برهمکنش فیتوپلاسماهای عامل بیماری‌های جاروک لیموترش و فیلودی کاهو در آلودگی مخلوط بوته‌های پروانش

نوع مقاله : مقاله کامل پژوهشی

نویسندگان

1 دانشجوی دکتری بیماری شناسی گیاهی، ، دانشکده کشاورزی، دانشگاه شیراز، ایران

2 استاد بخش گیاهپزشکی، دانشکده کشاورزی، دانشگاه شیراز، ایران

3 بخش گیاهپزشکی، دانشکده کشاورزی، دانشگاه شیراز، ایران

4 دانشیار بخش گیاهپزشکی، دانشکده کشاورزی، دانشگاه شیراز، ایران

چکیده

در این پژوهش اثر آلودگی مخلوط فیتوپلاسماهای جاروک لیموترش (16SrII-B) و فیلودی کاهو (16SrIX-D) بر روی علائم بیماری و تراکم  سلول‌های فیتوپلاسما در بوته‌های  پروانش (Catharanthus roseus (L.) G.Don) مطالعه شد. تعداد چهار بوته پروانش سالم  و هم‌سن  از طریق قطعات سس آلوده ابتدا با فیتوپلاسمای عامل فیلودی کاهو و پس از سه هفته با فیتوپلاسمای جاروک لیموترش مایه‌زنی شدند. علائم بیماری و جمعیت فیتوپلاسما در این بوته‌ها با علائم بیماری و جمعیت فیتوپلاسما در بوته‌هایی که تنها با یکی از این فیتوپلاسماها مایه‌زنی شده بودند مقایسه گردید. در بوته‌هایی که به صورت مخلوط مایه‌زنی شده بودند تنها کاهش خفیفی در اندازه برگ و فاصله میانگره‌ها ایجاد شد که با علائم شدید ناشی از هر یک از دو فیتوپلاسما، کاملا متفاوت بود. بوته‌هایی که به صورت مخلوط مایه‌زنی شده بودند در مقایسه با بوته‌هایی که تنها با یک فیتوپلاسما مایه‌زنی شده بودند طول عمر بیشتری داشتند. با استفاده از پی‌سی‌آر معمولی و آغازگرهای اختصاصی وجود هر دو فیتوپلاسما در بوته‌هایی که به صورت مخلوط مایه‌زنی شده بودند و همچنین در بوته‌های شاهد مثبت به اثبات رسید. آنالیز داده‌های حاصل از آزمون پی‌سی‌آر در زمان واقعی (Real-time PCR) نشان داد که جمعیت هر دو فیتوپلاسما در بوته‌هایی که به صورت مخلوط مایه‌زنی شده بودند در مقایسه با بوته‌های شاهد مثبت به طور معنی‌داری کاهش داشت. به نظر می‌رسد برهمکنش این دو فیتوپلاسما در آلودگی مخلوط از نوع آنتاگونیسم متقابل باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Interaction of witches’- broom disease of lime phytoplasma and lettuce phyllody phytoplasma in mixed infection of periwinkle plants

نویسندگان [English]

  • E. Salehi Abarghouei 1
  • K. Izadpanah 2
  • S. M. Taghavi 3
  • H. Hamzezarghani 4
  • A. Afsharifar 2
چکیده [English]

In this research the effect of mixed infection of witches’ broom disease of lime (WBDL) (16SrII-B) and lettuce phyllody (LP) (16SrIX-D) phytoplasmas on symptom expression and phytoplasma density  in periwinkle plants (Catharanthus roseus (L.) G. Don) were investigated. Four healthy periwinkle plants of the same age were first inoculated with LP phytoplasma and after three weeks with WBDL phytoplasma using detached fragments of inoculative dodder (Cuscuta campestris Yank.). Disease symptoms and phytoplasma population in plants inoculated with both phytoplasmas were compared with those in plants inoculated with either WBDL or LP phytoplasma. In doubly inoculated periwinkle plants the disease symptoms mitigated to mild reduction of leaf size and internodes which strongly differed from those in periwinkle plants singly inoculated with either WBDL or LP phytoplasma. Plants with mixed infection lived longer than those inoculated singly with either phytoplasma. Both WBDL and LP phytoplasmas could be detected in singly and doubly infected plants by conventional and real time PCR assays using specific primer pairs. Analysis of the data obtained by real-time PCR showed that the population of both WBDL and LP phytoplasmas in the mixed infection was significantly lower than that in plants singly inoculated with WBDL or LP phytoplasma. It seems that interaction of WBDL and LP phytoplasmas in mixed infection of periwinkle is of the mutual suppression type.

کلیدواژه‌ها [English]

  • Mixed infection
  • Mutual antagonism
  • Phytoplasma population
  • Real-time PCR
 
Alhudaib K., Arocha Y., Wilson M. and Jones P. 2009. Molecular identification, potential vectors and alternative hosts of the phytoplasma associated with a lime decline disease in Saudi Arabia. Crop Protection 28: 13-18.
Axtell M.J., Synder J.A. and Bartel D.P. 2007. Common functions for diverse
small RNAs of land plants. Plant Cell 19: 1750–1769
Bai, X., Correa V. R., Tourno T. Y., Ammar E. D., Kamoun S. and Hogenhout S. A. 2009. AY-WB Phytoplasma Secretes a Protein That Targets Plant Cell Nuclei. Molecular Plant- Microbe Interactions 22: 18-30.
Bertaccini A. and Duduk B. 2009. Phytoplasma and phytoplasma diseases: a review of recent research. Phytopathologia Mediterana 48: 355-378.
Bertaccini, A., Duduk, B., Paltrineri, S. and Contaldo, N. 2014. Phytoplasmas and Phytoplasmas Diseases: A Severe Threat to Agriculture. American Journal of Plant Sciences 1763-1788.
Carrillo-Tripp J., Lozoya-Gloriae. and Rivera-Bustamante R.F. 2007. Symptom remission and specific resistance of pepper plants after infection by Pepper golden mosaic virus. Phytopathology 97: 51-59.
Castelain C. Jullian J. P. Lemaire J. M. and Morvan G. 2007. Lutte biologique contre l’ECA: la prémunition. Arboriculture 611:33-38.                                                       
Chen J., Pu X., Deng X., Liu S., Li H. and Civerolo E. 2009. A phytoplasma related to ‘Candidatus phytoplasma asteris detected in citrus showing Huanglongbing (yellow shoot disease) symptoms in Guangdong, P. R. China. Phytopathology 99:236–242.
Das A. K., Nerkar S., Thakre N. and Kumar A. 2016. First report of 'Candidatus Phytoplasma trifolii' (16SrVIgroup) in Nagpur mandarin (Citrus reticulata) showing huanglongbing symptoms in central India. New Disease Reports 34: 15.
Eamens A., Wang M. B., Smith N. A. and Waterhouse P. M. 2008. RNA silencing in plants: yesterday, today, and tomorrow. Plant Physiology 456-468.
Freitag J. H. 1964. Interaction and mutual suppression among three strains of aster yellows virus. Virology24: 401-413.
Garrett K. A., Madden L. V., Hughes G. and Pfender W. F. 2004. New applications of statistical tools in plant pathology. Phytopathology 94(9): 999-1003
Kiss E. Seemüller E. and Sule S. 2013. Cross-protection Test with ‘Candidatus Phytoplasma mali’ 1/93 Strain against Closely and Distantly Related Phytoplasma Strains. Acta Phytopathologica et Entomologica Hungarica 207–218.
Koochakpour Z. and Fakheri B. 2014. RNA silencing and plant viral diseases. Iranian Journal of Fuzzy Systems 3: 946 -951.
 
Kunkel L. O. 1936. Immunological studies on the three peach diseases, yellows, rosette, and little peach. Phytopathology 26: 201-219.
Kunkel L. O. 1955. Cross protection between strains of aster yellows. Advances in Virus Research 3: 251-273.
Lou B., Bai X., Bai Y., Deng C., RoyChowdhury M., Chen C. and Song Y., 2014. Detection and molecular characterization of a 16SrIIA* phytoplasma in grape-fruit (Citrus paradise) with huanglongbing like symptoms in China. Journal of Phytopathology https://doi.org/10.1111/jph.12200.
MacIntyre D. L. Miyata S. T. Kitaoka M. and Pukatzki S. 2010. The Vibrio cholerae type VI secretion system displays antimicrobial properties. Proceedings of the National Academy of Sciences 107: 19520-19524.
McKinney H. H. (1929). Mosaic diseases in the Canary Islands, West Africa and Gibraltar. Journal of Agricultural Research39: 557-578.
Musetti R., Di Toppi L. S., Ermacora P. and Favali, M. A. 2004. Recovery in apple trees infected with the apple proliferation phytoplasma: an ultrastructural and biochemical study. Phytopathology 94: 203- 208.
Musetti R., Farhan K., De Marco F., Polizzotto R., Paolacci A., Ciaffi M., Ermacora P., Grisan S., Santi S. and Osler, R. 2013. Differentiallyregulated defence genes in Malus domestica during phytoplasma infection and recovery. European Journal of Plant Pathology 136: 13-19.
Musetti R., Paolacci A., Ciaffi M., Tanzarella  O. A., Polizzotto R., Tubaro F., Mizzau M., Ermacora P., Badiani M. and Osler, R. 2010. Phloem cytochemical modification and gene expression following the recovery of apple plants from apple proliferation disease. Phytopathology 100: 390-399.
Patui S., Bertolini A., Clincon, L., Ermacora P., Braidot E., Vianello A., and Zancani M. 2013. Involvement of plasma membrane peroxidases and oxylipin pathway in the recovery from phytoplasma disease in apple (Malus domestica). Physiologia Plantarum 148: 200-213.
Pfaffl M. W. 2004. Quantification strategies In: real-time PCR. Bustin S. A. (Ed). IUL.
California. USA  87-112.
Roggia M. F., Imai H., Shiraya T., Noda Y. and Ueta T. 2014. Protective Role of Glutathione Peroxidase 4 in Laser-Induced Choroidal Neovascularization in Mice. https://doi.org/10.1371/journal.pone.0098864.
Saberi E., Alavi A. M., Safaie N., Moslemkhany C. and Azadvar M. 2017. Bacterial pathogens associated with citrus huanglongbing-like symptoms in southern Iran. Journal of Crop Protection 6: 99-113.
Salaman R. N. 1933. Protective inoculation against a plant virus. Nature 131: 468.
Salehi M. Izadpanah K. Siampour M. Nejat N. and Siampour M. 2007. Partial characterization of phytoplasmas associated with lettuce and wild lettuce phyllodies in Iran. Plant Pathology 56: 669-676.
Salehi M. Izadpanah K. and Taghizadeh M. 2002. Witches broom disease of lime in Iran: New distribution areas, experimental herbaceous hosts and transmission trails: Proceeding of 15th Conference IOCV, IOCV, Riverside 293-296.
Sambrook J. Fritsch E.F. and Maniatis T. 1989. Molecular Cloning: A Laboratory Manual. II ed. Cold Spring Harbour Laboratory Press, Cold Spring Harbour, New York.
 
SAS. 1999. Statistical Analysis System User’ Guide Statistics. SAS Institute Inc. Cary NC 27513 USA.
Schneider B. Seemüller E. Smart C. D. and Kirkpatrick B. C. 1995. Phylogenetic Classification of Plant Pathogenic Mycoplasma like Organisms or PhytoplasmasMolecular and Diagnostic Procedures in Mycoplasmology369-380. In: Razin S. and Tully J.G. (Eds), Academic Press, San Diego.
Schneider B. Sule S. Jelkmann W. and Seemüller E. 2014. Suppression of aggressive strains of 'Candidatus Phytoplasma mali' by Mild Strains in Catharanthus roseus and Nicotiana occidentalis and identication of similar action in apple trees. Phytopathology 104: 453- 461.
Seemüller E. and Schneider B. 2004. ‘Candidatus Phytoplasma mali’, ‘Candidatus Phytoplasma pyri’ and ‘Candidatus Phytoplasma prunorum’, the causal agents of apple proliferation, pear decline and European stone fruit yellows, respectively. International Journal of Systematic and Evolutionary Microbiology 54: 1217–1226.
Seemüller E. and Schneider B. 2007. Differences in virulence and genomic features of strains of ‘Candidatus Phytoplasma mali’, the apple proliferation agent. Phytopathology 97: 964–970.
Seemüller E. Kampmann M. Kiss E. and Schneider B. 2011. HfIB gene-based phytopathogenic classification of Candidatus Phytoplasma mali strains and evidence that strain composition determines virulence in multiply infected apple trees. Molecular Plant-Microbe Interactions 24: 1258-1266.
Seemüller E. Kiss E. Sule S. and Schneider B. 2010. Multiple infection of apple trees by distinct strains of 'Candidatus Phytoplasma mali' and its pathological relevance. Phytopathology 100: 863-870.
Seemüller E. Kunze L. and Schaper U. 1984. Colonization behavior of MLO, and symptom expression of proliferation-diseased apple trees and decline-diseased pear trees over a period of several years. Z. Pflanzenkrankh. Pflanzenschutz91: 525-532.
Seemüller E. Sule S. Kube M. Jelkmann W. and Schneider B. 2013. The AAA+ ATPases and HflB/FtsH proteases of ‘Candidatus Phytoplasma mali’: Phylogenetic diversity, membrane topology, and relationship to strain virulence. Molecular Plant-Microbe Interactions Journal 26: 367- 376.
Sinclair W. A. and Griffiths, H. M. 2000. Variation in aggressiveness of
ash yellows phytoplasmas. Plant Disease 84:282-288.
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.
Teixeira D.C., Wulff N.A., Martins E.C., Kitajima E.W., Bassanezi R., Ayres A.J., Eveillard S., Saillard C. and Bove J.M. 2008. A phytoplasma closely related to the Pigeon Pea Witches-Broom Phytoplasma (16Sr IX) is associated with citrus huanglongbing symptoms in the state of S~ao Paulo, Brazil. Phytopathology 98:977–984.
Untergasser A. Cutcutache I. Koressaar T. Ye J. Faircloth B. C. Remm M. and  Rozen S.G. 2012. Primer3--new capabilities and interfaces. Nucleic Acids Res.
Valenta V. 1959. Interference studies with yellows-type plant viruses. II. Cross protection tests with European and American viruses. Acta Virologica 3: 145-152.
Zhang Y. Uyemoto J. K. and Kirkpatrick B. C. 1998. A small-scale procedure for extracting nucleic acids from woody plants infected with various phytopathogens for PCR assay. Journal of Virological Methods 71: 45-50.
Zreik L. Carle P. Bove J. M. and Garnier M. 1995. Characterization of the mycoplasmalike organism associated with witches’-broom disease of lime and proposition of a candidate’s taxon for the organism, ‘Candidatus Phytoplasma aurantifolia’. International Journal of Systematic Bacteriology 45: 449–453.