Primer Design for Verticillium Detection

1. Fig. S1 Banno et al CA9 549 cgcgtcccagagccaaactgcctctgggtcgtggatctgcctctgaattcacggcatctg V. longisporum 1429 cgcgtcccagagccaaactgcctctgggtcgtggatctgcctctgaattcacggcatctg V. dahliae 1773 ------------------------------------------------------------ CA9 609 cctccgaattcagggcaactgcctaacaggcggttggttctgattcaggaccagcctgag V. longisporum 1489 cctccgaattcagggcaactgcctaacaggcggttggttctgattcaggaccagcctgag V. dahliae 1773 ------------------------------------------------------------ CA9 669 tcacgagagatatggggaaggttcagagacttgacgggggtgggtgaattcactgctgct V. longisporum 1549 tcacgagagatatggggaaggttcagagacttgacgggggtgggtgaattcactgctgct V. dahliae 1773 ------------------------------------------------------------ CA9 729 gcaacaatataaatggggagagatcctcttcttcttcttccaacaaccaaaccaaaccac V. longisporum 1609 gcaacaatataaatggggagagatcctcttcttcttcttccaacaaccaaaccaaaccac V. dahliae 1773 ------------------------------------------------------------ CA9 789 aactgaacctcaaacaagacccacaagctcttcaaaatgtcccattcttctccctctcct V. longisporum 1669 aactgaacctcaaacaagacccacaagctcttcaaaatgtcccattcttctccctctcct V. dahliae 1773 ------------------------------------------------------------ CA9 849 gttctcgctaacgggagcgagtatgtcgtgagggatggaggttcgtctggtcaggcataa V. longisporum 1729 gttctcgctaacgggagcgagtatgtcgtgagggatggaggttcgtctggtcaggcataa CA9 909 ggaacgagaatgcagtggcgtggtttgcttaagataaagtccgggcttatgggaaaccat V. dahliae 1773 ------------------------------------------------------------ V. longisporum 1789 ggaacgagaatgcagtggcgtggtttgcttaagataaagtccgggcttatgggaaaccat V. dahliae 1773 ------------------------------------------------------------ CA9 969 aggtagctttgcaatcgaatgcatggccccaaaacgggagcctncggcttaatttgactc V. longisporum 1849 aggtagctttgcaatcgaatgcatggccccaaaacgggagcctgcggcttaatttgactc V. dahliae 1774 ------------------------------------ggagcctgcggcttaatttgactc CA9 1029 aacacggggaaactcaccaggtccagacacaacgaggattgacagattgagagctctttc V. longisporum 1909 aacacggggaaactcaccaggtccagacacaacgaggattgacagattgagagctctttc V. dahliae 1798 aacacggggaaactcaccaggtccagacacaacgaggattgacagattgagagctctttc CA9 1089 ttgattttgtgggtggtggtgcatg V. longisporum 1969 ttgattttgtgggtggtggtgcatggccgttcttagttggtggagtgatttgtctgctta CA9 1113 1858 V. dahliae ttgattttgtgggtggtggtgcatggccgttcttagttggtggagtgatttgtctgctta V. longisporum 2029 attgcgataacgaacgagaccttaacctgctaaatagtccgtattaccccggtagtacgc V. dahliae 1918 attgcgataacgaacgagaccttaacctgctaaatagtccgtattaccccggtagtacgc CA9 1113 V. longisporum 2089 cgacttcttagagggactatcggctcaagccgatggaagtttgaggcaataacaggtctg V. dahliae 1978 cgacttcttagagggactatcggctcaagccgatggaagtttgaggcaataacaggtctg Fig. S1 Nucleotide sequences of the group I intron in the 18S rDNA gene of Verticilliumlongisporum to design the detection primers. Grey arrowed underscores mark forward (VlspF2) and reverse (VdspR2) primers used for the first-round of quantitative nested real-time (QNRT) PCR. Black arrowed underscores mark forward (VlspF1) and reverse (VlspR4) primers used for the second-round of QNRT-PCR. GenBank accessions for the nucleotide sequences of the 18S rDNA region: V. longisporumCA9, AB585938; V. longisporum,AF153421; V. dahliae, AF104926.

2. Fig. S2 Banno et al CA9 52 cgttctgcgagcccgccggtacatcagtctctttatttataccaacgatacttctgagtg V. longisporum 51 cgttctgcgagcccgccggtacatcagtctctttatttataccaacgatacttctgagtg V. albo-atrum 72 cgttctgcgagcccgccggtacatcagtctctttatttataccaacgatacttctgagtg CA9 112 ttcttagcgaaccattaaaacttttaacaacggatctcttggctctagcatcgatgaaga V. dahliae 58 cgttctgcgagcccgccggtccatcagtctctctgtttataccaacgatacttctgagtg V. longisporum 111 ttcttagcgaaccattaaaacttttaacaacggatctcttggctctagcatcgatgaaga V. albo-atrum 132 ttcttagcgaactattaaaacttttaacaacggatctcttggctctagcatcgatgaaga V. dahliae 118 ttcttagcgaactattaaaacttttaacaacggatctcttggctctagcatcgatgaaga CA9 172 acgcagcgaaacgcgatatgtagtgtgaattgcagaattcagtgaatcatcgaatctttg V. longisporum 171 acgcagcgaaacgcgatatgtagtgtgaattgcagaattcagtgaatcatcgaatctttg V. albo-atrum 192 acgcagcgaaacgcgatatgtagtgtgaattgcagaattcagtgaatcatcgaatctttg V. dahliae 178 acgcagcgaaacgcgatatgtagtgtgaattgcagaattcagtgaatcatcgaatctttg CA9 232 aacgcacatggcgccttccagtatcctgggaggcatgcctgtccgag-cgtcgtttcaac V. longisporum 231 aacgcacatggcgccttccagtatcctgggaggcatgcctgtccgagacgtcgtttcaac V. albo-atrum 252 aacgcacatggcgccttccagtatcctgggaggcatgcctgtccgag-cgtcgtttcaac V. dahliae 238 aacgcacatggcgccttccagtatcctgggaggcatgcctgtccgag-cgtcgtttcaac CA9 291 cctcgagccccagtggcccggtgttggggatctacgtctgtaggcccttaaaagcagtgg V. longisporum 291 cctcgagccccagtggcccggtgttggggatctacgtctgtaggcccttaaaagcagtgg V. albo-atrum 311 cctcgagccccagtggcccggtgttggggatctacgtctgtaggcccttaaaagcagtgg V. dahliae 297 cctcgagccccagtggcccggtgttggggatctacgtctgtaggcccttaaaagcagtgg CA9 351 cggacccgcgtggcccttccttgcgtagtaattacagctcgcatcggagtcccgcaggca V. longisporum 351 cggacccgcgtggcccttccttgcgtagtaattacagctcgcatcggagtcccgcaggca V. albo-atrum 371 cggacccgcgtggcccttccttgcgtagtaattacagctcgcatcggagtcccgcaggca V. dahliae 357 cggacccgcgtggcccttccttgcgtagtagttacagctcgcatcggagtcccgcaggcg Fig. S2 Nucleotide differences in the 5.8S rDNA-ITS region among Verticilliumlongisporum, V. albo-atrumand V. dahliae to design the detection primers. Differences in the four sequences are indicated by black boxes. Black arrowed underscores mark forward (VaF1) and reverse (VaR1) primers used for V.longisporum 5.8S rDNA-ITS. Grey arrowed underscores mark forward (VdF1) and reverse (VdR1) primers used for V.dahliae 5.8S rDNA-ITS. GenBank accessions for nucleotide sequences of the 5.8S rDNA-ITS region: V. longisporumCA9, AB585937; V. longisporum,AY555951; V. albo-atrum, GU291258; V. dahliae, EU627007.