Bursa’da plum pox virus (Şarka)'ün yaygınlığının ve genetik çeşitliliğinin belirlenmesi

Öz

Plum pox virus (PPV) tarafından oluşturulan Şarka hastalığı, sert çekirdekli meyve türlerinin en önemli viral hastalığı olarak görülmektedir. Son yıllarda yapılan nükleotid dizileme çalışmaları ile PPV'nin Türkiye’de şehir merkezi ev bahçelerinde sanıldığından daha fazla yaygınlık ve genetik çeşitlilik gösterdiği belirlenmiştir. Ev bahçeleri Türkiye tipi PPV-D ve PPV-T ırkları ile bulaşık iken, kapama bahçeler Avrupa tipi PPV-M ırkı ile bulaşıktır. Bursa ili Türkiye’de önemli bir şeftali ve fidan üretim bölgesi olup, aynı zamanda Avrupa’dan gelen fidanların Türkiye içinde dağıtım merkezidir. Bu nedenle iyi izlenmesi gereken bir ildir. Bu çalışmada, Bursa ilinde surveyler yapılmış, tespit edilen PPV-pozitif örneklere ait izolatlar, Türkiye’nin diğer bölgelerinde ve Avrupa'da PPV izolatları ile karşılaştırılmıştır. Bursa ili ve civarında, 50 kapama bahçeden 118 örnek DASI-ELISA ve RT-PCR yöntemleri ile test edilmiş, 32 bahçeden alınan 64 örnek pozitif bulunmuştur. Bu örneklerin 664 nt uzunluğunda P3-6K1 bölgesi ve 969 nt uzunluğunda CT-3’UTR bölgesi dizilenmiş, GenBank'a kayıtlı 200 adet izolat ile birlikte analiz edilmiştir. Otuz şeftali bahçesinden alınan 56 izolatın, Türkiye kapama bahçelerinde tespit edilen Avrupa PPV-M izolatı olduğu belirlenmiştir. Bursa’da iki erik bahçesinden alınan 8 adet izolat ise PPV-D olarak tespit edilmiş olup, bu izolatlar Türkiye tipi PPV-D izolatları ile aynı grupta yer almıştır. Sonuç itibariyle, Bursa kapama şeftali bahçelerinin Avrupa tipi PPV-M ile yaygın olarak bulaşık olduğu ve bu izolatların PPV’nin yeni giriş yaptığı Aydın, Denizli, Çanakkale, Isparta ve Kayseri illeri kapama bahçe izolatları ile yakın genetik benzerlik gösterdiği belirlenmiştir. Bu sonuç, PPV-M ırkının Türkiye’de yayılmasında, Bursa-Türkiye üzerinden olan fidan trafiğinin rolüne işaret etmiştir.

Anahtar Kelimeler

• Şarka,Şeftali,Sert çekirdekli meyveler,PPV-Marcus,PPV-Dideron

Determination of prevalence and genetic diversity of plum pox virus (sharka) in peach orchards in Bursa

Öz

Plum pox virus (PPV) is the most important viral disease of stone fruit species. Recent nucleotide sequencing studies have revealed that PPV is prevalent and exhibits high genetic diversity in residential gardens of several provinces in Turkey. Trees in residential gardens have been found to be infected by Turkish type PPV-D and PPV-T while trees in professional orchards in several cities have been found to be affected by the European strain PPV-M. The province Bursa is main peach producer region in Turkey but moreover Bursa hosts big nursery industry and is the distribution center of seedlings imported from European countries. In this research, surveys were made in the orchards of Bursa and surrounding regions, and comparisons were made in order to investigate genetic relationships among the isolates of Bursa and other PPV isolates determined in Turkey as well as European PPV isolates. A total of  118 samples from 50 orchards were examined by DASI-ELISA and RT-PCR methods. Of the all, 64 samples from 32 orchards were found to be infected with PPV. The P3-6K1 gene region with 664 nt length and the CP-3’UTR with 970 nt of the 64 isolates were sequenced and analyzed along with 200 sequence records in the GenBank database. Fifty-six isolates were identified as European type PPV-M. Eighth isolates from two plums orchard in Bursa were identified as PPV-D and grouped with Turkish type PPV-D. These results indicate that Bursa-Turkey seedlings traffic pathway seems to be the source of PPV-M infection in the orchards in the country.

Anahtar Kelimeler

• Sharka,Peach,Stone fruits,PPV-Marcus,PPV-Dideron

Kaynakça

1. Akbaş, B., Değirmenci, K., Çiftçi, O., Kaya, A., Yurtmen, M., Uzunoğulları, N., Çelik, N., Türkölmez, Ş., 2011. Update on Plum pox virus distribution in Turkey. Phytopathology Mediterrian, 50: 75−83.
2. Cambra, M., Capote, N., Myrta, A,. Llácer, G., 2006. Plum pox virus and the estimated costs associated with sharka disease. EPPO Bull., 36: 202–204.
3. Candresse, T., Svanella-Dumas, L., Gentit, P., Çağlayan, K., Çevik, B., 2007. First report of the presence of Plum pox virus Rec strain in Turkey. Plant Dis., 91: 331.
4. Ceylan, A., Gürcan, K., Akbulut, M., 2015. Complete nucleotide sequence analysis of Plum Pox Virus 'Turkey' isolates. International Conference on Virus and Other Graft Transmissible Disease of Fruit Crops, S.50, 8-12 Haziran, Morioka, Japonya.
5. Ceylan, A., Gürcan, K., Akbulut, M., Ghaderi, M., 2014. Kayseri’de yüksek şarka enfeksiyonu. Erciyes Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 30(2): 80-85.
6. Ceylan, A., Gürcan, K., Ghaderi, M., Akbulut, M., Ulubaş Serçe, Ç., 2013. Transmission route of new plum pox virüs in Turkey. 2nd International Symposium on Plum Pox Virus, S.19, 3-6 September, Olomouc, Czech Republic.
7. Crescenzi, A., D’aquino, L., Comes, S., Nuzzaci, M., Piazzolla, P., 1997. Charecterisation of the sweet cherry isolate of Plum pox potyvirus, Plant Dis., 81(7): 711-714.
8. Çelik, N., Topkaya Kütük, B., 2013. Antalya ilinde şarka virüs hastalığının belirlenmesi. Derim, 30(2):1-10.

9. Çıtır, A., İlbağı, H., 2008. Serological identification of some important viruses on fruit trees and bushes in Tekirdağ province of Turkey, Proceedings of the Twentieth International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops, Fruit Tree Diseases’’, Antalya, Turkey. Acta Hortic., 781: 103-106.
10. Dallot, S., Glasa, M., Jevremovic, D., Kamenova, I., Paunovic, S., Labonne, G., 2011. Mediterranean and central-eastern European countries host viruses of two different clades of Plum pox virus strain M. Arch. Vir., 156: 539–542.
11. Deligöz, İ., Değirmenci, K., Sökmen, M., 2015. Samsun İlinde Sert Çekirdekli Meyve Türlerinde Şarka Hastalığı Etmeninin (Plum pox virus) Belirlenmesi. Anadolu Tarım Bilimleri Dergisi, 30(3):227-235.
12. Elibüyük, I.Ö. 2004. Current situation of sharka disease in Ankara, Turkey. Phytoparasitica, 32(4), 417–420.
13. Elibüyük, İ.Ö. 2006. Detection of Plum Pox Virus in Ornamental Prunus Cerasifera. Phytoparasitica, 34 (4): 347– 352.
14. EPPO, 2004. Standard PM 7/32 Plum pox potyvirus. Bulletin EPPO Bull., 34: 247–256.
15. García, J.A. Glasa M., Cambra, M., Candresse, T., 2014. Plum pox virus and sharka: a model potyvirus and a major disease. Molecular Plant Pathology, 15 (3): 226-41.
16. García, J. A., Cambra, M., 2007. Plum pox virus and sharka disease. Plant Viruses, 1:69–79.
17. Glasa, M., Marie-Jeanne, V., Moury, B., Kúdela, O., Quiot, J.B., 2002. Molecular variability 17 of the P3-6K1 genomic region among geographically and biologically distinct 18 isolates of Plum pox virus. Arch Virol., 147:563–575.
18. Glasa, M., Malinowski T., Predajna L., Pupola N., Dekena D., Michalczuk, L., Candresse, T., 2011. Sequence variability, recombination analysis, and specific detection of the W strain of Plum pox virus. Phytopathology, 101(8): 980–985.
19. Glasa, M., Prikhodko, Y., Predajna, L., Nagyova, A., Shneyder, Y., Zhivaeva, T., Subr, Z., Cambra, M., Candresse, T., 2013. Characterization of sour cherry isolates of Plum pox virus from the Volga basin in Russia reveals a new cherry strain of the virus. Phytopathology, 103(9): 972–979.
20. Glasa, M., Palkovics, L., Komínek, P., Labonne, G., Pittnerová, S., Kúdela, O., Candresse, T., Subr, Z., 2004. Geographically and temporally distant natural recombinant isolates of Plum pox virus are genetically very similar and form a unique PPV. Journal of General Virology, 85: 2671-81.
21. Gümüs, M., Paylan, I.C., Matic, S., Myrta, A., Sipahioglu H.M., Erkan, S., 2007. Occurrence and distribution of stone fruit viruses and viroids in commercial plantings of Prunus species in western Anatolia, Turkey, Journal of Plant Pathology, 89: 265–268.
22. Gürcan, K., Ceylan, A., 2016a. Full Genome Analysis of Plum pox virus-D isolates of Turkey. 3nd International Symposium on Plum Pox Virus, S. 19, 9-13 Mayıs, Antalya, Türkiye.
23. Gürcan, K., Ceylan, A., 2016b. Strain identification and sequence variability of plum pox virus in Turkey. Turkish Journal of Agriculture and Forestry. DOI: 10.3906/tar-1509-97.
24. Gürcan, K., Teber, S., Değirmenci, K., 2016. High Sequence variability of Plum pox virus: A new divergent group of Marcus strain. Romanian Biotechnological Letters (kabul edildi).
25. İlbağı, H., , Çıtır, A., 2014. Detection and partial molecular characterization of Plum pox virus on almond trees in Turkey. Phytoparasitica, 42(4): 485–491.
26. İlbağı, H., Cıtır, A., Bostan, H., 2008. Prunus spinosa L. A natural wild host of some important fruit viruses in Tekirdağ, Turkey, Proceedings of the Twentieth International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops, Fruit Tree Diseases, Antalya, Turkey, Acta Hortic., 781, 33-36.
27. James, D., Varga, A., Thompson, D., Hayes, S., 2003. Detection of a new and unusual isolate of Plum pox plum (Prunus domestica virus in). Plant Dis., 87: 1119–1124.
28. James, D., Varga, A., Sanderson, D., 2013. Genetic diversity of Plum pox virus: strains, disease and related challenges for control. Can. J. Plant Pathol., 35: 431–441.
29. Kalashyan, Y.A., Bilkey, N.D., Verderevskaya, T.D., Rubina, E.V., 1994. Plum pox virus on sour cherry in Moldava. EPPO Bull., 24: 645–649.
30. Kerlan, C., Dunez, J., 1979. Différenciation biologique et sérologique des souches du virus de la sharka. Annales de Phytopathologie, 11:241–250.
31. Koç, G., Baloğlu, S. 2006. First report of sharka in the Çukurova region of Turkey. J. of Plant Pathol., 88 (3 suppl.): 68.
32. Levy, L., Damsteegt, V., Welliver, R., 2000. First report of Plum pox virus (sharka disease) in Prunus persicae in the United States. Plant Dis, 84: 202.
33. Kurçman, S., 1973. Detection of sharka virus on apricot and plum trees in Ankara. Journal fo Turk Pyhtopatholgy, 2:124–129.
34. Maejima, K., Himeno, M., Komatsu, K., Takinami, Y., Hashimoto, M., Takahashi, S., Yamaji, Y., Oshima, K. Namba, S., 2011. Molecular epidemiology of Plum pox virus in Japan. Phytopathology, 101(5): 567–574.
35. Myrta, A., Boscia, D., Potere, O., Kolber, M., Nemeth, M., Terlizzi, B., Cambra, M., Savino, V., 2001. Existence of two serological subclusters of Plum pox virus, strain M. Eur. J. Plant Pathol., 107: 845–848.
36. Palmisano, F., Boscia, D., Minafra, A., Myrta, A., Candresse, T., 2012. An a typical albanian isolate of Plum pox virus could be the progenitor of the Marcusstrain..http://icvf.jki.bund.de/dokumente/upload/74140_
37. icvf2012abstracts.pdf. Accessed 15 October 2014.
38. Palmisano, F., Minafra, A., Myrta, A. Boscia, D., 2015. First Report Of Plum Pox Virus Strain PPV-T in Albania. J. Plant Pathol., 97(2): 391-403.
39. Roy, A.S., Smith, I.M., 1994. Plum pox situation in Europe. Bulletin OEPP/EPPO Bulletin. 24: 515-523.
40. Sahtiyancı, S., 1969. Virus de la Sharka chez la prunier. Bullet. Phytosan FAO, 69.
41. Scholthof, K.B., Adkins, S., Czosnek, H., Palukaitis, P., Jacquot, E., Hohn, T., Hohn, B., Saunders, K., Candresse, T., Ahlquist, P., Hemenway, C., Foster, G.D., 2011. Top 10 plant viruses in molecular plant pathology. Mol Plant P., 12: 938-954.
42. Kumar, S., 2013. MEGA6: Molecular 6 Evolutionary Genetics Analysis Version 6.0. Mol. Biol. Evol., 30:2725-2729.
43. Teber, S., Gürcan K., 2016. Recombination analysis of 51 PPV isolates including 10 genomes of PPV-M Istanbul, 3nd International Symposium on Plum Pox Virus, S. 33, 9-13, Antalya, Turkey.
44. Thompson, D., McCann, M., MacLeod, M., Lye, D., Green, M., James, D., 2001. First report of Plum Sertkaya, G., Ulubaş, Ç., Çağlayan, K. 2003. Detection and characterization of Plum pox potyvirus (PPV) by DAS-ELISA and RTPCR/ RFLP analysis in Turkey, Turk. J. Agric. For., 27: 213-220.
45. Tamura, K., Nei, M., Kumar, S., 2004. Prospects for inferring very large phylogenies by 4 using the neighbor-joining method. PNAS 101:11030-11035.
46. Tamura, K., Stecher, G., Peterson, D., Filipski, A., pox virus in Ontario, Canada. Plant Dis., 85 (11): 97.
47. Ulubaş Serçe, C., Candresse. T., Svanella-Dumas, L., Krizbai, L., Gazel, M., Çağlayan, K., 2009. Further characterization of a new recombinant group of Plum pox virus isolates, PPV-T, found in orchards in the Ankara province of Turkey. Virus Res., 142 (1-2):121–126.
48. Ulubaş Serçe Ç., Gazel, M., Çağlayan, K., 2011. Plum pox virus streynlerinin Türkiye’ deki Dağılımı. Türkiye IV. Bitki Koruma Kongresi Bildirileri, Kahramanmaraş s. 72.
49. Wetzel T., Candresse T., Ravelonandro M., Delbos R.P., Mazyad H., Aboul-Ata A.E., Dunez, J., 1991. Nucleotide sequence of the 3′ terminal region of the RNA from a widely divergent strain (El Amar) of Plum pox potyvirus. J. Gen. Vir., 72: 1741–1746.