干旱区生态环境知识资源中心(Arid): Identification of reliable reference genes for qRT-PCR in the ephemeral plant Arabidopsis pumila based on full-length transcriptome data

Arid

DOI	10.1038/s41598-019-44849-1
	Identification of reliable reference genes for qRT-PCR in the ephemeral plant Arabidopsis pumila based on full-length transcriptome data
	Jin, Yuhuan 1; Liu, Fang 1,2; Huang, Wei 1; Sun, Qi 1; Huang, Xianzhong 1
通讯作者	Huang, Xianzhong
来源期刊	SCIENTIFIC REPORTS
ISSN	2045-2322
出版年	2019
卷号	9
英文摘要	Arabidopsis pumila, an annual ephemeral plant, plays important roles in preventing wind and sand erosion, water and soil conservation, and microhabitat improvement in the North of Xinjiang, China. Studies of adaptive mechanisms in harsh desert environments at the genetic and genomic levels can be used to more effectively develop and protect this species. The quantitative real-time polymerase chain reaction (qRT-PCR) method is one of the essential means to achieve these goals, and the selection of an appropriate reference gene is the prerequisite for qRT-PCR. In this study, 10 candidate reference genes were identified from the full-length transcriptome data of A. pumila, and their expression stabilities under four abiotic stresses (drought, heat, cold and salt) and in seven different tissues (roots, hypocotyl, cotyledon, leaves, stems, flowers and siliques) were evaluated with four programmes geNorm, NormFinder, Bestkeeper and RefFinder. Although the most stable reference genes were variable under different treatments using different software, comprehensive ranking revealed that UEP and HAF1 under drought stress, UBQ9 and GAPDH under heat stress, UBC35 and GAPDH under cold stress, GAPDH and ACT1 under salt stress, and ACTT and GAPDH in different tissues were the most stable reference genes. Moreover, GAPDH and UBQ9 were the most suitable reference gene combinations for all samples. The expression pattern of the K+ uptake permease gene KUP9 further validated that the selected reference genes were suitable for normalization of gene expression. The identification of reliable reference genes guarantees more accurate qRT-PCR quantification for A. pumila and facilitates functional genomics studies of ephemeral plants.
类型	Article
语种	英语
国家	Peoples R China
开放获取类型	gold, Green Published
收录类别	SCI-E
WOS记录号	WOS:000470847700005
WOS关键词	TIME RT-PCR ; HOUSEKEEPING GENES ; SPRING EPHEMERALS ; QUANTITATIVE-PCR ; EXPRESSION ; NORMALIZATION ; VALIDATION
WOS类目	Multidisciplinary Sciences
WOS研究方向	Science & Technology - Other Topics
来源机构	中国农业大学
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/218790
作者单位	1.Shihezi Univ, Coll Life Sci, Special Plant Genom Lab, Shihezi 832000, Xinjiang, Peoples R China; 2.China Agr Univ, Coll Biol Sci, State Key Lab Plant Physiol & Biochem, Beijing 100193, Peoples R China
推荐引用方式 GB/T 7714	Jin, Yuhuan,Liu, Fang,Huang, Wei,et al. Identification of reliable reference genes for qRT-PCR in the ephemeral plant Arabidopsis pumila based on full-length transcriptome data[J]. 中国农业大学,2019,9.
APA	Jin, Yuhuan,Liu, Fang,Huang, Wei,Sun, Qi,&Huang, Xianzhong.(2019).Identification of reliable reference genes for qRT-PCR in the ephemeral plant Arabidopsis pumila based on full-length transcriptome data.SCIENTIFIC REPORTS,9.
MLA	Jin, Yuhuan,et al."Identification of reliable reference genes for qRT-PCR in the ephemeral plant Arabidopsis pumila based on full-length transcriptome data".SCIENTIFIC REPORTS 9(2019).