Evaluation of Reference Genes for Gene Expression Analysis in Eichhornia crassipes

Eichhornia crassipes is a notorious invasive aquatic weed, causing enormous ecological and economic losses worldwide. However, it has great potential in agriculture, industry, medical care, and other areas. While being such an important plant, it is poorly understood from the molecular perspective. Aiming to select suitable reference genes for gene expression quantification in E. crassipes, this study favors future research at the molecular level. In this work, 12 candidate reference genes were selected. Their expression stability in samples of different tissues, samples treated with various hormones, samples supplied with different levels of phosphorus (P), and pooled samples, were analyzed using GeNorm, NormFinder, BestKeeper, and RefFinder. Meanwhile, the optimal number of reference genes was calculated by GeNorm. The results showed that eIF and ElF1a were the two most stable reference genes in all samples and in tissue samples. In response to hormone treatments, Actin and eIF are the best choices of internal controls. In the case of P treatments, TUA and H2A are recommended to be used as reference genes. Overall, results from this work suggest different reference genes should be applied in qRT-PCR on E. crassipes, according to the specific experimental setup.

Automated summary

This study aims to select suitable reference genes for gene expression quantification in E. crassipes and analyze the expression stability of candidate reference genes in different tissue samples, hormone-treated samples, phosphorus-treated samples, and mixed samples. The results suggest that different reference genes should be selected according to the specific experimental setup.