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Transcriptome analysis of rice pathogen Acidovorax avenae subsp. avenae cultivated in vitro, in vivo and in co-culture with the rice rhizobacterium Burkholderia seminalis

ABSTRACT: Determining how a bacterial pathogen responds to its host and other bacterial species by altering gene expression is key to understand its pathogenesis and environmental adaption. Here, we used RNA-Seq to comprehensively and quantitatively assess the transcriptional response of the rice bacterial pathogen Acidovorax avenae subsp. avenae strain RS-1 cultivated in vitro, in vivo and in co-culture with rice rhizobacterium Burkholderia seminalis R456. Results revealed a surprisingly large number of regulatory differences between these conditions indicating adaptation of A. avenae subsp. avenae to specific ecological conditions. In particular, a number of potential virulence factors such as type 3 secretion system proteins were specifically expressed under in vivo conditions, whereas genes whose protein products are involved in inter-bacterial interaction such as auxin efflux carrier, small mechanosensitive ion channel protein, and ureidoglycolate hydrolase were among those specifically up-regulated under co-culture conditions. In addition, global genomic analysis of strain RS-1 identified 406 putative non-coding (nc) RNA genes. Interestingly, 8 ncRNA genes that were uniquely expressed under in vivo may be linked to pathogenicity while 4 ncRNA genes that were uniquely expressed under coculture conditions may be involved in adaption to co-cultivation with B. seminalis. Expression data obtained by RNA-Seq were also confirmed for selected genes by quantitative real-time PCR and two-dimensional gel electrophoresis as well as knockout analysis.

ORGANISM(S): Paracidovorax avenae RS-1

PROVIDER: GSE50522 | GEO | 2013/09/03

SECONDARY ACCESSION(S): PRJNA217854

REPOSITORIES: GEO

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Project description:Determining how a bacterial pathogen responds to its host and other bacterial species by altering gene expression is key to understand its pathogenesis and environmental adaption. Here, we used RNA-Seq to comprehensively and quantitatively assess the transcriptional response of the rice bacterial pathogen Acidovorax avenae subsp. avenae strain RS-1 cultivated in vitro, in vivo and in co-culture with rice rhizobacterium Burkholderia seminalis R456. Results revealed a surprisingly large number of regulatory differences between these conditions indicating adaptation of A. avenae subsp. avenae to specific ecological conditions. In particular, a number of potential virulence factors such as type 3 secretion system proteins were specifically expressed under in vivo conditions, whereas genes whose protein products are involved in inter-bacterial interaction such as auxin efflux carrier, small mechanosensitive ion channel protein, and ureidoglycolate hydrolase were among those specifically up-regulated under co-culture conditions. In addition, global genomic analysis of strain RS-1 identified 406 putative non-coding (nc) RNA genes. Interestingly, 8 ncRNA genes that were uniquely expressed under in vivo may be linked to pathogenicity while 4 ncRNA genes that were uniquely expressed under coculture conditions may be involved in adaption to co-cultivation with B. seminalis. Expression data obtained by RNA-Seq were also confirmed for selected genes by quantitative real-time PCR and two-dimensional gel electrophoresis as well as knockout analysis. Aaa strain RS-1 and B. seminalis strain R456 was isolated from diseased rice plants (Li et al., 2011; Xie et al., 2011) and rice rhizosphere (Zhang et al., 2007; Li et al., 2011), respectively, in our previous studies, and were stored in 20-30% sterile glycerol at -80°C. The samples of Aaa strain RS-1 for in vitro and in vivo analysis were prepared as described before (Ibrahim et al., 2012). The co-culture analysis of Aaa strain RS-1 with B. seminalis strain R456 were conducted according to Ruiz et al. (2009) and Di Cagno et al. (2009). Briefly, Aaa strain RS-1 and B. seminalis strain R456 was inoculated and incubated in chambers of a double culture vessel apparatus separated by a 0.4-μm membrane filter (Millipore Isopore™). In order to avoid the possible contamination during in vivo and co-culture operation, all bacterial samples were further confirmed based on the sequence analysis of 16S-rRNA (Li et al., 2011). Then samples were processed for RNA harvesting, mRNA purification and cDNA synthesis.

Project description:Background and study aims Colorectal cancer (CRC) is one of the most commoncancers among humans worldwide. Recent studies demonstrated that the composition of the bacterial community in the human gut, as well as inflammation occurring in the gut, are some of the factors that modify the risk of an individual to develop CRC. The human gut is home to more than 1000 bacterial species, including health-promoting species and disease-causing species. The consumption of rice bran, a by-product of rice milling, was previously shown to positively modify bacterial composition in the gut among healthy adults. The protective effect of a long-term rice bran consumption against CRC among individuals known to have higher risk of CRC, such as older individuals who are regular smokers and having a family history of CRC, needs to be established. This study aims to investigate whether the implementation of a 24-week dietary programme involving rice bran consumption among adults at high risk of CRC is feasible, and whether it has any effect in inducing a health-promoting modification of the bacterial community, as well as a reduction of inflammation, in the gut of these individuals. Who can participate? Chinese adults of either gender, who are aged 50 or above and are categorised to be in the high risk CRC group by the Asian-Pacific Colorectal Screening tool, in which classification is based on age, smoking status and family history of CRC. What does the study involve? After the recruited subjects were screened for eligibility of study participation and written informed consent had been obtained from them, they were randomly assigned into either Group A or Group B. Participants in Group A were given packets of rice bran and were asked to consume 30 grams of the rice bran at 24-hour intervals for 24 weeks. Participants in Group B were given packets of rice powder that has similar appearance and colour as the rice bran, and were asked to consume 30 grams of the rice powder, also at 24-hour intervals for 24 weeks. All participants were asked to provide a stool sample and blood sample at various time points during the study, namely just before rice bran consumption, as well as 6 weeks, 12 weeks and 24 weeks after the start of rice bran consumption. Laboratory tests were conducted on these samples. All participants were also instructed to complete a log book, detailing the date and time of rice bran or rice powder intake each day, and the amount consumed. The participants also completed a faecal diary where they documented the frequency of egestion, and the shape and amount of stool egested each day, as well as the occurrence of any abdominal discomfort or pain.

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Transcriptome analysis of rice pathogen Acidovorax avenae subsp. avenae cultivated in vitro, in vivo and in co-culture with the rice rhizobacterium Burkholderia seminalis

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