Selection and Validation of Appropriate Reference Genes for Real-Time Quantitative PCR Analysis in Needles of Larix olgensis under Abiotic Stresses
| Autor: | Chenghao Li, Dandan Li, Jia Yang, Sen Yu, Xiao Liu, Minzhen Zeng |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
Normalization (statistics) abiotic stress reference gene Quantitative PCR analysis Computational biology Biology 01 natural sciences law.invention 03 medical and health sciences law Reference genes Gene expression Gene Polymerase chain reaction 030304 developmental biology Abiotic component 0303 health sciences Abiotic stress Forestry lcsh:QK900-989 qPCR lcsh:Plant ecology gene expression Larix olgensis 010606 plant biology & botany |
| Zdroj: | Forests Volume 11 Issue 2 Forests, Vol 11, Iss 2, p 193 (2020) |
| ISSN: | 1999-4907 |
| DOI: | 10.3390/f11020193 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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