Sequencing and Analysis of the<i> Shewanella putrefaciens</i> WS13 Genome

Chen, Li; Yang, Shengping; Qian, Yun‐Fang; Xie, Jing

doi:10.1166/jbmb.2019.1850

Cited by 4 publications

(5 citation statements)

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“…Shewanella putrefaciens WS 13 strain used in this study was isolated from a shrimp in a putrefactive state (Litopenaeus vannamei) and stored in our laboratory (Chen et al, 2019a). The strain was maintained in Luria Broth (LB, Land Bridge Technology, Beijing, China) with 50% (v/v) glycerol at −80 • C. The strain was recovered in 9 mL of LB and incubated at 30 • C with shaking at 200 rpm for 12 h, and repeated the same operation.…”

Section: Bacterial Strains and Culture Preparationmentioning

confidence: 99%

Comparative Proteome Analysis of Shewanella putrefaciens WS13 Mature Biofilm Under Cold Stress

Yan

Xie

2020

Front. Microbiol.

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Section: Bacterial Strains and Culture Preparationmentioning

confidence: 99%

Comparative Proteome Analysis of Shewanella putrefaciens WS13 Mature Biofilm Under Cold Stress

Yan

Xie

2020

Front. Microbiol.

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“…S. putrefaciens WS13 strain was isolated from spoilage shrimp Litopenaeus vanname in refrigerator ( Chen et al., 2019 ). The isolate was maintained in Luria Broth (LB, Land Bridge Technology, Beijing, China) with 50% (v/v) glycerol at -80 °C freezer in our laboratory at Shanghai Ocean University, Shanghai, China.…”

Section: Methodsmentioning

confidence: 99%

Comparative Transcriptome Analysis of Shewanella putrefaciens WS13 Biofilms Under Cold Stress

Yan

Yang

Xie

2022

Front. Cell. Infect. Microbiol.

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Shewanella putrefaciens is a Gram-negative bacterium that can cause seafood spoilage under low-temperature conditions. The bacterium easily forms biofilms to enhance its survival in challenging environments. Our previous research revealed that the biofilm formed by S. putrefaciens WS13 under the low temperature (4 °C) has larger biomass and tighter structure than at an optimum growth temperature (30 °C). In this study, comparative transcriptome analysis was further performed to get insights into the global-level of gene expression in the biofilm formed by S. putrefaciens WS13 under the refrigerating and optimal temperatures using Illumina RNA-Sequencing technique. The results revealed that a total of 761 genes were differentially expressed, of which 497 were significantly up-regulated and 264 were significantly down-regulated (p<0.05). The qRT-PCR results of randomly selected differentially expressed genes (DEGs) confirmed the RNA sequencing results. Comparison of transcriptome data revealed 28 significantly changed metabolic pathways under the cold stress, including the down-regulated chemotaxis, and motility, and up-regulated tryptophan metabolism, histidine biosynthesis, and quorum sensing, which benefited the biofilm formation of S. putrefaciens WS13 under the adverse circumstance. This study provided useful data for better understanding of the biofilm formation of S. putrefaciens, and also laid a theoretical foundation for novel vaccine and drug targets against the severe spoilage bacterium under the cold stress.

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“…S. putrefaciens WS13 strain, isolated from spoiled Litopenaeus vannamei , which was preserved by our group (NCBI No. CP028435.1) ( 27 ) was applied in this experiment. The strain was preserved in LB medium (Land Bridge Technology, Beijing, China) with 50% (v/v) glycerol (Sinopharm., AR, China) in a −80°C refrigerator and was activated to 8–9 log CFU/mL (OD 600 value = 0.80) in fresh LB medium under 30°C for 150 rpm, and then, the culture was diluted at 0.1% ratio with LB medium for further use.…”

Section: Methodsmentioning

confidence: 99%

“…Quantitative real-time PCR was operated with the following conditions: 50°C for 2 min; 95°C for 10 min; 40 cycles of 95°C for 15 s; and 60°C for 1 min. Genes were selected ( 27 ), and the expression level of genes was analyzed by the 2 −ΔΔCT method in triplicate. Statistical analysis was performed by Origin Pro 2021, and error bars represent the SD from the mean of measurements.…”

Section: Methodsmentioning

confidence: 99%

The enhanced expression of genes encoding diguanylate cyclases under cold stress contributes to the adhesion and biofilm formation of Shewanella putrefaciens WS13

et al. 2022

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Shewanella putrefaciens is a special spoilage bacterium of seafood during cold storage, which is easy to form biofilm and bring serious hazard to the seafood quality. Life cycle of biofilm starts after bacterial adhesion, which is essential for the formation and development of biofilm. As a ubiquitous second messenger in bacteria, c-di-GMP regulates the conversion between bacterial planktonic state and biofilm state. In this study, the adhesion and biofilm formation of S. putrefaciens WS13 under 4°C were compared to those under 30°C. Atom force microscope and scanning electron microscope were used to study the bacterial adhesion. Biofilm was analyzed by Fourier transform infrared spectroscopy, Bradford assay and phenol-sulfuric acid method. High-performance liquid chromatographic-tandem mass spectrometric and quantitative real-time PCR were applied to study c-di-GMP level and genes encoding diguanylate cyclases in cells, respectively. Results showed that the swarming mobility of S. putrefaciens WS13 was weaker under 4°C, however, the adhesive force under 4°C was 4–5 times higher than that under 30°C. Biofilm biomass, extracellular polysaccharides and extracellular proteins were 2.5 times, 3 times, and 1.6 times more than those under 30°C, respectively, but biofilm composition formed under both temperatures were similar. c-di-GMP level in S. putrefaciens WS13 under 30°C was no more than half of that in the corresponding growth stage under 4°C. Quantitative real-time PCR analysis also showed that the expression of genes encoding diguanylate cyclases were significantly enhanced under 4°C than that under 30°C. S. putrefaciens WS13 adapted to the cold stress by enhancing the expression of genes encoding diguanylate cyclases to promote bacterial adhesion and biofilm formation. This study provides a theoretical foundation for the research on the cold adaptation mechanism of specific spoilage bacteria of seafood based on c-di-GMP, and also provides a new idea to control seafood quality from the perspective of microbial molecular biology.

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Sequencing and Analysis of the Shewanella putrefaciens WS13 Genome

Cited by 4 publications

References 0 publications

Comparative Proteome Analysis of Shewanella putrefaciens WS13 Mature Biofilm Under Cold Stress

Comparative Proteome Analysis of Shewanella putrefaciens WS13 Mature Biofilm Under Cold Stress

Comparative Transcriptome Analysis of Shewanella putrefaciens WS13 Biofilms Under Cold Stress

The enhanced expression of genes encoding diguanylate cyclases under cold stress contributes to the adhesion and biofilm formation of Shewanella putrefaciens WS13

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