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Validation of reliability for reference genes under various abiotic stresses in tea plant

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Abstract

Reference genes are frequently used as a normalization standard to obtain reliable data during quantitative real-time polymerase chain reaction (qRT-PCR). However, recent studies showed that most traditional reference genes were not stable under different treatments or environmental stresses, which may lead to misinterpret expression of the target genes. In this study, 7 candidate reference genes in tea plant (Camellia sinensis (L.) Kuntze cv. Yingshuang) were selected and their expression stability under different abiotic stresses was analyzed using geNorm, NormFinder, and BestKeeper methods. Our results suggest that TUA1 (alpha-1 tubulin) has the most stable expression under damage stresses according to 3 methods of analysis. For drought stresses, 18S rRNA, and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) were the most stable genes. For cold, Al, and NaCl stresses, GAPDH and TUA1 may be the alternative options. Our results may provide an insight for identification of the optimal reference genes for tea plants under various treatments.

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Abbreviations

Ct:

cycle threshold

CV:

percentage covariance

EF1-α:

elongation factor 1-alpha

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

MMCS:

maximum moisture content of soil

qRT-PCR:

quantitative real-time polymerase chain reaction

T m :

melting temperature

TUA1:

alpha-1 tubulin

TUA2:

alpha-2 tubulin

UBI:

ubiquitin

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Authors and Affiliations

  1. College of Horticulture, Nanjing Agricultural University, Nanjing, Jiangsu, China

    Q. P. Ma, X. Chen & X. H. Li

  2. Suzhou Polytechnic Institute of Agriculture, Suzhou, Jiangsu, China

    S. Hao

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  1. Q. P. Ma
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  2. S. Hao
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  3. X. Chen
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  4. X. H. Li
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Correspondence to X. H. Li.

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Ma, Q.P., Hao, S., Chen, X. et al. Validation of reliability for reference genes under various abiotic stresses in tea plant. Russ J Plant Physiol 63, 423–432 (2016). https://doi.org/10.1134/S1021443716030080

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