Abstract
Real-time fluorescent quantitative PCR (qRT-PCR) is often chosen as an effective experimental method for analyzing gene expression. However, an appropriate reference gene as a standard is needed to obtain accurate gene expression data. To date, no internal reference genes have been reported for research on cranberries. Expanding the selection of internal reference genes for cranberry will enable reliable gene expression analysis, and, at the same time, can also lay a solid foundation for revealing the biological characteristics of cranberry. Here, we selected ten candidate reference gene families and used three statistical software tools—geNorm, NormFinder and BestKeeper—to evaluate their expression stability under the influence of different experimental factors. The results showed that protein phosphatase 2A regulatory subunit (PP2A) or RNA helicase-like 8 (RH 8) was the best choice for an internal reference gene when analyzing different cranberry cultivars. In two sample sets comprising different cranberry organs and three abiotic stress treatments, sand family protein (SAND) was the best choice as a reference gene. In this study, we screened genes that are stably expressed under the influence of various experimental factors by qRT-PCR. Our results will guide future studies involving gene expression analysis of cranberry.
Highlights
Cranberry (Vaccinium macrocarpon Ait.) is an evergreen shrub with considerable cold resistance and is often found in cool and high-altitude areas in the northern hemisphere [1]
The experimental results showed that the RNA concentration of all plant materials was above 1000 ng/μL and the 260/280 nm optical density ratios were all in the range of 1.8–2.0, indicating the RNA was of high quality
Analysis of gene expression under different experimental conditions is a major aspect of gene function analysis. qRT-PCR technology has the advantages of high specificity, high sensitivity, good reproducibility, speed and efficiency, and can provide accurate results for gene expression analysis
Summary
Cranberry (Vaccinium macrocarpon Ait.) is an evergreen shrub with considerable cold resistance and is often found in cool and high-altitude areas in the northern hemisphere [1]. Studies have shown that cranberries have a high nutrient content and are rich in polyphenolic bioactive substances such as flavonols, resveratrol and anthocyanidins [2]. These bioactive substances have positive effects such as preventing urinary tract infection, clearing Helicobacter pylori infection, and anti-oxidation and anti-tumor effects [2]. These advantages make it become one of the fruit crops, which have a good prospect for development.
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