可可西里和阿坝地区土壤真菌区系及青霉、曲霉分子鉴定
发布时间：2018-09-13 16:03
【摘要】：土壤真菌区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区系中第三大类,其种类以无性型真菌最多。真菌对土壤pH值的要求比放线菌和细菌的低,可以认为真菌是酸性土壤中尤其是森林土壤中的主要分解者。青霉属及其相关有性型真菌是常见的一类腐生菌,其种类广泛分布于自然界,在土壤、水体、空气中均能找到它们,而且也存在于在人类、动物、植物体表或体内,甚至其它真菌上,是一类与日常生活和工业生产关系密切的真菌。曲霉广泛分布在谷物、空气、土壤及各种有机物品上,应用于发酵工业和食品加工业。在人类、动物、植物免疫力出现异常的时候,有些种类就会变为条件致病菌而危害健康甚至生命。曲霉属中还有些种能产生对人体有害的真菌毒素,或引起水果、蔬菜和粮食霉腐。因此,曲霉也与人们的生活息息相关。为了分析高原地区的土壤真菌区系,采集来自青海省可可西里和四川省阿坝地区不同生境的土样进行分类研究。采用稀释平板法(孟加拉红培养基作为选择性培养基)对采集来的土样进行菌株分离和相关分析。对分离得到的菌株采用CTAB法提取菌株的基因组DNA,并扩增这些菌株的核糖体DNA ITS1-5.8S-ITS2序列,p-微管蛋白(BenA)部分基因序列以及钙调蛋白(CaM)部分基因序列。在Genbank中进行比对,得到的结果如下：青海省可可西里地区采集到土样122份,分离到菌株143株,经过分子鉴定,初步鉴定出117株,属于12属,其中,青霉属Penicillium:已鉴定55株菌属于11个种,其中3个新记录种,6株菌待鉴定：曲霉属Aspergillus:已鉴定30株菌属于9个种,其中1个新记录种,2株菌待鉴定；踝节菌属Talaromyces:已鉴定1株菌属于1个种,是新纪录种：枝孢属Cladosporium：分离出2株菌分属2个种,2株菌待鉴定；镰孢菌属Fusarium:分离出4株菌属于2个种；链格孢属Alternaria:只有1株菌：木霉属Trichoderma:6株菌属于4个种：截盘多毛孢属Truncatella:3株菌待鉴定：异茎点霉属Paraphoma:1株菌待鉴定；埃里格孢属Embellisia:2株菌属于1个种；茎点霉属Phoma:1株菌：短梗蠕孢属Thchocladium:1株菌。四川阿坝地区采集到土样64份,分离到188个菌株,初步鉴定出162株,属于15属：青霉属Penicillium:已确定97株属于1 5个种,其中6个新记录种,27株菌待览定：曲霉属Aspergillus:己确定6株菌为4个种,其中1个新记录种,2株菌待鉴定：踝：诲菌属Talaromyces:已确定1株菌,是新纪录种；枝孢属Cladosporium:己确定3株菌属于2个种,4株菌待鉴定：镰孢菌属Fusarium:1株菌：链格孢属Alternaria:己鉴定1株菌,1株菌待鉴定；木霉属Trichoderma:2株菌属于1个种,1株菌待鉴定：截盘多毛孢属Truncatella:共6株菌,待鉴定：中国南方拟盘多毛孢属Pestalotiopsis:]株菌待鉴定；三毛孢属Robillarda:3株菌待鉴定；顶孢霉属Scytalidium:2株菌待鉴定；弯颈霉属Tolypocladium:1株菌待鉴定；沃德霉属Wardomyces:1株菌待鉴定；伞形霉属Umbelopsis:1株菌待鉴定；漆斑菌属Myrothecium:1株菌待鉴定。由于扩增钙调蛋白和p-微管蛋白基因部分序列的引物是特异性针对青霉和曲霉设计的,不适用于所有的土壤真菌,并且核糖体DNA ITS1-5.8S-ITS2序列无法单一鉴定菌株,故待鉴定的菌株尚有待进一步分析。本研究鉴定了一个中国曲霉新纪录种,日耳曼曲霉Aspergillus germanicus。米用形态学性状及钙调蛋白和β-微管蛋白基因部分序列及核糖体DNAITS1-5.8S-ITS2序列进行多相系统分类学分析,两株分离自中国山东泰山地区土壤的曲霉菌(AS3.15303和AS3.15304)与日耳曼曲霉A. germanicus模式菌株CBS123887的形态学和分子性状的比较分析,被鉴定为日耳曼曲霉A. germanicus。参考我国迄今已报道的曲霉物种,确定日耳曼曲霉A. germanicus是曲霉属焦曲霉组的一个中国新记录种。
[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.
【学位授予单位】：长江大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S154.38
本文编号：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.
【学位授予单位】：长江大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S154.38
【参考文献】
相关期刊论文 前3条
1 孔华忠,梁宗琦;分离自江西的一个青霉属新种(英文)[J];菌物系统;2003年01期
2 孔华忠,齐祖同;中国曲霉属的几个新记录和罕见分类群[J];植物研究;1985年02期
3 柳朔怡;吴尚为;;分子系统学在真菌分类命名中的应用与进展[J];微生物学免疫学进展;2015年01期
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