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

Table 2.

Primers and corresponding assays for the diagnosis of Diaporthe spp. on soybean.

Target Gene	Target Species (Specificity)	Primer/Probe (Combination)	Sequence (5′-3′)	T_m (°C)	Assay	Ref.
ITS	Diaporthe sp., but based on D. phaseolorum and D. longicolla	Phom.I	GAGCTCGCCACTAGATTTCAGGG	60	PCR	[15]
ITS		Phom.II	GGCGGCCAACCAAACTCTTGT	60	PCR	[15]
ITS	D. aspalathi	DphLe	TCGGCCTTGGAAGTAGAAGAC	60	PCR	[20]
ITS	D. aspalathi	DphRi	ACTGAATGCGTTGCGATTCT	60	PCR	[20]
ITS	D. caulivora	DPC-3F	TTTATGTTTATTTCTCAGAGTTTCAGTGTAA	60	qPCR	[21]
		DPC-3R	GGCGCACCCAGAAACC
		DPC-3P	FAM-CGGGCTGCTCCCCGTCTCC-TAMRA
ITS	D. longicolla	PL-3F	CAGAGATTCACTGTAGAAACAAGAGTTT	60	qPCR	[21]
		PL-3R	CCGGCCTTTTGTGACAAA
		PL-3P	FAM-CGGGCTGCTCCCTGTCTCCAG-TAMRA
ITS	D. longicolla, D. aspalathi, D. sojae	PL-5F	CGAGCTCGCCACTAGATTTCA	60	qPCR	[21]
		PL-5R	CCTCAAGCCTGGCTTGGTGATGG
		PL-5P	FAM-CCATCACCAAGCCAGGCTTGAGG-TAMRA
TEF	D. longicolla	DPCL-F	TGTCGCACCTTTACCACTG	60	qPCR ^a	[22]
		DPCL-R	GAACGATCCAAAAAGCTCTC
		DPCL-P	FAM-GCATCACTTTCATTCCCACTTTCTG-BMN-Q535
TEF	D. caulivora	DPCC-F	GCCTGCAAAACCCTGTTAC	60	qPCR ^a	[22]
		DPCC-R	CATCATGCTTTAAAAATGGGG
		DPCC-P	Cy5-CTCTTACCACACCTGCCGTCG-BMN-Q620
TEF	D. eres	DPCE-F	ACTCACTCAATCCTTGTCAC	60	qPCR ^a	[22]
		DPCE-R	GAGGGTCAGCATAATATTCG
		DPCE-P	ROX-CCATCAACCCCATCGCCTCTTTC-BMN-Q590
TEF	D. novem	DPCN-F	AAAACCCTGCTGGCATTAAC	60	qPCR ^a	[22]
		DPCN-R	TATTCTTGACAGTTCGTTTCG
		DPCN-P	HEX-TCTACCACTTTCAACCCTATCAATC-BMN-BMN-Q535

^a Probe based real-time PCR. These four primer-probe-combination were designed for use together in a quadruplex reaction.