分析与检测

基于PCR-DGGE技术分析宣威火腿中真菌群落结构

  • 邹颖玲 ,
  • 刘姝韵 ,
  • 王桂瑛 ,
  • 普岳红 ,
  • 葛长荣 ,
  • 廖国周
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  • 1(云南农业大学 食品科学技术学院,云南 昆明,650201)
    2(云南农业大学 云南省畜产品加工工程技术研究中心,云南 昆明,650201)
硕士研究生(王桂瑛副教授和廖国周教授为共同通讯作者,E-mail:kmwgy@126.com,liaoguozhou@ynan.edu.cn)

收稿日期: 2019-09-27

  网络出版日期: 2020-04-24

基金资助

云南省现代农业生猪产业技术体系专项(2018KJTX013);云南省科技计划项目国际科技合作专项(2018IA101)

Analysis of fungal community structure in Xuanwei ham by PCR-DGGE

  • ZOU Yingling ,
  • LIU Shuyun ,
  • WANG Guiying ,
  • PU YueHong ,
  • GE Changrong ,
  • LIAO Guozhou
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  • 1(College of Food Science and Technology, Agricultural University, Kunming 650201, China)
    2(Livestock Product Processing Engineering and Technology Research Center of Yunnan Province, Yunnan Agricultural University,Kunming 650201, China)

Received date: 2019-09-27

  Online published: 2020-04-24

摘要

该研究以不同加工年份(1、2和3年)的宣威火腿为对象,采用聚合酶链式反应-变性梯度凝胶电泳(polymerase chain reaction-denaturing gradient gel electrophoresis,PCR-DGGE)技术,通过PCR扩增、优势条带切胶、克隆测序等分析不同加工年份宣威火腿表面及内部的真菌群落结构及多样性,为进一步探讨宣威火腿品质提供理论依据。结果表明,Aspergillus pseudoglaucus(假灰绿曲霉)、Phialosimplex caninusAspergillus penicillioides(帚状曲霉)、Yamadazyma triangularisWallemia sebiCandida glucosophila等是宣威火腿中的主要真菌,其中假灰绿曲霉是宣威火腿表面和内部的优势菌种。多样性分析表明,加工3年的宣威火腿表面真菌群落多样性最高,加工2年火腿表面真菌群落多样性最低。加工2年火腿内部真菌群落多样性最高,加工3年火腿内部真菌群落多样性最低。

本文引用格式

邹颖玲 , 刘姝韵 , 王桂瑛 , 普岳红 , 葛长荣 , 廖国周 . 基于PCR-DGGE技术分析宣威火腿中真菌群落结构[J]. 食品与发酵工业, 2020 , 46(6) : 269 -274 . DOI: 10.13995/j.cnki.11-1802/ts.022394

Abstract

In this study, Xuanwei ham from different processing years (1, 2 and 3 years) was used as the object. The polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technology was used. The structure and diversity of fungal communities on the surface and inside of Xuanwei ham in different processing years were analyzed by PCR amplification, dominant band cutting, clone sequencing and so on, providing theoretical basis for further discussion of Xuanwei ham quality. The results showed that the main fungi in Xuanwei ham were Aspergillus pseudoglaucus, Physiosimplex caninus, Aspergillus penicillioides, Yamadazyma triangularis, Wallemia sebi, Candida glucosophila and so on. Among them, Aspergillus pseudogriseus was the dominant species on both the surface and the inside of Xuanwei ham. The diversity analysis showed that the fungi community on the surface of Xuanwei ham processed for 3 years was the highest, and the fungi community on the surface of ham processed for 2 years was the lowest. The fungi community diversity in ham was the highest after 2 years of processing, and the lowest after 3 years of processing.

参考文献

[1] 董寅初. 中国传统风味肉制品的现代化势在必行(上)[J]. 肉类研究,1998(2):3-6.
[2] 乔发东. 宣威火腿标准化生产与品质改良技术研究[D]. 北京: 中国农业大学,2004, 6:48-58.
[3] 王桥美, 杨瑞娟, 严亮. 微生物多样性与宣威火腿品质关系的研究进展[J]. 食品安全导刊,2016(33X):137-139.
[4] FIDEL TOLDRA, MONICA FLORESl, YOLANDA SANZ. Dry-cured hamflavour: enzymatic generation and process influence[J]. FoodChemistry, 1997, 59 (4): 523-530.
[5] 李晓燕,王卫,张佳敏,等. 发酵微生物提升传统腌腊肉制品风味和安全性研究进展[J]. 食品工业,2018,39(5):275-279.
[6] MUYZER G, WALL D E. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction amplified genes coding for 16SrRNA[J].Appl Environ Microbiol, 1993, 59(3): 695-700
[7] TEMMERMAN R, SCHEIRLINCK I, HUYS G, et al. Culture-independent analysis of probiotic products by denaturing gradient gel electrophoresis[J]. Appl Environ Microbiol, 2003,69(1): 220-226.
[8] ZHANG G, SADIQ F A, ZHU L, et al. Investigation of microbial communities of chinese sourdoughs using culture-dependent and dgge approaches[J]. Journal of Food Science, 2015, 80(11): M2 535-M2 542.
[9] NALEPA B, MARKIEWICZ L H. PCR-DGGE markers for qualitative profiling of microbiota in raw milk and ripened cheeses[J]. LWT - Food Science and Technology, 2017:S0023643817303729.
[10] 孙丹丹,卢士玲,李开雄,等. 贮藏温度对冷鲜羊肉微生物菌群生长变化的影响[J]. 食品工业科技,2017,38(4):327-331;341.
[11] 姚淑敏,陈璐,闫华文. 应用PCR-DGGE方法研究甜酒曲中真菌多样性[J]. 中国酿造,2016,35(11):44-48.
[12] AMPE F, BENOMAR N, MOIZAN C, et al. Polyphasic study of the spatialdistribution ofmicroorganisms in Mexican Pozo,l a fermented maizedough, demonstrates the need for cultivation-independent methods to investigate traditional fermentations[J]. Appl Environ Microbio, 1999, 65(12): 5 464-5 473.
[13] 郭继平,马光,张志强等. 喷施菌剂后葡萄叶片真菌多样性的PCR-DGGE分析[J]. 中国生物防治学报,2018,34(6):937-944.
[14] 郑艳,姚婷. PCR-DGGE分析甘薯酸浆自然发酵过程中细菌多样性[J]. 食品科学,2016,37(7):99-103.
[15] KUANG Y, TANI K, SYNNOTT A J, et al. Characterization of bacterial population of raw milk from bovine mastitis by culture-independent PCR-DGGE method[J]. Biochemical Engineering Journal, 2009, 45(1):76-81.
[16] 刘姝韵,尚思奇,孙灿,等. 基于PCR-DGGE分析云南牛干巴的细菌群落结构[J]. 肉类研究,2017,31(9):44-50.
[17] 龚治翔,马晓寒,任志广,等. 连作烤烟根际土壤细菌群落16S rDNA-PCR-DGGE分析[J]. 中国农业科技导报,2018,20(2):39-47.
[18] 李平兰,沈清武,吕燕妮,等. 宣威火腿成熟产品中主要微生物菌相构成分析[J]. 中国微生物态学杂志, 2003(5):262-263.
[19] 黄艾祥,周利,葛长荣,等. 速制云腿中霉菌的分离及作用[J]. 肉类工业,2002(7):22-23.
[20] 甄宗圆. 金华火腿微生物区系研究[D]. 重庆:西南农业大学,2004,6:33-38.
[21] 林克忠,杨耀寰,竺尚武,等. 金华火腿的质量和色香味形成与霉菌关系的研究[J]. 肉类研究,1992(2):10-16;21.
[22] 马萍,江东福,段若玲,等. 宣威火腿及其微生物作用菌群的研究[J]. 云南大学学报(自然科学版),1990,3(12):64-70.
[23] NUNEZ F, RODRIGUEZ M M, CODROBA J J, et al. Yeas tpopulation during ripening of dry-cured Iberian ham[J]. International journal of food microbiology, l996, 29: 271-280.
[24] MUHAMED L J, ITO H, WATANABE H, et al. The effect of substrate on the radiation resistance of yeasts isolated from sausage meat[J]. Lett App Microbiol, 1986, 22(1): 80-84.
[25] 黄艾祥. 云南干腌火腿品质特征形成与微生物作用研究[D]. 重庆:西南大学,2006,5:87-102.
[26] 夏耕,谢骏,王海英,等. 基于PCR-DGGE技术分析生物絮团的细菌群落结构[N]. 水产学报,2012,36(10):1 563-1 571.
[27] 罗湘南,梁英娟,付红霞,等. PCR-DGGE技术在微生物生态学中的应用[J]. 生物学杂志,2007(6):58-60.
[28] VALLAEYS T, TOPP E, MUYZER G, et al. Evaluation of denaturing gradient gel electrophoresis in the detection of 16S rDNA sequence variation in rhizobia and methanotrophs[J]. FEMS Microbiol Ecol, 1997, 24(3): 279-285.
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