İndirekt Enzyme-Linked Immunosorbent Assay ELISA ile Kayısı Yaprak Doku Diskierinde Plum Pox Potyvirus PPV 'nin Tanılanması

Abstract

Plum Pox Potyvirus PPV ile enfekteli kay ı sı yapraklar ı ndan bir mantar delici yard ımı yla 2, 3.5, 5 ve 6 mm çapl ı diskler kesilmiştir. Bu farkl ı çaptaki yaprak diskleri ELISA tabakalar ı ndaki kuyucuklara 1, 2, 3 ve 4'er tane olacak şekilde konulmuştur.Denemede, PPV ile enfekteli yapraklar ı n belirtili ve belirtisiz dokular ı kullan ı lmıştı r. Çal ışmada diskler için 37° C'de 4 saat ve 4° C'de bir gece olmak üzere iki inkübasyon s ı cakl ı k derecesi kullan ı lmışt ı r. Yaprak disk say ı sı ve büyüklüğ ü artt ı kça absorbans değerinin de artt ığı gözlenmiştir. Dört saat 37° C'lik bir inkübasyon peryoduna b ı rakı lan disklerin absorbans de ğerleri bir gece 4° C'de b ı rakı lanlardan biraz daha yüksek olarak bulunmu ştur. Ayr ı ca, belirtili yerlerden al ı nan disklerin belirtisizlerinkinden daha yüksek absorbans de ğeri verdiğ i görülmüştür. Bütün koşullarda enfekteli disklerin absorbans de ğerleri sağl ı kl ı ları nkinden en az iki kat daha yüksek olmu ştur

Keywords

• Plum Pox Potyvirus
• kay ı s ı
• yaprak doku diskleri

Detection of Plum Pox Potyvirus PPV in Leaf Tissue Disks of Apricot by Indirect Enzyme-Linked Immunosorbent Assay ELISA

Abstract

The Plum Pox Potyvirus PPV infected leaf disks of 2, 3.5, 5 and 6.5 mm in diameter were cut of the leaf by cork borer. Leaf disks in different diameters were placed into the wells in the ELISA plate in increased numbers, starting with one till 4 folds. In this assay, symptomatic and asymptomatic leaves of infected trees were used. In the assay, two incubation times for disks as 37 ° C at hr and 4° C in overnight was used. It is observed that as number and size of leaf disks increase, the absorbance values were also increased. Absorbance values of incubation period of 37 ° C was greater than that of overnight. In addition, absorbance values of disks from symptomatic area was all times found a bit greater than that asymptomatic disks. Results are found to be similar both leaf disks of apricots and plums. In all conditions, absorbance values of the infected disks were at least two times greater than that of healthy ones.

Keywords

• Plum Pox Potyvirus
• apricot
• leaf tissue disks

References

1. Baumgartnerova, H., H. Slovakova and W. Petrusova, 1998. Relationship between concentration of Plum Pox Virus and content of pigments in virus-infected symptomatic apricot leaves. Acta Virologica, 42 (4) 216-218.
2. Crescenzi, A., L. D'Aquino, S. Comes, M. Nuzzaci, and P. Piazzola, 1997. Charecterization of the sweet cherry isolate of Plum Pox Potyvirus. Plant Disease, 81: 711-714.
3. Desvignes, J. C. 1999. Virus Diseases of Fruit Trees. Editions Centre Technique Interprofessionnel des Fruits et Legumes, 202 pp.
4. Dunez, J., M. Ravelonandro and T. Candresse, 1994. Plum Pox: Advances in research on the disease and its causal agent, and possible means of control. Bulletin OEPP/EPPO Bulletin, 24: 537-542.
5. Elibüyük, İ . Ö. and G. Erdiller, 1991. Studies on incidence and identification of sharka disease on apricots, plums and peaches in Ankara Province. VI. Phytopathology Congress of Turkey, No: 6 411-415 pp.
6. Elibüyük, İ . Ö. and G. Erdiller, 1995. The susceptibility of some apricot and plum varieties to Plum Pox (sharka) virus. x. International symposium on apricot culture. Acta Horticulturae, No: 384, 549-552 pp.
7. Elibüyük, İ . Ö. and G. Erdiller, 1998. Researches on virus diseases of stone fruit trees in Malatya region. VIII. Phytopathology Congress of Turkey, Türkiye Fitopatoloji Derne ği Yay ı nları , 8: 8.
8. Erdiller, G. 1988. Investigation on the causes of fruit dropping of apricot and plum trees in Ankara province. Journal of Turkish Phytopathology, 17(3) 98.

9. Kurçman, S. 1973. Nachweis des Scharka-Virus an Aprikosen und Pflaumenbaumen in Ankara. J. Turkish Phytopath., 2(3). Marko, S. and S. Cohen, 1979. Rapid detection and titer evaluation of viruses in pepper by enzyme-linked immunosorbent assay. Phytopathology, 69, 1259-1265.
10. Menassa, R., K. M. Makkouk. and A. A. Abbasher, 1986. Detection of Zucchini Yellow Mosaic Virus in intact leaf disks and tissue extracts by enzyme-linked immunosorbent assay. J. Phytopathology, 115, 152-159.
11. Nemchinov, L. and A. Hadidi, 1996. Characterisation of the sour cherry strain of Plum Pox Virus. Phytopathology 86, 575- 580.
12. Ne-meth, M. 1986. Virus, Mycoplasma and Rickettsia Diseases of Fruit Trees. Martinus Nijhoff / Dr. W. Junk Publishers, 841 pp.
13. Nemeth, M. 1994. History and Importance of Plum Pox in StoneFruit Production. Bulletin OEPP/EPPO Bulletin, 24, 525-536.
14. Ostermann, W. D. and D. Hampe, 1986. Nachweis pflanzenpathogener Viren mit Hilfe des ELISA unter Verwendung von Blattgewebescheiben. Arch. Phytopathol. Pflanzenschutz, 22 (1-9) 9-17.
15. Polak, J. 1995. Reliability of detection and relative concentration of Plum Pox Virus determined by ELISA in a peach tree during the vegetation period. Journal of Plant Disease and Protection, 102, 16-22.
16. Rankovic, M. and S. Vuksanovic, 1985. Possibilities and problems in diagnosing sharka on apricot by the ELISA technique. Zastita Bilja, 36 (2) 161-166.
17. Romaine, C. P., S. R. Newhart and D. Anzola, 1981. Enzymelinked ı mmunosorbent assay for plant viruses in ı ntact leaf tissue disks. Phytopathology, 71, 308-312.
18. Roy, A. S. and I. M. Smith, 1994. Plum Pox Situation in Europe. Bulletin OEPP/EPPO Bulletin, 24, 515-523.
19. Sahtiyanc ı , Ş . 1969. Virus de la Sharka Ches la Prunier. Bulletin Phytosan, FAO, 17 (3) 69.
20. Uyemoto J. K, C. F. Luhn, W. Asai, R. Beede, J. A. Beutel and R. Fenton, 1989. Incidence of Ilarviruses in young peach trees in California. Pl. Dis., 73, 217-220.
21. Yürektürk, M. 1984. Marmara Bölgesinde Sert Çekirdekli Meyvelerde Görülen Sharka Virus Hastal ığı Üzerinde Araşt ı rmalar. Atatürk Bahçe Kültürleri Ara şt ı rma Enstitüsü, 37 s