蜂房哈夫尼亚菌对粘质沙雷氏菌群体感应信号分子产生与生物被膜形成的调控
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摘要
以腐败海鲈鱼中分离得到的2?株有群体感应现象的细菌为研究对象,经16S rRNA分子生物学鉴定其为蜂房哈夫尼亚菌(Hafnia alvei),命名为H. alvei-14;粘质沙雷氏菌(Serratia marcescens)产灵红素,命名为S.marcescens-01。研究不同培养条件下H.alvei-14与S.marcescens-01共培养后其信号分子N-酰基高丝氨酸内酯(N-acyl homoscrinc lactones,AHLs)分泌量和生物被膜产生量。结果表明:与H. alvei-14共培养能促进S.marcescens-01分泌AHLs和生物被膜的形成,这种影响可能与菌群间群体感应有关。环境条件的改变是影响细菌分泌AHLs和生物被膜形成的重要影响因子,较高盐浓度、pH值过低或过高均不利于微生物分泌AHLs和生物被膜形成,而限制性碳源及氮源等胁迫环境均可以刺激菌体群体感应系统,促进AHLs的释放和生物被膜的形成。
The two bacterial strains with quorum sensing isolated from spoiled sea bass were identi?ed based on 16 S rRNA gene sequence analysis as Hafnia alvei-14 and Serratia marcescens-01 with the ability to produce prodigiosin, respectively.The production of the quorum-sensing signal molecules N-acyl homoserine lactones(AHLs) and biofilm formation in S. marcescens-01 were investigated when co-cultured with H. alvei-14 under different culture conditions. The results showed that co-culture with H. alvei could promote AHLs secretion and bio?lm production in S. marcescens, which might be related to quorum sensing. Changes in environmental conditions were the important factors that could affect the secretion of AHLs and bio?lm formation. Higher salt concentration and too low or too high pH values were unfavorable to microbial secretion of AHLs and bio?lm formation, while restrictive carbon and nitrogen sources could stimulate the quorum sensing system and promote the release of AHLs and bio?lm formation.
The two bacterial strains with quorum sensing isolated from spoiled sea bass were identi?ed based on 16 S rRNA gene sequence analysis as Hafnia alvei-14 and Serratia marcescens-01 with the ability to produce prodigiosin, respectively.The production of the quorum-sensing signal molecules N-acyl homoserine lactones(AHLs) and biofilm formation in S. marcescens-01 were investigated when co-cultured with H. alvei-14 under different culture conditions. The results showed that co-culture with H. alvei could promote AHLs secretion and bio?lm production in S. marcescens, which might be related to quorum sensing. Changes in environmental conditions were the important factors that could affect the secretion of AHLs and bio?lm formation. Higher salt concentration and too low or too high pH values were unfavorable to microbial secretion of AHLs and bio?lm formation, while restrictive carbon and nitrogen sources could stimulate the quorum sensing system and promote the release of AHLs and bio?lm formation.
引文
[1]AN D,DANHORN T,FUQUA C,et al.Quorum sensing and motility mediate interactions between Pseudomonas aeruginosa and Agrobacterium tumefaciens in biofilm cocultures[J].Proceedings of the National Academy of Sciences of the United States of America,2006,103(10):3828-3233.DOI:10.1073/pnas.0511323103.
[2]崔方超,李婷婷,刘明爽,等.大菱鲆荧光假单胞菌的群体感应现象及不同碳源培养下的腐败特性研究[J].现代食品科技,2015,31(12):49-55.DOI:10.13982/j.mfst.1673-9078.2015.12.008.
[3]MILLER M B,SKORUPSKI K,LENZ D H,et al.Parallel quorum sensing systems converge to regulate virulence in Vibrio cholerae[J].Cell,2002,110(3):303-314.DOI:10.1016/s0092-8674(02)00829-2.
[4]PAPENFORT K,BASSLER B L.Quorum sensing signal-response systems in Gram-negative bacteria[J].Nature Reviews Microbiology,2016,14(9):576-588.DOI:10.1038/nrmicro.2016.89.
[5]ATKINSON S,WILLIAMS P.Quorum sensing and social networking in the microbial world[J].Journal of the Royal Society Interface,2009,6(40):959-978.DOI:10.1098/rsif.2009.0203.
[6]LIU L,GUI M,WU R Y,et al.Progress in research on biofilm formation regulated by LuxS/AI-2 quorum sensing[J].Food Science,2016,37(19):254-262.DOI:10.7506/spkx1002-6630-201619043.
[7]黄旭镇.大黄鱼特定腐败菌波罗的海希瓦氏菌QS系统的鉴定及茶多酚调控研究[D].杭州:浙江工商大学,2014:1-94.DOI:10.7666/d.Y2750834.
[8]张旭东,闫超杰,孙剑,等.冷鲜牛肉中蜂房哈夫尼亚菌(Hafnia alvei)的分离与鉴定[J].微生物学杂志,2010,30(3):106-108.DOI:10.3969/j.issn.1005-7021.2010.03.025.
[9]PADILLA D,REMUZGO-MARTíNEZ S,ACOSTA F,et al.Hafnia alvei and Hafnia paralvei.taxonomy defined but still far fromvirulence and pathogenicity[J].Veterinary Microbiology,2013,163(1/2):200-201.DOI:10.1016/j.vetmic.2012.11.041.
[10]张彩丽,朱素琴,汪映,等.一株沙雷氏菌的分离及其群体感应现象[J].生物技术通报,2014(7):150-155.DOI:10.13560/j.cnki.biotech.bull.1985.2014.07.032.
[11]ALMEIDA C,AZEVEDO N F,SANTOS S,et al.Correction:discriminating multi-species populations in biofilms with peptide nucleic acid fluorescence in situ hybridization(PNA FISH)[J].PLoSONE,2011,6(3):e14786.DOI:10.1371/journal.pone.0014786.
[12]羊扬.大肠杆菌I型群体感应功能研究[D].扬州:扬州大学,2014:1-135.DOI:10.7666/d.Y2631920.
[13]CHU W,VATTEM D A,MAITIN V,et al.Bioassays of quorum sensing compounds using Agrobacterium tumefaciens and Chromobacterium violaceum[J].Methods in Molecular Biology,2011,692:3-19.DOI:10.1007/978-1-60761-971-0_1.
[14]KABIR A H,ROY A G,ALAM M F,et al.Detection of quorum sensing signals in Gram-negative bacteria by using reporter strain CV026[J].Notulae Scientia Biologicae,2010,2(4):72-75.DOI:10.15835/nsb.2.4.4863.
[15]綦国红,董明盛,吴胜明,等.HPLC-MS法检测N-酰基-高丝氨酸内酯类信号分子[J].分析测试学报,2007,26(3):417-419.DOI:10.3969/j.issn.1004-4957.2007.03.030.
[16]郭秀春,郑立,张魁英,等.气相色谱-质谱法检测细菌中N-酰基高丝氨酸内酯类信号分子[J].分析测试学报,2012,31(3):347-350.DOI:10.3969/j.issn.1004-4957.2012.03.020.
[17]RAVN F L,CHRISTENSEN A B,MOLIN S,et al.Influence of food preservation parameters and associated microbiota on production rate,profile and stability of acylated homoserine lactones from food-derived Enterobacteriaceae[J].International Journal of Food Microbiology,2003,84(2):145-156.DOI:10.1016/S0168-1605(02)00405-1.
[18]GEISENBERGER O,GIVSKOV M,RIEDEL K,et al.Production of N-acyl-L-homoserine lactones by P.aeruginosa isolates from chronic lung infections associated with cystic fibrosis[J].FEMS Microbiology Letters,2000,184(2):273-278.DOI:10.1111/j.1574-6968.2000.tb09026.x.
[19]DANDEKAR A A,CHUGANI S,GREENBERG E P.Bacterial quorum sensing and metabolic incentives to cooperate[J].Science,2012,338:264-266.DOI:10.1126/science.1227289.
[20]JAHID I K,HA S D.A review of microbial biofilms of produce:future challenge to food safety[J].Food Science&Biotechnology,2012,21(2):299-316.DOI:10.1007/s10068-012-0041-1.
[21]刘晶晶,陈晶瑜.培养条件对金黄色葡萄球菌-大肠杆菌混合生物被膜的影响[J].中国酿造,2016,35(7):20-23.DOI:10.11882/j.issn.0254-5071.2016.07.005.
[22]张煜琛,李灿,孙力军,等.共存体系中对虾优势腐败菌对副溶血弧菌生物被膜形成的影响[J].西北农林科技大学学报(自然科学版),2016,44(9):42-48.DOI:10.13207/j.cnki.jnwafu.2016.09.007.
[23]励建荣,国竞文,李婷婷.细菌共培养及其系统中群体感应现象的研究进展[J].食品工业科技,2016,37(23):377-382.DOI:10.13386/j.issn1002-0306.2016.23.062.
[24]赵丹丹,刘琳,王迪,等.pH和NaCl质量浓度对发酵鱼糜中腐败菌Aeromonas veronii bv.veronii群体感应的影响[J].食品与发酵工业,2018,44(2):33-39.DOI:10.13995/j.cnki.11-1802/ts.015488.
[25]WANG M Z,ZHENG X,HE H Z,et al.Ecological roles and release patterns of acylated homoserine lactones in Pseudomonas sp.HF-1 and their implications in bacterial bioaugmentation[J].Bioresource Technology,2012,125:119-126.DOI:10.1016/j.biortech.2012.08.116.
[26]孔西曼,张公亮,王佳莹,等.环境因素对即食海参蜂房哈夫尼菌群体感应的影响[J].现代食品科技,2017,33(1):87-92;105.DOI:10.13982/j.mfst.1673-9078.2017.1.014.
[27]郑昕,汪美贞,沈东升.群体感应对生物强化过程影响[J].科技通报,2013,29(11):189-194.DOI:10.3969/j.issn.1001-7119.2013.11.042.
[28]BURM?LLE M,WEBB J S,RAO D,et al.Enhanced biofilm formation and increased resistance to antimicrobial agents and bacterial invasion are caused by synergistic interactions in multispecies biofilms[J].Applied&Environmental Microbiology,2006,72(6):3916-3923.DOI:10.1128/AEM.03022-05.
[29]BRINK H G,NICOL W.The influence of shear on the metabolite yield of Lactobacillus rhamnosus biofilms[J].New Biotechnology,2014,31(5):460-467.DOI:10.1016/j.nbt.2014.06.003.
[30]黄迅辰,戴贤君.荧光假单胞菌生物膜的定性观察及影响因素研究[J].食品科技,2015,40(6):14-18.DOI:10.13684/j.cnki.spkj.2015.06.004.
[2]崔方超,李婷婷,刘明爽,等.大菱鲆荧光假单胞菌的群体感应现象及不同碳源培养下的腐败特性研究[J].现代食品科技,2015,31(12):49-55.DOI:10.13982/j.mfst.1673-9078.2015.12.008.
[3]MILLER M B,SKORUPSKI K,LENZ D H,et al.Parallel quorum sensing systems converge to regulate virulence in Vibrio cholerae[J].Cell,2002,110(3):303-314.DOI:10.1016/s0092-8674(02)00829-2.
[4]PAPENFORT K,BASSLER B L.Quorum sensing signal-response systems in Gram-negative bacteria[J].Nature Reviews Microbiology,2016,14(9):576-588.DOI:10.1038/nrmicro.2016.89.
[5]ATKINSON S,WILLIAMS P.Quorum sensing and social networking in the microbial world[J].Journal of the Royal Society Interface,2009,6(40):959-978.DOI:10.1098/rsif.2009.0203.
[6]LIU L,GUI M,WU R Y,et al.Progress in research on biofilm formation regulated by LuxS/AI-2 quorum sensing[J].Food Science,2016,37(19):254-262.DOI:10.7506/spkx1002-6630-201619043.
[7]黄旭镇.大黄鱼特定腐败菌波罗的海希瓦氏菌QS系统的鉴定及茶多酚调控研究[D].杭州:浙江工商大学,2014:1-94.DOI:10.7666/d.Y2750834.
[8]张旭东,闫超杰,孙剑,等.冷鲜牛肉中蜂房哈夫尼亚菌(Hafnia alvei)的分离与鉴定[J].微生物学杂志,2010,30(3):106-108.DOI:10.3969/j.issn.1005-7021.2010.03.025.
[9]PADILLA D,REMUZGO-MARTíNEZ S,ACOSTA F,et al.Hafnia alvei and Hafnia paralvei.taxonomy defined but still far fromvirulence and pathogenicity[J].Veterinary Microbiology,2013,163(1/2):200-201.DOI:10.1016/j.vetmic.2012.11.041.
[10]张彩丽,朱素琴,汪映,等.一株沙雷氏菌的分离及其群体感应现象[J].生物技术通报,2014(7):150-155.DOI:10.13560/j.cnki.biotech.bull.1985.2014.07.032.
[11]ALMEIDA C,AZEVEDO N F,SANTOS S,et al.Correction:discriminating multi-species populations in biofilms with peptide nucleic acid fluorescence in situ hybridization(PNA FISH)[J].PLoSONE,2011,6(3):e14786.DOI:10.1371/journal.pone.0014786.
[12]羊扬.大肠杆菌I型群体感应功能研究[D].扬州:扬州大学,2014:1-135.DOI:10.7666/d.Y2631920.
[13]CHU W,VATTEM D A,MAITIN V,et al.Bioassays of quorum sensing compounds using Agrobacterium tumefaciens and Chromobacterium violaceum[J].Methods in Molecular Biology,2011,692:3-19.DOI:10.1007/978-1-60761-971-0_1.
[14]KABIR A H,ROY A G,ALAM M F,et al.Detection of quorum sensing signals in Gram-negative bacteria by using reporter strain CV026[J].Notulae Scientia Biologicae,2010,2(4):72-75.DOI:10.15835/nsb.2.4.4863.
[15]綦国红,董明盛,吴胜明,等.HPLC-MS法检测N-酰基-高丝氨酸内酯类信号分子[J].分析测试学报,2007,26(3):417-419.DOI:10.3969/j.issn.1004-4957.2007.03.030.
[16]郭秀春,郑立,张魁英,等.气相色谱-质谱法检测细菌中N-酰基高丝氨酸内酯类信号分子[J].分析测试学报,2012,31(3):347-350.DOI:10.3969/j.issn.1004-4957.2012.03.020.
[17]RAVN F L,CHRISTENSEN A B,MOLIN S,et al.Influence of food preservation parameters and associated microbiota on production rate,profile and stability of acylated homoserine lactones from food-derived Enterobacteriaceae[J].International Journal of Food Microbiology,2003,84(2):145-156.DOI:10.1016/S0168-1605(02)00405-1.
[18]GEISENBERGER O,GIVSKOV M,RIEDEL K,et al.Production of N-acyl-L-homoserine lactones by P.aeruginosa isolates from chronic lung infections associated with cystic fibrosis[J].FEMS Microbiology Letters,2000,184(2):273-278.DOI:10.1111/j.1574-6968.2000.tb09026.x.
[19]DANDEKAR A A,CHUGANI S,GREENBERG E P.Bacterial quorum sensing and metabolic incentives to cooperate[J].Science,2012,338:264-266.DOI:10.1126/science.1227289.
[20]JAHID I K,HA S D.A review of microbial biofilms of produce:future challenge to food safety[J].Food Science&Biotechnology,2012,21(2):299-316.DOI:10.1007/s10068-012-0041-1.
[21]刘晶晶,陈晶瑜.培养条件对金黄色葡萄球菌-大肠杆菌混合生物被膜的影响[J].中国酿造,2016,35(7):20-23.DOI:10.11882/j.issn.0254-5071.2016.07.005.
[22]张煜琛,李灿,孙力军,等.共存体系中对虾优势腐败菌对副溶血弧菌生物被膜形成的影响[J].西北农林科技大学学报(自然科学版),2016,44(9):42-48.DOI:10.13207/j.cnki.jnwafu.2016.09.007.
[23]励建荣,国竞文,李婷婷.细菌共培养及其系统中群体感应现象的研究进展[J].食品工业科技,2016,37(23):377-382.DOI:10.13386/j.issn1002-0306.2016.23.062.
[24]赵丹丹,刘琳,王迪,等.pH和NaCl质量浓度对发酵鱼糜中腐败菌Aeromonas veronii bv.veronii群体感应的影响[J].食品与发酵工业,2018,44(2):33-39.DOI:10.13995/j.cnki.11-1802/ts.015488.
[25]WANG M Z,ZHENG X,HE H Z,et al.Ecological roles and release patterns of acylated homoserine lactones in Pseudomonas sp.HF-1 and their implications in bacterial bioaugmentation[J].Bioresource Technology,2012,125:119-126.DOI:10.1016/j.biortech.2012.08.116.
[26]孔西曼,张公亮,王佳莹,等.环境因素对即食海参蜂房哈夫尼菌群体感应的影响[J].现代食品科技,2017,33(1):87-92;105.DOI:10.13982/j.mfst.1673-9078.2017.1.014.
[27]郑昕,汪美贞,沈东升.群体感应对生物强化过程影响[J].科技通报,2013,29(11):189-194.DOI:10.3969/j.issn.1001-7119.2013.11.042.
[28]BURM?LLE M,WEBB J S,RAO D,et al.Enhanced biofilm formation and increased resistance to antimicrobial agents and bacterial invasion are caused by synergistic interactions in multispecies biofilms[J].Applied&Environmental Microbiology,2006,72(6):3916-3923.DOI:10.1128/AEM.03022-05.
[29]BRINK H G,NICOL W.The influence of shear on the metabolite yield of Lactobacillus rhamnosus biofilms[J].New Biotechnology,2014,31(5):460-467.DOI:10.1016/j.nbt.2014.06.003.
[30]黄迅辰,戴贤君.荧光假单胞菌生物膜的定性观察及影响因素研究[J].食品科技,2015,40(6):14-18.DOI:10.13684/j.cnki.spkj.2015.06.004.