Selection of RNA isolation method for molecular detection of grapevine viruses
P. Komínekhttps://doi.org/10.17221/10463-PPSCitation:Komínek P. (2002): Selection of RNA isolation method for molecular detection of grapevine viruses. Plant Protect. Sci., 38: 267-270.
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