Next Article in Journal
Degradation of Ecosystem Services and Deforestation in Landscapes With and Without Incentive-Based Forest Conservation in the Ecuadorian Amazon
Previous Article in Journal
A Decision Support Tool for Assessing the Impact of Climate Change on Multiple Ecosystem Services
Open AccessArticle

Identification of Suitable Reference Genes for RT-qPCR Assays in Liriodendron chinense (Hemsl.) Sarg

1
College of Forestry, Nanjing Forestry University, Nanjing 210037, China
2
Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China
*
Author to whom correspondence should be addressed.
Forests 2019, 10(5), 441; https://doi.org/10.3390/f10050441
Received: 1 April 2019 / Revised: 30 April 2019 / Accepted: 17 May 2019 / Published: 22 May 2019
(This article belongs to the Section Forest Ecophysiology and Biology)
The precision and reliability of reverse transcription quantitative polymerase chain reaction (RT-qPCR) depend mainly on suitable reference genes; however, reference genes have not yet been identified for Liriodendron chinense (Hemsl.) Sarg. In this study, the expression stability of 15 candidate reference genes, ACT7, ACT97, UBQ1, eIF2, eIF3, HIS, BIG, AGD11, EFG, GAPDH, CYP, RPL25, UBC, RPB1, and TUB, was tested across multiple organs of L. chinense using four algorithms, geNorm, NormFinder, BestKeeper, and RefFinder. To understand the difference between the selected reference genes and the unsuitable candidate reference genes, the expression level of a target gene, LcPAT7, was normalized across various plant samples. ACT97 and eIF3 represented the best combination across all samples tested, while AGD11 and UBQ1 were unsuitable for normalization in this case. In the vegetative organ subset, ACT97, ACT7, and GAPDH showed the highest expression stability. For floral organs, UBC and eIF3 were the most stable reference genes. Unsuitable reference genes underestimated the expression levels of a target gene, LcPAT7. This study identified two reference genes (ACT97 and eIF3) for the precise and reliable normalization of L. chinense RT-qPCR data across various organs. Our work provides an effective framework for quantifying gene expression in L. chinense. View Full-Text
Keywords: accuracy of RT-qPCR; internal control genes; gene expression stability accuracy of RT-qPCR; internal control genes; gene expression stability
Show Figures

Figure 1

MDPI and ACS Style

Tu, Z.; Hao, Z.; Zhong, W.; Li, H. Identification of Suitable Reference Genes for RT-qPCR Assays in Liriodendron chinense (Hemsl.) Sarg. Forests 2019, 10, 441.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop