Guangyan Li, Xiaohui Sun, Xiaoping Zhu, Bin Wu, Hao Hong, Zhimei Xin, Xiangqi Xin, Jiejun Peng, Shanshan Jiang

Genes (Basel); 2023 Jul 19; 14(7):1477. doi: 10.3390/genes14071477.

Abstract

Quantitative real-time PCR (qRT-PCR) in sweet potatoes requires accurate data normalization; however, there are insufficient studies on appropriate reference genes for gene expression analysis. We examined variations in the expression of eight candidate reference genes in the leaf and root tissues of sweet potatoes (eight nonvirus-infected or eight virus-infected samples). Parallel analyses with geNorm, NormFinder, and Best-Keeper show that different viral infections and origin tissues affect the expression levels of these genes. Based on the results of the evaluation of the three software, the adenosine diphosphate-ribosylation factor is suitable for nonvirus or virus-infected sweet potato leaves. Cyclophilin and ubiquitin extension proteins are suitable for nonvirus-infected sweet potato leaves. Phospholipase D1 alpha is suitable for virus-infected sweet potato leaves. Actin is suitable for roots of nonvirus-infected sweet potatoes. Glyceraldehyde-3-phosphate dehydrogenase is suitable for virus-infected sweet potato roots. The research provides appropriate reference genes for further analysis in leaf and root samples of viruses in sweet potatoes.

See https://pubmed.ncbi.nlm.nih.gov/37510381/

Figure 3

Graphic representation of expression data in relation to candidate reference genes. (A) Expression of candidate reference genes in susceptible leaves. (B) Expression of candidate reference genes at the pathogenetic root. Univariate analysis of variance was used to indicate statistical significance with different letters. Eight candidate reference genes were shown as adenosine diphosphate-ribosylation factor (ARF), glyceraldehyde−3−phosphate dehydrogenase (GAP), phospholipase D1 alpha (PLD), Ubiquitin (UBI), actin (ACT), 18S ribosomal RNA (18S), alpha tubulin (ATUB), and cyclophilin (CYP).