Next Article in Journal
Identification and Expression of NAC Transcription Factors of Vaccinium corymbosum L. in Response to Drought Stress
Previous Article in Journal
Study of Gliding Arc Plasma Treatment for Bamboo-Culm Surface Modification
Open AccessArticle

Screening and Evaluation of Stable Reference Genes for Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Analysis in Chinese Fir Roots under Water, Phosphorus, and Nitrogen Stresses

1
Forestry College, Fujian Agriculture and Forestry University, Fuzhou 350002, China
2
Southern Swedish Forest Research Center, Faculty of Forest Science, Swedish University of Agricultural Sciences, PO Box 49, SE-230 53 Alnarp, Sweden
3
Fujian Provincial Colleges and University Engineering Research Center of Plantation Sustainable Management, Fuzhou 350002, China
*
Author to whom correspondence should be addressed.
Forests 2019, 10(12), 1087; https://doi.org/10.3390/f10121087
Received: 22 October 2019 / Revised: 20 November 2019 / Accepted: 27 November 2019 / Published: 1 December 2019
(This article belongs to the Section Forest Ecophysiology and Biology)
Chinese fir (Cunninghamia lanceolata) is an economical important timber species widely planted in southeastern Asia. Decline in yield and productivity during successive rotation is believed to be linked with abiotic stress, such as drought stress and nitrogen (N) and phosphorus (P) starvation. Molecular breeding could be an option to develop tolerant genotypes. For gene expression studies using quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR), stable reference genes are needed for normalization of gene expression under different experimental conditions. However, there is no internal reference genes identified for Chinese fir under abiotic stresses. Thus, nine internal reference genes based on transcriptome data were selected and analyzed in the root of Chinese fir under drought stress and N and P starvation. Data were analyzed using geNorm, NormFinder, and BestKeeper, to screen and identify the best reference genes. The results showed that the UBQ and GAPDH genes were the two most stable genes under drought stress and the Actin1 and GAPDH were the two most stable genes under P starvation. Further, it was discovered that the Actin1 and UBC were the two most stable genes under N starvation among nine candidate reference genes. The gene expression of drought stress induced expression protein 14-3-3-4, the P transporter gene ClPht1;3, and the nitrate transporter gene NRT1.1 were used to verify the stability of the selected reference genes under drought stress and P and N starvation, respectively, and the results revealed that the screened reference genes were sufficient to normalize expression of the target genes. In conclusion, the results demonstrate that the stability of reference genes was closely related to the external conditions and reference genes applied to the roots of Chinese fir under different abiotic stress treatments were different. Our data will facilitate further studies on stress ecology and gene function analysis in Chinese fir.
Keywords: Cunninghamia lanceolata; reference genes; qRT-PCR; geNorm; NormFinder; BestKeeper; gene expression Cunninghamia lanceolata; reference genes; qRT-PCR; geNorm; NormFinder; BestKeeper; gene expression
MDPI and ACS Style

Chen, R.; Chen, W.; Tigabu, M.; Zhong, W.; Li, Y.; Ma, X.; Li, M. Screening and Evaluation of Stable Reference Genes for Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Analysis in Chinese Fir Roots under Water, Phosphorus, and Nitrogen Stresses. Forests 2019, 10, 1087.

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