Isolation and characterization of native Bacillus thuringiensis isolates from Syrian soil and testing of their insecticidal activities against some insect pests

Bacillus thuringiensis was detected in 12.5% of soil samples collected from different regions in Syria and 25 B. thuringiensis isolates were found to be highly toxic to larvae of Ephestia kuehniella Zeller, Phthorimaea operculella Zeller, and Cydia pomonella L. (Lepidoptera), but not to the larvae of Culex quinquefasciatus (Diptera). Light microscopy investigation showed the presence of bipyramidal and cuboidal parasporal bodies produced by these isolates. The existence of different cry genes in the tested isolates was studied using a PCR strategy with a set of general primers recognizing some of the cry genes reported in the relevant literature. Primers corresponding to 2 types of cry genes (cry1 and cry2) successfully amplified DNA in all of the tested isolates. Moreover, the proteins encoded by these genes were detected in the SDS-PAGE of the purified parasporal bodies. The 50% lethal concentration of the spore-crystal mixture of the 25 isolates against E. kuehniella larvae varied from 8.4 to 97.6 µg g-1. A comparison of the LC50 values of the tested isolates with those of the reference strains B. thuringiensis kurstaki HD-1 and HD-73 (20.8 and 46 µg g-1, respectively) showed that some of these isolates have a higher toxicity potential. Moreover, flagellar serotyping revealed that 4 isolates, which were among the most toxic, belonged to serotype kurstaki. This study constitutes the first isolation and characterization of local B. thuringiensis isolates in Syria. Some of these isolates exhibit toxic potential and, therefore, could be adopted for future applications to control some important insect pests.

Isolation and characterization of native Bacillus thuringiensis isolates from Syrian soil and testing of their insecticidal activities against some insect pests

Bacillus thuringiensis was detected in 12.5% of soil samples collected from different regions in Syria and 25 B. thuringiensis isolates were found to be highly toxic to larvae of Ephestia kuehniella Zeller, Phthorimaea operculella Zeller, and Cydia pomonella L. (Lepidoptera), but not to the larvae of Culex quinquefasciatus (Diptera). Light microscopy investigation showed the presence of bipyramidal and cuboidal parasporal bodies produced by these isolates. The existence of different cry genes in the tested isolates was studied using a PCR strategy with a set of general primers recognizing some of the cry genes reported in the relevant literature. Primers corresponding to 2 types of cry genes (cry1 and cry2) successfully amplified DNA in all of the tested isolates. Moreover, the proteins encoded by these genes were detected in the SDS-PAGE of the purified parasporal bodies. The 50% lethal concentration of the spore-crystal mixture of the 25 isolates against E. kuehniella larvae varied from 8.4 to 97.6 µg g-1. A comparison of the LC50 values of the tested isolates with those of the reference strains B. thuringiensis kurstaki HD-1 and HD-73 (20.8 and 46 µg g-1, respectively) showed that some of these isolates have a higher toxicity potential. Moreover, flagellar serotyping revealed that 4 isolates, which were among the most toxic, belonged to serotype kurstaki. This study constitutes the first isolation and characterization of local B. thuringiensis isolates in Syria. Some of these isolates exhibit toxic potential and, therefore, could be adopted for future applications to control some important insect pests.

___

  • Arthur FH (1996) Grain protectants: current status and prospects for the future. J Stored Prod Res 32: 293-302. Bell CH (2000) Fumigation in the 21st century. Crop Prot 19: 563- 569.
  • Ben-Dov E, Wang Q, Zaritsky A, Manasherob R, Barak Z, Schneider B, Khamraev A, Baizhanov M, Glupov V, Margalith Y (1999) Multiplex PCR screening to detect cry9 genes in Bacillus thuringiensis strains. Appl Environ Microb 65: 3714-3716.
  • Ben-Dov E, Zaritsky A, Dahan E, Barak Z, Sinai R, Manasherob R, Khamraev A, Troitskaya E, Dubitsky A, Berezina N, Margalith Y (1997) Extended screening by PCR for seven cry-group genes from fi eld-collected strains of Bacillus thuringiensis. Appl Environ Microb 63: 4883-4890.
  • Bernhard K, Jarrett P, Meadows M, Butt J, Ellis DJ, Roberts GM, Pauli S, Rodgers P, Burges HD (1997) Natural isolates of Bacillus thuringiensis: worldwide distribution, characterization, and activity against insect pests. J Invertebr Pathol 70: 59-68.
  • Bravo A, Gill SS, Soberon M (2007) Mode of action of Bacillus thuringiensis Cry and Cyt toxins and their potential for insect control. Toxicon 49: 423-435.
  • Bravo A, Sarabia S, Lopez L, Ontiveros H, Abarca C, Ortiz A, Ortiz M, Lina L, Villalobos F, Peña G, Nuñes M, Soberón M, Quintero R (1998) Characterization of cry genes in a Mexican Bacillus thuringiensis strain collection. Appl Environ Microb 64: 4965-4972.
  • Brower JH, Smith L, Vail PV, Flinn PW (1995) Biological control. In: Integrated Management of Insects in Stored Products (Eds. B Subramanyam, DW Hagstrum), Marcel Dekker, New York, pp. 223-286.
  • Carozzi N, Karmer V, Warren G, Evola S, Koziel M (1991) Prediction of insecticidal activity of Bacillus thuringiensis strains by polymerase chain reaction product profi les. Appl Environ Microb 57: 3057-3061.
  • Chilcot CN, Wigley PJ (1993) Isolation and toxicity of Bacillus thuringiensis from soil and insect habitats in New Zealand. J Invertebr Pathol 61: 244-247.
  • Crickmore N (2006) Beyond the spore - past and future developments of Bacillus thuringiensis as a biopesticide. J Appl Microbiol 101: 616-619.
  • DeLucca AJ, Simonson JG, Larson AD (1981) Bacillus thuringiensis distribution in soils of the United States. Can J Microbiol 27: 865-870.
  • Dulmage HT, Aisawa K (1982) Distribution of Bacillus thuringiensis in nature. In: Microbial and Viral Pesticides (Ed. E Kurstak), Marcel Dekker, New York, pp. 209-237.
  • Federici BA, Park HW, Sakano Y (2006) Insecticidal protein crystals of Bacillus thuringiensis. In: Inclusions in Prokaryotes (Ed. JM Shively), Springer-Verlag, Berlin-Heidelberg, pp. 195-235.
  • Finney DJ (1971) Probit Analysis. Cambridge University Press, Cambridge.
  • Hossain MA, Ahmed S, Hoque S (1997) Abundance and distribution of Bacillus thuringiensis in the agricultural soil of Bangladesh. J Invertebr Pathol 70: 221-225.
  • Iriarte J, Bel Y, Ferrandis MD, Andrew R, Murillo JF, Caballero P (1998) Environmental distribution and diversity of Bacillus thuringiensis in Spain. Syst Appl Microbiol 21: 97-106.
  • Jacob TA, Cox PD (1977) Th e infl uence of temperature and humidity on the life-cycle of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae). J Stored Prod Res 13: 107-118.
  • Kaur S (2002) Potential for developing novel Bacillus thuringiensis strains and transgenic crops and their implications for Indian agriculture. Agr Biotech Net 4: 1-10.
  • Lambert B, Peferoen M (1992) Insecticidal promise of Bacillus thuringiensis. Bioscience 42: 112-122.
  • Lecadet MM, Frachon E, Dumanoir VC, Ripouteau H, Hamon S, Laurent P, Th iéry I (1999) Updating the H-antigen classifi cation of Bacillus thuringiensis. J Appl Microbiol 86: 660-672.
  • Makee H (2005) Factors infl uencing the parasitism of codling moth eggs by Trichogramma cacoeciae March. and T. principium Sug. et Sor. (Hymen. Trichogrammatidae). J Pest Sci 78: 31-39.
  • Maniatis T, Fritsch EF, Sambrook J (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, New York.
  • Marec F (1990) Genetic control of pest Lepidoptera: Induction of sexlinked recessive mutations in Ephestia kuehniella (Pyralidae). Act Entomol Bohemoslov 87: 445-458.
  • Martin P, Travers R (1989) Worldwide abundance and distribution of Bacillus thuringiensis isolates. Appl Environ Microb 55: 2437- 2442.
  • Moore D, Lord JC, Smith S (2000) Pathogens. In: Alternatives to Pesticides in Stored- Product IPM (Eds. BH Subramanyam, DW Hagstrum,), Kluwer Academic Publishers, Dordrecht, Netherlands, pp. 209-237.
  • Ohba M, Aizawa K (1986a) Distribution of Bacillus thuringiensis in soils of Japan. J Invertebr Pathol 47: 277-282.
  • Ohba M, Aizawa K (1986b) Insect toxicity of Bacillus thuringiensis isolated from soils of Japan. J Invertebr Pathol 47: 12-20.
  • Ohba M, Aratake Y (1994) Comparative study of the frequency and fl agellar serotype fl ora of Bacillus thuringiensis in soils and silkworm-breeding environments. J Appl Bacteriol 76: 203- 209.
  • Ohba M, Tsuchiyama A, Shisa N, Nakashima K, Lee D, Ohgushi A, Wasano N (2002) Naturally occurring Bacillus thuringiensis in oceanic islands of Japan, Daito-shoto and Ogasawara-shoto. Appl Entomol Zool 37: 477-480.
  • Porcar M, Juárez-Pérez V (2003) PCR-based identifi cation of Bacillus thuringiensis pesticidal crystal genes. FEMS Microbiol Rev 26: 419-432.
  • Rampersad J, Khan A, Ammons D (2002) Usefulness of staining parasporal bodies when screening for Bacillus thuringiensis. J Invertebr Pathol 79: 203-204.
  • Roh JY, Jae YC, Ming SL, Byung RJ, Yeon HE (2007) Bacillus thuringiensis as a specifi c, safe, and eff ective tool for insect pest control. J Microbiol Biotechnol 17: 547-559.
  • Saour G, Makee H (1997) Radiation induced sterility in male potato tuber moth Phthorimaea operculella. (Lep., Gelechiidae). J Appl Entomol 121: 411-415.
  • Strong D, Lawton J, Southwood R (1984) Insect on Plant Community Patterns and Mechanism. Blackwell Science, London.
  • Travers RS, Martin PAW, Reichelderfer CF (1987) Selective process for effi cient isolation of soil Bacillus species. Appl Environ Microb 53: 1263-1266.
  • Vásquez M, Parra C, Hubert E, Espinoza P, Th eoduloz C, Meza-Basso L (1995) Specifi city and insecticidal activity of Chilean strains of Bacillus thuringiensis. J Invertebr Pathol 66: 143-148.
  • Wu D, Chang F (1985) Synergism in the mosquitocidal activity of 26 and 65 kDa proteins from Bacillus thuringiensis subsp. fukuokaensis crystal proteins. Appl Environ Microb 57: 1075- 1081.
Turkish Journal of Agriculture and Forestry-Cover
  • ISSN: 1300-011X
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK
Sayıdaki Diğer Makaleler

Agroecological land use potential of Amik Plain, Turkey

Şeref KILIÇ

Field appraisal of seed priming to improve the growth, yield, and quality of direct seeded rice

Hafeez Ur REHMAN, Shahzad Maqsood Ahmed BASRA, Muhammad FAROOQ

Production of cellulase by Pleurotus ostreatus and Pleurotus sajor-caju in solid state fermentation of lignocellulosic biomass

Md. Ibrahim KHALIL, Mia Mahmudul HOQUE, Mafroz Ahmed BASUNIA, Nadia ALAM

Micropropagation of olive tree Olea europaea L. ‘Oueslati’

Anissa CHAARI RKHIS, Mohamed MAALEJ, Noureddine DRIRA, Alvaro STANDARDI

Effect of mini-plug container depth on root and shoot growth of four forest tree species during early developmental stages

Panagiota KOSTOPOULOU, Kalliopi RADOGLOU, Olympia DINI PAPANASTASI

Isolation and characterization of native Bacillus thuringiensis isolates from Syrian soil and testing of their insecticidal activities against some insect pests

Hassan AMMOUNEH, Muhanad HARBA, Emad IDRIS, Hayat MAKEE

Potential of Steinernema feltiae (Rhabditida: Steinernematidae) as a biological control agent against the cabbage maggot Delia radicum (Diptera: Anthomyiidae) in oilseed rape

İ. Alper SUSURLUK

Stability properties of certain durum wheat genotypes for major quality characteristics

Mehmet Ali SAKİN, Oral DÜZDEMİR, Abdulvahit SAYASLAN, Ferhat YÜKSEL

Field evaluation of effective microorganisms (EM) application for growth, nodulation, and nutrition of mung bean

Arshad JAVAID, Rukhsana BAJWA

Field evaluation of eff effective microorganisms (EM ) application for growth, nodulation, and nutrition of mung bean

Arshad JAVAID, Rukhsana BAJWA