Identification of suitable reference genes for quantitative reverse transcription PCR in Luffa (Luffa cylindrica)

Gangjun Zhao; Meng Wang; Yaqin Gan; Hao Gong; Junxing Li; Xiaoming Zheng; Xiaoxi Liu; Siying Zhao; Jianning Luo; Haibin Wu

doi:10.1007/s12298-022-01182-8

Identification of suitable reference genes for quantitative reverse transcription PCR in Luffa (Luffa cylindrica)

Physiol Mol Biol Plants. 2022 Apr;28(4):737-747. doi: 10.1007/s12298-022-01182-8. Epub 2022 May 3.

Authors

Gangjun Zhao^{1

2}, Meng Wang², Yaqin Gan², Hao Gong¹, Junxing Li¹, Xiaoming Zheng¹, Xiaoxi Liu¹, Siying Zhao², Jianning Luo¹, Haibin Wu^{1

3

2}

Affiliations

¹ Vegetable Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, 510640 China.
² Guangdong Key Laboratory for New Technology Research of Vegetables, Guangzhou, 510640 China.
³ Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou, 510642 China.

Abstract

Reverse transcription real-time quantitative PCR is widely used to quantify gene expression. Reference genes are usually used as internal controls to measure the target gene expression level. To date, there is no consensus on the use of systematically validated reference genes in different tissues of Luffa. This study evaluated the expression stability of 11 candidate reference genes in different tissues using five algorithms (BestKeeper, comparative delta-Ct method, GeNorm, NormFinder, and RefFinder). Protein phosphatase 2A was the most stable gene, while alpha Tubulin was the least stable. The relative expression of ethylene-related genes in different tissues was also analyzed to reveal their role in sex determination. This study provides the basis for using suitable reference genes to evaluate targeted gene expression.

Supplementary information: The online version contains supplementary material available at 10.1007/s12298-022-01182-8.

Keywords: Luffa; PP2A; RT-qPCR; Reference genes; Sex determination; Sponge gourd.