Screening of Suitable Reference Genes for qRT-PCR Normalization in Sisal

doi:10.3969/j.issn.1000-2561.2019.11.010

Abstract

Abstract:

In this study, eight reference genes (EF1α, MADH, ACT2, GAPDH, TUB, ACT54, CYP and EF1β) from the transcriptome database, were selected and analyzed in Rema No.1 including Phytophthora nicotianae Breda, MeJA, SA and ABA treatments. Statistical tools, including GeNorm, NormFinder and BestKeeper, were utilized to assess the suitability of reference genes based on the stability rankings for different species. The stability of the eight reference genes was significantly different under different treatments in Rema No.1. For Phytophthora nicotianae Breda and MeJA stresses, EF1α was identified as the most stable gene and EF1β was identified as the least stable gene. For SA stress, EF1α was identified as the most stable gene, and ACT2 was identified as the least stable gene. For ABA stress, GAPDH was identified as the most stable gene, and ACT54 was identified as the least stable gene. The results would provide reliable and optional available reference genes in gene expression analysis of sisal.

Key words: sisal, qRT-PCR, reference gene

ZHANG Yanmei,WANG Ruifang,YANG Ziping,LU Zhiwei,LI Junfeng,ZHAO Yanlong,LU Junying,ZHOU Wenzhao. Screening of Suitable Reference Genes for qRT-PCR Normalization in Sisal[J]. Chinese Journal of Tropical Crops, 2019, 40(11): 2166-2173.

Figures/Tables 8

References 30

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基因简称 Gene symbol	基因名称 Gene name	引物序列（5′-3′） Primer sequences (5′-3′)	产物长度 Amplicon length/bp
ACT2	肌动蛋白Actin2	F: CGTGTTGCACCTGAAGAACA R: CCATCACCCGAATCCAACAC	138
GAPDH	3-磷酸甘油醛脱氢酶Glyceraldehyde-3-phosphate dehydrogenase	F: GACACCGGTAGACTCCACAA R: AAGACCCTTCTCTTTGGCGA	115
EF1α	转录延伸因子Elongation factor 1-α	F: CAAGGGGTGGGTACTCAGAG R: AGCTTGAGAAGGAGCCCAAA	108
CYP	亲环蛋白Cyclophilin (peptidyl-prolylcis- trans isomerase)	F: GGTCACACACAAGGTTTAC R: TACCCACACCTTTCTCAC	100
TUB	β微管蛋白β-tubulin	F: CTAGACAATGAAGCTCTGTATG R: GCCGAGATCAAGTGATTAAG	100
MADH	苹果酸酶Malate dehydrogenase	F: GATTTCTGAGCGGTTGAAT R: GGACAGTTGCATGACTTAC	104
EF1β	转录延伸因子Elongation factor 1-β	F: GACAGAGGAGGACAAGAA R: CATCCAAGAGAACAGATGAC	100
ACT54	肌动蛋白Actin54	F: GGCTGTCTTCCCAAGTA R: CTTGGATTGGGCTTCATC	100

基因简称 Gene symbol	基因名称 Gene name	引物序列（5′-3′） Primer sequences (5′-3′)	产物长度 Amplicon length/bp
ACT2	肌动蛋白Actin2	F: CGTGTTGCACCTGAAGAACA R: CCATCACCCGAATCCAACAC	138
GAPDH	3-磷酸甘油醛脱氢酶Glyceraldehyde-3-phosphate dehydrogenase	F: GACACCGGTAGACTCCACAA R: AAGACCCTTCTCTTTGGCGA	115
EF1α	转录延伸因子Elongation factor 1-α	F: CAAGGGGTGGGTACTCAGAG R: AGCTTGAGAAGGAGCCCAAA	108
CYP	亲环蛋白Cyclophilin (peptidyl-prolylcis- trans isomerase)	F: GGTCACACACAAGGTTTAC R: TACCCACACCTTTCTCAC	100
TUB	β微管蛋白β-tubulin	F: CTAGACAATGAAGCTCTGTATG R: GCCGAGATCAAGTGATTAAG	100
MADH	苹果酸酶Malate dehydrogenase	F: GATTTCTGAGCGGTTGAAT R: GGACAGTTGCATGACTTAC	104
EF1β	转录延伸因子Elongation factor 1-β	F: GACAGAGGAGGACAAGAA R: CATCCAAGAGAACAGATGAC	100
ACT54	肌动蛋白Actin54	F: GGCTGTCTTCCCAAGTA R: CTTGGATTGGGCTTCATC	100