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Analysis of multiple transcriptomes of the African oil palm (Elaeis guineensis) to identify reference genes for RT-qPCR

文献类型： 外文期刊

作者： Xia, Wei 1 ; Mason, Annaliese S. 3 ; Xiao, Yong 1 ; Liu, Zheng 1 ; Yang, Yaodong 1 ; Lei, Xintao 1 ; Wu, Xiaoming; Ma, 1 ;

作者机构： 1.Chinese Acad Trop Agr Sci, Coconuts Res Inst, Hainan Key Lab Trop Oil Crops Biol, Wenchang 571339, Hainan, Peoples R China

2.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, MOA Key Lab Trop Crop Biol & Genet Resources Util, Haikou 571101, Hainan, Peoples R China

3.Univ Queensland, Sch Agr & Food Sci, Brisbane, Qld 4072, Australia

4.Univ Queensland, Ctr Integrat Legume R

关键词： Elaeis guineensis;Reference gene;Transcriptomes;RT-qPCR

期刊名称：JOURNAL OF BIOTECHNOLOGY （ 影响因子：3.307； 五年影响因子：3.778 ）

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收录情况： SCI

摘要： The African oil palm (Elaeis guineensis), which is grown in tropical and subtropical regions, is a highly productive oil-bearing crop. For gene expression-based analyses such as reverse transcription-quantitative real time PCR (RT-qPCR), reference genes are essential to provide a baseline with which to quantify relative gene expression. Normalization using reliable reference genes is critical in correctly interpreting expression data from RT-qPCR. In order to identify suitable reference genes in African oil palm, 17 transcriptomes of different tissues obtained from NCBI were systematically assessed for gene expression variation. In total, 53 putative candidate reference genes with coefficient of variation values <3.0 were identified: 18 in reproductive tissue and 35 in vegetative tissue. Analysis for enriched functions showed that approximately 90% of identified genes were clustered in cell component gene functions, and 12 out of 53 genes were traditional housekeeping genes. We selected and validated 16 reference genes chosen from leaf tissue transcriptomes by using RT-qPCR in sets of cold, drought and high salinity treated samples, and ranked expression stability using statistical algorithms geNorm, Normfinder and Bestkeeper. Genes encoding actin, adenine phosphoribosyltransferase and eukaryotic initiation factor 4A genes were the most stable genes over the cold, drought and high salinity stresses. Identification of stably expressed genes as reference gene candidates from multiple transcriptome datasets was found to be reliable and efficient, and some traditional housekeeping genes were more stably expressed than others. We provide a useful molecular genetic resource for future gene expression studies in African oil palm, facilitating molecular genetics approaches for crop improvement in this species

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