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Table 1 Genes and primer sets used for RT-qPCR

From: Genome-wide screening of novel RT-qPCR reference genes for study of GLRaV-3 infection in wine grapes and refinement of an RNA isolation protocol for grape berries

Gene abbreviation

Accession no.

NCBI gene description

Primer sequence (5′–3′)

Amplicon size (bp)

Amplification efficiency (E)

EF-Hand

NM_001280964.1

EF-hand calcium-binding protein

F: TTTGACAGGGACCGTAGTGG

R: GTCAGCCCCTTTACCGTGAG

188

98.25

GDT1

XM_019218145.1

GDT1-like protein 5

F: GGCTGCTCCAAACCTGTTGTC

R: ACCTTGCTATCCCCTTTGGC

192

104.68

CYSP

NM_001281060.1

Cysteine protease

F: AAAATCAGGGTTCGTGTGGGTC

R: GCAGTGTTCATCAGCCCACC

190

96.79

NDUFS8

XM_003631606.3

NADH dehydrogenase [ubiquinone] iron-sulfur protein 8 (mitochondrial)

F: CCGTAGAACGACCAGGTACGAC

R: GCAATCTCGGTTTCCCAGCG

189

94.66

EIF5A2

XM_002285469.4

Eukaryotic translation initiation factor 5A-2

F: CCGCAAGAACGGCTACATCG

R: CGGGTAACATGTGGAACATCAC

183

103.45

Gluc

XM_010651552.1

Endo-1,3;1,4-beta-d-glucanase

F: GCTTTTGCTGGGGTGCCAAG

R: TGTTTCACGAGTGCCGGTG

175

95.74

YSL8

XM_002283586.3

Thioredoxin-like protein YLS8

F: TCAGGCGTGAAGAGAGAAAGC

R: AGCCAGAACTTCATCCATCTGC

186

97.28

Actin

XM_002277287.4

Actin 1

F: ATCAGGAAGGACCTCTATGG

R: ATCCACATCTGCTGGAAGG

206

92.98

NAD5

GU585873.1

NADH dehydrogenase subunit 5

F: GATGCTTCTTGGGGCTTCTTGTT

R: CTCCAGTCACCAACATTGGCATAA

181

92.08

UFGT

XM_010659535.2

Anthocyanidin 3-O-glucosyltransferase 2

F: TCTTCCCTTCTGTGGTGCTTG

R: TTATTGAGCAGGGGTCCAACAG

187

99.10

  1. Seven novel reference genes candidates identified from our own RNA-Seq data were EF-hand, GDT1, CYSP, NDUFS8, EIF5A2, Gluc, YSL8. Two conventional reference genes used by a past study, Gutha et al. [4] were actin and NAD5. UFGT, shown previously as a responsive gene whose expression was influenced by GLRaV-3 infection, was chosen in this study for RT-qPCR validation