Welcome to Chinese Journal of Tropical Crops,

Chinese Journal of Tropical Crops ›› 2020, Vol. 41 ›› Issue (8): 1505-1512.DOI: 10.3969/j.issn.1000-2561.2020.08.001

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Screening of Stable Reference Genes for qRT-PCR Analysis in Macadamia integrifolia

YANG Qian1,2,3,YANG Ziping1,ZHOU Yali1,CHEN Dongquan1,LIU Heng1,2,3,*()   

  1. 1. South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Zhanjiang, Guangdong 524091, China
    2. Key Laboratory of Tropical Fruit Biology, Ministry of Agriculture and Rural Affairs, Zhanjiang, Guangdong 524091, China
    3. National Field Genebank for Tropical Fruits, Zhanjiang, Guangdong 524091, China
  • Received:2019-09-16 Revised:2019-11-08 Online:2020-08-25 Published:2020-09-17
  • Contact: LIU Heng


Macadamia nut is an important economics crop in subtropical areas of China. The selection of a suitable reference gene is an important prerequisite for successful gene expression analysis by real-time fluorescence quantitative PCR (RT-qPCR). In order to select the appropriate reference genes, we investigated the expression stability of 11 candidate genes (18S rRNA, Actin, CYP, EF1a, EF1b, GAPDH, MDH, TUBa, TUBb, UBQ, UBC) in RT-qPCR experiments in different tissues, including kernel, peel, roots, stem, leaf from Macadamia with geNorm, NormFinder, BestKeeper, ΔCt, RefFinder program software packages. As determined by geNorm, MDH/EF1b were the most stable reference genes, TUBa was the least stable gene. BestKeeper revealed that MDH was the most stables reference gene, and EF1b ranked the second. The rank in BestKeeper was similarly with that in geNorm. The result by NormFinder showed GAPDH was the most stable gene, CYP ranks the second, the least stable gene was TUBa. ΔCt algorithm demonstrated that18S was the most stable gene, GAPDH ranks the second, MDH and EF1b ranked the third and fourth. To obtain a consensus result of the most stable reference genes according to the RefFinder approach, the geometric mean of the four algorithms corresponding rankings for each candidate gene were calculated: MDH>18S>GADPH>EF1b>CYP>UBC>EF1a>Actin>TUBb>UBQ>TUBa. The result showed that MDH was the most suitable reference gene for macadamia in different tissues.

Key words: Macadamia integrifolia, reference genes, real-time fluorescence quantitative PCR, stability analysis

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