Reference Genes for Quantitative Real-time PCR Normalization of Cenostigma Pyramidale Roots Under Salt Stress and Mycorrhizal Association

Overview

Journal Genet Mol Biol

Specialty Genetics

Date 2021 Jun 1

PMID 34061138

Authors

Gabriella Frosi

Jose Ribamar Costa Ferreira-Neto

Joao Pacifico Bezerra-Neto

Lais Luana de Lima

David Anderson de Lima Morais

Valesca Pandolfi

Ederson Akio Kido

Leonor Costa Maia

Mauro Guida Santos

Ana Maria Benko-Iseppon

Affiliations

Soon will be listed here.

Abstract

Cenostigma pyramidale is a native legume of the Brazilian semiarid region which performs symbiotic association with arbuscular mycorrhizal fungi (AMF), being an excellent model for studying genes associated with tolerance against abiotic and biotic stresses. In RT-qPCR approach, the use of reference genes is mandatory to avoid incorrect interpretation of the relative expression. This study evaluated the stability of ten candidate reference genes (CRGs) from C. pyramidale root tissues under salt stress (three collection times) and associated with AMF (three different times of salinity). The de novo transcriptome was obtained via RNA-Seq sequencing. Three algorithms were used to calculate the stability of CRGs under different conditions: (i) global (Salt, Salt+AMF, AMF and Control, and collection times), (ii) only non-inoculated plants, and (iii) AMF (only inoculated plants). HAG2, SAC1, aRP3 were the most stable CRGs for global and AMF assays, whereas HAG2, SAC1, RHS1 were the best for salt stress assay. This CRGs were used to validate the relative expression of two up-regulated transcripts in Salt2h (RAP2-3 and PIN8). Our study provides the first set of reference genes for C. pyramidale under salinity and AMF, supporting future researches on gene expression with this species.

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