بررسی فنوتیپی و ژنوتیپی جدایه های Pseudomonas syringae pv. syringae عامل بلاست مرکبات در غرب مازندران و شرق گیلان

پژوهش های حفاظت گیاهان ایران

مقاله 1، دوره 30، شماره 3، آذر 1395، صفحه 359-367   

نوع مقاله: مقالات پژوهشی

شناسه دیجیتال (DOI): 10.22067/jpp.v30i3.30987

نویسندگان

سمانه سمیعی شیرکده¹؛ مرتضی گل محمدی^{* 2}؛ سید علی الهی نیا¹؛ سمانه بشیری³

¹دانشگاه گیلان

²وهشکده مرکبات و میوه‌های نیمه گرمسیری

³دانشگاه کردستان

چکیده

بیماری بلاست یکی از بیماری های شایع در مناطق گرمسیری و نیمه گرمسیری کشت مرکبات از جمله شمال ایران می باشد که عمدتاً به-وسیله جدایه های باکتری (Pss) Pseudomonas syringae pv. syringae ایجاد می شود. به منظور بررسی فنوتیپی و ژنوتیپی جدایه های Pss عامل بیماری، نمونه برداری از درختان مرکبات در غرب مازندران و شرق گیلان طی ماه های اسفند 1390 تا اوایل خرداد 1391 صورت گرفت. جدایه ها از نظر خصوصیات فنوتیپی، بیماری زایی مورد بررسی قرار گرفتند که تعداد 27 جدایه باکتری از بافت های آلوده بر اساس آزمون های فنوتیپی و بیماری زایی به‌عنوان Pss شناسایی و جدا شدند. جهت تأیید شناسایی جنس و گونه باکتری از روش مولکولی (PCR) با استفاده از جفت آغازگرهای اختصاصی باکتری مربوط به ژن اختصاصی تولید سیرینگومایسن استفاده گردید. تمامی جدایه ها با جفت آغازگر اختصاصی قطعه قابل انتظار 198 جفت بازی از ژن syrB را تولید کردند. انگشت نگاری ژنومی جدایه ها به روش rep-PCRبا استفاده از آغازگرهای ERIC و REP انجام شد. نتایج به دست آمده از دندروگرام تلفیق نقوش الکتروفورز حاصل از آغازگرهای ERIC و REP نشان داد که جدایه ها در سطح تشابه 75 درصد در 6 گروه قرار گرفتند. علی رغم وجود خصوصیات فنوتیپی مشابه، نتایج بیانگر وجود ناهمگونی از نظر ژنتیکی در بین جدایه های Pss، عامل بلاست مرکبات در غرب مازندران و شرق گیلان می باشد.

کلیدواژه‌ها

انگشت نگاری؛ تنوع ژنتیکی؛ Pss؛ rep-PCR

مراجع

1- Abbasi V., Rahimian H., Tajick-Ghanbari M.A., and Rezaian V. 2011. The assessment of genetic diversity of strains of Pseudomonas syringae pv.syringae causing bacterial canker in stone fruit in some northern province of IRAN. Iranian Journal of Plant Patholology, 47: 133-135. (in Persian with English abstract)

2- Aldaghi M., Rahimian H., and Mohamadi M. 2010. Comparison of phenotypic, serological and molecular characteristics of Pseudomonas syringae pv. syringae strains, the causal agent of bacterial canker of stone fruits and blight of cereals. Iranian Journal of Plant Pathology, 45(4): 91-93. (in Persian with English abstract)

3- Beigi F., Rahimian H., Goltapeh-mohammadi E., Shams-Bakhsh M., Barzegar A., Busquets A., Garcia- Valdes E., and Lalvkat G. 2012. Phenotypic and Pathogenecity Characteristics of the Agents Causing Citrus Blast Disease in the Northern Provinces of Iran. Iranian Journal of Plant Protection, 43(2):211-222. (in Persian)

4- Bradbury J.F. 1986. Pseudomonas syringae pv. syringae. p.175-177. In: Guide to Plant Pathogenic Bacteria. CAB International Mycological Institute, Kew, England.

5- Cheng G.Y., Legard D.E., Hanter J.E., and Barr T.J. 1989. Modified bean pod assay to detect strains of Pseudomonas syringae pv. syringae that cause bacterial brown spot snap bean. Plant Disease, 73: 419-423.

6- Cirvilleri G., Bonaccorsi A., Scuderi G., and Scortichni M. 2005. Potential biological control activity and genetic diversity of Pseudomonas syringae pv. syringae strains. Phytopathology, 153: 633-750.

7- Golmohammadi M. 2004. Study the distribution and identification of blast disease of citrus in the West of Mazandaran and Gilan East. Citrus Research Institute of the country. The final report of the research project.

8- Gross D.C., Cody Y.S., Proebsting E.L., Radamaker J.W., and Spots R.A. 1984. Ecotypes and pathogenicity of Ice- nucleation active Pseudomonas syringae isolated from deciduous fruit tree orchards. Phytopathology, 74: 241-284.

9- Gross D.C. 1991. Molecular and genetic analysis of toxin production by pathovars of Pseudomonas syringae. Annual Review of Phytopathology, 29:247-278.

10- Hassanzadeh N. 1995. Incidence and progress of different diseases incited by pathovars of Pseudomonas syringae. Journal of Agricultural Sciences Islamic Azad University, 1:5-14.

11- Hildebrand D.C., Schroth M.N., and Huisman O.C. 1982.The DNA homology matrix and non-random variation concepts as the basis for the taxonomic treatment of plant pathogenic and other bacteria. Annual Review of Phytopathology, 20: 235-256.

12- Hirano S.S., and Upper C.D. 2000. Bacteria in the leaf ecosystem with emphasis on Pseudomonas syringae pathogen, ice nucleus and epiphyte. Microbology and Molecular Biology Reviews, 624–653.

13- Jones J.B., Chase A.R., and Harris G.K. 1993. Evaluation of the Biolog GN Microplate system for identification of some plant-pathogenic bacteria. Plant Disease, 77:553–558.

14- Klement Z., Farkas G.L., and Loverkovich L. 1974. Hypersensitive reaction induced by phytopathogenic bacteria in tobacco leaf. Phytopathology, 64: 474-477.

15- Kovacs N. 1956. Identification of Pseudomonas pyocyanea by the oxidase reaction. Nature, London, 178-703.

16- Lilley A.K., Bailey M.J., and Fry J.C. 1996. Diversity of mercury resistance plasmid obtained by exogenous isolation from the bacteria of sugar beet in three successive seasons. FEMS Microbiology Ecology, 20: 211-227.

17- Little E.L., Bostock R.M., and Kirkpatrick B.C. 1998. Genetic characterization of Pseudomonas syringae pv. syringae strain from stone fruits in California. Applied and Environmental Microbiology, 64: 3818-3823.

18- Louws F.J., Fulbright D.W., Stephens C.T., and De Bruiin F.J. 1994. Specific genomic fingerprints of phytopathogenic Xanthomonas and Pseudomonas pathovars and strains generated with repetitive sequences and PCR. Applied and Environmental Microbiology, 80: 2286-2292.

19- Louws F.J., Rademarker J.W., and Brujin F.J. 1999. The three Ds of PCR-Base genoimic analysis of phytobacterial diversity, detection and diagnosis. Annual Review of Phytopathology, 37: 81-125.

20- Mosivand M., Rahimian H., and Shams-Bakhsh M. 2009. Phenotypic and genotypic relateness among Pseudomonas syringae pv. syringae strains isolates from sugarcane, stone fruits and wheat. Iranian Journal of Plant Patholology, 45: 75-85. (in Persian with English abstract)

21- Najafi Pour G., and Taghavi S.M. 2011. Comparison of P. syringae pv. syringae from different hosts based on pathogenicity and BOX-PCR in Iran. Journal of Agricultural Science and Technology, 13: 431-442.

22- Palleroni N.J., Kunisawa R., Contopoulou R., and Doudoroff M. 1973. Nucleic acid homologies in the genus Pseudomonas. International Journal of Systematic Bacteriology, 23:333-339.

23- Rademarker J.W., Hoste B., Louws F.J., kersters k., Swings J., Vauterine L., Vauterine P., and De bruijn F.J. 2000. Comparison of AFLP and rep-PCR genomic fingerprinting with DNA-DNA homology studies: Xanthomonas as a model system. International Journal of Systematic and Evolutionary Microbiology, 50: 665-677.

24- Rohlf F.J. 2000. NTSYSpc, Numerical taxonomy and multivariate analysis system. Version 2.1 Exete software. Applied Biostatistics INC., NY, USA. 493-505.

25- Schadd N.W., Jones J.B., and Chun W. 2001. Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3thed. American Phytopathology Society Press.

26- Shams-Bakhsh M., and Rahimian H. 1990. Identification agents of Citrus Blast in Mazandaran. p. 150. In Proceedings of the 9th Iranian Plant Protection Congress. 1990. Ferdowsi University of Mashhad, Mashhad, Iran.

27- Sorensen K.N., Kim K.H., and Takemoto J.Y. 1998. PCR Detection of cyclic lipodepsinonapeptide-producing Pseudomonas syringae pv. syringae and similarity of strains. Applied and Environmental Microbiology, 64: 226-230.

28- Versalovic J., Koeuth T., and Lupski J.R. 1991. Distribution of repetitive DNA sequences in Eubacteria and application fingerprinting bacterial genomes. Nucleic Acids Research, 19: 6823-6831.

29- Versalovic J., Scheider M., De bruijn F.J., and Lupski J.R. 1994. Genomic fingerprinting of bacteria using repetitive sequence based polymerase chain reaction. Methods in Molecular and Cellular Biology, 5: 25-40.

30- Weingart H., and Volksch B. 1997. Genetic fingerprinting of Pseudomonas syringae pathovars using ERIC, REP and IS50-PCR. Phytopathology, 87: 339-345.

31- Whiteside L.O., Garnesy S.M., and Timmer L.W. 1989. Campendium of citrus disease. 2thed. American Phytopathology Society Press.

32- Yessad S., Manceau C., and Luisetti J. 1992. A detached leaf assay to evaluate virulence and pathogenicity of strains of Pseudomonas syringae pv. syringae on pear. Plant Disease. 76(4):370-373.

33- Young J.M. 1991. Pathogenicity and identification of the lilac pathogen Pseudomonas syringae pv syringae Van Hall 1902. Annual Applied Biology, 118: 283-298.

34- Young J.M., Takikawa Y., Gardan L., and Stead D.E. 1992. Changing concepts in the taxonomy of plant pathogenic bacteria. Annual Review of Phytopathology, 30: 67-105.

35- Zhao Y., Damicone J.P., Demezas D.H., Rangaswamy V., and Bender C.L. 2000. Bacterial leaf spot diseases of leafly crucifers in Oklahama caused by Pseudomonas syringe pv. maculicola. Plant Disease, 84: 1015- 1020.