A highly effective and versatile technology for the isolation of RNAs from grapevines and other woody perennials for use in virus diagnostics

Table 2 RT-PCR and RT-qPCR detection of GRSPaV, GLRaV2, GLRaV3 and GRBaV from total RNA isolated from both leaf and cambium tissues of grapevines

Sample no.	GRSPaV		GLRaV-2		GLRaV-3		GRBaV		UBI
	L	C	L	C	L	C	L	C	L	C
Syrah:
SY-1	+	+	-	-	-	-	+	+	+	+
SY-2	+	+	-	-	+	+	-	-	+	+
SY-3	+	+	-	-	+?	-	+	+	+	+
SY-4	+	+	-	-	+?	-	+	+	+	+
SY-5	+	+	-	-	+	+	+	+	+	+
	32.3	28.1			18.3	15.2	19.0	14.7	25.2	22.0
SY-6	+	+	-	-	+	+	-	-	+	+
SY-7	+	+	-	-	-	-	+	+	+	+
	26.2	25.7			37.7	35.5	18.8	14.3	22.0	21.2
Gamay:
GA-1	+	+	-	-	-	-	-	-	+	+
GA-2	+	+	-	-	-	-	-	-	+	+
Cabernet Franc:
CF-1	+	+	-	-	-	-	-	-	+	+
CF-2	+	+	-	-	+	+	-	-	+	+
	30.4	28.4			17.1	4.5	ND	34.5	22.0	23.1
CF-3	+	+	-	-	-	-	-	-	+	+
Chardonnay:
CH-1	+	+	+	+	+	+	-	+?	+	+
	26.2	27.4			17.1	14.5	ND	32.8	22.0	22.6
CH-2	+	+	-	-	+	+	-	-	+	+
CH-3	-	-	-	-	+	+	+	+	+	+
	ND	ND			20.9	16.9	19.4	15.0	28.1	23.6
Gewurztraminer:
GE-1	-	-	-	-	+	+	-	-	+	+
GE-2	+	+	-	-	+	+	+?	-	+	+
Riesling:
RI-1	+	+	-	-	+	+	-	-	+	+
RI-2	+	+	-	-	+	+	-	-	+	+

Nineteen grapevine samples randomly selected from six varieties of Vitis vinifera were used to isolate total RNA from leaf or cambium tissue with the modified Sigma kit. + and -: positive and negative, respectively, in the detection of viruses by RT-PCR. The primers used were listed in Additional file 1: Table S1 with RSP35 and RSP36 being used for GRSPaV. Cq of RT-qPCR was given for select samples where qPCR was conducted. ?: weak DNA band after PCR amplification. ND: not determined

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