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Characterization of reference genes for RT-qPCR in the desert moss Syntrichia caninervis in response to abiotic stress and desiccation/rehydration.
Front Plant Sci. 2015; 6:38.FP

Abstract

Syntrichia caninervis is the dominant bryophyte of the biological soil crusts found in the Gurbantunggut desert. The extreme desert environment is characterized by prolonged drought, temperature extremes, high radiation and frequent cycles of hydration and dehydration. S. caninervis is an ideal organism for the identification and characterization of genes related to abiotic stress tolerance. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) expression analysis is a powerful analytical technique that requires the use of stable reference genes. Using available S. caninervis transcriptome data, we selected 15 candidate reference genes and analyzed their relative expression stabilities in S. caninervis gametophores exposed to a range of abiotic stresses or a hydration-desiccation-rehydration cycle. The programs geNorm, NormFinder, and RefFinder were used to assess and rank the expression stability of the 15 candidate genes. The stability ranking results of reference genes under each specific experimental condition showed high consistency using different algorithms. For abiotic stress treatments, the combination of two genes (α-TUB2 and CDPK) were sufficient for accurate normalization. For the hydration-desiccation-rehydration process, the combination of two genes (α-TUB1 and CDPK) were sufficient for accurate normalization. 18S was among the least stable genes in all of the experimental sets and was unsuitable as reference gene in S. caninervis. This is the first systematic investigation and comparison of reference gene selection for RT-qPCR work in S. caninervis. This research will facilitate gene expression studies in S. caninervis, related moss species from the Syntrichia complex and other mosses.

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Authors+Show Affiliations

Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography - Chinese Academy of Sciences Ürümqi, China.

Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography - Chinese Academy of Sciences Ürümqi, China.

Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography - Chinese Academy of Sciences Ürümqi, China.

Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography - Chinese Academy of Sciences Ürümqi, China.

Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography - Chinese Academy of Sciences Ürümqi, China.

Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography - Chinese Academy of Sciences Ürümqi, China.

Department of Plant Biology, Southern Illinois University Carbondale, IL, USA.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

25699066

Citation

Li, Xiaoshuang, et al. "Characterization of Reference Genes for RT-qPCR in the Desert Moss Syntrichia Caninervis in Response to Abiotic Stress and Desiccation/rehydration." Frontiers in Plant Science, vol. 6, 2015, p. 38.

Li X, Zhang D, Li H, et al. Characterization of reference genes for RT-qPCR in the desert moss Syntrichia caninervis in response to abiotic stress and desiccation/rehydration. Front Plant Sci. 2015;6:38.

Li, X., Zhang, D., Li, H., Gao, B., Yang, H., Zhang, Y., & Wood, A. J. (2015). Characterization of reference genes for RT-qPCR in the desert moss Syntrichia caninervis in response to abiotic stress and desiccation/rehydration. Frontiers in Plant Science, 6, 38. https://doi.org/10.3389/fpls.2015.00038

Li X, et al. Characterization of Reference Genes for RT-qPCR in the Desert Moss Syntrichia Caninervis in Response to Abiotic Stress and Desiccation/rehydration. Front Plant Sci. 2015;6:38. PubMed PMID: 25699066.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - Characterization of reference genes for RT-qPCR in the desert moss Syntrichia caninervis in response to abiotic stress and desiccation/rehydration. AU - Li,Xiaoshuang, AU - Zhang,Daoyuan, AU - Li,Haiyan, AU - Gao,Bei, AU - Yang,Honglan, AU - Zhang,Yuanming, AU - Wood,Andrew J, Y1 - 2015/02/05/ PY - 2014/10/20/received PY - 2015/01/15/accepted PY - 2015/2/21/entrez PY - 2015/2/24/pubmed PY - 2015/2/24/medline KW - NormFinder KW - RefFinder KW - Syntrichia caninervis KW - geNorm KW - quantitative real-time PCR KW - reference gene SP - 38 EP - 38 JF - Frontiers in plant science JO - Front Plant Sci VL - 6 N2 - Syntrichia caninervis is the dominant bryophyte of the biological soil crusts found in the Gurbantunggut desert. The extreme desert environment is characterized by prolonged drought, temperature extremes, high radiation and frequent cycles of hydration and dehydration. S. caninervis is an ideal organism for the identification and characterization of genes related to abiotic stress tolerance. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) expression analysis is a powerful analytical technique that requires the use of stable reference genes. Using available S. caninervis transcriptome data, we selected 15 candidate reference genes and analyzed their relative expression stabilities in S. caninervis gametophores exposed to a range of abiotic stresses or a hydration-desiccation-rehydration cycle. The programs geNorm, NormFinder, and RefFinder were used to assess and rank the expression stability of the 15 candidate genes. The stability ranking results of reference genes under each specific experimental condition showed high consistency using different algorithms. For abiotic stress treatments, the combination of two genes (α-TUB2 and CDPK) were sufficient for accurate normalization. For the hydration-desiccation-rehydration process, the combination of two genes (α-TUB1 and CDPK) were sufficient for accurate normalization. 18S was among the least stable genes in all of the experimental sets and was unsuitable as reference gene in S. caninervis. This is the first systematic investigation and comparison of reference gene selection for RT-qPCR work in S. caninervis. This research will facilitate gene expression studies in S. caninervis, related moss species from the Syntrichia complex and other mosses. SN - 1664-462X UR - https://www.unboundmedicine.com/medline/citation/25699066/Characterization_of_reference_genes_for_RT_qPCR_in_the_desert_moss_Syntrichia_caninervis_in_response_to_abiotic_stress_and_desiccation/rehydration_ DB - PRIME DP - Unbound Medicine ER -