title: Transcriptome-Wide Identification of Reference Genes for Expression Analysis of Soybean Responses to Drought Stress along the Day.
abstract: The soybean transcriptome displays strong variation along the day in optimal growth conditions and also in response to adverse circumstances, like drought stress. However, no study conducted to date has presented suitable reference genes, with stable expression along the day, for relative gene expression quantification in combined studies on drought stress and diurnal oscillations. Recently, water deficit responses have been associated with circadian clock oscillations at the transcription level, revealing the existence of hitherto unknown processes and increasing the demand for studies on plant responses to drought stress and its oscillation during the day. We performed data mining from a transcriptome-wide background using microarrays and RNA-seq databases to select an unpublished set of candidate reference genes, specifically chosen for the normalization of gene expression in studies on soybean under both drought stress and diurnal oscillations. Experimental validation and stability analysis in soybean plants submitted to drought stress and sampled during a 24 h timecourse showed that four of these newer reference genes (FYVE, NUDIX, Golgin-84 and CYST) indeed exhibited greater expression stability than the conventionally used housekeeping genes (ELF1-β and β-actin) under these conditions. We also demonstrated the effect of using reference candidate genes with different stability values to normalize the relative expression data from a drought-inducible soybean gene (DREB5) evaluated in different periods of the day.
authors.affiliation: Embrapa Soybean, Brazilian Agricultural Research Corporation, Londrina, Paraná, Brazil; Department of Biology, State University of Londrina, Londrina, Paraná, Brazil.
authors.forename: Juliana
authors.initials: J
authors.lastname: Marcolino-Gomes
authors.affiliation: Embrapa Soybean, Brazilian Agricultural Research Corporation, Londrina, Paraná, Brazil.
authors.forename: Fabiana Aparecida
authors.initials: FA
authors.lastname: Rodrigues
authors.affiliation: Embrapa Soybean, Brazilian Agricultural Research Corporation, Londrina, Paraná, Brazil.
authors.forename: Renata
authors.initials: R
authors.lastname: Fuganti-Pagliarini
authors.affiliation: Embrapa Soybean, Brazilian Agricultural Research Corporation, Londrina, Paraná, Brazil; Department of Crop Science, Federal University of Viçosa, Viçosa, Minas Gerais, Brazil.
authors.forename: Thiago Jonas
authors.initials: TJ
authors.lastname: Nakayama
authors.affiliation: Embrapa Soybean, Brazilian Agricultural Research Corporation, Londrina, Paraná, Brazil; Department of Biology, State University of Londrina, Londrina, Paraná, Brazil.
authors.forename: Rafaela
authors.initials: R
authors.lastname: Ribeiro Reis
authors.affiliation: Embrapa Soybean, Brazilian Agricultural Research Corporation, Londrina, Paraná, Brazil.
authors.forename: Jose Renato
authors.initials: JR
authors.lastname: Bouças Farias
authors.affiliation: Plant Gene Expression Center, ARS/USDA, Albany, California, United States of America; Department of Plant and Microbial Biology, University of California-Berkeley, Berkeley, California, United States of America.
authors.forename: Frank G
authors.initials: FG
authors.lastname: Harmon
authors.affiliation: Embrapa Agroenergy, Brazilian Agricultural Research Corporation, Brasília, Brazil.
authors.forename: Hugo Bruno
authors.initials: HB
authors.lastname: Correa Molinari
authors.affiliation: Embrapa Soybean, Brazilian Agricultural Research Corporation, Londrina, Paraná, Brazil; Department of Biology, State University of Londrina, Londrina, Paraná, Brazil.
authors.forename: Mayla Daiane
authors.initials: MD
authors.lastname: Correa Molinari
authors.affiliation: Embrapa Soybean, Brazilian Agricultural Research Corporation, Londrina, Paraná, Brazil.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Nepomuceno
mesh_terms: D003681:Dehydration / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods; D018506:Gene Expression Regulation, Plant / Q000235:genetics* / Q000502:physiology; D015723:Gene Library; D017343:Genes, Plant / Q000235:genetics* / Q000502:physiology; D016133:Polymerase Chain Reaction; D018749:RNA, Plant / Q000235:genetics / Q000502:physiology; D013025:Glycine max / Q000235:genetics* / Q000502:physiology; D059467:Transcriptome / Q000235:genetics / Q000502:physiology
chemical_list: D018749:RNA, Plant