Table 1. Primer sequences for qPCR analysis of target and reference genes.

Gene	Coffee source name	Name	Primer sequence (5'-3')	Primer length (bp)	Amplicon length (bp)	Ta (°C)	Tm (°C)	PCR efficiency
ICS2	Scaffold21359	Isochorismate synthase 2	Fw: TGCCATAGTACGAGAAAACA	20	124	60	79.0	94
			Rev: CCCAGAAAATCGACCATAAA	20
PALa	CaPAL F 13097	Phenylalanine ammonia-lyase	Fw: GCAGGTCCTACTCATTGTACAAG	23	166	60	82.0	89
			Rev: CCATTCCACTCTTTCAAACAATCC	24
NPR1	Scaffold33187	Non-expressor of PR1	Fw: AGGGCATTGGATTCTGACGA	20	126	60	81.5	88
			Rev: CTCTGTTGTGGTCTTTGCGT	20
PR1	Scaffold170607	Pathogenesis-related 1	Fw: GCCCGTAAAGTCACCTGT	18	177	60	86.5	91
			Rev: AACTACGCTGCCAAAATC	18
OPR3	Scaffold2739	12-oxoplytodienotae reductase 1-like	Fw: ATAACTCCCCACCTTCCAAC	20	198	58	81.5	91
			Rev: ACAGCCTTATCCCAACTCTAT	22
COI1	Scaffold40077	Coronatine insensitive 1	Fw: CTTAGCATCACCACCCACC	19	157	62	81.5	93
			Rev: TCCGATCCCCCATACCAAC	19
PR10b	CF589103	Pathogenesis-related 10	Fw: GCCACCATCCTTGAAGAGAA	20	151	55	80.0	99
			Rev: CAACTCTCTGCTTGGCAGTCT	21
ACS5	Scaffold82864	1-aminocyclepropane-1-carboxylic	Fw: AGGGCGTCCTGGTCACTAA	19	148	60	83.0	90
		acid synthase 5	Rev: CTCGGCGAGCTAAAAACTGT	20
ACO2	Scaffold57328	1-aminocyclepropane-1-carboxylic	Fw: AAAGTCAGCAATTACCCTCCA	21	144	58	87.0	93
		acid oxidase 2	Rev: ATCCACCCATTCACCATCCT	20
ETR1	Scaffold776	Ethylene resistant 1	Fw: GCCCCCAAGATATTCCTAAG	20	92	60	79.5	89
			Rev: TGCAAGACCAAGACCACTAC	20
EIN2	Scaffold3828	Ethylene insensitive 2	Fw: GTTACTTCTCCAAAACCTACT	21	134	60	78.5	93
			Rev: TCCCATTTACCACTCTTATCT	21
CTR1	Scaffold16054	Constitutive triple response 1	Fw: GCAGCTGTGGGTTTCAAGG	19	162	60	83.0	90
			Rev: AGTGGGGGAGGGTTTAGTC	19
ERF1	C312112	Ethylene-responsive factor 1	Fw: TGGCTGGGCACATTTGAC	18	85	58	84.0	90
			Rev: GGATTGCTGCTTGACCTC	18
IDEc	isotig10635	Insuline Degrading Enzyme	Fw: TGATCTAAGCTGGTGGAAAGC	21	91	55	76.3	99
			Rev: TCAGGTGCATCAGGATGATT	20
S24c	SNG-U349723*	Ribosomal protein S24	Fw: GCCCAAATATCGGCTTATCA	20	92	60	7.6	95
			Rev: TCTTCTTGGCCCTGTTCTTC	20
β-Tub9c	isotig08544	Tubulin beta-9	Fw: ACCCTCCAGCAAACTGATGA	20	100	55	77.3	92
			Rev: AGGATGCCACTGCTGATGAT	20

bp–base pairs, Ta–Annealing temperature, Tm–Melting temperature

* Unigene accession number according to the SOL Genomics Network.

^a Fernandez et al., (unpublished).

^b Ramiro et al., 2009 [45].

^c Figueiredo et al., 2013 [14].