Molecular detection of Cry genes in the Bacillus thuringiensis native isolates from the soils of tobacco fields of Mazandaran and Golestan provinces
Subject Areas : Plant Pests
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Keywords: Molecular investigation, isolate, Cry genes, Bacillus thuringiensis, PCR,
Abstract :
In recent years, the continuous use of chemical pesticides has led to the emergence of resistance in pests and environmental pollution. Hence, the need for the use of environmentally friendly and safe insecticides has become necessary. One of the most successful microbial agents in this field is Bacillus thuringiensis (Bt). Bt is a gram-positive, aerobic, soil-dwelling, spore-forming, that, following sporulation, forms crystal proteins encoded by Cry genes. For this purpose, in this study, 90 native isolates of Bt from soils of tobacco fields of Mazandaran and Golestan provinces were isolated and molecularly examined based on the Cry gene, a factor producing effective proteins against pests, based on the PCR method using seven pairs of specific primers for the Cry I, Cry 1Ab, Cry 1Ac, Cry 1F, Cry 1I, Cry 2 and Cry 9 genes. The results of bands obtained showed that the frequency of the genes in different isolates was very different, so that some genes such as Cry 1Ab had the highest frequency in the isolates studied, while the Cry I and Cry 1I genes had the lowest frequency. In some isolates, the genes were amplified in different sizes than expected. The results of this study can be .useful for tracking native Bt isolates that have Cry genes effective on insects
توحیدی، ف.، ناظمی، ع. و خاتمی¬نژاد، م.ر. 1390. جداسازی و شناسایی مولکولی انواع ژن¬های Cry در باسیلوس تورینجینسیس¬های جدا شده از خاک¬های غرب مازندران. مجله زیست¬فناوری دانشگاه آزاد اسلامی 3(11): 1-6.
جهانگيري، ر.، صالحی¬جوزانی، غ.، سعید، خ.ع.، مصدق، ح. و نظریان، ا. 1387. شناسايي ژن¬های جدید Cry در جدایه¬های بومی باکتری Bacillus thuringiensis ایران. خلاصه مقالات دهمين كنگره ژنتيك ايران. 463 صفحه.
ریاضی، ا.، زرگری، ک. و کشاورزی، م. 1388. شناسایی انواع ژن¬های Cry1 در نمونه¬های Bacillus thuringiensis جمع¬آوری شده از استان¬های خراسان و همدان. نشریه دانش زیستی ایران 4(2): 19-28.
سلطانی¬نژاد، پ.، مهرخو، ف. و راشکی، م. 1399. شناسایی ژن¬های Cry ویژه بالپولکداران در سویه¬های ایرانی Bacillus thuringiensis. بیست و یکمین کنگره ملی و نهمین کنگره بین¬المللی زیست¬شناسی ایران.
سیفی¬نژاد، ع.، بی¬همتا، م.ر.، امیدی، م. و صالحی¬جوزانی، غ.ر. 1386. ژنومیکس عملکردی باکتری باسیلوس تورینجینسیس (توکسین¬های Cry). نشریه ژنتیک نوین 3(10): 24-17.
شازده¬احمدی، م.، سجادی، ا. و شهادتی مقدم، ز. 1398. بررسی برخی خصوصیات ژنی جدایه¬های بومی باکتری Bacillus thuringiensis از خاک¬های مناطق جنگلی استان گلستان. مجله گیاه¬پزشکی کاربردی 8(1): 25-37.
صادقی، م. صادقی، ا. و کریمی، ا. 1395. مروری بر باکتری Bacillus thuringiensis (Bt) به¬عنوان آفت¬کش زیستی. مجله ایمنی زیستی 9(2): 99-114.
گرایلی مرادی، ف.، مـحـمـدی شـریف، م.، هـادی¬زاده، ع. و بـابـایی¬زاد، و. 1393 الف. تنـوع باکـتری Bacillus thuringiensis از خاک¬های دارای پوشش گیاهی مختلف. مجله محیط زیست طبیعی، منابع طبیعی ایران 67 (3): 321–313.
گرایلی مرادی، ف.، محمدی شریف، م.، هادی¬زاده، ع. و بابایی¬زاد، و. 1393 ب. ردیابی مولکولی جدایه¬های بومی باکتری Bacillus thuringiensis از خاک¬های دارای پوشش گیاهی مختلف. مجله مهندسی ژنتیک و ایمنی زیستی 3(1): 54–41.
محمودی، ر.، ایرجیان، غ.ر.، جوادی، ا. و پناهی¬کوخدان، ا. 1396. بررسی وجود ژن Cry در باسیلوس تورینجینسیس¬های جدا شده از خاک. مجله ارمغان دانش 22(2): 280–271.
Ammouneh, H., Ismail, H., Al-bedda, A., Abou Baker, N. and Harba, M. 2013. Characterization of lepidopteran-active Bacillus thuringiensis isolates recovered from infected larvae. Biocontrol Science and Technology 23: 607-623.
Apaydin, O., Yenidunya, A., Harsa, S. and Gunes, H. 2005. Isolation and Characterization of Bacillus thuringiensis strains from different grain habitats in Turkey. World Journal of Microbiology and Biotechnology 21: 285-292.
Aramideh, Sh., Safaralizadeh, M.H., Pourmirza, A.A., Rezazadeh Bari, M., Keshavarzi, M. and Mohseniazar, M. 2010. Characterization and pathogenic evaluation of Bacillus thuringiensis isolates from west Azerbaijan province of Iran. African Journal of Microbiology Research 4: 1224-1229.
Carrozi, N.B., Kramer, V.C., Warren, G.W., Evola, S. and Koziel, M. 1991. Prediction of insecticidal activity of Bacillus thuringiensis strains by Polymerase Chain Reaction product profiles. Applied and Environmental Microbiology 57: 3057-3061.
Cinar, C., Apaydin, O., Yenidunya, A.F., Harsa, S. and Gunes, H. 2008. Isolation and characterization of Bacillus thuringiensis strains from olive-related habitats in Turkey. Journal of Applied Microbiology 104: 515-525.
Hannay, C.L. and Fitz, J.P. 1955. The protein crystals of Bacillus thuringiensis Ber. Canadian Journal of Microbiology 1: 694-710.
Hongyu, Z., Ziniu, Y. and Wangxi, D. 2000. Isolation, distribution and toxicity of Bacillus thuringiensis from warehouses in China. Crop Protection 19: 449-454.
Juarez- Perez, V.M., Ferrandis, M.D. and Frutus, R. 1997. PCR- based approach for detection of novel Bacillus thuringiensis Cry genes. Applied and Environmental Microbiology 63(8): 2997-3002.
Kaur, S. 2006. Molecular approaches for identification and construction of novel insecticidal genes for crop protection. World Journal of Microbiology and Biotechnology 22: 233-253.
Keshavarzi, M. 2008. Isolation, Identification and Differentiation of Local Bacillus thuringiensis Strains. Journal of Agriculture and Science Technology 10: 493-499.
Kim, H.S., Lee, D.W., Woo, S.D., Yu, Y.M. and Kang, S.K. 1998. Distribution, serological identification, and PCR analysis of Bacillus thuringiensis isolated from soils of Korea. Current Microbiology 37: 195-200.
Martinez, C., Ibarra, J.E and Caballero, P. 2005. Association analysis between serotype, cry gene content, and toxicity to Helicoverpa armigera larvae among Bacillus thuringiensis isolates native to Spain. Journal of Invertebrate Pathology 90: 91-97.
Martínez, C. and Caballero, P. 2002. Contents of cry genes and insecticidal toxicity of Bacillus thuringiensis strains from terrestrial and aquatic habitats. Journal of Applied Microbiology 92: 745-752.
Nariman, A.H. 2007. PCR Detection of cry Genes in Local Bacillus thuringiensis Isolates. Australian Journal of Basic and Applied Sciences 1(4): 461-466.
Porcar, M. and Juarez-Perez, V. 2003. PCR- based identification of Bacillus thuringiensis pesticidal crystal genes. FEMS Microbiology Reviews 26: 419-432.
Pooja, A.S., Krishnaraj, P.U. and Prashanthi, S.K. 2013. Profile of cry from native Bacillus thuringiensis isolates and expression of cry1I. African Journal of Biotechnology 12: 3545-3562.
Rolle, R.L., Ejiofor, A. and Johnson, T.L. 2005. Determination of the plasmid size and location of δ-endotoxin genes of Bacillus thuringiensis by pulse field gel electrophoresis. African Journal of Biotechnology 4: 580-585.
Salehi Jouzani, G., Pourjan Abad, A., Seifinejad, A., Marzban, R., Kariman, K. and Maleki, B. 2008. Distribution and diversity of Dipteran-specific cry and cyt genes in native Bacillus thuringiensis strains obtained from different ecosystems of Iran. Journal of Industrial Microbiology and Biotechnology 35: 83-94.
Seifinejad, A., Salehi Jouzani, G.R. and Abdmishani, C. 2008. Characterization of Lepidoptera– active Cry and Vip genes in Iranian Bt strain collection. Biological Control 44: 216-226.
Travers, R.S., Martin, P.A.W. and Reichelderfer, C.F. 1987. Selective process for efficient isolation of soil Bacillus spp. Applied and Environmental Microbiology 53: 1263-1266.
Tuba Etinkaya, F. 2002. Isolation of Bacillus thuringiensis and Investigation of Its Crystal Protein Genes. Izmir Institute of Technology İzmir, Turkey.