Plant Protect. Sci., 2002, 38(11):267-270 | DOI: 10.17221/10463-PPS

Selection of RNA isolation method for molecular detection of grapevine viruses

P. Komínek
Research Institute of Crop Production, 161 06 Prague-Ruzyně, Czech Republic

Grapevines infected with Grapevine leafroll-associated virus-1 (GLRaV-1) and Grapevine leafroll-associated virus-3 (GLRaV-3) were selected. Total RNA was isolated from grapevine phloem tissue scrapped from dormant canes by three different methods: extraction with urea buffer followed with phenol-chloroform extraction, method using ConcertTM reagent (Invitrogen) followed with chloroform-isopropylalcohol extraction, and procedure using RNeasy Plant Mini Kit (Qiagen). The highest yield of RNA was obtained using ConcertTM reagent. If this RNA was used in RT-PCR, GLRaV-1 and GLRaV-3 were easily detected. From RNA isolated by other two methods these viruses were not detected.

Keywords: Grapevine leafroll-associated virus-1; Grapevine leafroll-associated virus-3; grapevine; ELISA; RT-PCR

Published: December 31, 2002  Show citation

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Komínek P. Selection of RNA isolation method for molecular detection of grapevine viruses. Plant Protect. Sci. 2002;38(SI 2 - 6th Conf EFPP):267-270. doi: 10.17221/10463-PPS.
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    References

    1. B OSCIA D., G REIF C., G UGERLI P., M ARTELLI G.P., WALTER B., GONSALVES D. (1995): Nomenclature of grapevine leafroll-associated putative closteroviruses. Vitis, 34: 171-175.
    2. GOOD X., MONIS J. (2001): Partial genome organization, identification of the coat protein gene, and detection of Grapevine leafroll-associated virus-5. Phytopathology, 91: 274-281. Go to original source... Go to PubMed...
    3. MACKENZIE D.J., MCLEAN M.A., MUKERJI S., GREEN M. (1997): Improved RNA extraction from woody plants for the detection of viral pathogens by reverse transcription-polymerase chain reaction. Plant Dis., 81: 222-226. Go to original source... Go to PubMed...
    4. MINAFRA A., HADIDI A. (1994): Sensitive detection of grapevine virus A, B or leafroll-associated III from viruliferous mealybugs and infected tissue by cDNA amplification. J. Virol. Meth., 47: 175-188. Go to original source... Go to PubMed...
    5. MONIS J., BESTWICK R.K. (1996): Detection and localization of grapevine leafroll associated closteroviruses in greenhouse and tissue culture grown plants. Am. J. Enol. Viticult., 47: 199-205. Go to original source...
    6. ROUTH G., ZHANG Y.-P., SALDARELLI P., ROWHANI A. (1998): Use of degenerate primers for partial sequencing and RT-PCR-based assays of grapevine leafroll-associated viruses 4 and 5. Phytopathology, 88: 1238-1243. Go to original source... Go to PubMed...
    7. WOODHAM R.C., ANTCLIFF A.J., KRAKE L.R., TAYLOR R.H. (1984): Yield differences between sultana clones related to virus status and genetic factors. Vitis, 23: 73-83.

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