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.
Highlights
Moringa oleifera Lam., belonging to a single-genus family Moringaceae, is a fast-growing tree species and widely distributed in the tropical and subtropical regions [1]
To evaluate the usefulness of the validated reference genes, we examined the expressions of three isotypes of the superoxide dismutase (SOD) gene under NaCl and PEG water stresses, including copperzinc SOD (Cu/Zn-SOD), iron SOD (Fe-SOD), and manganese SOD (Mn-SOD)
PCR amplification efficiency ranged from 90.28% for MDH2 to 104.18% for ubiquitin extension protein (UEP), which was within the range from 90% to 110%
Summary
Moringa oleifera Lam., belonging to a single-genus family Moringaceae, is a fast-growing tree species and widely distributed in the tropical and subtropical regions [1]. This species, with a great economic value for food and medical industry [1, 2, 3, 4], has gained interest globally, especially in the developing countries because of its rich nutrition in various organs (in particular, the mostly used leaves and seeds). Besides the physiological and medical studies, researchers start to investigate genetic diversity of M. oleifera using molecular markers, and to develop marker-assistant selection (MAS) for genetic improvement [5]. The transcriptomes from M. oleifera leaves were sequenced in our lab, which provided a well-assembled and annotated sequence database for gene function research
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
