Arid
DOI10.1007/s00299-012-1245-9
Reference gene selection for qPCR in Ammopiptanthus mongolicus under abiotic stresses and expression analysis of seven ROS-scavenging enzyme genes
Shi, Jing; Liu, Meiqin; Shi, Junna; Zheng, Guangshun; Wang, Yanping; Wang, Jinyu; Chen, Yuzhen; Lu, Cunfu; Yin, Weilun
通讯作者Lu, Cunfu
来源期刊PLANT CELL REPORTS
ISSN0721-7714
EISSN1432-203X
出版年2012
卷号31期号:7页码:1245-1254
英文摘要

Ammopiptanthus mongolicus, the only evergreen broadleaf shrub endemic to the northwest desert of China, is a valuable species for plant abiotic stress research. No report has so far described the selection of reference genes to get stringent normalization for qPCR in A. mongolicus. This work identified reliable reference genes for normalization of qPCR data in A. mongolicus under abiotic stresses from 14 reference gene candidates (UBQ, Tub1, Tub2, Abc1, Ubc1, Ubc2, Ubc4, Ubc5, eIF1, eIF2, eIF3, eIF4, EF1, EF2), and used the most suitable combination of reference genes to normalize the expression profiles of seven ROS-scavenging enzyme genes (AmSOD, AmAPX, AmGPX, AmCAT, AmGLR, AmPrx, and AmTrx). We set a series of 22 experimental samples covering the control and different time points under cold, dry, salt, and heat stresses. According to geNorm and NormFinder, the combination of eIF1 and eIF3 was best for accurate normalization across all the treatments, confirmed by normalizing qPCR data with AmHsp90. In contrast, these data show that Tub1, Abc1, and EF1 are not suitable reference gene candidates. After being normalized against eIF1 and eIF3, the seven ROS-scavenging enzyme genes exhibited differentially up- or down-regulated expression patterns. AmSOD and AmGPX were up-regulated by all four treatments, indicating that they may participate in an anti-oxidative mechanism under abiotic stresses in A. mongolicus. AmCAT exhibited a much higher expression level than AmAPX, AmPrx, and AmGPX, suggesting a principle role in detoxifying excessive H2O2. AmSOD, AmGPX and AmAPX showing the most abundant transcripts under heat, AmCAT and AmGLR under drought, and AmPrx under salt, were observed. Expression patterns of the seven ROS-scavenging enzyme genes suggest different antioxidant protection roles of these genes under abiotic stresses. These results are valuable for future research on gene expression and abiotic stress tolerance in A. mongolicus.


英文关键词Ammopiptanthus mongolicus ROS-scavenging enzyme qPCR Reference gene
类型Article
语种英语
国家Peoples R China
收录类别SCI-E
WOS记录号WOS:000305219900009
WOS关键词REAL-TIME PCR ; QUANTITATIVE RT-PCR ; INTERNAL CONTROL ; OXIDATIVE STRESS ; NORMALIZATION ; VALIDATION ; PLANT ; L. ; MODEL
WOS类目Plant Sciences
WOS研究方向Plant Sciences
来源机构北京林业大学
资源类型期刊论文
条目标识符http://119.78.100.177/qdio/handle/2XILL650/174449
作者单位Beijing Forestry Univ, Natl Engn Lab Tree Breeding, Key Lab Genet & Breeding Forest Trees & Ornamenta, Coll Biol Sci & Biotechnol,Minist Educ, Beijing 100083, Peoples R China
推荐引用方式
GB/T 7714
Shi, Jing,Liu, Meiqin,Shi, Junna,et al. Reference gene selection for qPCR in Ammopiptanthus mongolicus under abiotic stresses and expression analysis of seven ROS-scavenging enzyme genes[J]. 北京林业大学,2012,31(7):1245-1254.
APA Shi, Jing.,Liu, Meiqin.,Shi, Junna.,Zheng, Guangshun.,Wang, Yanping.,...&Yin, Weilun.(2012).Reference gene selection for qPCR in Ammopiptanthus mongolicus under abiotic stresses and expression analysis of seven ROS-scavenging enzyme genes.PLANT CELL REPORTS,31(7),1245-1254.
MLA Shi, Jing,et al."Reference gene selection for qPCR in Ammopiptanthus mongolicus under abiotic stresses and expression analysis of seven ROS-scavenging enzyme genes".PLANT CELL REPORTS 31.7(2012):1245-1254.
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