干旱区生态环境知识资源中心(Arid): Systematic selection and validation of suitable reference genes for quantitative real-time PCR normalization studies of gene expression in Nitraria tangutorum

Arid

DOI	10.1038/s41598-020-73059-3
	Systematic selection and validation of suitable reference genes for quantitative real-time PCR normalization studies of gene expression in Nitraria tangutorum
	Wang, Bo; Duan, Huirong; Chong, Peifang; Su, Shiping; Shan, Lishan; Yi, Dan; Wang, Lirong; Li, Yi
通讯作者	Li, Y
来源期刊	SCIENTIFIC REPORTS
ISSN	2045-2322
出版年	2020
卷号	10 期号:1
英文摘要	Suitable reference genes can be used to calibrate the error in quantitative real-time PCR (qPCR) experiments, making the results more credible. However, there are no reference genes suitable for multiple species and under different experimental conditions. Nitraria tangutorum Bobr. is a typical plant native to desert areas. It is drought-resistant, saline-alkali resistant, extreme temperatures-resistant, and has strong adaptability. To date, the importance of this germplasm has not been sufficiently understood; therefore, it is still unclear which genes can be used as reference genes to calibrate qPCR data of N. tangutorum. In this study we analyzed the expression levels of 10 candidate reference genes (ACT, GAPDH, TUA, TUB, CYP, UBC, His, PP2A, HSP, and EF1-alpha) in N. tangutorum seedlings under a series of experimental conditions, including in different organs (root, stem, and leaf) and under abiotic stresses (salt, drought, heat, and cold) and hormone stimuli (abscisic acid) by qPCR. Three software programs (geNorm, NormFinder, and BestKeeper) were used to evaluate the expression stability of the ten genes. Comprehensive analysis showed that EF1-a and His had the best expression stability, whereas HSP was the least suitable as a reference gene. The expression profile of NtCER7, a gene related to the regulation of cuticular wax biosynthesis in N. tangutorum, verified the accuracy of the experimental results. Based on this study, we recommend EF1-a and His as suitable reference genes for N. tangutorum. This paper provides the first data on stable reference genes in N. tangutorum, which will be beneficial to studying the gene expression of N. tangutorum and other Nitraria species in the future.
类型	Article
语种	英语
开放获取类型	Green Published, gold
收录类别	SCI-E
WOS记录号	WOS:000577346400006
WOS关键词	SUPERIOR REFERENCE GENES ; RT-PCR ; TRANSCRIPT NORMALIZATION ; ACCURATE NORMALIZATION ; HOUSEKEEPING GENES ; SALINITY ; IDENTIFICATION ; METABOLISM ; BOBR. ; RICE
WOS类目	Multidisciplinary Sciences
WOS研究方向	Science & Technology - Other Topics
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/326759
作者单位	[Wang, Bo; Chong, Peifang; Su, Shiping; Shan, Lishan; Yi, Dan; Li, Yi] Gansu Agr Univ, Coll Forestry, Lanzhou 730070, Peoples R China; [Wang, Lirong] Qinghai Nationalities Univ, Coll Ecol Environm & Resources, Xining 810007, Peoples R China; [Wang, Lirong] Qinghai Nationalities Univ, Inst Ecol & Environm Qinghai Tibet Plateau, Xining 810007, Peoples R China; [Wang, Lirong] Qinghai Nationalities Univ, Key Lab Biotechnol & Anal & Test Qinghai Tibet Pl, Xining 810007, Peoples R China; [Wang, Lirong] Qinghai Nationalities Univ, Lab Resource Chem & Ecoenvironm Protect Tibetan P, Xining 810007, Peoples R China; [Duan, Huirong] Chinese Acad Agr Sci, Lanzhou Inst Husb & Pharmaceut Sci, Lanzhou 730000, Peoples R China
推荐引用方式 GB/T 7714	Wang, Bo,Duan, Huirong,Chong, Peifang,et al. Systematic selection and validation of suitable reference genes for quantitative real-time PCR normalization studies of gene expression in Nitraria tangutorum[J],2020,10(1).
APA	Wang, Bo.,Duan, Huirong.,Chong, Peifang.,Su, Shiping.,Shan, Lishan.,...&Li, Yi.(2020).Systematic selection and validation of suitable reference genes for quantitative real-time PCR normalization studies of gene expression in Nitraria tangutorum.SCIENTIFIC REPORTS,10(1).
MLA	Wang, Bo,et al."Systematic selection and validation of suitable reference genes for quantitative real-time PCR normalization studies of gene expression in Nitraria tangutorum".SCIENTIFIC REPORTS 10.1(2020).