延胡索稀有内生放线菌的筛
发布时间：2019-03-20 16:53
【摘要】：植物内生放线菌是指在植物体内生活又对植物不产生明显危害的一类放线菌。长期与植物共生过程使植物内生放线菌形成了相对特殊的生理与代谢特点,这使植物内生放线菌成为了一种极有价值的微生物资源。随着酶、抗生素等一系列新的具有极强活性的化合物不断从植物内生放线菌中被分离出,植物内生放线菌的重要性日益引起生物学家、化学家以及农学家的兴趣与注意。本实验以采自黑龙江省五常市凤凰山风景区的一种传统的药用植物延胡索样品为分离源,使用高氏一号、HV和改良高氏二号等十种培养基,以浓度为50mg/L的萘啶酮酸、浓度为20mg/L放线菌酮组合为抑制剂,总共分离到放线菌78株。使用十字交叉法点接测定78株放线菌对十种真菌的抑菌活性,发现14株放线菌菌对至少4种真菌具有抑制作用。通过在培养基上的形态特征的比较,发现YHS9、NEAU-YHS12、NEAU-YHS15、YHS37和YHS39的培养形态比较特殊,对这五株菌株的16S rRNA进行了基因序列测定并使用Ez Taxon-e网站进行比对,得出结论YHS9属于链霉菌属,YHS37、YHS39属于小单孢菌属,NEAU-YHS12与NEAU-YHS15属于球包囊菌属,对NEAU-YHS12与NEAU-YHS15进行了多相分类学鉴定。Ez Taxon-e网站比对显示NEAU-YHS12和NEAU-YHS15属于球包囊菌属,与NEAU-YHS12和NEAU-YHS15相似度最高的菌株分别为为Sphaerosporangium cinnabarinum JCM 3291T(98.7%,98.6%),Sphaerisporangium flaviroseum YIM 48771T(98.6%,98.6%),Sphaerosporangium melleum JCM 13064T(98.5%,98.4%)和Sphaerisporangium dianthi NEAU-CY18T(98.4%,98.4%)。基因对比发现NEAU-YHS12和NEAU-YHS15的16s r RNA序列相似度为99.8%。菌株NEAU-YHS12和NEAU-YHS15与球包囊菌属的所有典型菌株以邻位连接法(Neighbour-Joining)与最大似然法(Maximum-likelihood)两种方法构建系统发育树,得到两株进化树拓扑结构一致,菌株NEAU-YHS12、NEAU-YHS15与第二高相似度的Sphaerisporangium flaviroseum YIM 48771T形成独立分支,然而其分支上的低靴值不支持此分支。菌株NEAU-YHS12和NEAU-YHS15的表型特征、化学分类特征与系统进化特征基本相同,同时又都符合球包囊菌株的特点。但NEAU-YHS12和NEAU-YHS15的DNA杂交值大于70%。故而菌株NEAU-YHS12和NEAU-YHS15确定为球包囊菌属的一个新种,命名为Sphaerisporangium corydalis,代表菌株为NEAU-YHS15,同时分别保藏于中国科学院微生物研究所(CGMCC 4.7148T)和德国不伦瑞克市微生物菌种保藏中心(DSM 46732T)。
[Abstract]:Endophytic actinomycetes are actinomycetes that live in plants and do not cause obvious harm to plants. Long-term symbiosis with plants makes endophytic actinomycetes form relatively special physiological and metabolic characteristics, which makes endophytic actinomycetes a valuable microbial resource. As a series of new active compounds such as enzymes and antibiotics have been isolated from endophytic actinomycetes, the importance of endophytic actinomycetes has attracted increasing interest and attention from biologists, chemists and agronomists. In this experiment, a traditional medicinal plant, Corydalis corydalis, collected from the Fenghuangshan Scenic spot of Wuchang City, Heilongjiang Province, was isolated by using 10 kinds of medium, such as Gao Shi No. 1, HV and modified Gao Shi No. 2, in which the concentration of 50mg/L was naphtholone acid. A total of 78 strains of actinomycetes were isolated at the concentration of 20mg/L combined with actinomyces as inhibitors. The inhibitory activity of 78 strains of actinomycetes against 10 kinds of fungi was determined by cross-crossing method. It was found that 14 strains of actinomycetes had inhibitory effects on at least 4 kinds of fungi. Through the comparison of morphological characteristics on the medium, it was found that the culture morphology of YHS9,NEAU-YHS12,NEAU-YHS15,YHS37 and YHS39 was special. The 16s rRNA of the five strains were sequenced and compared with the Ez Taxon-e website. Conclusion YHS9 belongs to Streptomyces, YHS37,YHS39 belongs to Micromonospora, and NEAU-YHS12 and NEAU-YHS15 belong to the genus Balloon. The polyphase taxonomy of NEAU-YHS12 and NEAU-YHS15 was carried out. The comparison of Ez Taxon-e website showed that NEAU-YHS12 and NEAU-YHS15 belonged to the genus Botrytis, and the strains with the highest similarity to NEAU-YHS12 and NEAU-YHS15 were Sphaerosporangium cinnabarinum JCM 3291T (98.7%), and NEAU-YHS15 (98.7%) with the highest similarity to NEAU-YHS12 and NEAU-YHS15, respectively. 98.6%), Sphaerisporangium flaviroseum YIM 48771T (98.6%, 98.6%), Sphaerosporangium melleum JCM 13064T (98.5%, 98.4%) and Sphaerisporangium dianthi NEAU-CY18T (98.4%, 98.4%). The 16s r RNA sequence similarity between NEAU-YHS12 and NEAU-YHS15 was 99.8%. The phylogenetic trees of strains NEAU-YHS12 and NEAU-YHS15 were constructed by two methods of Neighbour-Joining and Maximum-likelihood, and the topological structure of two phylogenetic trees was identical. Strain NEAU-YHS12, was found to have the same topological structure. NEAU-YHS15 forms an independent branch with the second high similarity Sphaerisporangium flaviroseum YIM 48771T, but the low boot value on the branch does not support this branch. The phenotypic characteristics, chemical taxonomic characteristics and phylogenetic characteristics of the strains NEAU-YHS12 and NEAU-YHS15 were basically the same as those of the Balloon strains at the same time. But the DNA hybrid value of NEAU-YHS12 and NEAU-YHS15 was more than 70%. Therefore, strains NEAU-YHS12 and NEAU-YHS15 were identified as a new species of the genus Balloon, named as NEAU-YHS15, as the representative strain of Sphaerisporangium corydalis,. At the same time, they were stored in the Institute of Microbiology, Chinese Academy of Sciences (CGMCC 4.7148T) and the Center for the Preservation of microbial strains (DSM 46732T) in Brunswick, Germany.
【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S182
本文编号：2444413
[Abstract]:Endophytic actinomycetes are actinomycetes that live in plants and do not cause obvious harm to plants. Long-term symbiosis with plants makes endophytic actinomycetes form relatively special physiological and metabolic characteristics, which makes endophytic actinomycetes a valuable microbial resource. As a series of new active compounds such as enzymes and antibiotics have been isolated from endophytic actinomycetes, the importance of endophytic actinomycetes has attracted increasing interest and attention from biologists, chemists and agronomists. In this experiment, a traditional medicinal plant, Corydalis corydalis, collected from the Fenghuangshan Scenic spot of Wuchang City, Heilongjiang Province, was isolated by using 10 kinds of medium, such as Gao Shi No. 1, HV and modified Gao Shi No. 2, in which the concentration of 50mg/L was naphtholone acid. A total of 78 strains of actinomycetes were isolated at the concentration of 20mg/L combined with actinomyces as inhibitors. The inhibitory activity of 78 strains of actinomycetes against 10 kinds of fungi was determined by cross-crossing method. It was found that 14 strains of actinomycetes had inhibitory effects on at least 4 kinds of fungi. Through the comparison of morphological characteristics on the medium, it was found that the culture morphology of YHS9,NEAU-YHS12,NEAU-YHS15,YHS37 and YHS39 was special. The 16s rRNA of the five strains were sequenced and compared with the Ez Taxon-e website. Conclusion YHS9 belongs to Streptomyces, YHS37,YHS39 belongs to Micromonospora, and NEAU-YHS12 and NEAU-YHS15 belong to the genus Balloon. The polyphase taxonomy of NEAU-YHS12 and NEAU-YHS15 was carried out. The comparison of Ez Taxon-e website showed that NEAU-YHS12 and NEAU-YHS15 belonged to the genus Botrytis, and the strains with the highest similarity to NEAU-YHS12 and NEAU-YHS15 were Sphaerosporangium cinnabarinum JCM 3291T (98.7%), and NEAU-YHS15 (98.7%) with the highest similarity to NEAU-YHS12 and NEAU-YHS15, respectively. 98.6%), Sphaerisporangium flaviroseum YIM 48771T (98.6%, 98.6%), Sphaerosporangium melleum JCM 13064T (98.5%, 98.4%) and Sphaerisporangium dianthi NEAU-CY18T (98.4%, 98.4%). The 16s r RNA sequence similarity between NEAU-YHS12 and NEAU-YHS15 was 99.8%. The phylogenetic trees of strains NEAU-YHS12 and NEAU-YHS15 were constructed by two methods of Neighbour-Joining and Maximum-likelihood, and the topological structure of two phylogenetic trees was identical. Strain NEAU-YHS12, was found to have the same topological structure. NEAU-YHS15 forms an independent branch with the second high similarity Sphaerisporangium flaviroseum YIM 48771T, but the low boot value on the branch does not support this branch. The phenotypic characteristics, chemical taxonomic characteristics and phylogenetic characteristics of the strains NEAU-YHS12 and NEAU-YHS15 were basically the same as those of the Balloon strains at the same time. But the DNA hybrid value of NEAU-YHS12 and NEAU-YHS15 was more than 70%. Therefore, strains NEAU-YHS12 and NEAU-YHS15 were identified as a new species of the genus Balloon, named as NEAU-YHS15, as the representative strain of Sphaerisporangium corydalis,. At the same time, they were stored in the Institute of Microbiology, Chinese Academy of Sciences (CGMCC 4.7148T) and the Center for the Preservation of microbial strains (DSM 46732T) in Brunswick, Germany.
【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S182
【参考文献】
相关期刊论文 前1条
1 黄晓辉;李珊;谭周进;谢丙炎;伍参荣;杨友才;;植物内生放线菌研究进展[J];生物技术通报;2008年01期
，本文编号：2444413
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