Determination and characterization of several virus diseases on almond trees in the Eastern Mediterranean Region

Abstract

Recently, production of almond has tended to increase in the Mediterranean countries as well as in Turkey. As in all plants, almond also suffer from viral diseases leading to qualitative and quantitative losses. Distribution of Prune dwarf virus (PDV), Prunus necrotic ring spot virus (PNRSV), Plum pox virus (PPV), Apple chlorotic leaf spot virus (ACLSV), Apple mosaic virus (ApMV) and Cherry leaf roll virus (CLRV) diseases on almond trees in the Eastern Mediterranean Region were investigated and its molecular and biological characterization were performed with this study. During the surveys conducted between 2012-2016, 605 almond leaf samples were collected and tested by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) from 5 provinces of the Eastern Mediterranean Region. 169 almond samples (27.93%) of 605 were found to be infected by at least one virus. Of these, 26 of 129 samples (20.16%) in Adana, 90 of 299 samples (30.10%) in Mersin, 11 of 71 samples (15.49%) in Kahramanmaraş, 34 of 89 samples (38.20%) in Hatay and 8 of 17 samples (47.06%) in Osmaniye were found to be infected at least one virus. While PNRSV, PDV, ACLSV and mixed infection were detected at rate 64%, 18%, 11% and 7% respectively, ApMV, PPV, and CLRV were not found in tested samples. Genomic regions containing the coat protein (CP) gene was reproduced by RT-PCR, resulting in bands of 616 bp for PNRSV, 862 bp for PDV and 358 bp for ACLSV. As they are similar in nucleotide level with 97-99% of the world PNRSV isolates registered to NCBI, 87-96% with PDV isolates, and 81-96% with ACLSV isolates of almond, it is considered that these pathogens spread due to the trade of plant materials.

Keywords

• almond,virus,ELISA,PCR

Doğu Akdeniz Bölgesinde badem ağaçlarında görülen bazı virüs hastalıklarının saptanması ve karakterizasyonu

Öz

Son yıllarda Akdeniz ülkelerinde olduğu gibi ülkemizde de badem üretimi yıldan yıla artış eğilimine girmiştir. Tüm bitkilerde olduğu gibi bademlerde de virüs hastalıklarından dolayı kayıplar meydana gelmektedir. Bu çalışma ile Doğu Akdeniz Bölgesindeki badem alanlarında ekonomik olarak zarar yapan Prune dwarf virus (PDV), Prunus necrotic ringspot virus (PNRSV), Plum pox virus (PPV), Apple chlorotic leaf spot virus (ACLSV), Apple mosaic virus (ApMV) ve Cherry leaf roll virus (CLRV)’un bölgemizdeki dağılımı araştırılarak, moleküler ve biyolojik karakterizasyonu yapılmıştır. 2012-2016 yılları arasında yapılan sürvey çalışmalarında, Doğu Akdeniz Bölgesindeki 5 ilden toplam 605 adet badem örneği toplanıp DAS-ELISA testine tabi tutulmuştur. Toplanan 605 badem örneğinden 169 tanesi (% 27,93) en az bir virüsle enfekteli bulunmuştur. Bunlardan, Adana ilinde toplanan 129 badem örneğinin 26’sı (%20,16), Mersin’de 299 örneğin 90’ı (%30,10), Kahramanmaraş’ta 71 örneğin 11’i (%15,49), Hatay’da 89 örneğin 34’ü (%38,20) ve Osmaniye’de 17 örneğin 8’i (%47,06) en az bir virüs ile enfekteli bulunmuştur. Test edilen örneklerde PNRSV %64, PDV %18, ACLSV %11 ve karışık enfeksiyon %7 oranında tespit edilmiş, ApMV, PPV ve CLRV ise tespit edilememiştir. Enfekteli badem izolatlarından, kılıf protein genini içeren gen bölgesi RT-PCR ile çoğaltılarak PNRSV için 616 bp, PDV için 862 bp ve ACLSV için 358 bp’lik bantlar elde edilmiştir. Badem izolatlarının NCBI’a kayıtlı dünya PNRSV izolatları ile %97-99, PDV izolatları ile %87-96, ACLSV izolatları ile %81-96 oranında nükleotit düzeyde benzerlik gösterdiğinden bu etmenlerin bitki materyallerinin ticaretinden dolayı yayıldığı düşünülmektedir.

Anahtar Kelimeler

• badem,virüs,ELISA,PCR

Kaynakça

1. Anonymous,2007.http://www.tarimkutuphanesi.com/BADEM_YETISTIRICILIGI_00009.html
2. Anonymous, 2002. http://nrsp5.prosser.wsu.edu/nrspidp.html, National Research Support Project 5, Supporting Research And Development Of ‘’Virus Free’’ Pome & Stone Fruit Trees.
3. Barba, M. and Riccıoni, L., 1993. Improvement of Diognostic Methods to Detect Plum Pox Virus in Apricot Plants. Agriculture, 139- 141.
4. Birişik, N., 2009. Yumuşak Çekirdekli Meyve Ağaçlarında Gövde Zararlanmalarına Neden Olan Viral Etmenlerin Biyolojik, Serolojik ve Moleküler Yöntemlerle Saptanması ve Karakterizasyonu. Ç.Ü. Fen Bilimleri Enstitüsü Doktora Tezi. 158 S.
5. Bora, T. ve Karaca, İ., 1970. Kültür Bitkilerinde Hastalığın ve Zararın Ölçülmesi, Ege Üniversitesi, Ziraat Fakültesi Ders Kitabı, No.167. İzmir.
6. Boscia, D., A.M.D’onghıa, B. Di Terlızzı, F. Fagıolı and Osler R., 1999. Accertamenti fitosanitari sul materiale di propagazione. Atti del Convegno Nazionale su Certificazione delle Produzioni Vivaistiche. Eds. V. Savino, P. La Notte, M. Saponari, L. Cavone, A. Bazzoni:99-153.
7. Candresse, T., M. Lanneau, F. Revers, N. Grasseau, G. Macquaire, S. German, T. Malinowski and J. Dunez. 1995. An immuno-capture PCR assay adapted to the detection and the analysis of the molecular variability of the apple chlorotic leaf spot virus. Acta Hort. 386: 136-147.
8. Clark, M. F. ve Adams, A. N., 1977. Characteristics Of The Micro-Plate Method Of Enzime Linked Immuno Sorbent Assay For The Detection Of Plant Viruses. Journal Of General Virology. 34, 475-483.

9. Dal Zotto, A., Nome, S.F., Di Rienzo, J.A. ve Docampo, D.M., 1999. Fluctuations of Prunus necrotic ringspot virus (PNRSV) at various phenological stages in peach cultivars. Plant Disease, volume 83, pages 1055-1057, (1999).
10. Desvignes, J. C., 1999. Virus Diseases of Fruit Trees, CTIFL, Paris, France.
11. Diekmann, M., ve Putter, C.A.J., 1996. Stone Fruits. FAO/IPGRI Technical Guidelines for the Safe Movement of Germplazm. Publication Office IPGRI, Rome.
12. Digiaro, M., Savino, V., Di Terlizzi, B., 1992. Ilarviruses in apricot and plum polen. Acta Horticulture, 309:93-98.
13. Elibüyük, İ. Ö., 2004. Ankara’da Şeftali Ağaçlarında Görülen Sharka Hastalığı Üzerinde Araştırmalar. Tarım Bilimleri Dergisi 2005, 11 (3) 236-243.
14. FAO, 2017. Statistical Databases, faostat-agriculture. http://faostat.fao.org/faostat/ =agriculture
15. Grieco, F., Alkowni, R., Saponari, M., Savino, V. ve Martelli, G.P., 2000. Molecular detection of olive viruses. Bulletin OEPP/EPPO Bulletin. 30: 469-473.
16. Hadidi, A. ve Levy, L., 1994. Accurate identification of Plum pox potyvirus and its differentiation from Asian prunus latent potyvirus in Prunus germplasm. Eur. Plant Prot. Organ. Bull., 24: 633-643.
17. Hassan, M., Myrta A., Polak J., 2006: Simultaneous detection and identification of four pome fruit viruses by one-tube pentaplex RT-PCR. Journal of Virological Methods 133, 124-129.
18. Helguera, P.R., Docampo, D.M., Nome, S.F. and Ducasse, D.A., 2002.Enhanced Detection of Prune Dwarf Virus in Peach Leaves by İmmunocapture- Reverse Transcription Polymerase Chain Reaction with Nested Polymerase Chain Reaction (IC-RT-PCR Nested PCR). J.Phytopathology 150, 94-96. Blackwell Wissenschafts-Verlag, Berlin.
19. ISHS, 1983. Detection Of Virus And Virus-like Diseases Of Fruit Trees Acta Horticulturae, No. 130, 319-326.
20. Karabacak, M., İlbagi, H., 2013. Description and Detection of Almond Virus Diseases in The Trakya Region of Turkey. Türkiye Fitopatoloji Derneği Dergisi, 1-2-3, (2011), 33-40.
21. Kahn, R.P., 1976. Quarantine and the detection of stone fruit viruses in plant importations.Virus Diseases and Noninfectious Disorders of Stone Fruits in North America. U.S.Dept.Agric.Handbook. No. 437, 23-32 p.
22. Kundu, J.K., 2002. The application of RT-PCR Assay for the Detection of Apple Stem Pitting Virus and Apple Stem Grooving Virus in Four Apple Cultivars. Plant Protection Science. Vol.38-1:13-17.
23. Levy, L., Damsteegt, V., Scorza, R. ve Kölber, M., 2000. Plum pox potyvirus Disease of Stone Fruits, http:// www. apsnet. org/ online/ feature/PlumPox/, The American Phytopathological Society.
24. Maliogka, V.I., Charou, A., Efthimiou, K., Katsiani, A.T., 2010. Chatzivassiliou, E.K., Katis, N.I.1 21st International Conference on Virus and other Graft Transmissible Diseases of Fruit Crops. Julius-Kühn-Archiv, 427, 2010.
25. Micke, W.C., 1996. Almond Production Manual. Publication no: 3364, Division of Agriculture and Natural Resources, University of California, Oakland (US).
26. Milusheva, S.A., Borisova, A.Z., 2005. The İncidence of Prunus necrotic ringspot and Prune dwarf viruses in Prunus Species in South Bulgaria, Biotechnol Eq.19/2005/2, 42-45.
27. Myrta, A., Matic, S., Malinowski, T., Pasquini, G. ve Candresse, T., 2011. Apple chlorotic leaf spot in stone fruits. In A. Hadidi, M.Barba, T. Candresse and W.Jelkman (Eds.) Virus and Virus-Like Deseases of Stone Fruits. (pp.85-90) st. Paul, MN: APS Press.
28. Nemeth, M., 1986. Virus, Mycoplasma and Rickettsia Diseases of Fruit trees, Akademia Kiado, Budapest, 841.
29. Ogawa M. J., Zehr, I. E., Bird, W. G., Ritchie, F. D., Uriu, K. ve Uyemoto, K.J., 1995. Compendium Of Stone Fruit Diseases, Page: 69, The American Phytopathological Society Press.
30. Öztürk, Y., 2015. Kiraz Üretim Alanlarında Görülen Virüslerin Serolojik ve Moleküler Yöntemlerle Teşhisi ve Kılıf Protein Gen Dizilimlerinin Belirlenmesi. TAGEM_Projeler.
31. Öztürk, Y. ve Çevik, B., 2015. Genetic Diversity in the Coat Protein Genes of Prune dwarf virus Isolates from Sweet Cherry Growing in Turkey. Plant Pathol J. 2015 Mar; 31(1): 41–49.
32. Presting, G.G., Smith, O.P. ve Brown, C.r. 1995. Resistance to potato leafroll virüs in potato plants transformed with the coat protein gene or with vector control constructs. Phytopathology 85:436-442.
33. Rana, T., Chandel, V., Hallan, V., ve Zaidi, A.A, 2008. Characterisation of Apple chlorotic leaf spot virus infecting almonds in India. Australasian Plant Disease Notes, 2008, 3, 65—67.
34. Reyes, F., Fiore, N., Reyes, M. A., Sepulveda, P., Paredes, V. And Prieto, H., 2002. Biological Behavior And Partial Molecular Characterization Of Six Chilean Isolates Of Plum Pox Virus, Plant Disease, Vol.87, No.1, P.15-20
35. Saraçoğlu, K. ve Erkan, S, 2016. Fasulye Tohumlarındaki Viral Etmenlerin Saptanmasında Tanı Yöntemlerinin Duyarlılıklarının İncelenmesi, Ege Üniv. Ziraat Fak. Derg., 2016, 53 (3):309-315.
36. Spiegel, S., Scott, S. W., Bowman-Vance, V., Tam, Y., Galiakparov, N. N. ve Rosner, A., 1996. Improved Detection Of Prunus necrotic ringspot virus by Polymerase Chain Reaction. Europhean Journal Of Plant Pathology, (102). 681-685.
37. Ulubas Serce C, Ertunc F, Öztürk A., 2009. Identification and Genomic Variability of Prune dwarf virus Variants Infecting Stone Fruit Trees in Turkey. J Phytopathol. 2009;157:298–305.
38. Ulubaş, Ç., ve Ertunc, F., 2004. RT-PCR Detection and Molecular Characterization of Prunus necrotic ringspot virüs Isolates Occuring in Turkey. Journal of Phytopathology, 2004;152:498–502.
39. Vaskova, D., Petrzik, K. ve Spak, J. 2000. Molecular variability of the capsid protein of the Prune dwarf virus. Eur. J. Plant Pathol. 106:573-580.
40. Youssef SA, Shalaby AA, Mazyad HM, Hadidi A., 2002. Detection and identification of Prune dwarf virus and Plum pox virus by standard and multiplex RT-PCR probe capture hybridization (RT-PCR-ELISA) J Plant Pathol. 2002;84: 113–119.