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Volume 35 Issue 11
Nov.  2020
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Article Contents
ZHANG L Z, HAN X Y, WU J H, et al. Reference Gene Selection for RT-qPCR Analysis on Cucumis melo [J]. Fujian Journal of Agricultural Sciences,2020,35(11):1179−1187 doi: 10.19303/j.issn.1008-0384.2020.11.002
Citation: ZHANG L Z, HAN X Y, WU J H, et al. Reference Gene Selection for RT-qPCR Analysis on Cucumis melo [J]. Fujian Journal of Agricultural Sciences,2020,35(11):1179−1187 doi: 10.19303/j.issn.1008-0384.2020.11.002

Reference Gene Selection for RT-qPCR Analysis on Cucumis melo

doi: 10.19303/j.issn.1008-0384.2020.11.002
  • Received Date: 2020-07-29
  • Rev Recd Date: 2020-10-04
  • Available Online: 2020-11-13
  • Publish Date: 2020-11-28
  •   Objective  In search for internal reference genes of Cucumis melo L. that could stably express in different tissues and under stress conditions to warrant accuracy and reliability of the RT-qPCR analysis on target gene expression.   Method   Expression stabilities of 9 candidate genes, 18srRNA, TUA, EF1a, Actin1, Actin2, Actin3, Actin4, CYC, and UBI-ep, from the roots, leaves, seeds, and fruits of Xinyinhui melon being treated by water, cinnamic acid, saline alkali or ABA were determined by RT-qPCR and analyzed using the BestKeeper, NormFinder, and geNorm software.  Result   In different tissues, the top 5 choice genes identified by BestKeeper ranked as CYC18s rRNAUBI-epEF1aTUA, those by NormFinder EF1aUBIepActin4CYCActin3, and those by geNorm Actin4=Actin3Actin1EF1aUBI-ep. Under various stresses, they were 18s rRNAActin3Actin4EF1aUBI-ep as ranked by BestKeeper, EF1aUBI-epActin4CYC18s rRNA by NormFinder, and EF1a=UBI-epActin4CYCActin by geNorm. Overall, EF1a appeared to be most stable among the 9 genes. Insofar as variety of tissues is concerned, Actin4, Actin3, Actin1, and EF1a were more stable than the others; and, under stress, EF1a and UBI-ep tended to be superior.   Conclusion   Stably expressed in the tissues under the stresses as tested, EF1a was selected as the reference gene for studies on C. melo to reduce experimental errors. To further ensure accuracy, application of dual reference genes in RT-qPCR analysis might be considered.
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