تعیین ویژگی‌های فتوتیپی و ژنوتیپی باکتری عامل بیماری خوشه صمغی گندم در جنوب استان کرمان

نوع مقاله : مقاله کامل پژوهشی

نویسندگان

1 دانشجوی کارشناسی‌ارشد بیماری شناسی‌گیاهی، گروه گیاه‌پزشکی، دانشکده کشاورزی، دانشگاه جیرفت، جیرفت، ایران

2 استادیار، گروه گیاه‌پزشکی، دانشکده کشاورزی، دانشگاه جیرفت، جیرفت، ایران.

3 استادیار پژوهشی، بخش تحقیقات گیاه‌پزشکی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی جنوب استان کرمان، سازمان تحقیقات، آموزش و ترویج کشاورزی، جیرفت، ایران.

4 مربی، گروه گیاه‌پزشکی، دانشکده کشاورزی، دانشگاه جیرفت، جیرفت، ایران.

چکیده

بیست‌و‌یک جدایه باکتری از توده‌های گندم محلی جنوب استان کرمان دارای نشانه‌های بیماری خوشه‌صمغی، جداسازی شد. ویژگی‌های فنوتیپی و افتراقی جدایه‌ها مطابق روش‌های متداول باکتری‌شناسی انجام شد. شناسایی مولکولی و تنوع ژنتیکی جدایه‌ها با تکثیر و تعیین توالی نوکلئوتیدی قطعه‌هایی از ژن‌های 16S rRNA، gyrB و recA بررسی شد. با استفاده از واکنش زنجیره‌ای پلیمراز قطعه‌هایی به‌طول 700 و 558 جفت‌باز به‌ترتیب مربوط به ژن‌های 16S rRNA و gyrB از تمام جدایه‌ها تکثیر شد. در واکنش زنجیره‌ای پلیمراز با استفاده از آغازگرهای اختصاصی ژن recA، هیچ قطعه‌ای در نمونه‌های مورد بررسی و جدایه شاهد Rathayibacter tritici تکثیر نشد، در حالی‌که در جدایه شاهد R. iranicus قطعه‌ای به‌طول 600 جفت‌باز تکثیر گردید. توالی نوکلئوتیدی نواحی ژنی تکثیر شده با یکدیگر، با جدایه R. tritici و با استرین‌های این باکتری موجود در بانک ژن یکسان بود. نتایج آزمون‌های فنوتیپی و بررسی داده‌های حاصل از تکثیر و تعیین توالی جایگاه‌های ژنی یاد شده نشان داد که باکتری R. tritici عامل بیماری خوشه‌صمغی گندم در جنوب استان کرمان است. این اولین گزارش از وجود این گونه باکتری در استان کرمان است.

کلیدواژه‌ها


عنوان مقاله [English]

Phenotypic and genotypic characterization of wheat spike blight bacterium in southern Kerman province

نویسندگان [English]

  • M. Azadi Moghadam 1
  • Z. Azami Sardooei 2
  • M. Azadvar 3
  • K. Salari 4
چکیده [English]

Twenty-one bacterial strains were isolated from wheat showing spike blight disease symptoms in southern Kerman province. Differential phenotypic features of the strains were studied using common bacteriological methods. Molecular identification of the isolates was studied by amplification and partial sequence analyses of 16S rRNA, gyrB and recA genes. PCR assays using specific primer sets amplified 700 and 558 bp amplicons of 16S rRNA and gyrB genes from all strains, respectively. PCR assay with specific primer set for recA gene amplified a 600 bp fragment from Rathayibacter iranicus, but not from R. tritici strain and the isolated strains from wheat in southern Kerman province, as well. Sequences of the amplicons obtained from each gene locus were identical to those sequences for R. tritici reference strains as well as the R. tritici strains available in GenBank. Results obtained from phenotypic tests and analyses of the data from amplification and sequencing of the studied gene loci indicated that R. tritici is the causal agent of wheat spike blight disease in southern Kerman province. To the best of our knowledge, this is the first report of R. tritici on wheat in Kerman province.

کلیدواژه‌ها [English]

  • Gummy spike
  • Housekeeping genes
  • Rathayibacter
  • wheat
Akhtar A. 1987. Bacterial gumming disease of wheat. FAO Plant Protection Bulletin 35:102.
Amani B. 1969. Gummy spike blight disease. Iranian Journal of Plant Pathology 1:15-24. (in Persian with English Summary)
Azadvar M., Sharifi M., Rooshan K. A., Amiri M., Azadi Moghaddam M. and Azami Sardooei Z. 2017. First report of gummy spike blight disease of wheat in southern Kerman.First International and Second National Conference of Agriculture, Environment and Food Security, University of Jiroft. Jiroft. Iran. (in Persian with English Summary)
Babaeizad V. and Rahimian H. 2002. Identity and distribution of Rathayibacter species causing gummy spike blight of wheat in some wheat growing areas of Iran. Iranian Journal of Plant Pathology 38:47-55. (in Persian with English Summary)
Carne W. M. 1926. Earcockle (Tylenchus tritici) and bacterial disease (Pseudomonas tritici) of wheat. Journal of Agriculture Western Australia 3:508-512.
Collins M. D. and Bradbury J. F. 1986. Plant pathogenic species of Corynebacterium, pp. 1276-1284. in: P. H. A. Sneath, N. S. Mair, M. E. Sharpe and J. G. Holt (eds). Bergey’s Manual of Systematic Bacteriology.Vol. 2. Baltimore, MD: The Williams & Wilkins Co.
Dorofeeva L. V., Evtushenko L. I., Krausova V. I., Karpov A. V., Subbotin S. A. and Tiedje J. M. 2002. Rathayibacter caricis sp. nov. and Rathayibacter festucae sp. nov., isolated from the phyllosphere of Carex sp. and the leaf gall induced by the nematode Anguina graminis on Festuca rubra L., respectively. International Journal of Systematic and Evolutionary Microbiology 52:1917–1923.
Dorofeeva L. V., Starodumova I. P., Krauzova V. I., Prisyazhnaya N. V., Vinokurova N. G., Lysanskaya V. Y., Tarlachkov S. V. and Evtushenko L. I. 2018. Rathayibacter oskolensis sp. nov., a novel actinobacterium from Androsace kosopoljanskii Ovcz. (Primulaceae) endemic to the central Russian Upland. International Journal of Systematic and Evolutionary Microbiology 68:1442-1447.
Ebadzadeh H. R., Ahmadi K., Mohammadnia Afroozi S., Abbas Taghani R., Abbasi M. and Yari S. 2017. Agricultural statistics1395. (Vol. 1) Ministry of Jihad -e- Agriculture. Tehran, Iran.
Gupta P. and Swarup G. 1968. On the ear cockle and yellow ear rot diseases of wheat. 1. Symptoms and histopathology. Indian Phytopathology 21:318-323.
Hall T. A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucleic Acids Symposium series 41:95-98.
Hutchinson O. M. 1917. A bacterial disease of wheat in the Punjab. Memoirs of the Department of Agriculture in India. Bacteriological Series 17:75-169.
Klement Z., Farkas G. L. and Lovrekovich L. 1964. Hypersensitive reaction induced by phytopathogenic bacteria in tobacco leaf. Phytopathology 54:474.
Kovacs N. 1956. Identification of Pseudomonas solanacearum by the oxidase reaction. Nature 178:703.
Misaghi I. and Grogan R. G. 1969. Nutritional and biochemical comparisons of plant pathogenic and saprophytic fluorescent pseudomonas. Phytopathology 59:1436-1450.
Nikakhtar S., Rahimian H. and Babaeizad V. 2016. Evaluation of the phenotypic and genotypic diversity of the bacterial species causing gummy spike blight of wheat and barley. Journal of Agricultural Biotechnology 8:139 -156. (in Persian with English Summary)
Paruthi I. J. and Gupta D. C. 1987. Incidence of ‘tundu’ in barley and kanki in wheat field infested with Anguina tritici. Haryana Agricultural University Journal of Research 17:78–79.
Postnikova E., Agarkova I., Altundag S., Eskandari F., Sechler A., Karahan A., Vidaver A. K., Schneider W., Ozakman M. and Schaad N. W. 2009. Rathayibacter iranicus isolated from symptomless wheat seeds in Turkey. Plant Pathology 58:796.
Richert K., Brambilla E. and Stackebrandt E. 2005. Development of PCR primers specific for the amplification and direct sequencing of gyrB genes from microbacteria order Actinomycetales. Current Microbiology 60:115–123.
Riley I. T., Swart A., Postnikova E., Agarkova I., Vidaver A. and Schaad N. 2004. New association of a toxigenic Rathayibacter sp. and Anguina woodi in Ehrharta villosa var.villosa in South Africa. Phytopathology 94:S88.
Sasaki J., Chijimatsu M. and Suzuki K. 1998. Taxonomic significance of 2,4-diaminobutyric acid isomers in the cell wall peptidoglycan of actinomycetes and reclassification of Clavibacter toxicus as Rathayibacter toxicus comb. nov. International Journal of Systematic Bacteriology 48:403–410.
Schaad N. W., Jones, J. B., and Chun, W. 2001. Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed., APS Press., St. paul, Minnesota, USA. 373 p.
Scharif G. 1961. Corynebacterium iranicum sp. nov. on wheat (Triticum vulgare L.) in Iran, and a comparative study of it with C. tritici and C. rathayi. Entomologie et Phytopathologie Appliques 19:1–24.
Singh S., Goswami P., Singh R. and Heller K. J. 2009. Application of molecular identification tools for Lactobacillus, with a focus on discrimination between closely related species: a review. LWT-Food Science and Technology 42:448–457.
Skerman V. B. D., McGowan V. and Sneath P. H. A. 1980. Approved lists of bacterial names. International Journal of Systematic Bacteriology 30:225-420.
Soltani M., Rahimian H. and Babaeizad V. 2011. The assessment of genetic diversity of strains of Rathayibacter iranicus (the causal agent of gummy spike blight in wheat) in Isphahan and Fars using REP-PCR. 7th National Biotechnology Congress of Islamic Republic of Iran. Niroo Research Institute, Tehran, Iran. 5 pages.
Swarup G., Sethi C. L. and Goten N. 1993. Ear cockle (nematode gall) and yellow slime, pp. 153-161. In: S. B. Mathur and B. M. Cunfer (Eds). Seed borne diseases and seed health testing of wheat. Jordbrug Forlaget, Frederiksberg, Denmark.
Tamura K., Peterson D., Peterson N., Stecher G., Nei M., and Kumar S. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution 28:2731-2739.
Vasilenko O. V., Starodumova I. P., Tarlachkov S. V., Dorofeeva L. V., Avtukh A. N. and Evtushenko L. I. 2016. Draft genome sequence of Rathayibacter tanaceti strain VKM Ac-2596 isolated from Tanacetum vulgare infested by a foliar nematode. Genome Announcements 4:e00512-16. doi:10.1128/genomeA.00512-16.
Wilson K. 1997. Preparation of Genomic DNA from Bacteria, 2.4.1-2.4.5. In: F. M. Ausubel, R. Brent, R. E. Kingston, D. D. Moore, J. G. Seidman, J. A. Smith and K. Struhl (Eds). Current Protocols in Molecular Biology. Wiley & Sons, New York.
Zgurskaya H. I., Evtushenko L. I., Akimov V. N. and Kalakoutskii L. V. 1993. Rathayibacter gen, nov., including the species Rathayibacter rathayi comb, nov., Rathayibacter tritici comb, nov., Rathayibacter iranicus comb, nov. and six strains from annual grasses. International Journal of Systematic Bacteriology 43:143-149.