Prunus Yapraklarında Prunus Necrotic Ring Spot PNRSV ve Apple Chlorotic Leaf Spot ACLSV Virüslerinin Dağılımı

Abstract

Prunus necrotic ringspot virus PNRSV ’ü ile infekteli Prunus mahaleb ve Apple chlorotic leafspot virus ACLSV ’ü ile infekteli şeftali P. persica L. yapraklarının farklı bölgelerinden alınan doku diskleri bu virüslerin yaprak dokusundaki dağılımlarını belirlemek amacı ile sırasıyla enzyme-linked immunosorbent assay ELISA ve reverse transcriptase polymerase chain reaction RT-PCR teknikleri ile analiz edilmişlerdir. Gerçekleştirilen ELISA testleri sonucunda her iki virüsünde yaprak ayasında yaprak sapı bölgesinde daha konsantre oldukları ve konukçu yapraklarında düzensiz bir dağılım gösterdikleri tespit edilmiştir. Aynı yaprak bölgelerinin kullanıldığı RT-PCR testlerinde ise her iki virüsün genetik materyalinin tüm yaprak bölgeleri için birbirine yakın ölçülerde amplifikasyon ürünleri oluşturduğu belirlenmiş ve testlenen yaprak bölgeleri arasında viral konsantrasyon bakımından bariz farklılıkların olmadığı saptanmıştır. RT-PCR testi sonuçlarından elde edilen kesin, net ve dengeli teşhisi ifade eden bantlar, ACLSV ve PNRSV virüslerinin Prunus yapraklarının testlenen tüm bölgelerinde homojen bir dağılım sergilediğini göstermiştir. Her iki virüs, kullanılan test yöntemine göre konukçularında farklı dağılım sergilemişlerdir. Elde edilen bulgular ışığında PNRSV ile ACLSV’nin konukçularındaki dağılımını belirlemede ELISA testi ile PCR testi arasında bir korelasyon saptanmamıştır

Keywords

• Prunus necrotic ringspot virüs
• Apple chlorotic leaf spot virüs
• ELISA
• RT-PCR
• PNRSV
• ACLSV

Distribution of Prunus Necrotic Ringspot PNRSV and Apple Chlorotic Leaf Spot Viruses ACLSV in Prunus Leaves

Abstract

Leaf discs taken from different canopy and leaf sites of Prunus necrotic ringspot virus PNRSV infected Prunus mahaleb and Apple chlorotic leafspot virus ACLSV infected peach P. persica tree were analyzed for the determination of virus distribution in leaf tissues by enzyme-linked immunosorbent assay ELISA and reverse transcriptase polymerase chain reaction RT-PCR techniques. The ELISA results suggest high virus concentration of PNRSV and ACLSV at the basal leaf section of the lamina and uneven virus distribution in their host leaves. Clear and relatively balanced amplification bands were obtained when the same leaf sections were analyzed by RT-PCR. No conspicuous differences were found in terms of viral concentrations among the leaves tested. Amplification bands of RT-PCR test results suggest homogeneous distribution of both viruses in tested Prunus leaves. Both viruses were exhibited different patterns of distribution in their hosts according to the detection method used. No correlation was found between ELISA and RT-PCR tests in determining the distribution of both viruses

Keywords

• Prunus necrotic ringspot virus
• Apple chlorotic leaf spot virus
• ELISA
• RT-PCR
• PNRSV
• ACLSV

References

1. Foissac, X., L. Savalle-Dumas, P. Gentit, M. J. Dulucq and T. Candresse, 2000. Polyvalent detection of Fruit tree Tricho, Capillo and Faveaviruses by Nested RT-PCR Using degenerated and inosine containing primers (PDO-RT- PCR). Acta Horticulturae, 357: 52-59.
2. Knapp, E., A. Camara-Machado, H. Puhringer, Q. Wang, V. Hanzer, B. Weiss, H. Katinger and M. Lamier Camara Machado, 1995. Localization of fruit tree viruses by immuno tissue printing in infected shoots of Malus sp. and Prunus sp. J. of Virological Methods, 55: 157-173.
3. Menzel, W., W. Jelkmann and E. Maiss, 2002. Detection of four apple viruses by multiplex RT-PCR assays with coamplification of plant mRNA as internal control. Journal of Virological Methods, 99: 81-92.
4. Myrta, A, O. Potere, F. Ismaeli and D. Boscia, 2003. PPV Distribution on Prunus: Experiences of Diagnosis With ELISA. Options Mediterraneennes, 45: 107-110.
5. Nemeth, M. 1986. Virus, mycoplasma and rickettsia diseases of fruit trees. Akademia Kiado, Budapest 841 s.
6. Pallas, V., M. L. Badenes and G. Llacer, 1998. Requirements for the stone fruit certification programme in Spain. Options Mediterraneennes, 19: 129-133.
7. Parakh, D. R., A. M. Shamloul, A. Hadidi, S. W. Scott, H. E. Waterworth, H. E. Howell and G. I. Mink, 1995. Detection of Prune dwarf Ilarvirus from infected fruits using reverse transcription-polymerase chain reaction. Acta Horticulturae, 386: 421-430
8. Rosner, A., Y. Shiboleth, S. Spiegel, L. Krisbai and M. Kölber, 1998. Evaluating The Use of İmmunocapture and Sap- Dilution PCR for The Detection of Prunus necrotic ringspot virus. Acta Horticulture, 472. 17th Int. Symp. on Fruit Virus Diseases.

9. Rowhani, A., L. Maningas, S. Lile, D. Daubert and A. Golino, 1995. Deveplopment of Detection System for Viruses of Woody Plants based on PCR Analysis of Immmobilized Virions. The American Phtopathological Society, Vol.85 No.3.
10. Spiegel, S., S. W. Scott, V. Bowman-Vance, Y. Tam, N. N. Galiakparov and A. Rosner, 1996. Improved detection of Prunus necrotic ringspot virus by the polymerase cahain reaction. European Journal of Plant Pathology,102: 681- 685.
11. Spiegel, S., Y. Tam, L. Maslenin, M. Kolber, M. Nemeth and A Rosner, 1999. Typing prunus necrotic ringspot virus isolates by serology and restriction endonuclease analysis of PCR products. Annals of Applied Biology, 135(1):395-400. Symposium on Apricot Culture and Decline 10th September 2001 Avignon, France.
12. Ulubaş,Ç., F. Ertunç, 2004. RT-PCR detection and molecular characterisation of Prunus necrotic ringspot virus isolates occuring in Turkey. J. Phytopathology, 152: 498-502.