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Molecules
Volume 23
Issue 1
10.3390/molecules23010172
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Article

Reference Gene Selection for Quantitative Real-Time Reverse-Transcriptase PCR in Annual Ryegrass (Lolium multiflorum) Subjected to Various Abiotic Stresses

by
Qiuxu Liu
1,†,
Xiao Qi
2,†,
Haidong Yan
3,
Linkai Huang
1,
Gang Nie
1 and
Xinquan Zhang
1,*
1
Department of Grassland Science, Animal Science and Technology College, Sichuan Agricultural University, Chengdu 611130, China
2
National Animal Husbandry Service, Ministry of Agriculture, Beijing 100000, China
3
Department of Horticulture, Virginia Polytechnic Institute and State University, 490 West Campus Dr., Blacksburg, VA 24061, USA
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Molecules 2018, 23(1), 172; https://doi.org/10.3390/molecules23010172
Received: 23 November 2017 / Revised: 12 January 2018 / Accepted: 13 January 2018 / Published: 16 January 2018
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Abstract

To select the most stable reference genes in annual ryegrass (Lolium multiflorum), we studied annual ryegrass leaf tissues exposed to various abiotic stresses by qRT-PCR and selected 11 candidate reference genes, i.e., 18S rRNA, E2, GAPDH, eIF4A, HIS3, SAMDC, TBP-1, Unigene71, Unigene77, Unigene755, and Unigene14912. We then used GeNorm, NormFinder, and BestKeeper to analyze the expression stability of these 11 genes, and used RefFinder to comprehensively rank genes according to stability. Under different stress conditions, the most suitable reference genes for studies of leaf tissues of annual ryegrass were different. The expression of the eIF4A gene was the most stable under drought stress. Under saline-alkali stress, Unigene14912 has the highest expression stability. Under acidic aluminum stress, SAMDC expression stability was highest. Under heavy metal stress, Unigene71 expression had the highest stability. According to the software analyses, Unigene14912, HIS3, and eIF4A were the most suitable for analyses of abiotic stress in tissues of annual ryegrass. GAPDH was the least suitable reference gene. In conclusion, selecting appropriate reference genes under abiotic stress not only improves the accuracy of annual ryegrass gene expression analyses, but also provides a theoretical reference for the development of reference genes in plants of the genus Lolium. View Full-Text
Keywords: reference gene; annual ryegrass; abiotic stress; qRT-PCR; Lolium reference gene; annual ryegrass; abiotic stress; qRT-PCR; Lolium
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MDPI and ACS Style

Liu, Q.; Qi, X.; Yan, H.; Huang, L.; Nie, G.; Zhang, X. Reference Gene Selection for Quantitative Real-Time Reverse-Transcriptase PCR in Annual Ryegrass (Lolium multiflorum) Subjected to Various Abiotic Stresses. Molecules 2018, 23, 172. https://doi.org/10.3390/molecules23010172

AMA Style

Liu Q, Qi X, Yan H, Huang L, Nie G, Zhang X. Reference Gene Selection for Quantitative Real-Time Reverse-Transcriptase PCR in Annual Ryegrass (Lolium multiflorum) Subjected to Various Abiotic Stresses. Molecules. 2018; 23(1):172. https://doi.org/10.3390/molecules23010172

Chicago/Turabian Style

Liu, Qiuxu, Xiao Qi, Haidong Yan, Linkai Huang, Gang Nie, and Xinquan Zhang. 2018. "Reference Gene Selection for Quantitative Real-Time Reverse-Transcriptase PCR in Annual Ryegrass (Lolium multiflorum) Subjected to Various Abiotic Stresses" Molecules 23, no. 1: 172. https://doi.org/10.3390/molecules23010172

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