裸子植物内生真菌Xylaria sp. NCY2和Tubercularia sp. TF5次生代谢产物的研究
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摘要
植物内生菌指定殖在植物组织内部而又不引起直接和明显病症的一类微生物,是植物微生态系统中的天然组成成分。植物内生菌在进化过程中与宿主建立了和谐的关系,其次生代谢产物十分丰富,是强生理活性和新结构类型天然产物的丰富来源。
裸子植物是地球上最古老的木本植物,其中不乏药用植物,我国是裸子植物种类最多,资源最丰富的国家。对其内生真菌的研究将有利于我国微生物药物的开发和珍稀植物资源的保护。本文以福建省两种珍稀药用裸子植物红豆杉和长叶榧为对象,对其内生真菌次生代谢产物进行研究,旨在发现具有较高生物活性的结构新颖的化合物,为开发新的微生物药物奠定基础。
从福建邵武和沙县的南方红豆杉和长叶榧树皮中分离得到内生真菌92株。采用抗菌活性筛选模型和细胞毒筛选模型对这些真菌的发酵液进行检测,抑菌实验显示,48.9%的菌株对一种或多种指示菌有抑制作用,19.6%的菌株具有高抗菌活性;抗肿瘤实验显示,16.3%的菌株对HeLa,KB或HL-60细胞有抑制作用(ID_(50)≤1:50),ID_(50)在200或200倍以上的高活性菌株占总供测菌株的5.4%,表明植物内生真菌中存在着广泛的抗真菌和抗肿瘤化学成分资源。
通过使用形态学和分子生物学方法,将NCY2菌株鉴定为炭角菌(Xylariasp.NCY2)。
基于发酵产物的活性筛选和TLC分析结果以及菌株的分类学地位,从92株内生真菌中选择了两株NCY2和TF5,进行了较详尽的次生代谢产物研究。从NCY2菌株PD培养基液体静置发酵产物中分离鉴定了21个化合物,其中新化合物15个,包括H19、H21、NC、H53、H41、H59、H60七个新倍半萜化合物和hp2、H63、H50-2、H51-1、H51-2、H48、H40、hp7八个新聚酮化合物。从TF5菌株PDA培养基发酵产物中分离鉴定了一个新二萜苷化合物T9和8个已知化合物,包括两个二萜和三个松孢菌素类化合物。
对分离得到的16个新化合物进行了抗菌和抗肿瘤活性测定,结果表明,他们只有微弱的抗菌和抗肿瘤的活性,10μg/mL浓度时对一种或几种指示菌的抑制率大约为20%-40%,对肿瘤细胞株抑制率大约为20%-30%,深入的活性研究值得进一步开展。
本论文的研究结果表明,裸子植物内生真菌是结构新颖和新的生理活性物质的重要来源,是开发新药的宝贵资源。
Endophytes are the microbes that colonize living, internal tissues of plants without causing any immediate, overt negative effects. They are natural constituent of microecosystem inside plants and have established a harmonious relationship with their hosts in the long-term evolution process. Endophytes represent a dependable source of bioactive and chemically novel compounds since their secondary metabolites are abundant.
Gymnosperm plants are the most ancient xylophyte, many of them are pharmaceutical plants. China has very rich gymnosperm plants. Studies on endophytic fungi will promote the development of microbial medicine and help the protection of the valuable and rare plants. We have studied the secondary metabolites of the endophytic fungi from gymnosperm plants in Fujian province, Taxus mairei and Torreya jackii Chun, in order to discover the structure novel compounds with potential for new drugs.
92 strains of endophytic fungi were obtained from the bark of Taxus mairei and Torreya jackii Chun collected in Shaowu and Sha countries, Fujian province. The antitumor and antimicrobial activities of the fermentation broths of these endophytic fungi were tested. The results showed that the fermentation broths of 45 strains (about 48.9% of the total strains isolated) show noticeable growth inhibition against one or more indicator fungi, and about 19.6% of them have high activities against fungi; 15 (about 16.3%) of them have inhibition activities against HeLa, KB and/or HL-60 tumor cells with the ID_(50)≤1:50, and 5 (about 5.4%) of them have the high activities with the ID_(50)≤1:200.
The strain NCY2 was identified by morphological and molecule biological methods to be Xylaria sp.
According to the results of bioactivity screening, TLC analysis and Phylogeny among the isolates, two strains namely NCY2 and TF5, were chosen for further chemical research. Seven new sesquiterpenoids (H19, H21, NC, H53, H41, H59, H60) and eight new polyketides (hp2, H63, H50-2, H51-1, H51-2, H48, H40, hp7), along with 6 known compounds were isolated from liquid still culture of strain NCY2. A new diterpene glucoside compound (T9) and 8 known compounds were isolated from the fermentation culture of the strain TF5 on agar medium.
The antitumor and antimicrobial activities of the 16 new compounds were measured and they showed weak activities, the further researches on their bioactivities were valuable.
The results of this study indicated that endophytic fungi from gymnosperm plants are important source of novel chemically compounds, they are precious resources with potential for exploitation new drugs.
裸子植物是地球上最古老的木本植物,其中不乏药用植物,我国是裸子植物种类最多,资源最丰富的国家。对其内生真菌的研究将有利于我国微生物药物的开发和珍稀植物资源的保护。本文以福建省两种珍稀药用裸子植物红豆杉和长叶榧为对象,对其内生真菌次生代谢产物进行研究,旨在发现具有较高生物活性的结构新颖的化合物,为开发新的微生物药物奠定基础。
从福建邵武和沙县的南方红豆杉和长叶榧树皮中分离得到内生真菌92株。采用抗菌活性筛选模型和细胞毒筛选模型对这些真菌的发酵液进行检测,抑菌实验显示,48.9%的菌株对一种或多种指示菌有抑制作用,19.6%的菌株具有高抗菌活性;抗肿瘤实验显示,16.3%的菌株对HeLa,KB或HL-60细胞有抑制作用(ID_(50)≤1:50),ID_(50)在200或200倍以上的高活性菌株占总供测菌株的5.4%,表明植物内生真菌中存在着广泛的抗真菌和抗肿瘤化学成分资源。
通过使用形态学和分子生物学方法,将NCY2菌株鉴定为炭角菌(Xylariasp.NCY2)。
基于发酵产物的活性筛选和TLC分析结果以及菌株的分类学地位,从92株内生真菌中选择了两株NCY2和TF5,进行了较详尽的次生代谢产物研究。从NCY2菌株PD培养基液体静置发酵产物中分离鉴定了21个化合物,其中新化合物15个,包括H19、H21、NC、H53、H41、H59、H60七个新倍半萜化合物和hp2、H63、H50-2、H51-1、H51-2、H48、H40、hp7八个新聚酮化合物。从TF5菌株PDA培养基发酵产物中分离鉴定了一个新二萜苷化合物T9和8个已知化合物,包括两个二萜和三个松孢菌素类化合物。
对分离得到的16个新化合物进行了抗菌和抗肿瘤活性测定,结果表明,他们只有微弱的抗菌和抗肿瘤的活性,10μg/mL浓度时对一种或几种指示菌的抑制率大约为20%-40%,对肿瘤细胞株抑制率大约为20%-30%,深入的活性研究值得进一步开展。
本论文的研究结果表明,裸子植物内生真菌是结构新颖和新的生理活性物质的重要来源,是开发新药的宝贵资源。
Endophytes are the microbes that colonize living, internal tissues of plants without causing any immediate, overt negative effects. They are natural constituent of microecosystem inside plants and have established a harmonious relationship with their hosts in the long-term evolution process. Endophytes represent a dependable source of bioactive and chemically novel compounds since their secondary metabolites are abundant.
Gymnosperm plants are the most ancient xylophyte, many of them are pharmaceutical plants. China has very rich gymnosperm plants. Studies on endophytic fungi will promote the development of microbial medicine and help the protection of the valuable and rare plants. We have studied the secondary metabolites of the endophytic fungi from gymnosperm plants in Fujian province, Taxus mairei and Torreya jackii Chun, in order to discover the structure novel compounds with potential for new drugs.
92 strains of endophytic fungi were obtained from the bark of Taxus mairei and Torreya jackii Chun collected in Shaowu and Sha countries, Fujian province. The antitumor and antimicrobial activities of the fermentation broths of these endophytic fungi were tested. The results showed that the fermentation broths of 45 strains (about 48.9% of the total strains isolated) show noticeable growth inhibition against one or more indicator fungi, and about 19.6% of them have high activities against fungi; 15 (about 16.3%) of them have inhibition activities against HeLa, KB and/or HL-60 tumor cells with the ID_(50)≤1:50, and 5 (about 5.4%) of them have the high activities with the ID_(50)≤1:200.
The strain NCY2 was identified by morphological and molecule biological methods to be Xylaria sp.
According to the results of bioactivity screening, TLC analysis and Phylogeny among the isolates, two strains namely NCY2 and TF5, were chosen for further chemical research. Seven new sesquiterpenoids (H19, H21, NC, H53, H41, H59, H60) and eight new polyketides (hp2, H63, H50-2, H51-1, H51-2, H48, H40, hp7), along with 6 known compounds were isolated from liquid still culture of strain NCY2. A new diterpene glucoside compound (T9) and 8 known compounds were isolated from the fermentation culture of the strain TF5 on agar medium.
The antitumor and antimicrobial activities of the 16 new compounds were measured and they showed weak activities, the further researches on their bioactivities were valuable.
The results of this study indicated that endophytic fungi from gymnosperm plants are important source of novel chemically compounds, they are precious resources with potential for exploitation new drugs.
引文
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