Data/hora: 05/06/2024 18:22:03

Biblioteca(s): | Embrapa Agrobiologia.
Data corrente: | 24/11/2016
Data da última atualização: | 23/10/2018
Autoria: | SILVA, P. R. A. da; VIDAL, M. S.; SOARES, C. de P.; POLESE, V.; ARAUJO, J. L. S. de; BALDANI, J. I.
Afiliação: | PAULA RENATA ALVES DA SILVA, UFRRJ; MARCIA SOARES VIDAL, CNPAB; CLEITON DE PAULA SOARES, BOLSISTA DA EMBRAPA AGROBIOLOGIA; VALERIA POLESE, UFRRJ; JEAN LUIZ SIMOES DE ARAUJO, CNPAB; JOSE IVO BALDANI, CNPAB.
Título: | Selection and evaluation of reference genes for RT-qPCR expression studies on Burkholderia tropica strain Ppe8, a sugarcane-associated diazotrophic bacterium grown with different carbon sources or sugarcane juice
Ano de publicação: | 2016
Fonte/Imprenta: | Antonie van Leeuwenhoek, v. 109, n. 111, p. 1493-1502, 2016.
DOI: | 10.1007/s10482-016-0751-0
Idioma: | Inglês
Notas: | Epub, 17 Aug., 2016.

Conteúdo:

Among the members of the genus Burkholderia, Burkholderia tropica has the ability to fix nitrogen and promote sugarcane plant growth as well as act as a biological control agent. There is little information about how this bacterium metabolizes
carbohydrates as well as those carbon sources found in the sugarcane juice that accumulates in stems during plant growth. Reverse transcription quantitative PCR (RT-qPCR) can be used to evaluate changes in gene expression during bacterial growth on different carbon sources. Here we tested the expression of six reference genes, lpxC, gyrB, recA, rpoA, rpoB, and rpoD, when cells were grown with glucose, fructose, sucrose, mannitol, aconitic acid, and sugarcane juice as carbon sources. The lpxC, gyrB, and recA were selected as the most stable reference genes based on geNorm and NormFinder software analyses. Validation of these three reference genes during strain Ppe8 growth on the same carbon sources showed that genes involved in
glycogen biosynthesis (glgA, glgB, glgC) and trehalose biosynthesis (treY and treZ) were highly expressed when Ppe8 was grown in aconitic acid relative to other carbon sources, while otsA expression (trehalose biosynthesis) was reduced with all carbon
sources. In addition, the expression level of the ORF_6066 (gluconolactonase) gene was reduced on sugarcane juice. The results confirmed the stability of the three selected reference genes (lpxC, gyrB, and recA) during the RT-qPCR and also their robustness
by evaluating the relative expression of genes involved in glycogen and trehalose biosynthesis when strain Ppe8 was grown on different carbon sources and sugarcane juice.

Palavras-Chave: | Carbon metabolism; Diazotrophic bacteria; Normalizing genes; Real time PCR; Relative expression.
Categoria do assunto: | S Ciências Biológicas
Registro original: | Embrapa Agrobiologia (CNPAB)
Exemplares: | CNPAB - (40369-1) - UPC - SP - PP - 2016.00161 -  - 2016.00161 - DIS