Park, Sojung
(Department of Molecular Science and Technology, Ajou University)
,
Nam, Eun woo
(Department of Molecular Science and Technology, Ajou University)
,
Kim, Yeeun
(Department of Molecular Science and Technology, Ajou University)
,
Lee, Seohyeon
(Department of Applied Chemistry and Biological Engineering, Ajou University)
,
Kim, Seul I
(Department of Molecular Science and Technology, Ajou University)
,
Yoon, Hyunjin
(Department of Molecular Science and Technology, Ajou University)
Salmonellosis is a form of gastroenteritis caused by Salmonella infection. The main transmission route of salmonellosis has been identified as poorly cooked meat and poultry products contaminated with Salmonella. However, in recent years, the number of outbreaks attributed to contaminated raw produc...
Salmonellosis is a form of gastroenteritis caused by Salmonella infection. The main transmission route of salmonellosis has been identified as poorly cooked meat and poultry products contaminated with Salmonella. However, in recent years, the number of outbreaks attributed to contaminated raw produce has increased dramatically. To understand how Salmonella adapts to produce, transcriptomic analysis was conducted on Salmonella enterica serovar Virchow exposed to fresh-cut radish greens. Considering the different Salmonella lifestyles in contact with fresh produce, such as motile and sessile lifestyles, total RNA was extracted from planktonic and epiphytic cells separately. Transcriptomic analysis of S. Virchow cells revealed different transcription profiles between lifestyles. During bacterial adaptation to fresh-cut radish greens, planktonic cells were likely to shift toward anaerobic metabolism, exploiting nitrate as an electron acceptor of anaerobic respiration, and utilizing cobalamin as a cofactor for coupled metabolic pathways. Meanwhile, Salmonella cells adhering to plant surfaces showed coordinated upregulation in genes associated with translation and ribosomal biogenesis, indicating dramatic cellular reprogramming in response to environmental changes. In accordance with the extensive translational response, epiphytic cells showed an increase in the transcription of genes that are important for bacterial motility, nucleotide transporter/metabolism, cell envelope biogenesis, and defense mechanisms. Intriguingly, Salmonella pathogenicity island (SPI)-1 and SPI-2 displayed up- and downregulation, respectively, regardless of lifestyles in contact with the radish greens, suggesting altered Salmonella virulence during adaptation to plant environments. This study provides molecular insights into Salmonella adaptation to plants as an alternative environmental reservoir.
Salmonellosis is a form of gastroenteritis caused by Salmonella infection. The main transmission route of salmonellosis has been identified as poorly cooked meat and poultry products contaminated with Salmonella. However, in recent years, the number of outbreaks attributed to contaminated raw produce has increased dramatically. To understand how Salmonella adapts to produce, transcriptomic analysis was conducted on Salmonella enterica serovar Virchow exposed to fresh-cut radish greens. Considering the different Salmonella lifestyles in contact with fresh produce, such as motile and sessile lifestyles, total RNA was extracted from planktonic and epiphytic cells separately. Transcriptomic analysis of S. Virchow cells revealed different transcription profiles between lifestyles. During bacterial adaptation to fresh-cut radish greens, planktonic cells were likely to shift toward anaerobic metabolism, exploiting nitrate as an electron acceptor of anaerobic respiration, and utilizing cobalamin as a cofactor for coupled metabolic pathways. Meanwhile, Salmonella cells adhering to plant surfaces showed coordinated upregulation in genes associated with translation and ribosomal biogenesis, indicating dramatic cellular reprogramming in response to environmental changes. In accordance with the extensive translational response, epiphytic cells showed an increase in the transcription of genes that are important for bacterial motility, nucleotide transporter/metabolism, cell envelope biogenesis, and defense mechanisms. Intriguingly, Salmonella pathogenicity island (SPI)-1 and SPI-2 displayed up- and downregulation, respectively, regardless of lifestyles in contact with the radish greens, suggesting altered Salmonella virulence during adaptation to plant environments. This study provides molecular insights into Salmonella adaptation to plants as an alternative environmental reservoir.
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문제 정의
In this study, transcriptome analysis provided a comprehensive understanding of Salmonella adaptation to plant environments. Salmonella reprogrammed the transcription of a myriad of genes in contact with plant tissues, and the repertoire of genes with altered expression levels was influenced by the bacterial lifestyles between planktonic and epiphytic types.
가설 설정
The data from three independent tests were averaged. (B) Bacterial enumeration in the presence of radish greens. The numbers of planktonic cells (CFU/ml) and epiphytic cells (CFU/g) adhering to plant tissues were counted using XLD agar plating.
제안 방법
For each gene, the ΔCt values of PBS-treated cells were subtracted from the ΔCt values of planktonic or epiphytic cells, and the mean values of three independent tests were plotted with their SDs.
For each gene, the ΔCt values of PBS-treated cells were subtracted from the ΔCt values of planktonic or epiphytic cells, and the mean values of three independent tests were plotted with their standard deviations (SDs).
RNA-Seq was conducted using the Illumina Hiseq 2500 platform (Illumina) generating single-end reads approximately 100 bp in length (Chunlab, Korea). The RNA sequencing reads were mapped to the FORC_038 genome sequence (GenBank accession number CP015574) using the CLRNAseq program (Chunlab) and normalized using Reads Per Kilobase of transcript per Million mapped reads (RPKM), Relative Log Expression (RLE), and Trimmed Mean of M-value (TMM) [20-22]. To standardize mRNA abundance in response to plant tissues, log2[FC] (log2[fold change between the RNA samples]) values were computed by dividing the TMMnormalized reads of planktonic or epiphytic cells by the TMM-normalized reads of PBS-treated cells.
데이터처리
The Student’s t-test was used to produce p-values for determining the significance.
성능/효과
Although the physiological role of the periplasmic nitrate reductase is yet to be defined, the expression of napABC genes is induced in response to anaerobic growth conditions, and Salmonella lacking the periplasmic nitrate concentrations [42]. S. Virchow FORC_038 in contact with radish greens decreased the expression of the narZYWV operon regardless of lifestyle between planktonic and sessile cells, but remarkably increased the transcription of narGHJI and napABC operons in its planktonic status, implicating anaerobic nitrate respiration for energy production in planktonic cells (Fig. 4A). Genes required for nitrite reduction were accordingly induced in planktonic cells to detoxify or further reduce nitrite to ammonia, including the genes of nrfABCDEGG and nirBDC operons.
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