Evaluation of the expression of internal control transcripts by real-time RT-PCR analysis during tomato flower abscission

Abstract

Many investigations of the abscission mechanisms in plants are based on gene expression studies. Quantitative real-time RT-PCR (qRT-PCR) is widely and increasingly used for mRNA quantification, and the results are typically referenced to an internal control gene to avoid bias. We investigated the suitability of twelve tomato (Lycopersicon esculentum Mill. var. Liaoyuanduoli) housekeeping genes during hormone, high salt and temperature-induced abscission. The free software-based applications NormFinder and qBase PLUS were used to statistically identify the best internal controls for a given set of biological samples. The expression stability of a number of housekeeping genes were validated during tomato abscission. The two most suitable reference genes for the commonly used treatments of the major hormones related to abscission are TBP and RPL8. In some cases, more than three reference genes may be required, depending on the type of samples being compared. Four suitable reference genes (TBP/DNAj/RPL8/EXPRESSION) are recommended for more complex analysis, such as hormone and bio-stress induced abscission samples.

Keywords: Abscission, housekeeping gene, Tomato, Lycopersicon esculentum, TBP, DNAj, RPL8, EXPRESSION