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. 2023 May 18;9(5):587. doi: 10.3390/jof9050587

Table 11.

Primers and corresponding assays for diagnosis of P. pachyrhizi and P. meibomiae on soybean.

Target Gene Target Species (Specificity) Primer/Probe (Combination) Sequence (5′-3′) Tm (°C) Fragment Length (bp) Assay Ref.
ITS P. pachyrhizi Ppm1 GCAGAATTCAGTGAATCATCAAG 65 a 60 b 141 PCR/qPCR [150]
Ppa2 GCAACACTCAAAATCCAACAAT
ITS P. meibomiae Ppm1 GCAGAATTCAGTGAATCATCAAG 139
Pme2 GCACTCAAAATCCAACATGC
ITS Phakopsora Ppm1 GCAGAATTCAGTGAATCATCAAG 77
Ppm2 CTCAAACAGGTGTACCTTTTGG
ITS Phakopsora FAM-probe c FAM-CCAAAAGGTACACCTGTTTGAGTGTCA-TAMRA
VIC-probe d VIC-TGAACGCACCTTGCACCTTTTGGT-TAMRA
ITS P. pachyrhizi ITS1rustF4a e GAGGAAGTAAAAGTCGTAACAAGGTTTC 60 nested qPCR [151,152]
ITS1rustF10d TGAACCTGCAGAAGGATCATTA
ITS1rustR3d TGTGAGAGCCTAGAGATCCATTG
ITS1PhpFAM1 FAM-TCATTGAT-TGATAAGATCTTTGGGCAATGG-3IABlkFQ

a With standard PCR. b With qPCR. c For use with Ppm1/Ppa2 and Ppm1/Pme2. d For use with Ppm1/Ppm2. e Combined with Ppa2 from [150] in first round of nested PCR.