Selection and Validation of Appropriate Reference Genes for Real-Time Quantitative PCR Analysis in Needles of Larix olgensis under Abiotic Stresses

Li, Dandan; Yu, Sen; Zeng, Minzhen; Liu, Xiao; Yang, Jia; Li, Chenghao

doi:10.3390/f11020193

Submit to this Journal Review for this Journal Edit a Special Issue

Article Menu

...

Open AccessArticle

Selection and Validation of Appropriate Reference Genes for Real-Time Quantitative PCR Analysis in Needles of Larix olgensis under Abiotic Stresses

^†,

^†,

and

State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040, Heilongjiang, China

Author to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Forests 2020, 11(2), 193; https://doi.org/10.3390/f11020193

Received: 15 January 2020 / Revised: 4 February 2020 / Accepted: 5 February 2020 / Published: 10 February 2020

(This article belongs to the Special Issue Forest Genetics and Tree Improvement)

View Full-Text Download PDF

Browse Figures

Citation Export

Abstract

Larix olgensis Henry is an important afforestation species in northeastern China because of its fast juvenile growth, high-quality timber, and significant economic and ecological values. The selection of appropriate reference genes is necessary for the normalization of gene expression determination during quantitative real-time polymerase chain reaction (qRT-PCR) experiments. In this study, qRT-PCR was used to study gene expression. Three software packages geNorm, NormFinder, BestKeeper were used, and a comprehensive ranking of candidate reference genes was produced based on their output to evaluate the expression stability of 16 candidate reference genes from L. olgensis under drought, salt, cold, and heat stress. PP2A-1 and GAPDH ranked as the most stable reference genes under drought and cold stress, PP2A-1 and UBQ10 were most stable under salt stress, and TIP41 and ACT2 were most stable under heat stress. The least stable gene was ADP, which ranked the last under all treatments. Expression profile analysis of the antioxidant gene CAT using the two most stable and the single least stable reference genes under each stress further verified that the selected reference genes were suitable for gene expression normalization. This study provides an important foundation for the selection of suitable reference genes for the normalization and quantification of L. olgensis gene expression under abiotic stress conditions. View Full-Text

Keywords: Larix olgensis; reference gene; qPCR; abiotic stress; gene expression

▼ Show Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited

Supplementary Material

Supplementary File 1:
ZIP-Document (ZIP, 702 KiB)

Share and Cite

MDPI and ACS Style

Li, D.; Yu, S.; Zeng, M.; Liu, X.; Yang, J.; Li, C. Selection and Validation of Appropriate Reference Genes for Real-Time Quantitative PCR Analysis in Needles of Larix olgensis under Abiotic Stresses. Forests 2020, 11, 193. https://doi.org/10.3390/f11020193

AMA Style

Li D, Yu S, Zeng M, Liu X, Yang J, Li C. Selection and Validation of Appropriate Reference Genes for Real-Time Quantitative PCR Analysis in Needles of Larix olgensis under Abiotic Stresses. Forests. 2020; 11(2):193. https://doi.org/10.3390/f11020193

Chicago/Turabian Style

Li, Dandan, Sen Yu, Minzhen Zeng, Xiao Liu, Jia Yang, and Chenghao Li. 2020. "Selection and Validation of Appropriate Reference Genes for Real-Time Quantitative PCR Analysis in Needles of Larix olgensis under Abiotic Stresses" Forests 11, no. 2: 193. https://doi.org/10.3390/f11020193

Find Other Styles

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Metrics

Abstract views Pdf views Html views

Article Menu

Selection and Validation of Appropriate Reference Genes for Real-Time Quantitative PCR Analysis in Needles of Larix olgensis under Abiotic Stresses

Abstract

Supplementary Material

Share and Cite

Article Metrics

Article Access Statistics

Article Access Map by Country/Region

Further Information

Guidelines

MDPI Initiatives

Follow MDPI