【摘要】：錦州小菜是遼寧錦州的特色醬腌菜,其品種繁多、味道鮮脆爽口,是當(dāng)?shù)匕傩毡貍涞牟妥佬〔?其產(chǎn)品遠(yuǎn)銷全國(guó)并出口至日本等國(guó)家。本試驗(yàn)首次利用Illumina高通量測(cè)序?qū)ψ匀话l(fā)酵錦州小菜中的微生物多樣性進(jìn)行研究,以利于進(jìn)一步挖掘、保護(hù)和利用自然發(fā)酵錦州小菜中的微生物資源,提高錦州小菜品質(zhì)和開(kāi)發(fā)優(yōu)良發(fā)酵菌種提供依據(jù)。首先對(duì)傳統(tǒng)發(fā)酵錦州小菜進(jìn)行理化檢驗(yàn);其次,采用聚合酶鏈?zhǔn)椒磻?yīng)結(jié)合變性梯度凝膠電泳(PCR-DGGE)法對(duì)傳統(tǒng)發(fā)酵錦州小菜中生物多樣性進(jìn)行分析,最后通過(guò)Illumina高通量測(cè)序方法對(duì)小菜多樣性指數(shù)和群落結(jié)構(gòu)進(jìn)行對(duì)比研究,結(jié)果如下:1.傳統(tǒng)發(fā)酵小菜總酸含量為0.20～1.40%;芥菜絲氨基氮含量最高:0.81g/100mL,蝦油、及蝦油腌菜制品的氨基酸態(tài)氮含量較高,分別在0.21～0.34g/100mL;沈陽(yáng)早市什錦菜的亞硝酸含量最高為17.184mg/Kg;醬菜類及蝦油含鹽量最高,在10.79～25.74 g/100mL之間。錦州早市樣品菌落數(shù)在105cfu/g～5×105cfu/g之間;而沈陽(yáng)早市,農(nóng)貿(mào)市場(chǎng)以及部分超市的小菜的菌落計(jì)數(shù)在106cfu/g以上。2.利用DGGE技術(shù)對(duì)10份具有地域性代表的錦州小菜進(jìn)行多樣性分析,傳統(tǒng)發(fā)酵錦州小菜中可能存在的優(yōu)勢(shì)菌有植物乳桿菌(Lactobacillus plantarum)、魏斯氏菌屬(Weissella sp.)、熱帶假絲酵母(Candida tropicalis)。其中,7份樣品檢測(cè)到植物乳桿菌;6份樣品檢測(cè)到食竇魏斯氏菌;共有3份檢測(cè)到希臘魏斯氏菌;2份樣品檢測(cè)到fabifermentans(Lactobacillus fabifermentans);此外,1 份樣品檢測(cè)到假單細(xì)胞菌(Psychromonas arctica);5份樣品檢測(cè)到熱帶假絲酵母菌;4份樣品檢測(cè)到彎曲粘膜菌(Campylobacter mucosalis strain);1份樣品檢測(cè)到白色念珠菌(Candida albicans)。3.通過(guò)Illumina高通量測(cè)序法對(duì)27份傳統(tǒng)發(fā)酵錦州小菜多樣性進(jìn)行分析:錦州樣品中細(xì)菌物種相對(duì)于沈陽(yáng)早市的小菜樣品具有菌群共性及差異性;錦州小菜中細(xì)菌在門(mén)水平上共檢測(cè)到5已知細(xì)菌門(mén)。其中厚壁菌門(mén)(Firmicutes)為主要的細(xì)菌優(yōu)勢(shì)菌門(mén);在屬水平上共檢測(cè)到53種細(xì)菌屬。其中,乳桿菌屬(Lac tobacillu)、藍(lán)細(xì)菌屬(Cyanoiacter)、魏斯氏菌屬(Weissella)可能是該采集時(shí)間段中優(yōu)勢(shì)菌屬;此外,還有Halanaerobium屬、葡萄糖桿菌屬(Gluconobacter)、腸球菌屬(Enterobacteriaceae)、不動(dòng)桿菌屬(Acinetobacter)、醋酸桿菌屬(Acetobacter)等39種細(xì)菌菌屬均在不同樣品中檢測(cè)出。22份傳統(tǒng)發(fā)酵錦州小菜中真菌在門(mén)水平上共檢測(cè)到子囊菌門(mén)(Ascomycota)、擔(dān)子菌門(mén)(Basidiomycota)等4種已知真菌門(mén);在屬水平上,共檢測(cè)到20種真菌菌屬。其中,酵母菌目屬(Saccharomycetales)、Hyphopichia菌屬(Hyphopichia)、畢赤酵母屬(Pichia)在此采集發(fā)酵階段成為主要優(yōu)勢(shì)菌群。另外,所有樣品均檢測(cè)到了青霉菌屬(Penicillium),比例為0.009%～14.740%。由此推測(cè)錦州地區(qū)的小菜存在豐富的細(xì)菌微生物發(fā)酵菌群,賦予了錦州小菜獨(dú)特風(fēng)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.
【學(xué)位授予單位】：沈陽(yáng)農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TS255.1

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