Table S1 Primer combinations used for cloning and RT-qPCR
Designation of
primers
sequence
usage
WSMV5#
WS3-2000*
WS5-1970
WS3-4040
WS5-3880
WS3-5550
WS5-5280
WS3-7500
WS5-7250
WS5-7305
WSMV3
AATTAAACCAACCCAAATCGATCTGAC
CAATATCCGTTCTTCGGCACATA
GAAGATTTCATTATGTGCCGAAGAA
GAATGTTCTGTGATCCATAACCGGC
GCAGAGATGCTAGTGTTTTACAACTG
CTGGTGCATGATTCATATTCTCTTG
GATGAATTCATATTGGAGAAGCG
GCCATGAAGTATTGCCGTATGTT
GCGAACATTCACATCAGCACCAATTG
CAAGGCTGTGGTTGATGAGTTTAAC
TTTTTTTTTTGTAGCGTCGCCCTAACCCACAC
PCR
PCR and cDNA
PCR
PCR
PCR
PCR and cDNA
PCR
PCR and cDNA
PCR
PCR
PCR and cDNA
WS3-8910
WS5-7750
WS3-7895
as7855WSEuFam
(with WS5-7750
and WS3-7895)
GCATAATGGCTCGAAGTGATG
CTTATCAATGCCGACACAAAGGA
GCTTCATGAATGTGTGTGACATGTA
TCGATGCTACGATACGCTTCATCAAATGAA
as7850WSUSAtto
CTACGGTAAGCCTCGTCAAATGAGTAA
cDNA for RT-qPCR
qPCR,
qPCR
qPCR-FAM-BHQ1-labelled
probe, specific for Cluster B
isolates, antisense
qPCR-Atto-BHQ1-labelled
probe, specific for cluster A
isolates, antisense
(with WS5-7750
and WS3-7895)
5#-reverse primers, 3- forward primers (used also for cDNA synthesis),* numbering denotes the 5'-proximal
position of the nucleotide in relation to the complete genome.
Fig. S1 1 to 4 Inoculations with WSMV-PV57 (1 KS03HW12, 2 KS96HW10-3, 3 RonL, 4 Alcedo), 5 to 8
inoculations with WSMV-Hoym (5 KS03HW12, 6 KS96HW10-3, 7 RonL, 8 Alcedo)
Fig. S2 Features of the sequences of the investigated WSMV isolates
A – Changes in the cleavage site P1/HCPro
B – Characteristic ClaI restriction site (underlined) in the coat protein gene of European isolates
A
B
Fig. S3 Standard curves for RT-qPCR using a dilution series of plasmid DNA containing nucleotides 7305 to
9381 of WSMV Austria (A) or nucleotides 7750 to 7895 of USA PV57(B).
A405nm
Fig. S4 Challenge of German winter wheat cultivars with WSMV isolate Hoym. Five plants of each cultivar
were inoculated and after 28 days tested by DAS ELISA.