Biodiversity of Lecanosticta pine-needle blight pathogens suggests a Mesoamerican Centre of origin

IMA Fungus

Table 2 Primers used for PCR amplification and sequencing in this study

Locus	Primer name	Direction	Primer sequence 5′ to 3’	Annealing temperature used (°C)	Amplification success	Reference
BT1	Bt1a	Forward	TTC CCC CGT CTC CAC TTC TTC ATG	50	87.4%	Glass and Donaldson 1995
	Bt1b	Reverse	GAC GAG ATC GTT CAT GTT GAA CTC	50		Glass and Donaldson 1995
BT2^a	T1	Forward	AAC ATG CGT GAG ATT GTA AGT	52	–	O’Donnell and Cigelnik 1997
	β-Sandy-R	Reverse	GCR CGN GGV ACR TAC TTG TT	52		Stukenbrock et al. 2012
	Bt2a	Forward	GGT AAC CAA ATC GGT GCT GCT TTC	52	–	Glass and Donaldson 1995
	Bt2b	Reverse	ACC CTC AGT GTA GTG ACC CTT GGC	52		Glass and Donaldson 1995
TEF1	EF1-728F	Forward	CAT CGA GAA GTT CGA GAA GG	52	88.2%	Carbone and Kohn 1999
	EF-2	Reverse	GGA RGT ACC AGT SAT CAT GTT	52		O’Donnell et al. 1998
ITS	ITS1	Forward	GAA GTA AAA GTC GTA ACA AGG	56	100%	White et al. 1990
	ITS4	Reverse	TCC TCC GCT TAT TGA TAT GC	56		White et al. 1990
MS204	MS204F.cerato	Forward	AAG GGC ACC CTC GAG GGC CAC	55	71.7%	Fourie et al. 2015
	MS204R.cerato	Reverse	GAT GGT RAC GGT GTT GAT GTA	55		Fourie et al. 2015
RPB2	RPB2-5f2	Forward	GGG GWG AYC AGA AGA AGG C	56	82.7%	Sung et al. 2007
	fRPB2-7cR	Reverse	CCC ATR GCT TGY TTR CCC AT	56		Liu et al. 1999

^aBT2 amplification success using all primer combinations was very low and abandoned

ISSN: 2210-6359