Ordu ili kivi bahçelerinde görülen kök-ur nematodu Meloidogyne incognita'nın moleküler teşhisi

Kök-ur nematodları, kivi bitkisi (Actinidia deliciosa)’nde zararlı olan önemli nematod gruplarından birisidir. Bu çalışma, Ordu ili kivi bahçelerinde görülen kök-ur nematodlarının, PCR (Polymerase Chain Reaction) yöntemi ile teşhisini gerçekleştirmek için yapılmıştır. Bu amaçla 2011-2012 yıllarında Ordu ili Merkez, Perşembe, Ünye, Gülyalı, İkizce, Ulubey ilçelerini kapsayacak şekilde 17 farklı kivi bahçesinden bulaşık kök örnekleri toplanmıştır. Kök-ur nematodu ile enfekteli kivi köklerinden elde edilen dişiler doğrudan teşhis için kullanılmıştır. Moleküler teşhisler, teşhis anahtarı kapsamında sunulan 5S-18S genleri arasındaki rDNA bölgesi IGS-2 için kullanılan primerler ve türe özgü SCAR primerlerinin kullanılması suretiyle gerçekleştirilmiştir. Ordu ili kivi bahçelerinden elde edilen bütün örneklerin teşhisi sonucunda tespit edilen tür Meloidogyne incognita olmuş ve SEC1F/SEC1R türe özgü primerleri 502 bp değerinde amplicon vermiştir. Bu çalışma, kivi bitkisi üzerinde ilk detaylı çalışma niteliğindedir

Anahtar Kelimeler:

Actinidia deliciosa, Kivi, Meloidogyne incognita, rDNA, Root-knot nematodes, SCAR

Molecular identification of root-knot nematode Meloidogyne incognita from kiwi fruit orchards in Ordu province, Turkey

Root-knot nematodes (Meloidogyne spp.) are one of the major groups of nematodes identified on kiwi fruit (Actinidia deliciosa Chev.). This investigation was conducted to identify root-knot nematodes encountered the in kiwi fruit orchards in the Ordu province of Turkey by means of Polymerase Chain Reaction (PCR). For this purpose, infected plant root samples were collected from 17 kiwi fruit orchards located in Center, Perşembe, Ünye, Gülyalı, İkizce and Ulubey districtsof the province during the years 2011-2012.Females directly obtained from the infected roots of orchard populations were used for the molecular identifications. It was carried out with an identification key using ribosomal DNA (rDNA) region, Intergenic Spacer (IGS-2) region between 5S-18S genes, and the speciesspecific Sequence Characterized Amplified Region (SCAR) markers. All samples from the districts of Ordu province were identified as Meloidogyne incognita by the exact size of 502 bp amplicons by SEC1F/SEC1R scar primers. This study was the first detailed investigation both in the province and country on kiwi fruit

Keywords:

Actinidia deliciosa, Kiwi, Meloidogyne incognita, rDNA , Root-knot nematodes, SCAR,

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Adam, M. A. M., M. S., Phillips & V.C., Blok, 2007. Molecular diagnostic key for identification of single juveniles of seven common and economically important species of root-knot nematode (Meloidogyne spp.). Plant Pathology, 56: 190-197.
Anonymous, 2013. Turkish Statistical Institute. http://rapor.tuik.gov.tr. (Date accessed 29.03.2013)
Anwar, S. A. & M. V., McKenry, 2010. Incidence and reproduction of Meloidogyne incognita on vegetable crop genotypes. Pakistan Journal of Zoology, 42: 135-141.
Blok, V. C., M. S., Phillips & M., Fargette, 1997. Comparison of sequences from the ribosomal DNA intergenic region of Meloidogyne mayaguensis and other major tropical root-knot nematodes. Journal of Nematology 29: 16- 22.
Carneiro, R. M. D. G., Randig, O., M. R. A., Almeida & A. C. M. M., Gomes, 2004. Additional information on Meloidogyne ethiopica whitehead, 1968 (Tylenchida: Meloidogynidae), a root-knot nematode parasitising kiwi fruit and grape-vine from Brazil and Chilei. Nematology, 6: 109–123.
Cenis, J. L. 1993. Identification of four major Meloidogyne spp. by random amplified polymorphic DNA (RAPDPCR). Phytopathology, 83: 76-78.
Devran, Z. & M. A., Söğüt, 2009. Distribution and identification of root-knot nematodes from Turkey. Journal of Nematology, 41: 128-133.
El-Borai, F. E. & L. W., Duncan 2005. Nematode Parasites of Subtropical and Tropical Fruit Tree Crops: Plant Parasitic Nematodes in Subtropical and Tropical Agriculture, 2nd Edition, Ed: Luc, M., Sikora, R.A., Bridge, J., CABI Publishing, UK, pp: 46-492.
Elekçioğlu, I. H., & N., Uygun, 1994. Occurrence and distributionof plant-parasitic nematodes in cash crop in Eastern Mediterraneanregion of Turkey. Proceeding of 9th Congress of The MediterraneanPhytopathological Union, Kuşadası -Turkey 409–410.
Elekçioğlu, I. H., B., Ohnesorge, G., Lung, & N., Uygun, 1994. Plant parasitic nematodes in the Mediterranean region of Turkey.Nematologia Mediterranea 22:59–63.
Guzman-Plazola, R. A., J. D. J., Navas, E., Caswell-Chen, E., Zavaleta-Mejía & I. C., del Prado-Vera, 2006. Spatial distribution of Meloidogyne species and races in the tomato (Lycopersicon esculentum mill.) producing region of morelos, Mexico. Nematropica, 36: 215-229.
Haygood R. A., J. A., Saunders & R. W., Miller, 1990. Widespread occurrence of Meloidogyne incognita on kiwifruit in the coastal areas of South Carolina. Plant Disease 74. pp:81. Disease Note. http://www.apsnet.org/publications/PlantDisease/BackIssues/Documents/1990Abstracts/PD_74_81F.htm).
Hill, M. G., N. A., Mauchline & K. A., Stannard, 2008. Predicting armoured scale insect (Homoptera: Diaspididae) phenology on kiwifruit (Actinidia sp.). New Zealand Journal of Crop and Horticultural Science, 36: 253-262.
Hussey R. S., 1990. Biochemical and molecular methods of identifying Meloidogyne species: Symposium introduction. Journal of Nematology, 22: 8-9.
Hussey, R. S., & G. J. W. Janssen, 2002. “Root-Knot Nematodes: Meloidogyne Species, 43-70”. In: Plant Resistance to Parasitic Nematodes (Eds: J.L. Starr, R. Cook & J. Bridge). CABI publishing, Printed and bound in the UK by Biddles Ltd, 258 pp.
Kaşkavalcı, G. & C. Öncüer, 1999. Aydın İli’nin yazlık sebze yetiştirilen önemli bölgelerinde bulunan Meloidogyne Goeldi, 1887 (Tylenchida: Meloidogynidae) türlerinin yayılışları ve ekonomik önemleri üzerinde araştırmalar. Türkiye Entomoloji Dergisi, 23(2): 149-160.
Kepenekci, I. & G. Öztürk 1999. New species nematoda fauna of the Turkey belonging to Scutylenchus (Dolichodoroidea: Tylenchida) genus. Plant Protection Bulletin, 39: 23-33.
Kepenekci, I. & G. Öztürk 2000. Two new species belonging to Pratylenchoides Winslow, 1958 (Nematoda: Pratylenchidae) genus for the Nematoda fauna of Turkey. Turkish Journal of Entomology, 24: 125-132.
Katı, T., 2006. Research on Distribution and İnfestation Ratio with Species and Race Determination of Root-Knot Nematodes (Nematoda: Meloidogynidae: Meloidogyne spp.) of Greenhouses in Bafra and Çarşamba Province in Samsun. Ondokuz Mayıs Univesity, Msc thesis, Department of Palnt Protection, Samsun. 71 pp.
Kayani, M. Z., T., Mukhtar, M. A., Hussain & Ul-Haque, M. I. 2013. Infestation assessment of root-knot nematodes (Meloidogyne spp.) associated with cucumber in the pothowar region of Pakistan. Crop Protection 47: 49-54.
Khan, H., & R., Ahmad, 2000. Geographical distribution and frequency of occurrence of root-knot nematodes in Punjab–Pakistan. International Journal of Agriculture and Biology, 2: 354-355.
Ma, K. C., Y. S., Jo, B. H., Kim & D. G., Lim, 2007. Seasonal occurrence and aspects of root-knot nematodes in major kiwifruit cultivation areas of Korea. 753: VI International Symposium on Kiwifruit, Rotorua (New Zealand). Acta Horticulturae (ISHS) 753:719-724.
Mennan, S. & O. Ecevit, 1996. Studies on biology, distrubitionand the ratio of infestation of root-knot nematodes (Meloidogyne spp.) in summer vegetable growing area in Bafra and Cxarsxamba plains. Proceedings of the Third Turkish National Congress of Entomology 700–705.
Mckenna, C. E., S. J., Dobson & J. M., Phare, 2009. The insect pest complex of Actinidia arguta kiwifruit, New Zealand, Plant Protection 62: 262-267.
Moens, M., R. N. & Perry, J. L., Starr, 2009. “Meloidogyne species a Diverse Group of Novel and İmportant Plant Parasites” pp: 1-17. In: Root Knot Nematodes (Eds: R. N., Perry, M., Moens, J. L., Starr, MPG Books Group, UK. 488 pp.
Örümlü, A. E., 2003. Investigations on the nematological problems of fruit nurseries in Bademli (Ödemiş-İzmir) district. Phd thesis, Ege University, Graduate school of natural and applied sciences, İzmir. Pp. 69.
Özarslandan, A., 2009. Identification of Meloidogyne Species Collected From Different Parts of Turkey and Determination of Virulence of Some Root-Knot (Meloidogyne spp.) Populations. Phd Thesis, University of Çukurova, Institute of Natural applied sciences, Adana. 84 pp.
Philippi, I., B. A., Latorre, G. F., Perez & L. Castillo, 1996. Identification of the root-knot nematodes (Meloidogyne spp.) on kiwifruit by isoenzyme analysis in Chile. Fitopatologia 31, 96–101.
Ploetz, R. C. 2011. Tropical Fruit Crops and the diseases that affect their production. Tropical Biology and Conservation Management,Vol:III. (http://www.eolss.net/ebooks/Sample%2°Chapters/C20/E6-142-TA- 05.pdf (Date accessed 25.01.2012).
Söğüt, M. A. & I. H., Elekçioğlu, 2000. Determination of Meloidogyne spp. Goeldi, 1892 (Nemata: Heteroderidae) species races found in vegetable growing areas of the Mediterranean region of Turkey.Turkish Journal of Entomology 24:33–40.
Steven, D., Valenzuela, L. & R. H., Gonzalez, 1997. Kiwifruit pests in chile. III International Symposium on Kiwifruit, Thessaloniki (Greece). Acta Horticulturae, (ISHS) 444: 773-778.
Tesarova, B., M., Zouhar & P., Rysanek, 2003. Development of PCR for specific determination of root-knot nema- tode Meloidogyne incognita. Plant Protection Science, 39: 23–28.
Tomkins, A. R., 1996. Pest control on kiwifruit with an insecticidal soap. New Zealand Plant Protection Society, Proc. 49th N.Z. Plant Protection Conference, Nelson, New Zealand, 49: 6-11.
Wishart, J., M. S. Phillips & V. C., Blok, 2002. Ribosomal intergenic spacer: A polymerase chain reaction diagnostic for Meloidogyne chitwoodi, M. fallax, and M. hapla. Phytopathology, 92: 884-892.
Yüksel, H., 1974. Kök-ur nematodlarının (Meloidogyne spp) Türkiye’deki durumu ve bunların populasyon problemleri üzerinde düsünceler. Atatürk Üniversitesi Ziraat Fakültesi Dergisi, 5: 83-105.
Zijlstra, C., D. T. H. M., Donkers-venne & M., Fargette, 2000. Identification of Meloidogyne incognita, M. javanica and M. arenaria using sequence characterised amplified region (SCAR) based PCR assays. Nematology, 2: 847- 853.
Zijlstra C. & R. A., Van Hoof, 2006. A multiplex real time polymerase chain reaction (TaqMan) assay for the simultaneous detection of Meloidogyne chitwoodi and M. fallax. Phytopathology, 96: 1255-1262.
ZhengWang, J., H., HongWen, Z., CaiHong, , Z., ZhongHui & W., ShengMei, 2009. The development of the chinese kiwifruit industry. IX Convegno Nazionale dell'Actinidia, Viterbo-Latina, Italia, 6-8 Ottobre 2009. Italus Hortus, 16: 245-251.