Validation of Reference Genes for Quantifying Changes in Physiological Gene Expression in Apple Tree under Cold Stress and Virus Infection

Abstract

Quantitative reverse transcription PCR is used for gene expression analysis as the accurate and sensitive method. To analyze quantification of gene expression changes in apple plants, 10 housekeeping genes (ACT, CKL, EF-1α, GAPDH, MDH, PDI, THFs, UBC, UBC10, and WD40) were evaluated for their stability of expression during infection by Apple stem grooving virus (ASGV) or in cold-stress apple plant buds. Five reference-gene validation programs were used to establish the order of the most stable genes for ASGV as CKL>THFs>GAPDH>ACT, and the least stable genes WD40<UBC10<EF-1α for infection by ASGV. For treatment by three different temperatures, the most stable genes were WD40>CKL>UBC10, and the least stable genes were ACT<UBC<THFs. To validate our findings, using two of the most stable and the one least stable validated reference genes, one defense responsive gene and one cold-stress-responsive gene were examined to compare their relative changes in gene expression. In conclusion, our results provide a useful framework for choice of suitable reference genes according to different biotic or abiotic stress conditions in plant species.

Index system of rural human settlement in rural revitalization under the perspective of China

“…Biegańska et al 13 focus on changing of demographic and social aspects caused by development around the city in rural areas in Latvia, Poland, and Germany, a number of rural regions face persistent population decline. In Korea experience 14 – 16 , It discussed rural development policies currently upgraded by the South Korean in terms of questions such as the decrease of agricultural productivity, the depopulation in rural areas. Faradiba et al 17 aim to look for the impact of climate, disaster, and social community on rural development.…”