Abstract
Eremosparton songoricum (Litv.) Vass. (E. songoricum) is a rare and extremely drought-tolerant desert plant that holds promise as a model organism for the identification of genes associated with water deficit stress. Here, we cloned and evaluated the expression of eight candidate reference genes using quantitative real-time reverse transcriptase polymerase chain reactions. The expression of these candidate reference genes was analyzed in a diverse set of 20 samples including various E. songoricum plant tissues exposed to multiple environmental stresses. GeNorm analysis indicated that expression stability varied between the reference genes in the different experimental conditions, but the two most stable reference genes were sufficient for normalization in most conditions. EsEF and Esα-TUB were sufficient for various stress conditions, EsEF and EsACT were suitable for samples of differing germination stages, and EsGAPDHand EsUBQ were most stable across multiple adult tissue samples. The Es18S gene was unsuitable as a reference gene in our analysis. In addition, the expression level of the drought-stress related transcription factor EsDREB2 verified the utility of E. songoricum reference genes and indicated that no single gene was adequate for normalization on its own. This is the first systematic report on the selection of reference genes in E. songoricum, and these data will facilitate future work on gene expression in this species.
Highlights
The drought-tolerant plant Eremosparton songoricum (Litv.) Vass. is a leafless perennial clone-forming semi-shrub [1]
Our data show that expression stability varied between reference genes in different experimental setups and that no single reference gene is suitable for accurate normalization of Quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR)
(1) For all the tested samples, ACT and β-TUB1 were the most stable reference genes, and seven genes were needed for accurate qRT-PCR normalization for the experimental design of all samples (V7/8 = 0.163)
Summary
The drought-tolerant plant Eremosparton songoricum (Litv.) Vass. is a leafless perennial clone-forming semi-shrub [1]. E. songoricum is a rare endemic plant of central Asia and is the only species of the genus in China [2], where it is found only in the Gurbantunggut desert (Xinjiang, China [3]). As a rare and extremely drought-tolerant Leguminosae shrub, E. songoricum is being developed as a model organism for investigating the morphological, biochemical, physiological and molecular adaptations and responses to water deficit. The DREB (dehydration-responsive element binding protein) gene plays a crucial role in plant stress response and signal transduction [4,5]. The DREB2 gene family is responsive to dehydration stress [5], with many plant species accumulating elevated amounts of its transcript in response to drought and water deficit stress [6,7,8,9]
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