PCR-DGGE指纹技术研究复合保鲜剂对冷藏带鱼贮藏期间微生物变化的影响

食品科学 ›› 2012, Vol. 33 ›› Issue (14): 255-260.

PCR-DGGE指纹技术研究复合保鲜剂对冷藏带鱼贮藏期间微生物变化的影响

蓝蔚青，谢晶*

上海海洋大学食品学院，上海水产品加工及贮藏工程技术研究中心

收稿日期:2011-06-03 修回日期:2012-06-19 出版日期:2012-07-25 发布日期:2012-07-27
通讯作者: 谢晶 E-mail:jxie@shou.edu.cn
基金资助:
“十二五”国家支撑计划项目“冷链食品物流产业技术创新战略联盟”——易腐特色农产品冷链物流核心技术与装备研发及其示范;上海市教育委员会重点学科建设项目

PCR-DGGE Analysis of the Effect of Natural Composite Preservative on Dynamic Changes of Microorganisms in Cutlassfish (Trichiurus haumela) during Cold Storage

Received:2011-06-03 Revised:2012-06-19 Online:2012-07-25 Published:2012-07-27

摘要/Abstract

摘要： 为探讨复合保鲜剂的作用机理和延长水产品货架期，应用聚合酶链式反应(PCR)-变性梯度凝胶电泳(DGGE)指纹技术研究复合保鲜剂对冷藏带鱼贮藏期间微生物种群多样性演替规律的影响。采用SDS(sodium dodecyl sulfate)化学裂解法，分别从对照组与复合保鲜剂处理组的冷藏带鱼中提取总细菌基因组DNA，同时进行16S rDNA的V3可变区PCR扩增，再通过DGGE得到动态变化的指纹图谱。DGGE图谱表明，产品在贮藏初期具有丰富的微生物群落，说明污染微生物的多样性，但经过一段时间后，只有少数种类细菌存活并最终成为主导菌群；通过对冷藏带鱼DGGE图谱上的主要条带进行测序分析，获得12种细菌。对照组与复合保鲜剂处理组在腐败后期的特定腐败微生物种类具有高度的相似性。嗜冷杆菌为带鱼贮藏初期的优势菌，随着贮藏时间的延长，希瓦氏菌与假单胞菌的比例逐渐增加，在贮藏过程中成为主要优势菌，荧光假单胞菌与弧菌在贮藏期间占有较大比例。同时，冷藏带鱼经复合保鲜剂处理后，其中的希瓦氏菌与假单胞菌受到明显抑制。

关键词: 带鱼, 复合保鲜剂, PCR-DGGE, 微生物, 动态变化

Abstract: To clarify the mechanism of action of natural composite preservative (NCP) and prolong the shelf life of aquatic products, Polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) fingerprinting was used to study the effect of an NCP consisting of chitosan, lysozyme and tea polyphenols in 1.0% acetic acid solution at respective concentrations of 10.0 g/L, 6 × 106 U/L and 3.0 g/L on the evolution of diversity of microbial populations in cutlassfish (Trichiurus haumela) during cold storage. SDS pyrolysis method was used for total genomic DNA extraction from control and NCP-treatment samples of cutlassfish, and the V3 variable regions of 16 S rDNA were subjected to PCR amplification for DGGE fingerprinting of dynamic changes. The results of DGGE fingerprinting indicated that cutlassfish possessed abundant microbial communities during the early part of the storage period (16 d). However, with increasing storage time, only few bacterial species kept alive and became dominant at last. The main brands in the DGGE patterns of cutlassfish were sequenced to identify 12 bacterial species. The specific spoilage organisms (SOSs) of control and NCP-treatment samples of cutlassfish in the final stages of spoilage showed high similarity. Psychrobacter sp. was dominant in the early stages, whereas the proportions of Shewanella sp. and Pseudomonas sp. increased gradually with the extension of storage time, and became dominant. Moreover, both Pseudomonas fluorescens and Vibrio sp. were highly proportional. In addition, NCP treatment had no obvious inhibitory effect on Shewanella sp. or Pseudomonas sp.

Key words: Trichiurus haumela, natural composite preservative, polymerase chain reaction-denaturing gradient gel

中图分类号:

蓝蔚青，谢晶*. PCR-DGGE指纹技术研究复合保鲜剂对冷藏带鱼贮藏期间微生物变化的影响[J]. 食品科学, 2012, 33(14): 255-260.

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