ردیابی و شناسایی فیتوپلاسمای همراه با بیماری زوال گلابی و پسیل گلابی (Cacopsylla pyricola) در استان‌های فارس و خراسان رضوی

نوع مقاله : مقاله کامل پژوهشی

نویسندگان

1 دانش‌آموخته کارشناسی ارشد گروه بیماری شناسی گیاهی دانشگاه زابل.

2 استاد پژوهشی بیماری‌شناسی گیاهی

3 پزوهشیار بیماری شناسی گیاهی گروه بیماری شناسی گیاهی دانشگاه زابل

4 استادیار پژوهشی حشره‌شناسی کشاورزی مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی فارس.

چکیده

در بازدیدهای سال‌های 1386 تا 1391 در باغ‌های گلابی استان‌های خراسان رضوی و فارس علائم زوال گلابی مشاهده گردید. در آزمون‌های PCR دو مرحله‌ای با استفاده از جفت آغازگر P1/P7 در دور اول و جفت آغازگرهای R16F2n/R16R2 و fO1/rO1 در دور دوم، در درختان دارای علائم زوال در باغ‌های مشهد و چناران در استان خراسان رضوی و سه منطقه از فارس شامل باجگاه، دشت ارژن و استهبان، پسیل(Cacopsylla pyricola)‌های جمع‌آوری‌شده از روی درختان دارای علائم در باغ‌های ناسیونال مشهد و باجگاه شیراز و گیاهان مایه‌زنی‌شده با عامل زوال گلابی به ترتیب قطعاتی به اندازه تقریبی 1200 و 1050 جفت باز از ژن آر. ان. ای. ریبوزومی16S  تکثیر شد. آزمون‌های RFLP واقعی و مجازی نشان داد که فیتوپلاسما‌های ردیابی‌شده در درختان گلابی، حشرات پسیل و گیاهان مایه‌زنی‌شده، با یکدیگر و با 'Candidatus Phytoplasma pyri' ، عامل زوال گلابی یکسان می‌باشند. آنالیز فیلوژنتیکی نیز تعلق فیتوپلاسماهای ردیابی‌شده در درختان گلابی و نمونه‌های پسیل به'Ca. Phytoplasma pyri'  را تأیید کرد. فیتوپلاسمای عامل بیماری زوال گلابی از باغ‌های ناسیونال مشهد و باجگاه به ترتیب به عنوان نماینده‌های عامل زوال گلابی در استان‌های فارس و خراسان رضوی از طریق پیوند به نهال‌های گلابی و "به" و از طریق سس به پروانش انتقال داده شد. این اولین گزارش از وجود فیتوپلاسمای عامل زوال گلابی در پسیل گلابی در ایران و اولین گزارش از بیماری زوال گلابی در استان خراسان رضوی و نقاط جدیدی از استان فارس می‌باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Detection and identification of the phytoplasma associated with pear decline disease and the pear psylla, Cacopsylla pyricola in Fars and Khorasan-e-Razavi provinces*

نویسندگان [English]

  • S.M. Sherafat 1
  • M. Salehi 2
  • M. Salari Salari 3
  • N. Rastegari Rastegari 4
چکیده [English]

During 2007-2012 surveys, pear decline (PD) symptoms was observed in different pear orchards in Mashhad andChenaran (Khorasan-e-Razavi province), and 11 areas in Fars province. In nested PCR assays using P1/P7 primer pair in the first round and R16F2n/R16R2 and fO1/rO1 primer pairs in the second round, expected fragments of 1250 and 1050 bp, respectively, were amplified from symptomatic naturally infected pear trees, pear psyllid (Cacopsylla pyricola) insects collected on PD affected trees and experimentally inoculated plants. Real and virtual RFLP analyses showed that phytoplasmas detected in naturally decline affected pear trees, inoculated plants and psyllid insects were identical and belonged to 'Candidatus Phytoplasma pyri'. Phylogenetic analysis also confirmed that the detected phytoplasmas  are closely related to  'Ca. Phytoplasma pyri'. As representatives of PD phytoplasmas detected in Fars and Khorasan-e-Razavi provinces, respectively, pear decline agents from Bajgah and National orchard were transmitted from PD affected trees to pear and quince young trees by grafting and to periwinkle via dodder inoculation. This is the first report of association of 'Ca. Phytoplasma pyri' with PD disease and psylla insects in Khorasan-e-Razavi and some areas in Fars province, Iran.

کلیدواژه‌ها [English]

  • 'Candidatus Phytoplasma pyri'
  • Molecular analysis
  • Phytoplasma
  • 16SrX-C
 
.
Abou-Jawdah Y., Karakashian A., Sobh H., Martini M. and Lee I.-M. 2002. An epidemic of almond witches’-broom in Lebanon: classification and phylogenetic relationship of the associated phytoplasma. Plant Disease 86:477–484.
Ahmadi A., Mehrvar A., Lotfalizadeh H., Gharekhani G. and Burckhardt D. 2012. Determination of the species diversity of Psylloidea superfamily (Hemiptera) in East-Azarbaijan province. Journal of Sustainable Agriculture and Production Science 22:193-212. (In Farsi With English Summary).
Avinent L., Llacer G., Almasellas J. and Tora R. 1997. Pear decline in Spain. Plant Pathology 46:694–698.
Bertaccini A. and Duduk B. 2009. Phytoplasma and phytoplasma diseases: a review of recent research. Phytopathologia Mediterranea 2009 48:355–78.
Burckhardt D. and Hodkinson I. D. 1986. A revision of the west Palaearctic pear psyllids (Hemiptera: Psyllidae). Bulletin of Entomological Research 76:119-132.
Carraro L., Loi N., Ermacora P., Gregoris A. and Osler R. 1998. Transmission of pear decline by using naturally infected Cacopsylla pyri. Acta Horticulture 472:665-668.
Davies D. L., Guise C. M., Clark M. F. and Adams N. A. 1992. Parry’s disease of pears is similar to pear decline and associated with mycoplasma-like organisms transmitted by Cacopsylla pyricola. Plant Pathology 41: 194–203.
Deng S. and Hiruki D. 1991. Amplification of 16S rRNA genes from culturable and nonculturable mollicutes. Journal of Microbiological Methods 14:53–61.
Emami M. S. 2016. Field evaluation of two biorational compounds in the control of pear psylla, Cacopsylla pyricola (Förster), on pear trees.Archives of Phytopathology and Plant Protection 49:11-18.
Errea P., Aguelo V. and Hormaza1 J. I. 2002. Seasonal variations in detection and transmission of pear decline phytoplasma. Journal of Phytopathology 150:439–443.
Garcia-Chapa M., Lavina A., Sanchez I., Medina V. and Batlle A. 2003. Occurrence, symptom expression and characterization of phytoplasma associated with pear decline disease in Catalonia (Spain). Journal of Phytopathology151: 584–590.
Gazel M., Ulubasserce C., Caglayan K. and Ozturk H. 2007. Detection of Candidatus Phytoplasma pyri in Turkey. Bulletin of Insectology 60:125-126.
Gundersen D. E. and Lee I. M. 1996. Ultrasensitive detection of phytoplasmas by nested-PCR assays using two universal primer pairs. Phytopathologia Mediterranea 35:144-151.
Hashemi-Tameh M., Bahar M. and Zirak L. 2014. Molecular characterization of phytoplasmas related to apple proliferation and aster yellows groups associated with pear decline disease in Iran. Journal of Phytopathology 162:660-669
Hsiu-Lin L., Ching-Chung  C. and Chan-Pin L. 2007. Detection and identification of the phytoplasma associated with pear decline in Taiwan. European Journal of Plant Pathology 117:281–291.
Jensen D. D., Griggs W. H., Gonzales C. Q. and Schneider H. 1964. Pear decline virus transmission by pear psylla. Phytopathology 54:1346-1351.
Lee I. M., Gundersen-Rindal D. E., Davis R. E. and Bartoszyk I. M. 1998. Revised classification scheme of phytoplasmas based on RFLP analyses of16S rRNA and ribosomal protein gene sequences. International Journal of Systematic Bacteriology 48:1153–1169.
Lemoine J.1991. Deperissement du poirier: röle de Psylla pyri dans sa dissemination. arboriculture fruitière 442:28-32.

Lepka P., Stitt M., Moll E. and Seemüller E .1999. Effect of phytoplasmal infection on concentration and translocation of carbohydrates and amino acids in periwinkle and tobacco. Physiological and Molecular Plant Pathology 55:59-68.

 
Lorenz K. H., Schneider B., Ahrens U., and Seemuller E. 1995. Detection of the apple proliferation and pear decline phytoplasmas by PCR amplification of ribosomal and non ribosomal DNA. Phytopathology 85:771-776.
Marcone C., Hergenhahn F., Ragozzino A. and Seemuller E. 1999. Dodder transmission of pear decline, European stone fruit yellows, rubus stunt, picris echinoids yellows and cotton phyllody phytoplasmas to periwinkle. Journal of Phytopathology147:187-192.
Musetti R., Favali  M. and Pressacco L. 2000. Histopathology and polyphenol content in plants infected by phytoplasmas. Cytobios 102:133-147.
Németh M. 1986. Virus, Mycoplasma and Rickettsia Diseases of Fruit Trees. Lancaster, Boston, USA/Dordrecht, Netherlands: M. Nijhoff Publishers. 841 pp.
Salehi M., Izadpanah K., Taghavi S M. and Rahimian H. 2008. Characterization of a phytoplasma associated with pear decline in Iran. Journal of Phytopathology 156:493–495.
Sambrook, J., Fritsch E. F. and Maniatis T. 1989. Molecular cloning: a laboratory manual. vol. 3. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
Schneider B., Seemeider E., Smart C. D. and Kirkpatrick B. C. 1995. Phylogenetic classification of plant pathogenic mycoplasma-like organisms or phytoplasmas, pp. 369-380. In: S. Razin and J. G. Tully (Eds). Molecular and Diagnostic Procedures in Mycoplasmology, Vol.1. Academic Press, San Diego, CA, USA.
Seemuler E. 1989. Pear decline, pp: 188-201. In:P. R. Fridlund, (Ed). Virus and Virus like Diseases of Pome Fruits and Simulating Noninfectious Disorders. Cooperative Extension College of Agriculture and Horne Economics, Washington State University, Pullman.
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., Schneider B. and Jarausch B. 2011. Pear Decline Phytoplasma, pp. 77-.84. In: A. Hadidi, M. Barba, T.  Candresse and W. Jelkmann (Eds). Virus and Virus-Like Diseases of Pome and Stones Fruits. The American Phytopathological Society, St. Paul, Minnesota, USA.
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.
Wei W., Davis R. E., Lee I. M., and Zhao Y. 2007. Computer-simulated RFLP analysis of 16S rRNA genes: identification of ten new phytoplasma groups. International Journal of Systematic and Evolutionary Microbiology 57:1855-67.
Weintraub P. G. and Beanland L. 2006. Insect vectors of phytoplasmas. Annual. Review of Entomology 51:91–111.
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.
Zhao Y., Wei W., Lee I.-M., Shao J., Suo X. and Davis R. E. 2009. Construction of an interactive online phytoplasma classiffication tool, iPhyClassifier, and its application in analysis of the peach X-disease phytoplasma group (16SrIII). International Journal of Systematic and Evolutionary Microbiology 59:2582-2593.