title: Screening Reliable Reference Genes for RT-qPCR Analysis of Gene Expression in Moringa oleifera.
abstract: Moringa oleifera is a promising plant species for oil and forage, but its genetic improvement is limited. Our current breeding program in this species focuses on exploiting the functional genes associated with important agronomical traits. Here, we screened reliable reference genes for accurately quantifying the expression of target genes using the technique of real-time quantitative polymerase chain reaction (RT-qPCR) in M. oleifera. Eighteen candidate reference genes were selected from a transcriptome database, and their expression stabilities were examined in 90 samples collected from the pods in different developmental stages, various tissues, and the roots and leaves under different conditions (low or high temperature, sodium chloride (NaCl)- or polyethyleneglycol (PEG)- simulated water stress). Analyses with geNorm, NormFinder and BestKeeper algorithms revealed that the reliable reference genes differed across sample designs and that ribosomal protein L1 (RPL1) and acyl carrier protein 2 (ACP2) were the most suitable reference genes in all tested samples. The experiment results demonstrated the significance of using the properly validated reference genes and suggested the use of more than one reference gene to achieve reliable expression profiles. In addition, we applied three isotypes of the superoxide dismutase (SOD) gene that are associated with plant adaptation to abiotic stress to confirm the efficacy of the validated reference genes under NaCl and PEG water stresses. Our results provide a valuable reference for future studies on identifying important functional genes from their transcriptional expressions via RT-qPCR technique in M. oleifera.
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangdong, 510642, China.|Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University, Guangdong, 510642, China.|Guangdong Province Research Center of Woody Forage Engineering Technology, South China Agricultural University, Guangdong, 510642, China.|College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, Guangdong, 510642, China.
authors.forename: Li-Ting
authors.initials: LT
authors.lastname: Deng
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangdong, 510642, China.|Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University, Guangdong, 510642, China.|Guangdong Province Research Center of Woody Forage Engineering Technology, South China Agricultural University, Guangdong, 510642, China.|College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, Guangdong, 510642, China.
authors.forename: Yu-Ling
authors.initials: YL
authors.lastname: Wu
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangdong, 510642, China.|Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University, Guangdong, 510642, China.|Guangdong Province Research Center of Woody Forage Engineering Technology, South China Agricultural University, Guangdong, 510642, China.|College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, Guangdong, 510642, China.
authors.forename: Jun-Cheng
authors.initials: JC
authors.lastname: Li
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangdong, 510642, China.|Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University, Guangdong, 510642, China.|Guangdong Province Research Center of Woody Forage Engineering Technology, South China Agricultural University, Guangdong, 510642, China.|College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, Guangdong, 510642, China.
authors.forename: Kun-Xi
authors.initials: KX
authors.lastname: OuYang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangdong, 510642, China.|Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University, Guangdong, 510642, China.|Guangdong Province Research Center of Woody Forage Engineering Technology, South China Agricultural University, Guangdong, 510642, China.|College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, Guangdong, 510642, China.
authors.forename: Mei-Mei
authors.initials: MM
authors.lastname: Ding
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangdong, 510642, China.|Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University, Guangdong, 510642, China.|Guangdong Province Research Center of Woody Forage Engineering Technology, South China Agricultural University, Guangdong, 510642, China.|College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, Guangdong, 510642, China.
authors.forename: Jun-Jie
authors.initials: JJ
authors.lastname: Zhang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangdong, 510642, China.|Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University, Guangdong, 510642, China.|Guangdong Province Research Center of Woody Forage Engineering Technology, South China Agricultural University, Guangdong, 510642, China.|College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, Guangdong, 510642, China.
authors.forename: Shu-Qi
authors.initials: SQ
authors.lastname: Li
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangdong, 510642, China.|Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University, Guangdong, 510642, China.|Guangdong Province Research Center of Woody Forage Engineering Technology, South China Agricultural University, Guangdong, 510642, China.|College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, Guangdong, 510642, China.
authors.forename: Meng-Fei
authors.initials: MF
authors.lastname: Lin
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangdong, 510642, China.|Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University, Guangdong, 510642, China.|Guangdong Province Research Center of Woody Forage Engineering Technology, South China Agricultural University, Guangdong, 510642, China.|College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, Guangdong, 510642, China.
authors.forename: Han-Bin
authors.initials: HB
authors.lastname: Chen
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangdong, 510642, China.|Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University, Guangdong, 510642, China.|Guangdong Province Research Center of Woody Forage Engineering Technology, South China Agricultural University, Guangdong, 510642, China.|College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, Guangdong, 510642, China.
authors.forename: Xin-Sheng
authors.initials: XS
authors.lastname: Hu
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangdong, 510642, China.|Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University, Guangdong, 510642, China.|Guangdong Province Research Center of Woody Forage Engineering Technology, South China Agricultural University, Guangdong, 510642, China.|College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, Guangdong, 510642, China.
authors.forename: Xiao-Yang
authors.initials: XY
authors.lastname: Chen
mesh_terms: D000222:Adaptation, Physiological / Q000235:genetics*; D020869:Gene Expression Profiling; D029588:Moringa oleifera / Q000235:genetics* / Q000254:growth & development; D018515:Plant Leaves / Q000235:genetics* / Q000254:growth & development; D010940:Plant Proteins / Q000235:genetics*; D018517:Plant Roots / Q000235:genetics* / Q000254:growth & development; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012015:Reference Standards; D013312:Stress, Physiological / Q000235:genetics*; D059467:Transcriptome
chemical_list: D010940:Plant Proteins