Genome-wide identification of internal reference genes for normalization of gene expression values during endosperm development in wheat.

Genome-wide identification of internal reference genes for normalization of gene expression values during endosperm development in wheat.
J Appl Genet. 2019 Jul 11;:
Authors: Mu J, Chen L, Gu Y, Duan L, Han S, Li Y, Yan Y, Li X
Abstract
Internal reference genes that are stably expressed are essential for normalization in comparative expression analyses. However, gene expression varies significantly among species, organisms, tissues, developmental stages, stresses, and treatments. Therefore, identification of stably expressed reference genes in developmental endosperm of bread wheat is important for expression analysis of endosperm genes. As the first study to systematically screen for reference genes across different developmental stages of wheat endosperm, nine genes were selected from among 76 relatively stable genes based on high-throughput RNA sequencing data. The expression stability of these candidate genes and five traditional reference genes was assessed by real-time quantitative PCR combined with three independent algorithms: geNorm, NormFinder, and BestKeeper. The results showed that ATG8d was the most stable gene during wheat endosperm development, followed by Ta54227, while the housekeeping gene GAPDH, commonly used as an internal reference, was the least stable. ATG8d and Ta54227 together formed the optimal combination of reference genes. Comparative expression analysis of glutenin genes indicated that credible quantific...

Source: J Appl Genet - July 10, 2019 Category: Genetics & Stem Cells Authors: Mu J, Chen L, Gu Y, Duan L, Han S, Li Y, Yan Y, Li X Tags: J Appl Genet Source Type: research