两种植物及其根际微生物的化学研究
文献类型：学位论文
| 作者 | 范黎明 |
| 学位类别 | 博士 |
| 答辩日期 | 2005 |
| 授予单位 | 中国科学院昆明植物研究所 |
| 授予地点 | 中国科学院昆明植物研究所 |
| 导师 | 郝小江 |
| 关键词 | 粉花绣线菊 滑桃树 二砧生物碱 根际微生物 相互作用 |
| 其他题名 | Approach to Chemical Study of Two Plants and Their Rhizosphere Microbes |
| 学位专业 | 植物学 |
| 中文摘要 | 本论文以植物和根际微生物次生代谢产物之间的相关性为切入点，以粉花绣线菊和滑桃树及其根际微生物为研究材料．，从五个产地的粉花绣线菊二i陆生物碱成分、粉花绣线菊根际微生物及其化学成分、滑桃树根际微生物及其化学成分三个方面开展实验。为了完善粉花绣线菊（spiraea japonica）复合群采样的系统性和全面性，对五个产地的粉花绣线菊生物碱进行分析，共分离鉴定了13个C20型二菇生物碱，其中6-hydroxylspirasine Iv（3）、spiroqine（11）、6-hydroxylspiraqille（12）和生物碱季胺盐N-amilloethyl sPil．adille A cllloride（13）为新化合物。结果表明，云南维西的粉花绣线菊中得到的生物碱为。tisine型，云南腾冲、湖北神农架、浙江天目山和浙江括苍山的粉花绣线菊中得到的生物碱为hetisine型。此外，使用LC-ESI-MS方法，在浙江括苍山的粉花绣线菊中也检测到。tiine型二帖生物碱。在同一种粉花绣线菊材料中含有。tisine型和hetisine型两种类型的绣线菊生物碱，表明两者在生源合成上有相关性。结合染色体倍性数据，认为粉花绣线菊中生物碱类型及其染色体倍性都与地理分布有直接关系，这利1关系很可能是粉花绣线菊复合群在演化过程中适应环境的结果，而且提示atisile型和hetisile型生物碱的生源合成具有相关性。绣线菊二菇生物碱的抗真菌活性初探不口酿酒酵母对。tisille型二褚生物碱sniramineA/B的生物转化实验表明，在所使用的实验条件下，所试真菌对绣线菊生物碱不敏感。采用梯度稀释法，从绣线菊和滑桃树（Trewia nudiflora）的根际土壤中分离得到21株真菌和130多株放线菌，对这些菌株的抗橙色青霉活性进行了初筛，其中4株真菌和23株放线菌表现出中等强度的抗真菌活性，1株放线菌（Tll48）有很强的活性。绣线菊根际真菌Cw菌株和F16菌株（鉴定为绣线菊盾壳霉）、滑桃树根际放‘线困Tn48菌株(根据16SrDNA序列鉴定为链霉菌属)进行了次生代谢产物的分离定。从CW菌株的发酵提取物中分离鉴宁了化合物acumminatopyrone、麦角甾醇过氧化物和恩镰抱素类抗生素，从绣线菊盾壳霉的发酵提取物中分离鉴定了7个化合物，包括3个麦角澎醇类化合物，一个不饱和长链脂肪酸，一个脑营脂以及两个二酮毗嗓化合物coniospiromideA（23）和coniospiramideB（24），没有发现与二菇生物碱化学结构相关的化合物。从Tll48菌株的发酵提取物中分离得到nonactic acid系列化合物（25-29）以及放线菌酮cycloheximide（30），没有发现美登木素类相关化合物。通过部分绣线菊根际菌株对绣线菊二菇生物碱的生物转化研究、“趋化性”实验、共培养实验，没有发现对绣线菊生物碱非常敏感的根际微生物菌株。但是发现腾冲绣线菊提取物中的非生物碱部分可以明显影响绣线菊盾壳霉的麦角形醇含量。推测绣线菊盾壳霉与粉花绣线菊之间的关系是类似病原菌与宿主的关系。绣线菊化学成分对绣线菊盾壳霉的麦角错醇有明显影响，可能是植物对潜在的病原菌进行化学防御的表现。而绣线菊盾壳霉的麦角品醇虽然受到影响，却没有在菌丝体微观形态和菌落外部形态等方面发生明显改变，体现出对绣线菊防御反应的适应性。滑桃树根际放线菌Tn48菌株产生具有强抗真菌活性的放线菌酮。Tn48菌株能够同时产生nonactic acid和cycloheximide两种类型的抗生素，表明微生物菌株产生次生代谢物受环境等诸多因素的影响。通过抗肿瘤活性测试，表明Tn48菌株的次生代谢产物具有强效抗肿瘤活性。选择了10种培养基对Tn48菌株进行定量的对比发酵，提取物进行LC-MS检测分析、抗橙色青霉活性测试以及抗肿瘤活性测试。 |
| 英文摘要 | To investigate the relativity between the secondary metabolites of plants and their rhizosphere microbes, Spiraea japonica Linn, f., the rhizosphere microbes of S. japonica and Trewia nudiflora L. were used as experimental material, and researches about the chemical components of S. japonica, the rhizosphere microbes of S. japonica and T. nudiflora as well as the influence of the chemical components of S. japonica on its rhizosphere strains were carried out. Thirteen diterpenoid alkaloids were isolated from S. aponica collected from different area, among which 6-hydroxylspirasine IV (3), spiraqine (11), 6-hydroxylspiraqine (12) and N-aminoethyl spiradine A chloride (13) were new ones. My experimental data indicated that alkaloids of S. japonica var. indsa from Wei-Xi were atisine-type, while the alkaloids of S. japonica vax.fortunei from Teng-Chong, 5. japonica from Shen-Nong-Jia, S, japonica var. glabra from Tian-Mu-Shan and Kuo-Cang-Shan were hetisine-type. However, LC-ESI-MS analysis revealed that both atisine- and hetisine-type diterpenoid alkaloids were detected in the extract of S. japonica var. glabra collected from Kuo-Cang-Shan, Zhejiang Province. The co-ocurrence of two types of Spiraea alkaloids in the same S. japonica material indicated the biosynthesis con-elation between them. Combined with the karyotype data, it's suggested that the structure types of alkaloids and the karyotypes of S. japonica complex were related directly to the geographical contribution of S.japonica and such a relationship may be a result of adaptation to the natural environments by S. japonica complex during evolution, and indicated the biosynthetic relationship between the atisine- and hetisine-types of Spiraea alkaloids. The primary antifungal experiments of Spiraea alkaloids and the biotransfonnation of atisin-type spiramine A/B by Saccharomyces cerevisiae showed that on our experiment conditions, the fungi used were no susceptible to Spiraea alkaloids. The rhizosphere fungi and actinomyces of S. japonica and T. nudiflora were isolated by gradient dilution method. The primary antifungal screening of such isolates were carried out against Penicillium avellaneum UC-4376. The chemical components of rhizosphere fungi strains CW, F16 (identified as Coniothyrium spiraeae Miyake) of S. japonica and rhizosphere actinomyces strain Tn48 (identified as Streptomyces spp. based on 16S rDNA) of T. nudiflora were analyzed. Four compounds including acuminatopyrone, ergosterol peroxide and enniatins were isolated from the extract of strain CW. Seven compounds including ergosterols, unsarurated fatty acid, cerebroside and diketopiperazines were isolated from the extract of strain C. spiraeae. No Spiraea alkaloids-similared compounds were isolated or detected. Six compounds consisting of nonactic acid series and cycloheximide were isolated from the extract of strain Tn48. No maytansinoid were detected. During the experiments of biotransformation of Spiraea alkaloids by parts of Spiraea rhizosphere microbes and chemotaxis-similar experiment, no alkaloids-susceptible strains were found. The non-alkaloid fraction of S. japonica from Teng-chong could, however, affect obviously the content of ergosterol in C. spiraeae. It's supposed that such effect represent the chemical defense of plant against potential pathogen. C. spiraeae didn't show exterior symptoms under the absence of ergosterol, suggested the adaptation to defense of S. japonica. On the other hand, the rhizosphere actinomyces strain Tn48 of T. nudijlora produced cycloheximide with strong antifungal activity which may be contributed to the plant defense against other pathogen. These facts could be considered as the proof supporting the interaction and interadaptation between plants and their rhizosphere microbes. Two types of antibiotics including nonactic acid series and cycloheximide produced by strain Tn48 indicated that lots of factors including environment affect the production of secondary metabolites. The antitumor results showed that secondary metabolites of strain Tn48 exhibit strong antitumor activity. The extracts of Tn48 growing on 10 different media were analyzed through LC-MS, antifungal and antitumor test. |
| 语种 | 中文 |
| 公开日期 | 2011-10-25 |
| 页码 | 129 |
| 源URL | [http://ir.kib.ac.cn/handle/151853/776]  |
| 专题 | 昆明植物研究所_昆明植物所硕博研究生毕业学位论文 |
| 推荐引用方式 GB/T 7714 | 范黎明. 两种植物及其根际微生物的化学研究[D]. 中国科学院昆明植物研究所. 中国科学院昆明植物研究所. 2005. |
入库方式： OAI收割
来源：昆明植物研究所
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