可可西里和阿壩地區(qū)土壤真菌區(qū)系及青霉、曲霉分子鑒定
發(fā)布時間：2018-09-13 16:03
【摘要】：土壤真菌區(qū)系是指生活在土壤中完成生活史或半生活史的所有真菌。土壤真菌是土壤微生物區(qū)系中第三大類,其種類以無性型真菌最多。真菌對土壤pH值的要求比放線菌和細(xì)菌的低,可以認(rèn)為真菌是酸性土壤中尤其是森林土壤中的主要分解者。青霉屬及其相關(guān)有性型真菌是常見的一類腐生菌,其種類廣泛分布于自然界,在土壤、水體、空氣中均能找到它們,而且也存在于在人類、動物、植物體表或體內(nèi),甚至其它真菌上,是一類與日常生活和工業(yè)生產(chǎn)關(guān)系密切的真菌。曲霉廣泛分布在谷物、空氣、土壤及各種有機(jī)物品上,應(yīng)用于發(fā)酵工業(yè)和食品加工業(yè)。在人類、動物、植物免疫力出現(xiàn)異常的時候,有些種類就會變?yōu)闂l件致病菌而危害健康甚至生命。曲霉屬中還有些種能產(chǎn)生對人體有害的真菌毒素,或引起水果、蔬菜和糧食霉腐。因此,曲霉也與人們的生活息息相關(guān)。為了分析高原地區(qū)的土壤真菌區(qū)系,采集來自青海省可可西里和四川省阿壩地區(qū)不同生境的土樣進(jìn)行分類研究。采用稀釋平板法(孟加拉紅培養(yǎng)基作為選擇性培養(yǎng)基)對采集來的土樣進(jìn)行菌株分離和相關(guān)分析。對分離得到的菌株采用CTAB法提取菌株的基因組DNA,并擴(kuò)增這些菌株的核糖體DNA ITS1-5.8S-ITS2序列,p-微管蛋白(BenA)部分基因序列以及鈣調(diào)蛋白(CaM)部分基因序列。在Genbank中進(jìn)行比對,得到的結(jié)果如下：青海省可可西里地區(qū)采集到土樣122份,分離到菌株143株,經(jīng)過分子鑒定,初步鑒定出117株,屬于12屬,其中,青霉屬Penicillium:已鑒定55株菌屬于11個種,其中3個新記錄種,6株菌待鑒定：曲霉屬Aspergillus:已鑒定30株菌屬于9個種,其中1個新記錄種,2株菌待鑒定；踝節(jié)菌屬Talaromyces:已鑒定1株菌屬于1個種,是新紀(jì)錄種：枝孢屬Cladosporium：分離出2株菌分屬2個種,2株菌待鑒定；鐮孢菌屬Fusarium:分離出4株菌屬于2個種；鏈格孢屬Alternaria:只有1株菌：木霉屬Trichoderma:6株菌屬于4個種：截盤多毛孢屬Truncatella:3株菌待鑒定：異莖點(diǎn)霉屬Paraphoma:1株菌待鑒定；埃里格孢屬Embellisia:2株菌屬于1個種；莖點(diǎn)霉屬Phoma:1株菌：短梗蠕孢屬Thchocladium:1株菌。四川阿壩地區(qū)采集到土樣64份,分離到188個菌株,初步鑒定出162株,屬于15屬：青霉屬Penicillium:已確定97株屬于1 5個種,其中6個新記錄種,27株菌待覽定：曲霉屬Aspergillus:己確定6株菌為4個種,其中1個新記錄種,2株菌待鑒定：踝：誨菌屬Talaromyces:已確定1株菌,是新紀(jì)錄種；枝孢屬Cladosporium:己確定3株菌屬于2個種,4株菌待鑒定：鐮孢菌屬Fusarium:1株菌：鏈格孢屬Alternaria:己鑒定1株菌,1株菌待鑒定；木霉屬Trichoderma:2株菌屬于1個種,1株菌待鑒定：截盤多毛孢屬Truncatella:共6株菌,待鑒定：中國南方擬盤多毛孢屬Pestalotiopsis:]株菌待鑒定；三毛孢屬Robillarda:3株菌待鑒定；頂孢霉屬Scytalidium:2株菌待鑒定；彎頸霉屬Tolypocladium:1株菌待鑒定；沃德霉屬Wardomyces:1株菌待鑒定；傘形霉屬Umbelopsis:1株菌待鑒定；漆斑菌屬M(fèi)yrothecium:1株菌待鑒定。由于擴(kuò)增鈣調(diào)蛋白和p-微管蛋白基因部分序列的引物是特異性針對青霉和曲霉設(shè)計的,不適用于所有的土壤真菌,并且核糖體DNA ITS1-5.8S-ITS2序列無法單一鑒定菌株,故待鑒定的菌株尚有待進(jìn)一步分析。本研究鑒定了一個中國曲霉新紀(jì)錄種,日耳曼曲霉Aspergillus germanicus。米用形態(tài)學(xué)性狀及鈣調(diào)蛋白和β-微管蛋白基因部分序列及核糖體DNAITS1-5.8S-ITS2序列進(jìn)行多相系統(tǒng)分類學(xué)分析,兩株分離自中國山東泰山地區(qū)土壤的曲霉菌(AS3.15303和AS3.15304)與日耳曼曲霉A. germanicus模式菌株CBS123887的形態(tài)學(xué)和分子性狀的比較分析,被鑒定為日耳曼曲霉A. germanicus。參考我國迄今已報道的曲霉物種,確定日耳曼曲霉A. germanicus是曲霉屬焦曲霉組的一個中國新記錄種。
[Abstract]:Soil fungi flora refers to all fungi that have completed their life or half-life cycles living in the soil. Soil fungi are the third largest group of soil microbial flora with the largest number of asexual fungi. Fungi have lower pH requirements than actinomycetes and bacteria. Fungi can be considered as the dominant species in acidic soil, especially in forest soil. Penicillium and its related fungi are common saprophytic fungi. They are widely distributed in nature and can be found in soil, water and air. They also exist in human, animal, plant surface or body, and even other fungi. They are fungi closely related to daily life and industrial production. Fungi are widely distributed in cereals, air, soils, and various organic substances, and are used in fermentation and food processing industries. When the immunity of humans, animals, and plants is abnormal, some species become conditional pathogens that endanger health and even life. Some species of Aspergillus can produce or cause mycotoxins that are harmful to human beings. Fruit, vegetable and grain are moldy rot. Therefore, Aspergillus is also closely related to people's lives. In order to analyze the soil fungal flora in the plateau area, soil samples from different habitats in Kekexili, Qinghai Province and Aba, Sichuan Province were collected and classified. The genomic DNA of the isolated strains was extracted by CTAB method, and the sequence of ribosomal DNA ITS1-5.8S-ITS2, partial gene sequence of p-tubulin (BenA) and partial gene sequence of calmodulin (CaM) were amplified. 122 soil samples were collected from Kekexili area of Hainan Province and 143 strains were isolated. After molecular identification, 117 strains belonging to 12 genera were preliminarily identified. Among them, Penicillium: 55 strains belonged to 11 species, of which 3 were newly recorded and 6 strains were to be identified. Aspergillus: 30 strains belonged to 9 species, 1 new recorded species, 2 new recorded species. One strain belongs to one species and is a new record species: Cladosporium: 2 strains belong to 2 species, 2 strains to be identified; Fusarium: 4 strains belong to 2 species; Alternaria: only 1 strain: Trichoderma: 6 strains belong to 4 species. Three strains of Polytrichospora truncatella are to be identified: Paraphoma: 1 strain is to be identified; Embellisia: 2 strains are to be identified; Phoma: 1 strain is to be identified; Thchocladium: 1 strain is to be identified. 64 soil samples were collected from Aba area, Sichuan Province, and 188 strains were identified as 162 strains, belonging to 15 strains. Genus: Penicillium: 97 strains of Penicillium have been identified to belong to 15 species, of which 6 are newly recorded and 27 strains are to be identified: Aspergillus: 6 strains of Aspergillus have been identified as 4 species, 1 new record species and 2 strains to be identified: Ankle: Talaromyces: 1 strain has been identified as a new record species; Cladosporium: 3 strains of Cladosporium have been identified as belonging to 2 species. Four species and four strains of fungi to be identified: Fusarium: 1 strain: Alternaria: Alternaria: 1 strain has been identified, 1 strain to be identified; Trichoderma: 2 strains belong to 1 species, 1 strain to be identified: Truncatella: a total of 6 strains of fungi, to be identified: Pestalotiopsis: Pachymonas from southern China to be identified; 3. Trichospora Robillarda: 3 strains to be identified; Scytalidium: 2 strains to be identified; Tolypocladium: 1 strain to be identified; Wardomyces: 1 strain to be identified; Umbelopsis: 1 strain to be identified; Myrothecium: 1 strain to be identified. Due to the amplification of calmodulin and p-tubulin genes Some of the primers were designed specifically for Penicillium and aspergillus, and were not suitable for all soil fungi. Moreover, the ribosomal DNA ITS1-5.8S-ITS2 sequence could not be used to identify strains alone. Therefore, the strains to be identified need further analysis. Morphological and molecular characters of Aspergillus spp. (AS3.15303 and AS3.15304) and Aspergillus GermaniCus model strain CBS123887 isolated from the soil of Taishan Mountains, Shandong Province, China were analyzed by using morphological characters, partial sequences of calmodulin and beta-tubulin genes and ribosomal DNA ITS1-5.8S-ITS2 sequences. A. GermaniCus was identified as Aspergillus GermaniCus by comparative analysis. Referring to the species of Aspergillus reported so far in China, A. GermaniCus was identified as a new record species of Aspergillus pyrogena in China.
【學(xué)位授予單位】：長江大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S154.38
本文編號：2241663
[Abstract]:Soil fungi flora refers to all fungi that have completed their life or half-life cycles living in the soil. Soil fungi are the third largest group of soil microbial flora with the largest number of asexual fungi. Fungi have lower pH requirements than actinomycetes and bacteria. Fungi can be considered as the dominant species in acidic soil, especially in forest soil. Penicillium and its related fungi are common saprophytic fungi. They are widely distributed in nature and can be found in soil, water and air. They also exist in human, animal, plant surface or body, and even other fungi. They are fungi closely related to daily life and industrial production. Fungi are widely distributed in cereals, air, soils, and various organic substances, and are used in fermentation and food processing industries. When the immunity of humans, animals, and plants is abnormal, some species become conditional pathogens that endanger health and even life. Some species of Aspergillus can produce or cause mycotoxins that are harmful to human beings. Fruit, vegetable and grain are moldy rot. Therefore, Aspergillus is also closely related to people's lives. In order to analyze the soil fungal flora in the plateau area, soil samples from different habitats in Kekexili, Qinghai Province and Aba, Sichuan Province were collected and classified. The genomic DNA of the isolated strains was extracted by CTAB method, and the sequence of ribosomal DNA ITS1-5.8S-ITS2, partial gene sequence of p-tubulin (BenA) and partial gene sequence of calmodulin (CaM) were amplified. 122 soil samples were collected from Kekexili area of Hainan Province and 143 strains were isolated. After molecular identification, 117 strains belonging to 12 genera were preliminarily identified. Among them, Penicillium: 55 strains belonged to 11 species, of which 3 were newly recorded and 6 strains were to be identified. Aspergillus: 30 strains belonged to 9 species, 1 new recorded species, 2 new recorded species. One strain belongs to one species and is a new record species: Cladosporium: 2 strains belong to 2 species, 2 strains to be identified; Fusarium: 4 strains belong to 2 species; Alternaria: only 1 strain: Trichoderma: 6 strains belong to 4 species. Three strains of Polytrichospora truncatella are to be identified: Paraphoma: 1 strain is to be identified; Embellisia: 2 strains are to be identified; Phoma: 1 strain is to be identified; Thchocladium: 1 strain is to be identified. 64 soil samples were collected from Aba area, Sichuan Province, and 188 strains were identified as 162 strains, belonging to 15 strains. Genus: Penicillium: 97 strains of Penicillium have been identified to belong to 15 species, of which 6 are newly recorded and 27 strains are to be identified: Aspergillus: 6 strains of Aspergillus have been identified as 4 species, 1 new record species and 2 strains to be identified: Ankle: Talaromyces: 1 strain has been identified as a new record species; Cladosporium: 3 strains of Cladosporium have been identified as belonging to 2 species. Four species and four strains of fungi to be identified: Fusarium: 1 strain: Alternaria: Alternaria: 1 strain has been identified, 1 strain to be identified; Trichoderma: 2 strains belong to 1 species, 1 strain to be identified: Truncatella: a total of 6 strains of fungi, to be identified: Pestalotiopsis: Pachymonas from southern China to be identified; 3. Trichospora Robillarda: 3 strains to be identified; Scytalidium: 2 strains to be identified; Tolypocladium: 1 strain to be identified; Wardomyces: 1 strain to be identified; Umbelopsis: 1 strain to be identified; Myrothecium: 1 strain to be identified. Due to the amplification of calmodulin and p-tubulin genes Some of the primers were designed specifically for Penicillium and aspergillus, and were not suitable for all soil fungi. Moreover, the ribosomal DNA ITS1-5.8S-ITS2 sequence could not be used to identify strains alone. Therefore, the strains to be identified need further analysis. Morphological and molecular characters of Aspergillus spp. (AS3.15303 and AS3.15304) and Aspergillus GermaniCus model strain CBS123887 isolated from the soil of Taishan Mountains, Shandong Province, China were analyzed by using morphological characters, partial sequences of calmodulin and beta-tubulin genes and ribosomal DNA ITS1-5.8S-ITS2 sequences. A. GermaniCus was identified as Aspergillus GermaniCus by comparative analysis. Referring to the species of Aspergillus reported so far in China, A. GermaniCus was identified as a new record species of Aspergillus pyrogena in China.
【學(xué)位授予單位】：長江大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S154.38
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