延胡索稀有內(nèi)生放線菌的篩
發(fā)布時間：2019-03-20 16:53
【摘要】：植物內(nèi)生放線菌是指在植物體內(nèi)生活又對植物不產(chǎn)生明顯危害的一類放線菌。長期與植物共生過程使植物內(nèi)生放線菌形成了相對特殊的生理與代謝特點,這使植物內(nèi)生放線菌成為了一種極有價值的微生物資源。隨著酶、抗生素等一系列新的具有極強活性的化合物不斷從植物內(nèi)生放線菌中被分離出,植物內(nèi)生放線菌的重要性日益引起生物學(xué)家、化學(xué)家以及農(nóng)學(xué)家的興趣與注意。本實驗以采自黑龍江省五常市鳳凰山風(fēng)景區(qū)的一種傳統(tǒng)的藥用植物延胡索樣品為分離源,使用高氏一號、HV和改良高氏二號等十種培養(yǎng)基,以濃度為50mg/L的萘啶酮酸、濃度為20mg/L放線菌酮組合為抑制劑,總共分離到放線菌78株。使用十字交叉法點接測定78株放線菌對十種真菌的抑菌活性,發(fā)現(xiàn)14株放線菌菌對至少4種真菌具有抑制作用。通過在培養(yǎng)基上的形態(tài)特征的比較,發(fā)現(xiàn)YHS9、NEAU-YHS12、NEAU-YHS15、YHS37和YHS39的培養(yǎng)形態(tài)比較特殊,對這五株菌株的16S rRNA進(jìn)行了基因序列測定并使用Ez Taxon-e網(wǎng)站進(jìn)行比對,得出結(jié)論YHS9屬于鏈霉菌屬,YHS37、YHS39屬于小單孢菌屬,NEAU-YHS12與NEAU-YHS15屬于球包囊菌屬,對NEAU-YHS12與NEAU-YHS15進(jìn)行了多相分類學(xué)鑒定。Ez Taxon-e網(wǎng)站比對顯示NEAU-YHS12和NEAU-YHS15屬于球包囊菌屬,與NEAU-YHS12和NEAU-YHS15相似度最高的菌株分別為為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%)�；�?qū)Ρ劝l(fā)現(xiàn)NEAU-YHS12和NEAU-YHS15的16s r RNA序列相似度為99.8%。菌株NEAU-YHS12和NEAU-YHS15與球包囊菌屬的所有典型菌株以鄰位連接法(Neighbour-Joining)與最大似然法(Maximum-likelihood)兩種方法構(gòu)建系統(tǒng)發(fā)育樹,得到兩株進(jìn)化樹拓?fù)浣Y(jié)構(gòu)一致,菌株NEAU-YHS12、NEAU-YHS15與第二高相似度的Sphaerisporangium flaviroseum YIM 48771T形成獨立分支,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、化學(xué)分類特征與系統(tǒng)進(jìn)化特征基本相同,同時又都符合球包囊菌株的特點。但NEAU-YHS12和NEAU-YHS15的DNA雜交值大于70%。故而菌株NEAU-YHS12和NEAU-YHS15確定為球包囊菌屬的一個新種,命名為Sphaerisporangium corydalis,代表菌株為NEAU-YHS15,同時分別保藏于中國科學(xué)院微生物研究所(CGMCC 4.7148T)和德國不倫瑞克市微生物菌種保藏中心(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.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S182
本文編號：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.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S182
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 黃曉輝;李珊;譚周進(jìn);謝丙炎;伍參榮;楊友才;;植物內(nèi)生放線菌研究進(jìn)展[J];生物技術(shù)通報;2008年01期
，本文編號：2444413
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