Data/hora: 16/05/2024 14:56:47

Biblioteca(s): 	Embrapa Agrobiologia.	
Data corrente: 	27/12/2017	
Data da última atualização: 	03/01/2018	
Autoria: 	POLESE, V.; SOARES, C. de P.; SILVA, P. R. A. da; ARAUJO, J. L. S. de; BALDANI, J. I.; VIDAL, M. S.	
Afiliação: 	VALÉRIA POLESE, UFRRJ; LEITON DE PAULA SOARES, UFRRJ; PAULA RENATA ALVES DA SILVA, UFRRJ; JEAN LUIZ SIMOES DE ARAUJO, CNPAB; JOSE IVO BALDANI, CNPAB; MARCIA SOARES VIDAL, CNPAB.	
Título: 	Selection and validation of reference genes for RT-qPCR indicates that juice of sugarcane varieties modulate the expression of C metabolism genes in the endophytic diazotrophic Herbaspirillum rubrisubalbicans strain HCC103.	
Ano de publicação: 	2017	
Fonte/Imprenta: 	Antonie van Leeuwenhoekv, 110, n. 12, p. 1555-1568, 2017.	
ISSN: 	1572-9699	
DOI: 	https://doi.org/10.1007/s10482-017-0906-7	
Idioma: 	Inglês	
Conteúdo: 	Quantitative reverse transcription PCR (RT-qPCR) is an important tool for evaluating gene expression. However, this technique requires that specific internal normalizing genes be identified for different experimental conditions. To date, no internal normalizing genes are available for validation of data analyses for Herbaspirillum rubrisubalbicans strain HCC103, an endophyte that is part of the sugarcane consortium inoculant. This work seeks to identify and evaluate suitable reference genes for gene expression studies in HCC103 grown until middle log phase in sugarcane juice obtained from four sugarcane varieties or media with three different carbon sources. The mRNA levels of five candidate genes (rpoA, gyrA, dnaG, recA and gmK) and seven target genes involved in carbon metabolism (acnA, fbp, galE, suhB, wcaA, ORF_0127.0101 and _0127.0123) were quantified by RT-qPCR. Analysis of expression stability of these genes was carried out using geNorm and Normfinder software. The results indicated that the HCC103 dnaG and gyrA genes are the most stable and showed adequate relative expression level changes among the different sugarcane juices. The highest expression level was seen for ORF_0127.0101, which encodes a sugar transporter, in juice from sugarcane variety RB867515 and glucose as the carbon source. The suhB gene, encoding SuhB inositol monophosphatase, had a higher relative expression level on 0.5% glucose, 100% sugarcane juice from variety RB867515 and 0.5% aconitate. Together the results suggest that dnaG and gyrA genes are suitable as reference genes for RT-qPCR analysis of strain HCC103 and that juice from different sugarcane varieties modulates the expression of key genes involved in carbon metabolism.	
Palavras-Chave: 	Internal normalizing; Organic acids; Relative expression; Sugar transport.	
Thesaurus Nal: 	nitrogen fixation.	
Categoria do assunto: 	S Ciências Biológicas	
Registro original: 	Embrapa Agrobiologia (CNPAB)	
Exemplares: 	CNPAB - (40665-1) - UPC - AP - DD - 2017.00161 -  - 2017.00161 - DIS