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24: 1596-1599.
Thiery I, Frachon E (1997) Identification, isolation, culture and preservation of enthomopathogenic bacteria. In: Manual of Techniques in Insect Pathology (Ed. AL Lacey), Academic Press, London, pp. 55-73.
Turchaninova LP (1972) The bacterial flora of healthy Agrotis segetum Schiff in various developmental stages. Mikrobiol Zh 34: 119- 20.
Turchaninova LP (1973) Pathogenic bacterial flora of the winter cutworm (Agrotis segetum Schiff). Mikrobiol Zh 35: 576-80.
William GW, Susan MB, Dale AP, David JL (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173: 697- 703.
Xiang H, Wei GF, Jia S, Huang J, Miao XX, Zhou Z, Zhao LP, Huang YP (2006) Microbial communities in the larval midgut of laboratory and field populations cotton bollworm (Helicoverpa armigera). Can J Microbiol 52: 1085-1092.
Yilmaz H, Sezen K, Kati H, Demirbag Z (2006) The first study on the bacterial flora of the European spruce bark beetle, Dendroctonus micans (Coleoptera: Scolytidae). Biologia 61: 679-686.
Yun JS, Kim HH, Kim DW, Lee SM, Kim DS, Lee DW (2004) Pathogenicities of entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae against lepidopterous insect pests, Agrotis segetum, Artogeia rapae, Mamestra brassicae, Plutella xylostella, Spodoptera exigua and Spodoptera litura. Kor Turfgrass Sci 18: 221-229.
Zethner O (1980) Control of Agrotis segetum (Lepidoptera: Noctuidae) root crops by granulosis virus. Entomophaga 25: 27- 35.