The small non-coding RNA rli106 contributes to the environmental adaptation and pathogenicity of Listeria monocytogenes

Guo, Yun; Ji, Chunhui; Wang, Lixia; Ning, Chengcheng; Li, Na; Li, Zhiyuan; Shang, Yunxia; Li, Yaling; Sun, Yaoqiang; Huang, Xiaoxing; Li, Jie; Cai, Xuepeng; Meng, Qingling; Qiao, Jun

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The small non-coding RNA rli106 contributes to the environmental adaptation and pathogenicity of Listeria monocytogenes

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Mar 17, 2023

Journal of Veterinary Research

Volume 67 (2023): Issue 1 (March 2023)

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Published Online: Mar 17, 2023

Page range: 67 - 77

Received: Apr 01, 2022

Accepted: Mar 03, 2023

DOI: https://doi.org/10.2478/jvetres-2023-0013

Keywords
sRNA, environmental adaptation, pathogenicity, target gene.

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Growth curves of LM strains under different environmental stresses. LM EGD-e – parental strain of Listeria monocytogenes; LM-△rli106 – rli106 deletion strain of Listeria monocytogenes; CLM-△rli106 – rli106 complementation strain of Listeria monocytogenes
A–F – Growth curves of LM EGD-e, LM-△rli106 and CLM-△rli106 under the conditions of 42°C, pH 9, 5% NaCl and 8% NaCl, 3.8% ethanol and 5 mM H2O2. Bars indicate the standard error of the mean — Growth curves of LM strains under different environmental stresses. LM EGD-e – parental strain of Listeria monocytogenes; LM-△rli106 – rli106 deletion strain of Listeria monocytogenes; CLM-△rli106 – rli106 complementation strain of Listeria monocytogenes A–F – Growth curves of LM EGD-e, LM-△rli106 and CLM-△rli106 under the conditions of 42°C, pH 9, 5% NaCl and 8% NaCl, 3.8% ethanol and 5 mM H2O2. Bars indicate the standard error of the mean

Determination and comparisons of the motility of different strains of LM. LM EGD-e – parental strain of Listeria monocytogenes; LM-△rli106 – rli106 deletion strain of Listeria monocytogenes; CLM-△rli106 – rli106 complementation strain of Listeria monocytogenes
A – Kinetics of LM EGD-e (1), LM-△rli106 (2) and CLM-△rli106 (3) in semi-solid medium; B – Kinetics in tablets;
C – Diameter size of the bacterial colony on the plate; ns – not significant, P > 0.05. Bars indicate the standard error of the mean — Determination and comparisons of the motility of different strains of LM. LM EGD-e – parental strain of Listeria monocytogenes; LM-△rli106 – rli106 deletion strain of Listeria monocytogenes; CLM-△rli106 – rli106 complementation strain of Listeria monocytogenes A – Kinetics of LM EGD-e (1), LM-△rli106 (2) and CLM-△rli106 (3) in semi-solid medium; B – Kinetics in tablets; C – Diameter size of the bacterial colony on the plate; ns – not significant, P > 0.05. Bars indicate the standard error of the mean

Effects of rli106 gene deletion on biofilm formation of LM
A – Formation of biofilm by the parental strain of Listeria monocytogenes (LM EGD-e), rli106 deletion strain of Listeria monocytogenes (LM-△rli106) and rli106 complementation strain of Listeria monocytogenes (CLM-△rli106) at 12 h, 24 h and 48 h; B – Comparison of biofilm formation at 12 h, 24 h and 48 h. Bars indicate the standard error of the mean (SE); ** – P < 0.01 — Effects of rli106 gene deletion on biofilm formation of LM A – Formation of biofilm by the parental strain of Listeria monocytogenes (LM EGD-e), rli106 deletion strain of Listeria monocytogenes (LM-△rli106) and rli106 complementation strain of Listeria monocytogenes (CLM-△rli106) at 12 h, 24 h and 48 h; B – Comparison of biofilm formation at 12 h, 24 h and 48 h. Bars indicate the standard error of the mean (SE); ** – P < 0.01

Determination of adhesion, invasion and intracellular viability in RAW264.7 macrophages for the parental strain of Listeria monocytogenes (LM EGD-e), rli106 deletion strain of Listeria monocytogenes (LM-△rli106) and rli106 complementation strain of Listeria monocytogenes (CLM-△rli106). A – Adhesion rate of LM EGD-e, LM-△rli106 and CLM-△rli106 in RAW264.7 cells; B – Invasion rate of LM EGD-e, LM-△rli106 and CLM-△rli106 in RAW264.7 cells; C – Determination of intracellular viability in macrophages. Bars indicate the standard error of the mean (SE); ** – P < 0.01

Effects of rli106 gene deletion on virulence of LM. A – Survival curves of mice infected with the parental strain of Listeria monocytogenes (LM EGD-e), rli106 deletion strain of Listeria monocytogenes (LM-△rli106) and rli106 complementation strain of Listeria monocytogenes (CLM-△rli106); B and C – Bacterial loads in the livers and spleens of mice infected by LM EGD-e, LM-△rli106 and CLM-△rli106; bars indicate the standard error of the mean (SE); * – P < 0.05; ** – P < 0.01 (LM EGD-e)

Histopathological examination of liver, spleen and kidney of mice infected by LM strains (haematoxylin and eosin staining, ×400). LM EGD-e – parental strain of Listeria monocytogenes rli106; LM-△rli106 – rli106 deletion strain of Listeria monocytogenes; CLM-△rli106 – rli106 complementation strain of Listeria monocytogenes

Verification of interaction between rli106 sRNA and mRNA of target gene DegU. A – Complementary pairing position of DegU target gene and rli106; B and C – Bacterial lawns of BTH101 co-transformed by pUT18C and pMR-LacZ-DegU, and BTH101 co-transformed by pUT18C-rli106 and pMR-LacZ-DegU; D – OD420 nm of bacterial lawns; E – mRNA level of DegU target gene in the parental strain of Listeria monocytogenes (LM EGD-e), rli106 deletion strain of Listeria monocytogenes (LM-△rli106) and rli106 complementation strain of Listeria monocytogenes (CLM-△rli106); F and G – protein level of DegU in LM EGD-e, LM-△rli106 and CLM-△rli106

Primers used in the study

Primer name	Primer sequences (5′→3′)	Product size (bp)
rli106F	GCCTCTTTGCAATCGAAA
rli106R	ACTACTATTACTTTAGACTAGTTTTCAATT	225
P1	GGTACC AAATAGCCGACTGCTCCAGC
P2	AAATTCGTGTTATAATAAAATTGGTGAAATTAGATAAGTG	351
P3	CACTTATCTAATTTCACCAATTTTATTATAACACGAATTT
P4	CTGCAG GATCTCTAGCATTGGAAAGCCA	383
P5	AAATAGCCGACTGCTCCAGC
P6	GATCTCTAGCATTGGAAAGCCA	722
P7	CCGTGCTTGATTGCCGTTAC
P8	AACTAGGCCGCCTGCACC	1391/1166
CF	AAGCTTGCCTCTTTGCAATCGAAA
CR	CGGGATCCACTACTATTACTTTAGACTAG	239
rli106F1	GGATCCCAGTTGGCATATTAATATC
rli106R1	CGGGGTACCGGTTAAGCGGTTTTGC	271
DegUF	CAAGCTTAAAAGAGAAATGTTTAATT
DegUR	GGTACCACTGTTTTTTCACTAATGA	721
16S rRNAF	GATGCATAGCCGACCTGAGA
16S rRNAR	TGCTCCGTCAGACTTTCGTC	116
hemCF	CGGATCCATGACGTTATCTAAAGGA
hemCR	CCAAGCTTTTAAATTCGTTCTTCTACA	1122
DegUF1	GGATCCATGGCACTCAAAATCATGATTG
DegUF2	CAAGCTTTTAGCGAATGTATACCCAGCC	687

Language:: English

Publication timeframe:: 4 times per year
Journal Subjects:: Life Sciences, Molecular Biology, Microbiology and Virology, Life Sciences, other, Medicine, Veterinary Medicine

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The small non-coding RNA rli106 contributes to the environmental adaptation and pathogenicity of Listeria monocytogenes

Published Online: Mar 17, 2023

Page range: 67 - 77

Received: Apr 01, 2022

Accepted: Mar 03, 2023

DOI: https://doi.org/10.2478/jvetres-2023-0013

Keywords
sRNA, environmental adaptation, pathogenicity, target gene.

© 2023 Y. Guo et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1

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Fig. 7

Primers used in the study

The small non-coding RNA rli106 contributes to the environmental adaptation and pathogenicity of Listeria monocytogenes

Yun Guo

Chunhui Ji

Lixia Wang

Chengcheng Ning

Na Li

Zhiyuan Li

Yunxia Shang

Yaling Li

Yaoqiang Sun

Xiaoxing Huang

Jie Li

Xuepeng Cai

Qingling Meng

Jun Qiao

Published Online: Mar 17, 2023

Page range: 67 - 77

Received: Apr 01, 2022

Accepted: Mar 03, 2023

DOI: https://doi.org/10.2478/jvetres-2023-0013

KeywordssRNA, environmental adaptation, pathogenicity, target gene.

© 2023 Y. Guo et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

Fig. 7

Primers used in the study

Keywords
sRNA, environmental adaptation, pathogenicity, target gene.