锦州小菜主要理化指标及微生物多样性分析

发布时间：2018-11-18 18:37

【摘要】：锦州小菜是辽宁锦州的特色酱腌菜,其品种繁多、味道鲜脆爽口,是当地百姓必备的餐桌小菜,其产品远销全国并出口至日本等国家。本试验首次利用Illumina高通量测序对自然发酵锦州小菜中的微生物多样性进行研究,以利于进一步挖掘、保护和利用自然发酵锦州小菜中的微生物资源,提高锦州小菜品质和开发优良发酵菌种提供依据。首先对传统发酵锦州小菜进行理化检验;其次,采用聚合酶链式反应结合变性梯度凝胶电泳(PCR-DGGE)法对传统发酵锦州小菜中生物多样性进行分析,最后通过Illumina高通量测序方法对小菜多样性指数和群落结构进行对比研究,结果如下:1.传统发酵小菜总酸含量为0.20～1.40%;芥菜丝氨基氮含量最高:0.81g/100mL,虾油、及虾油腌菜制品的氨基酸态氮含量较高,分别在0.21～0.34g/100mL;沈阳早市什锦菜的亚硝酸含量最高为17.184mg/Kg;酱菜类及虾油含盐量最高,在10.79～25.74 g/100mL之间。锦州早市样品菌落数在105cfu/g～5×105cfu/g之间;而沈阳早市,农贸市场以及部分超市的小菜的菌落计数在106cfu/g以上。2.利用DGGE技术对10份具有地域性代表的锦州小菜进行多样性分析,传统发酵锦州小菜中可能存在的优势菌有植物乳杆菌(Lactobacillus plantarum)、魏斯氏菌属(Weissella sp.)、热带假丝酵母(Candida tropicalis)。其中,7份样品检测到植物乳杆菌;6份样品检测到食窦魏斯氏菌;共有3份检测到希腊魏斯氏菌;2份样品检测到fabifermentans(Lactobacillus fabifermentans);此外,1 份样品检测到假单细胞菌(Psychromonas arctica);5份样品检测到热带假丝酵母菌;4份样品检测到弯曲粘膜菌(Campylobacter mucosalis strain);1份样品检测到白色念珠菌(Candida albicans)。3.通过Illumina高通量测序法对27份传统发酵锦州小菜多样性进行分析:锦州样品中细菌物种相对于沈阳早市的小菜样品具有菌群共性及差异性;锦州小菜中细菌在门水平上共检测到5已知细菌门。其中厚壁菌门(Firmicutes)为主要的细菌优势菌门;在属水平上共检测到53种细菌属。其中,乳杆菌属(Lac tobacillu)、蓝细菌属(Cyanoiacter)、魏斯氏菌属(Weissella)可能是该采集时间段中优势菌属;此外,还有Halanaerobium属、葡萄糖杆菌属(Gluconobacter)、肠球菌属(Enterobacteriaceae)、不动杆菌属(Acinetobacter)、醋酸杆菌属(Acetobacter)等39种细菌菌属均在不同样品中检测出。22份传统发酵锦州小菜中真菌在门水平上共检测到子囊菌门(Ascomycota)、担子菌门(Basidiomycota)等4种已知真菌门;在属水平上,共检测到20种真菌菌属。其中,酵母菌目属(Saccharomycetales)、Hyphopichia菌属(Hyphopichia)、毕赤酵母属(Pichia)在此采集发酵阶段成为主要优势菌群。另外,所有样品均检测到了青霉菌属(Penicillium),比例为0.009%～14.740%。由此推测锦州地区的小菜存在丰富的细菌微生物发酵菌群,赋予了锦州小菜独特风味。
[Abstract]:Jinzhou vegetable is a special pickle in Jinzhou, Liaoning Province. It has a variety of varieties, fresh and crisp taste, and is a necessary table dish for local people. Its products are exported to Japan and other countries. In this experiment, Illumina high-throughput sequencing was used for the first time to study the microbial diversity in the natural fermenting vegetable of Jinzhou in order to further excavate, protect and utilize the microbial resources in the natural fermentation of Jinzhou. To improve the quality of Jinzhou vegetables and to develop good fermentation bacteria to provide the basis. Firstly, physical and chemical tests were carried out on the traditional fermented vegetables of Jinzhou. Secondly, polymerase chain reaction (PCR) combined with denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the biological diversity of traditional fermented vegetables in Jinzhou. Finally, the diversity index and community structure were compared by Illumina high throughput sequencing. The results were as follows: 1. The content of amino nitrogen in mustard silk was the highest: 0.81g / 100mL, 0.81 g / 100mL, 0.21g / 100mL, 0.21g / 100mL, respectively. The highest content of nitrite was 17.1844 mg / kg and the salt content of pickles and shrimp oil was the highest between 10.79 g/100mL and 25.74 g/100mL. The colony number of samples in Jinzhou early market was between 105cfu/g~5 脳 105cfu/g, while that of vegetables in Shenyang early market, farmers' market and some supermarkets was above 106cfu/g. 2. DGGE technique was used to analyze the diversity of 10 samples of Jinzhou vegetables with regional representation. The dominant bacteria in the traditional fermentation of Jinzhou vegetables were Lactobacillus plantarum, (Lactobacillus plantarum), Weiss, (Weissella sp.),. Candida tropicalis (Candida tropicalis). Of these, 7 detected Lactobacillus plantarum; 6 detected Weiss's bacteria in the sinuses; 3 detected Weiss's bacteria in Greece; 2 detected fabifermentans (Lactobacillus fabifermentans); and 1 detected pseudomonas (Psychromonas arctica); in 1 sample. Candida tropicalis was detected in 5 samples and Candida albicans (Candida albicans). 3 was detected in 4 samples (Campylobacter mucosalis strain);. The diversity of 27 traditional fermented vegetables in Jinzhou was analyzed by Illumina high-throughput sequencing method. The bacterial species in Jinzhou samples were compared with those in Shenyang early market. A total of 5 known bacteria phylum were detected at gate level. The dominant phylum was (Firmicutes), and 53 species of bacteria were detected at the generic level. Among them, Lactobacillus, (Lac tobacillu), cyanobacteria, (Cyanoiacter), Weiss's bacteria, (Weissella) may be the dominant genus in this collection period. In addition, there are Halanaerobium, (Gluconobacter), (Gluconobacter), Enterococcus, (Enterobacteriaceae), Acinetobacter, (Acinetobacter),. 39 strains of Acetobacter, including (Acetobacter), were detected in different samples. Four known fungi phylum, (Ascomycota), basidiomycetes, (Basidiomycota), were detected in 22 samples of traditional fermentative vegetable of Jinzhou. A total of 20 species of fungi were detected at genus level. Among them, Saccharomyces (Saccharomycetales), Hyphopichia, (Hyphopichia), Pichia pastoris (Pichia) became the main dominant flora in this collection and fermentation stage. In addition, the ratio of Penicillium (Penicillium), was 0. 009 and 14. 740%. It is inferred that there are abundant microbial fermentative microflora in the vegetable in Jinzhou area, which endows the special flavor of the small dish in Jinzhou.
【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TS255.1

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