| Primer name . | Nucleotide sequence (5′–3′) . | Purpose . |
|---|---|---|
| PlActionF | ACTGCTGAACGGGAAATT | Actin primers |
| PlActionR | ATGGCTGGAACAGGACTT | |
| PlWRKY65qF | TTTGCCGAAGAGAGAAGCGT | Specific primer for qRT-PCR |
| PlWRKY65qR | TATACCTCCGTCGCTCCTCC | |
| PlWRKY65F | ATGGACAGTCTATACAAT | Full-length DNA and cDNA amplification |
| PlWRKY65R | TCACGAGGTGGTCCCACAC | |
| PlWRKY65(B)F | CGGGATCCATGGACAGTCTATAC | Vector construction for GFP |
| PlWRKY65(K)R | GGGGTACCCGAGGTGGTCCCAC | |
| PlWRKY65(E)F | CGGAATTCAACCACCATGCCACC | Vector construction for VIGS |
| PlWRKY65(K)R | GGGGTACCCGAGGTGGTCCCACA | |
| pTRV1F | TTACAGGTTATTTGGGCTAG | Molecular detection of TRV |
| pTRV1R | CCGGGTTCAATTCCTTATC | |
| pTRV2F | TTTATGTTCAGGCGGTTCTTGTG | |
| pTRV2R | CAAACGCCGATCTCAAACAGTC | |
| PR1F | TACCCAGAGACGGTTCGACT | Primers for pathogenesis-related genes |
| PR1R | CACACGAGTTGGACCGGTAA | |
| PR2F | TGGCCAAAGGGGTCTCTAGA | |
| PR2R | TCCCATTTACGGCAAGCGTA | |
| PR4BF | ATCCCGCTCAACACTCTTGG | |
| PR4BR | TCCACAGAAAGCAGTCCACC | |
| PR5F | CAGTCTTCCCTCAGGCAAGG | |
| PR5R | GGTTTCACATGCGGGTTTCC | |
| PR10F | CCGGCAAGGATTTTCAAGGC | |
| PR10R | TTATCTTGATGGTCCCGGCG |
| Primer name . | Nucleotide sequence (5′–3′) . | Purpose . |
|---|---|---|
| PlActionF | ACTGCTGAACGGGAAATT | Actin primers |
| PlActionR | ATGGCTGGAACAGGACTT | |
| PlWRKY65qF | TTTGCCGAAGAGAGAAGCGT | Specific primer for qRT-PCR |
| PlWRKY65qR | TATACCTCCGTCGCTCCTCC | |
| PlWRKY65F | ATGGACAGTCTATACAAT | Full-length DNA and cDNA amplification |
| PlWRKY65R | TCACGAGGTGGTCCCACAC | |
| PlWRKY65(B)F | CGGGATCCATGGACAGTCTATAC | Vector construction for GFP |
| PlWRKY65(K)R | GGGGTACCCGAGGTGGTCCCAC | |
| PlWRKY65(E)F | CGGAATTCAACCACCATGCCACC | Vector construction for VIGS |
| PlWRKY65(K)R | GGGGTACCCGAGGTGGTCCCACA | |
| pTRV1F | TTACAGGTTATTTGGGCTAG | Molecular detection of TRV |
| pTRV1R | CCGGGTTCAATTCCTTATC | |
| pTRV2F | TTTATGTTCAGGCGGTTCTTGTG | |
| pTRV2R | CAAACGCCGATCTCAAACAGTC | |
| PR1F | TACCCAGAGACGGTTCGACT | Primers for pathogenesis-related genes |
| PR1R | CACACGAGTTGGACCGGTAA | |
| PR2F | TGGCCAAAGGGGTCTCTAGA | |
| PR2R | TCCCATTTACGGCAAGCGTA | |
| PR4BF | ATCCCGCTCAACACTCTTGG | |
| PR4BR | TCCACAGAAAGCAGTCCACC | |
| PR5F | CAGTCTTCCCTCAGGCAAGG | |
| PR5R | GGTTTCACATGCGGGTTTCC | |
| PR10F | CCGGCAAGGATTTTCAAGGC | |
| PR10R | TTATCTTGATGGTCCCGGCG |
Note: The underlined ‘GGATCC’, ‘GAATTC’, and ‘GGTACC’ nucleotides are the added restriction enzyme recognition sites for BamHI, EcoRI, and KpnI, respectively. Primers for the pTRV1 vector were designed within RNA-dependent RNA polymerase elements (PCR product size of 647 bp in theory), and primers for the pTRV2 vector were designed between MCS elements (PCR product size of 372 bp in theory)
| Primer name . | Nucleotide sequence (5′–3′) . | Purpose . |
|---|---|---|
| PlActionF | ACTGCTGAACGGGAAATT | Actin primers |
| PlActionR | ATGGCTGGAACAGGACTT | |
| PlWRKY65qF | TTTGCCGAAGAGAGAAGCGT | Specific primer for qRT-PCR |
| PlWRKY65qR | TATACCTCCGTCGCTCCTCC | |
| PlWRKY65F | ATGGACAGTCTATACAAT | Full-length DNA and cDNA amplification |
| PlWRKY65R | TCACGAGGTGGTCCCACAC | |
| PlWRKY65(B)F | CGGGATCCATGGACAGTCTATAC | Vector construction for GFP |
| PlWRKY65(K)R | GGGGTACCCGAGGTGGTCCCAC | |
| PlWRKY65(E)F | CGGAATTCAACCACCATGCCACC | Vector construction for VIGS |
| PlWRKY65(K)R | GGGGTACCCGAGGTGGTCCCACA | |
| pTRV1F | TTACAGGTTATTTGGGCTAG | Molecular detection of TRV |
| pTRV1R | CCGGGTTCAATTCCTTATC | |
| pTRV2F | TTTATGTTCAGGCGGTTCTTGTG | |
| pTRV2R | CAAACGCCGATCTCAAACAGTC | |
| PR1F | TACCCAGAGACGGTTCGACT | Primers for pathogenesis-related genes |
| PR1R | CACACGAGTTGGACCGGTAA | |
| PR2F | TGGCCAAAGGGGTCTCTAGA | |
| PR2R | TCCCATTTACGGCAAGCGTA | |
| PR4BF | ATCCCGCTCAACACTCTTGG | |
| PR4BR | TCCACAGAAAGCAGTCCACC | |
| PR5F | CAGTCTTCCCTCAGGCAAGG | |
| PR5R | GGTTTCACATGCGGGTTTCC | |
| PR10F | CCGGCAAGGATTTTCAAGGC | |
| PR10R | TTATCTTGATGGTCCCGGCG |
| Primer name . | Nucleotide sequence (5′–3′) . | Purpose . |
|---|---|---|
| PlActionF | ACTGCTGAACGGGAAATT | Actin primers |
| PlActionR | ATGGCTGGAACAGGACTT | |
| PlWRKY65qF | TTTGCCGAAGAGAGAAGCGT | Specific primer for qRT-PCR |
| PlWRKY65qR | TATACCTCCGTCGCTCCTCC | |
| PlWRKY65F | ATGGACAGTCTATACAAT | Full-length DNA and cDNA amplification |
| PlWRKY65R | TCACGAGGTGGTCCCACAC | |
| PlWRKY65(B)F | CGGGATCCATGGACAGTCTATAC | Vector construction for GFP |
| PlWRKY65(K)R | GGGGTACCCGAGGTGGTCCCAC | |
| PlWRKY65(E)F | CGGAATTCAACCACCATGCCACC | Vector construction for VIGS |
| PlWRKY65(K)R | GGGGTACCCGAGGTGGTCCCACA | |
| pTRV1F | TTACAGGTTATTTGGGCTAG | Molecular detection of TRV |
| pTRV1R | CCGGGTTCAATTCCTTATC | |
| pTRV2F | TTTATGTTCAGGCGGTTCTTGTG | |
| pTRV2R | CAAACGCCGATCTCAAACAGTC | |
| PR1F | TACCCAGAGACGGTTCGACT | Primers for pathogenesis-related genes |
| PR1R | CACACGAGTTGGACCGGTAA | |
| PR2F | TGGCCAAAGGGGTCTCTAGA | |
| PR2R | TCCCATTTACGGCAAGCGTA | |
| PR4BF | ATCCCGCTCAACACTCTTGG | |
| PR4BR | TCCACAGAAAGCAGTCCACC | |
| PR5F | CAGTCTTCCCTCAGGCAAGG | |
| PR5R | GGTTTCACATGCGGGTTTCC | |
| PR10F | CCGGCAAGGATTTTCAAGGC | |
| PR10R | TTATCTTGATGGTCCCGGCG |
Note: The underlined ‘GGATCC’, ‘GAATTC’, and ‘GGTACC’ nucleotides are the added restriction enzyme recognition sites for BamHI, EcoRI, and KpnI, respectively. Primers for the pTRV1 vector were designed within RNA-dependent RNA polymerase elements (PCR product size of 647 bp in theory), and primers for the pTRV2 vector were designed between MCS elements (PCR product size of 372 bp in theory)
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