Knowledge Resource Center for Ecological Environment in Arid Area
| 中国适碱真菌多样性及其酶活性测定 | |
| 其他题名 | The diversity of alkali-adapted fungi in China and their enzymes screening |
| 黄盼盼 | |
| 出版年 | 2017 |
| 学位类型 | 硕士 |
| 导师 | 刘杏忠 |
| 学位授予单位 | 中国科学院大学 |
| 中文摘要 | 碱环境包括碱湖、沙漠土壤和碱泉等，pH值通常长时间保持在10左右。适碱菌是生活在碱性环境下具有独特的生理机能和遗传基因的一类微生物，根据其碱性的耐受范围可将其划分为耐碱菌和嗜碱菌。适碱菌包括适碱真菌和适碱细菌。相对而言，在研究初期学者们对于适碱细菌的研究较多，而对适碱真菌的研究较少。随着研究领域的不断拓宽和研究方法的不断深入，关于适碱真菌的研究和报道在逐渐增加，它们在农业、生物修复、工业酶制剂等领域的开发应用潜力也不断被发掘。特别是适碱真菌能够产生一些碱性酶如碱性蛋白酶、碱性淀粉酶、以及碱性纤维素酶具有广泛的应用价值，在食品业、纺织业、制革工业和日化用品制造业都有应用。本课题研究旨在对中国典型盐碱地中的适碱真菌资源进行系统性调查，分析其多样性并检测其中的优势类群的酶活，为了解和开发我国适碱真菌资源提供理论依据，同时也为丰富我国的微生物资源、推动干旱及半干旱地区微生物资源的保护、开发和利用提供理论依据。本研究采集东北地区（吉林省和黑龙江省）和西北地区（宁夏回族自治区和甘肃省）典型盐碱地土壤样品作为研究材料，采用稀释涂布平板法来进行适碱真菌的分离，结合形态学和DNA序列分析进行物种多样性研究；利用真菌多相分类技术，鉴定我国适碱真菌，描述新物种，最终初步明确我国盐碱土壤中适碱真菌的多样性。然后对获得的适碱真菌中的优势类群进行8个不同pH（pH 4–11）的生长曲线测定，同时对其进行蛋白酶、淀粉酶、纤维素酶、木聚糖酶和脂肪酶的酶活性测定，以便加深对中国适碱真菌的了解。本研究通过可培养方法分离到盐碱土壤中的适碱真菌共1823株，这些菌株分布在424个种、188个属、33个目里。其中以肉座菌目为主，占所有菌株的44％；其次是格孢菌目、散囊菌目、Glomerellales等，分别占14％、10％和6％。优势种主要有Sarocladium kiliense、Sarocladium strictum、Gliomastix murorum、Acrostalagmus luteoalbus、Gibellulopsis nigrescens、Fusarium acuminatum、Fusarium equiseti等。通过生长曲线测定试验发现分离自同一地区、同一个种属的不同菌株的生长曲线趋势基本一致，其中Purpureocillium、Fusarium、Gibellulopsis和Plectosphaerella的成员大多是耐碱真菌；Sarocladium、Myrothecium、Gliomastix、Scopulariopsis和Acrostalagmus的成员大多是嗜碱真菌。本研究发现了两个新种，分别是Aspergillus jilinensis和Sarocladium jilinensis。 酶活测定结果显示，大部分适碱真菌都产生碱性蛋白酶和碱性木聚糖酶。其中Purpureocillium大部分菌株具有很强的碱性蛋白酶活性，但未显示出其他酶的活性；Sarocladium大部分菌株具有很强的碱性蛋白酶和碱性木聚糖酶活性；Myrothecium没有显示任何一种酶活；Gliomastix的菌株显示出碱性蛋白酶和碱性木聚糖酶活性；Fusarium中能检测出很强的碱性蛋白酶和碱性木聚糖酶活性以及一定的淀粉酶、纤维素酶活性，大部分菌株具有其中的两种或三种酶活；Gibellulopsis的菌株具有碱性蛋白酶、碱性淀粉酶以及很强的碱性木聚糖酶活性；Scopulariopsis的大部分菌株具有很强的碱性蛋白酶和碱性木聚糖酶活性；Plectosphaerella中部分菌株具有很强的碱性蛋白酶活性，大部分具有碱性木聚糖酶活性及一定的碱性脂肪酶活性；Acrostalagmus中的菌株都具有很强的碱性蛋白酶活性，少数还具有碱性淀粉酶和较强的木聚糖酶活性。大部分菌株都没有碱性淀粉酶活性和碱性纤维素酶活性。大部分菌株都没有脂肪酶活性，少数Fusarium、Gibellulopsis、Scopulariopsis和Plectosphaerella中的菌株具有一定的碱性脂肪酶活性。这为今后碱性真菌在工业酶制剂的应用提供了一定的理论基础。 |
| 英文摘要 | Alkaline environments include alkaline lakes, desert soils and alkaline springs, the pH of which is usually maintained at around 10 for a long time. Alkali-adapted microbes living in the alkaline environment have unique physiological property and genome. They can be divided into alkali-tolerant microbes and alkalophilic microbes according to their tolerance pH range. The alkali-adapted microbes include alkali-adapted bacteria and alkali-adapted fungi. The alkali-adapted bacteria have been deeply studied by previous studies. Recently, the development and application of alkali-adapted fungi in agriculture, bioremediation, industrial enzyme preparation have been continuously explored. In particular, alkali-adapted fungi can produce some alkaline enzymes such as alkaline protease, amylase and cellulase, which have a wide range of industrial applications in the food, textile, leather and daily chemical products. The aim of this study is to systematically investigate the diversity of alkali-adapted fungal resources in typical saline-alkali soils in China, analyze their diversity and the enzyme activities of dominant groups. The study provides a foundation of understanding and developing the alkaline fungal resources in China; and promoting the protection, development, utilization of microbial resources in arid and semi-arid areas.In this study, soil samples of typical alkaline environments in northeastern (Jilin Province and Heilongjiang Province) and northwestern China (Ningxia Hui Autonomous Region and Gansu Province) were collected as research materials, and the alkaline fungi were isolated by dilution plating method. The microbial diversities were investigated by the morphology and DNA sequence analyses. The fungal multi-phase classification technologies were applied to identify the alkali-adapted fungi in China and describe the new species to ensure the alkaline fungi diversity in China. Then, the growth curves of the dominant species in 8 pH values (pH 4–11) were determined. And the enzyme activities of protease, amylase, cellulase, xylanase and lipase were also screened in order to deepen the understanding of the alkali-adapted fungi.In this study, a total of 1823 strains of alkali-adapted fungi were isolated, which were distributed in 424 species, 188 genera and 33 orders. Among them, fungal species in Hypocreales were dominant species, accounting for 44%; followed by Pleosporales, Eurotiales, Glomerellales, accounting for 14%, 10% and 6%, respectively. The growth curves of different strains were similar in the same geographical region. The dominant species were Sarocladium kiliense, Sarocladium strictum, Gliomastix murorum, Acrostalagmus luteoalbus, Gibellulopsis nigrescens, Fusarium acuminatum, Fusarium equiseti and so on. The isolated alkaline-tolerant fungi in this study mostly belong to the genera of Purpureocillium, Fusarium, Gibellulopsis and Plectosphaerella. And most isolated alkalophilic fungi belong to Sarocladium, Myrothecium, Gliomastix, Scopulariopsis and Acrostalagmus. Two new species were found in this study, including Aspergillus jilinensis and Sarocladium jilinensis. The enzymes screening results showed that most alkali-adapted fungi produced alkaline protease and xylanase activities. Purpureocillium had strong alkaline protease activity but did not show the activities of other enzymes. Most Sarocladium strains had strong alkaline protease and xylanase activities. Strains of Myrothecium did not show any enzyme activity. Gliomastix spp. had alkaline protease and xylanase activities. Fusarium spp. showed strong alkaline protease, xylanase activities, and some amylase, cellulase activities, some strains showed two or three of these enzymes activities simultaneously. Gibellulopsis strains had alkaline protease, amylase activities and strong alkaline xylanase activity. Most of Scopulariopsis strains had strong alkaline protease and xylanase activities. Some strains of Plectosphaerella had alkaline protease activity and most strains had alkaline xylanase activity and some lipase activity. Strains of Acrostalagmus had strong alkaline protease activity, some of them also had some alkaline amylase activity and strong xylanase activity. Most strains did not show lipase activity, except some of Fusarium, Gibellulopsis, Scopulariopsis and Plectosphaerella strains, which provides a theoretical foundation for the application of alkali-adapted fungi in enzyme industry. |
| 中文关键词 | 中国 ; 适碱真菌 ; 多样性 ; 酶活 ; 测定 |
| 英文关键词 | China Alkali-adapted fungi Diversity Enzyme activity Screening |
| 语种 | 中文 |
| 国家 | 中国 |
| 来源学科分类 | 微生物学 |
| 来源机构 | 中国科学院微生物研究所 |
| 资源类型 | 学位论文 |
| 条目标识符 | http://119.78.100.177/qdio/handle/2XILL650/287927 |
| 推荐引用方式 GB/T 7714 | 黄盼盼. 中国适碱真菌多样性及其酶活性测定[D]. 中国科学院大学,2017. |
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