7 9 Vol. 7 No. 9 2016 9 Journal of Food Safety and Quality Sep., 2016 : * 胥伟, 吴丹, 姜依何, 朱旗 (1., 410128; 2., 410128) 摘要 : (),,,, (Aspergillus)(Penicillium)(Eurotium),,,,,,,, 关键词 : ; ; ; ; Microbial research on black tea: the community composition and food safety analysis XU Wei, WU Dan, JIANG Yi-He, ZHU Qi * (1. Key Laboratory of Tea Science of Ministry of Education, College of Horticulture and Landscape, Hunan Agriculture University, Changsha 410128, China; 2. National Research Center of Engineering Technology for Utilization of Botanical Functional Ingredients, Changsha 410128, China) ABSTRACT: Microorganism in the process of pile-fermentation (or Fahua-fermentation) which form the main quality of dark tea come from natural environment, and there are great differences in microbial diversity in different areas of dark tea. This article summarized several international research results of the microbial diversity in dark tea recently, microbial community and safety analysis in order to discuss on the microbial community characteristics of dark tea. The results showed that, Aspergillus, Penicillium and Eurotium mainly composed of microbial communities in dark tea. Aspergillus fungi presented in all aspects of the dark tea processing, and Penicillium fungi was rarely found during the beginning of pile fermentation process, but widespread in storage. The sample of Pu-er tea in pile fermentation were investigated by metagenomic technology, which found that bacteria was the main part of microbial, so it was necessary to further study on the relationship of bacteria and the quality of Pu-er tea, especially its functional components. The health risk of foodborne bacteriotoxin in dark tea products was rarely. Although there were a large number of fungi, mycotoxin was detected rarely in dark tea which was stored safely, so there was no 基金项目 : (31571802)(14A066)(CX2016B283) Fund: Supported by the National Natural Science Foundation of China (31571802), the Key Project of Hunan Provincial Education Bureau (14A066) and Hunan Provincial Innovation Foundation For Postgraduate (CX2016B283) * 通讯作者 :,,, E-mail: 1965994459@qq.com *Corresponding author: ZHU Qi, Professor, Ph.D. Supervisor, College of Horticulture and Landscape Architecture, Hunan Agricultural University, Changsha 410128, China. E-mail:1965994459@qq.com
3542 7 need to worry about drinking dark tea produced by regular manufacturers. KEY WORDS: Fu Brick tea; Kang Brick tea; Pu-Er tea; Liupao tea; microbes 1 引言,,,,,,,,,, 2 黑茶的发展 特性与现状, -- - [1],,,,,, 2014 28, 13.4%, 2013 4, 26%, [2], 2014 [3], 2014 80 [4] 3 黑茶微生物多样性分析 3.1 黑茶微生物多样性分析技术的发展, [5],,,,,,,,, ;,,,,, : ITS (PCR-denatured gradient gel electrophoresis, PCR-DGGE)16S/18S [6] [7] 18S, 248 6 18S 10/28, 23/28 18S, 3.2 黑茶微生物多样性分析, 3.2.1 黑茶初制过程微生物多样性分析, [8], [9],, [10] [11] [12] : (Aspergillus niger)(penicillium)(rhizopus) (Aspergillus glaucus)(yeast), (Aspergillus niger), 80%, (Saccharomyces)
9, : : 3543 (Aspergillius gloucus)(rhizopus)(penicillium),, [13],, (Saccharomyces cerevisiae)(penicillium glabrum) [14] (Aspergillus wentii Wegner var. fumeus Qi &Sun)(Aspergillus penicilliodes Speg.) (Aspergillus aureolatus Munt. -Cvet.&Bata) (Aspergillus egyptiacus Moub.&Moust)(Aspergillus foetidus Thom&Raper) (Aspergillus japonicus Saito var. japonicus)(aspergillus restrictus Smith) [15], (Bacillus coagulans) (Bacillus subtilis)(bacillus licheniformis) (Bacillus thermoamylovorans) (Geobacillus thermoleovorans) (Pediococcus acidilactici) (Lactobacillus plantarum), (Bacillus spp.) (Candida spp.) (Actinomyces) (Actinomyces) (Aspergillus)(Mucor)(Thamnidium) (Rhizopus) [16,17] ITS, (Aspergillus niger) (Aspergillus japonicus Saito var. aculeatus) (Aspergillus tubingensis Moss.) (Aspergillus foetidus) (Aspergillus awamori) (Aspergillus carbonarius)(aspergillus japonicus) (Aspergillus fumigatus)(eurotium amstelodami) (Rhizopus Ehrenberg) (Thamnidium simplex Brefeld)(M. racemosus Fres.) [17, 18] DNA,,, PCR-DGGE 16/18S [19,20] PCR-DGGE, (Nocardia) (Novosphingobium)(Brevundimonas) (Pseudomonas spp.)(clostridium ultunense) (Klebsiella) (Lactobacillus)ε-(ε-Proteus Hauser) (Saprospiraceae)(Myxovoccus) 6 ; (Wallemiasebi) (Candida sp.)(candida tropicalis) (Lodderomyces spp.)(debaryomyces hansenii) (Pichia kudriavzevii) (Saccharomyces cerevisiae) (Cryptococcussp.) (Rhodotorula graminis) (Eurotium amstelodami) (Aspergillus glaucus) (Rhizomucor miehei)(rhizomucor pusillus)(galactomyces geotrichum) (Candida ontarioensis) (Trichosporondermatis)(Penicillium decumbens) (Aspergillus penicillioides), 22, 7, 2 Abe [21] DGGE, (Aspergillus niger) (Blastobotrys adeninivorans), [22] [23] [24] ITS 16S, (Rhizomucor pusilus) (Aspergillus tubingensis)(candida tropicalis) (Talaromyces emersonii); (Bacillaceae)(Klebsiella) (Sphingobacteriuln)(Brevibacterium) (Rhodococcus)(Pseudomonas)(A. baumanii), (Bacillaceae)Zhao [25] 16S (0~20 d), 5 10 0~10 d (Enterobacteriaceae), 15~20 d (Bacillaceae), (B.coagulans) Lv [26], 76.26%, 16.35%, 7.16% (Actinobacteria) (Proteobacteria)(Firmicutes), (Ascomycota),,,, Zhao [27],,,
3544 7 : 17.82%(Pseudomonas)12.33% (Lactococcus) 9.53% (Unclassified-Cyanobacteria) 7.69% (Unclassified-Enterobacteriaceae) 7.36% (Curtobacterium)4.74%(Achromobacter) 3.99% (Acinetobacter), 2.97% (Rhodococcus)2.56%(Stenotrophomonas) 2.46% (Enterococcus) 2.17% (Erwinia)2.10%(Streptomyces)1.51% (Enterobacter)1.33%(Paenibacillus) 1.27% (Flavobacterium) 1.18% (Bacillus)1.15%(Microbacterium)1% (Klebsiella) 16.85% : 94.98%(Aspergillus), 4.88% (Rhizomucor) 0.14% 1 3.2.2 黑茶精制过程微生物多样性分析,, : --------, [28] 20 80,,, (Eurotium cristatum) [29-32],, [33] [34] (Eurotium cristatum), [35] 2,, (Aspergillus) (Penicillium) (Cladosporium)(Mucor)(Fusarium) (Trichoderma)(Eurotium cristatum) Table 1 表 1 黑茶初制过程微生物多样性的研究成果 Results of microbial diversities during the primary processing of dark tea (Wallemiasebi); (Brevundimonas) (Nocardia) (Novosphingobium) (Clostridium ultunense) (Pseudomonas spp.) (Lactobacillus) (Klebsiella) ε-(ε-proteus Hauser) (Saprospiraceae) (Myxovoccus) (Rhizobium) (Eurotium) (Rhizomucor) (Aspergillus) (Debaryomyces) (Penicillium) (Candida spp.); (Candida tropicalis); (Lodderomyces spp.); (Debaryomyces hansenii); (Pichia kudriavzevii); (Saccharomyces cerevisiae); (Cryptococcu ssp.); (Rhodotorula graminis); (Eurotium amstelodami); (Aspergillus glaucus) ; (Rhizomucor miehei); (Rhizomucor pusillus); (Galactomyces geotrichum); (Candida ontarioensis); (Trichosporondermatis); (Penicillium decumbens); (Aspergillus penicillioides)
9, : : 3545 续表 1 (Klebsiella) (sphingobacteriuln) (Brevibacterium) (Acinetobacter) (Enterobacteriaceae) (Bacillaceae) (Pseudomonas) (Lactococcus) (Curtobacterium) (Achromobacter) (Acinetobacter) (Rhodococcus) (Stenotrophomonas) (Enterococcus) (Erwinia) (Streptomyces) (Paenibacillus) (Flavobacterium) (Bacillus) (Microbacterium) (Klebsiella) (Aspergillus) (Rhizomucor) (Aspergillus niger) (Rhizomucor pusilus) (Aspergillus tubingensis) (Candida tropicalis) (Talaromyces emersonii) (Bacillus coagulans) (Bacillus subtilis) (Bacillus licheniformis) (Bacillus thermoamylovorans) (Geobacillus thermoleovorans) (Pediococcus acidilactici) (Lactobacillus plantarum) (A. baumanii) (Bacillaceae) (Candidasp.) (Actinomyces) (Actinomyces) (Aspergillus) (Mucor) (Thamnidium) (Rhizopus) (Aspergillus niger) (Aspergillus japonicus Saito var. aculeatus) (Aspergillus tubingensis Moss.) (Aspergillus foetidus) (Aspergillus awamori) (Aspergillus carbonarius) (Aspergillus japonicus) (Aspergillus fumigatus) (Eurotium amstelodami) (Rhizopus ehrenberg) (Thamnidium simplex Brefeld) (M. racemosus Fres.)
3546 7 ;, [36] 18 15, 8 3 2 1 3 (), [37] [38], (Eurotium) (Aspergillus) [39], (Eurotium cristatum), (Aspergillus cristatum) [40] Table 2 表 2 黑茶精制过程微生物多样性的研究成果 Results of microbial diversities during the refining process of dark tea (Eurotium) (Debaryomyces) (Aspergillus) (Cladosporium) (Mucor) (Fusarium) (Trichoderma) (Verticillium) (Pichia) (Pestalotiopsis) (Rhizomucor) (Beauveria) (Brevundimonas); (Nocardia); (Novosphingobium); (Clostridium ultunense); (Pseudomonas spp.); (Lactobacillus); (Klebsiella); ε-(ε-proteus Hauser); (Saprospiraceae); (Myxovoccus); (Rhizobium); (Aspergillus niger) (Eurotium cristatum) (Eurotiumchevalieri) (Eurotium costiforme) (Eurotium amstelodami) (Penicillium) / / / / (Aspergillus); (Eurotium); (Penicillium); / : /
9, : : 3547 DNA, (denatured gradient gel electrophoresis, DGGE) 16S/18S Xu [41] PCR-DGGE (Eurotium)(Debaryomyces) (Aspergillus)(Verticillium)(Pichia) (Pestalotiopsis)(Rhizomucor) (Beauveria), (Eurotium) (Debaryomyces)(Aspergillus) [42], : (Brevundimonas)(Nocardia) (Novosphingobium)(Clostridium ultunense) (Pseudomonas spp.) (Lactobacillus)(Klebsiella) ε- (ε-proteus Hauser)(Saprospiraceae) (Myxovoccus)(Rhizobium) 6,, (Aspergillus) (Eurotium)(Penicillium) [43] 2 3.2.3 黑茶贮存过程微生物多样性分析,, : [44], (Aspergillus)(Penicillium) (Rhizopus) : (Aspergillus niger) (Penicillium glabrum) (Penicillium roqueforti)(penicillium baen) (Rhizopus) [45], (Eurotium cristatum),, Xu [41] PCR-DGGE, (Beauveria), [46-49] (Aspergillus niger) [46] (Aspergillus ochraceus) [47] (Penicillium oxalicum) [48] (Penicillium chrysogenum) [49], [50] ITS, (Eurotium niveoglaucum) [22], 8 : (Aspergillus) (Eurotium) (Penicillium)BlastobotrysGuehomycesRasamsonia Rossbeevera (Emericella), (Aspergillus)(Eurotium) 3 3.2.4 黑茶微生物多样性研究,, Table 3 表 3 黑茶贮存过程微生物多样性研究成果 Results of microbial diversities during the storage of dark tea (Eurotium); (Beauveria); (Aspergillus) (Penicillium) (Aspergillus niger) (Aspergillus ochraceus) (Penicillium oxalicum) (Penicillium chrysogenum) (Aspergillus) (Penicillium) (Rhizopus) (Aspergillus niger) (Penicillium glabrum) (Penicillium roqueforti) (Penicillium baen) (Rhizopus)
3548 7 续表 3 (Aspergillus); (Eurotium); (Penicillium); Blastobotrys; Guehomyces; Rasamsonia; Rossbeevera; (Emericella) (Aspergillus niger) (Aspergillus tubingensis Moss.) (Aspergillus fumigatus) (Aspergillus oryzae) (Aspergillus sydowii) (Aspergillus ochraceus) (Aspergillus tamarii) (Sclerotium aspergillus) (Penicillium citrinum) (Penicillium chrysogenum) (Penicillium oxalicum) Penicillium mallochii (Penicillium chermesinum) (Penicillium meleagrinum) Penicillium brocae (Paecilomyces varioti) (Eurotium amstelodami) (Stephanoascus ciferrii) (Eurotium niveoglaucum), DGGE 16S, ε-, 75%,,,,,,,,, 4 黑茶的微生物毒素风险分析 [51],, [52] (mycotoxins) [52, 53] 300 [54], 25% [55]
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