| 云南德钦优良野生酵母菌的筛选及其生物防治性能研究 | |
| Alternative Title | Screening of novel wild yeasts from Yunnan Deqin and investigation of their bio-control performance |
| 张永玲 | |
| Subtype | 硕士 |
| Thesis Advisor | 龙丹凤 |
| 2020-05-31 | |
| Degree Grantor | 兰州大学 |
| Place of Conferral | 兰州 |
| Degree Name | 公共卫生硕士 |
| Degree Discipline | 公共卫生 |
| Keyword | 云南产区 酵母筛选 耐受性 拮抗酵母 生物防治 |
| Abstract | 目的：酵母菌在葡萄酒发酵过程中起主要作用，可直接影响葡萄酒的品质、香气等。但随着商业酵母的广泛应用，致使葡萄酒产品同质化现象日益严重，缺乏地域特色。我国葡萄酒产区气候、地理环境复杂多变，蕴含其中的酵母菌种类也丰富多样。本土酵母菌的开发和应用对于酿造出风格独特且兼有地域特色的产地葡萄酒具有重要的应用价值。另外，拮抗酵母菌用于采后果蔬的生物防治潜力巨大，筛选具有拮抗性能的野生酵母菌对于保护环境，提升食品的安全性具有重要意义。 方法：本研究以云南德钦产区赤霞珠和霞多丽葡萄浆果、叶片和土壤为试验材料，通过分离及鉴定，明确酵母菌的种类根据酵母菌对酒精、葡萄糖、pH和SO2的耐受性，筛选出抗性表现优良的野生酵母菌将筛选出的优良菌株进行炭黑曲霉、灰葡萄孢以及青霉菌的拮抗性能研究。 结果：（1）研究共分离筛选出791株野生酵母菌，鉴定为9属14种，包括葡萄汁有孢汉逊酵母（Hanseniaspora uvarum）、Hanseniaspora lachancei、耐渗透压有孢汉逊酵母（Hanseniaspora osmophila）、牧草红酵母（Rhodotorula graminis）、铁杉假丝酵母（Candida railenensis）、喜橄榄假丝酵母（Candida oleophila）、涎沫假丝酵母（Candida zeylanoides）、加利福尼亚假丝酵母（Candida californica）、Papiliotrema flavescens、发酵毕赤氏酵母（Pichia fermentans）、克鲁维毕赤酵母（Pichia kluyveri）、异常威克汉姆酵母（Wickerhamomyces anomalus）、Solicoccozyma aeria、湖北克鲁维酵母（Kluyveromyces hubeiensis），体现了云南德钦葡萄酒产区野生酵母菌的多样性。（2）研究分离的主要为非酿酒酵母，且大多可在18%的酒精度、60%的葡萄糖、2.5的pH和360mg/L的SO2环境下生长，其中葡萄汁有孢汉逊酵母（Hanseniaspora uvarum）耐高酒精度、高葡萄糖以及高SO2的能力均优于其他酵母菌，但在pH为7.0的环境生长不佳。综合所有抗逆性试验结果，共筛选出62株优良酵母菌。（3）研究筛选出73株可减缓或抑制炭黑曲霉（A. carbonarius）、灰葡萄孢（B. cinerea）和青霉菌（P. expansum）生长的酵母菌。且抑菌能力随酵母细胞浓度的增加而增强。另外，产挥发性化合物以及成膜能力强的菌株对霉菌的抑制作用也优于其他菌株。而Fe3+的浓度变化会影响酵母拮抗灰葡萄孢（B. Cinerea）产生抑菌圈的大小，但效果不明显。 结论：云南德钦葡萄酒产区野生酵母菌的多样性较丰富，共鉴定为9属14种根据抗逆性试验，筛选出62株优良酵母菌，主要为葡萄汁有孢汉逊酵母（Hanseniaspora uvarum）筛选出73株可减缓或抑制炭黑曲霉（A. carbonarius）、灰葡萄孢（B. cinerea）和青霉菌（P. expansum）生长的酵母菌，抑菌能力随酵母细胞浓度的增加而增强。另外，产挥发性化合物以及成膜能力强的菌株对霉菌的抑制作用优于其他菌株。 |
| Other Abstract | Objective: Yeast plays a major role during wine fermentation, which can directly affect wine quality and aroma. However, with the widespread application of commercial yeast, the phenomenon of homogenization of wine products has become increasingly serious, lacking regional characteristics. The climate and geographical environment of China's wine-producing regions are complex and changeable, which makes the yeasts also diverse. The development and application of local yeasts have important value for brewing wines with unique style and regional characteristics. In addition, the potential for biological control of antagonistic yeasts for harvesting fruits and vegetables is huge, and is of great significance for protecting the environment and improving food safety. Methods: In this study, the Cabernet Sauvignon and Chardonnay grape berries, leaves and soil were used as experimental materials. The representative strains were identified by combining the morphological and sequence analysis. According to the tolerance of yeast to alcohol, glucose, pH and SO2, the wild yeasts with excellent resistance were screened out. The selected novel strains were used to explore the antagonistic properties of Aspergillus carbonarius, Botrytis cinerea and Penicillium expansum. Results: (1) A total of 791 wild yeast strains were isolated and screened, identified as 9 genera and 14 species, including Hanseniaspora uvarum, Hanseniaspora lachancei, Hanseniaspora osmophila, Rhodotorula graminis, Candida railenensis, Candida oleophila, Candida zeylanoides, Candida californica, Papiliotrema flavescens, Pichia fermentans, Pichia kluyveri, Wickerhamomyces anomalus, Solicoccozyma aeria and Kluyveromyces hubeiensis, showing the diversity of wild yeast in the Deqin wine regions of yunnan. (2) The isolates are mainly non-Saccharomyces cerviae, most of them were able to grow in the environment of 18% alcohol content, 60% glucose, pH 2.5 and 360mg/L SO2. Among them, Hanseniaspora uvarum was superior to other yeasts in the ability of the above resistance, but it could not grow well in the environment of pH 7.0. Based on the results of all the stress resistance tests, a total of 62 excellent strains were screened. (3) The study screened out 73 strains of yeasts that could slow down or inhibit the growth of A. carbonarius, B. cinerea and P. expansum. And the bacteriostatic ability increases with the increasing of yeast cell concentration. In addition, the strains that produced volatile compounds and have strong film-forming capacity against mold are also superior to other strains. The change in the concentration of Fe3+ will affect the size of the inhibition zone produced by yeast against B. cinerea, but the effect was not obvious. Conclusion: The diversity of wild yeasts in Deqin wine region of Yunnan province was rich, with 14 species belonging to 9 genera. According to the stress resistance test, 62 excellent yeast strains were selected, mainly Hanseniaspora Uvarum. Seventy-three yeast strains were selected to slow down or inhibit the growth of A. carbonarius, B. cinerea and P. expansum. And the bacteriostatic ability increases with the increasing of yeast cell concentration. In addition, the strains with strong volatile compounds and membrane forming ability had better inhibitory effect on mold than other strains. |
| Pages | 75 |
| URL | 查看原文 |
| Language | 中文 |
| Document Type | 学位论文 |
| Identifier | https://ir.lzu.edu.cn/handle/262010/464821 |
| Collection | 公共卫生学院 |
| Affiliation | 公共卫生学院 |
| First Author Affilication | School of Public health |
| Recommended Citation GB/T 7714 | 张永玲. 云南德钦优良野生酵母菌的筛选及其生物防治性能研究[D]. 兰州. 兰州大学,2020. |
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