微杆菌Sneb159杀线虫活性物质的分离与鉴定

【目的】有关Microbacterium maritypicum在植物线虫生物防治方面的研究较少,探究菌株M. maritypicum Sneb159的杀线虫活性,明确菌株发酵液中具有杀线虫活性的物质,为生物农药的开发提供理论依据。【方法】本研究检测了菌株Sneb159发酵液对大豆胞囊线虫二龄幼虫的触杀活性;并以触杀活性为追踪,采用有机溶剂萃取、硅胶柱层析及半制备高效液相色谱等技术对菌株Sneb159发酵滤液中的活性物质进行分离纯化;采用核磁共振波谱对纯化物进行结构鉴定。【结果】菌株Sneb159具有杀线虫活性,在24 h和48 h处理组中线虫的死亡率均显著高于对照组。从菌株Sneb159发酵滤液中分离得到杀线虫活性物质A6,经结构鉴定确定该物质为苯乙酰胺。【结论】首次发现M. maritypicum对大豆胞囊线虫的触杀作用,并且明确活性物质为苯乙酰胺。结果表明菌株M. maritypicum Sneb159和苯乙酰胺在大豆胞囊线虫的生物防治方面具有较好的应用潜力。     

杀松材线虫细菌的分离、鉴定及其培养条件研究

为了对松萎蔫病进行安全、无污染的生物防治,从青岛大学校园内采集的样品中分离出120株菌。采用浸渍法从中筛选出1株具有较高杀线虫活性的细菌G8-1。杀线虫活性测定结果表明,该菌株发酵上清液48 h后的杀线虫率为90.81%。通过16S rRNA的序列分析,将其鉴定为Flectobacillus rhizosphaerae。通过单因素分析法和正交实验对该菌株的培养条件进行研究,确定最佳培养条件:最适初始pH为8.0,最适培养温度为35℃,最佳接种菌龄为9 h,最佳培养时间为3 d,菌株G8-1发酵上清液的杀线虫率可达到96.31%。对菌株产生的活性物质的稳定性进行初步分析,结果表明该菌株的活性物质都具有较好的热稳定性,并且在弱碱性条件下具有较高的杀线虫活性稳定性。本研究为松材线虫的生物防治提供参考。     

杀线虫放线菌DA09202的鉴定及其活性物质的解析

从海南热带植物园土壤样品中分离获得一株具有较强杀线虫活性的放线菌菌株DA09202,通过形态特征、生理生化特征、16SrDNA序列测定及其系统发育分析,初步鉴定为金色链霉菌。菌株DA09202发酵液采用溶媒萃取、硅胶柱层析、Sephadex LH-20凝胶过滤和制备薄层板层析,从中分离得到杀线虫活性化合物A23-1和A46-2。化合物A23-1经光谱和波谱分析(UV、1H-NMR、13C-NMR、DEPT、1H-1HCOSY、HMBC、HSQC)以及文献对照,鉴定为4′,7-二羟基异黄酮,化合物A46-2的结构正在鉴定中。     

1株根结线虫拮抗放线菌新种的分离与鉴定

从海南东寨港红树林底泥中分离到206株放线菌。利用24孔板法进行抗线虫活性筛选,获得根结线虫拮抗菌HAlll66。该菌株16SrDNA序列与Nocardiopsis trehalosi VKM Ac-942^T相似性最高（98．3％）,并在系统发育树上聚为同一分支,二者DNA—DNA杂交率38．8％。形态和生理生化特征也表现出一定差异。基于表型与分子特征,鉴定菌株HA11166为Nocardiopsis属新种,命名为东寨港拟诺卡氏菌（Nocardiopsis dongzhaigangensis sp. nov. ）。     

龙柏茎叶中抑菌活性成分的分离与鉴定

采用活性追踪和柱层析法,以苹果腐烂病菌、葡萄白腐病菌等为靶标菌,提取、分离、鉴定了龙柏茎叶中抑菌活性成分.结果表明,对龙柏茎叶乙醇提取物的石油醚、乙酸乙酯和正丁醇萃取物活性测定中,石油醚萃取物的活性最高.对石油醚萃取物进行硅胶柱层析,得到一种活性较高的化合物A,对苹果腐烂病菌的EC_(50)为0.862 4 mg/mL.经氢谱(1H-NMR)、碳谱(~(13)C-NMR)和红外等确定了化合物A的结构为4-松香酸.     

滇产与粤产山柰精油化学成分的研究

应用气相色谱/质谱/计算机联用并结合制备衍生物的方法,分别对滇产和粤产山奈(Kaempferiagalanga L.)进行了研究,共鉴定了桂皮酸乙酯、对甲氧基桂皮酸乙酯、龙脑等31个化学成分。除滇产山奈含油量低于粤产山奈外,其精油的化学成分基本相同,只是各化学成分的含量略有不同。     

对叶榕榕果内生细菌分离鉴定及抑菌活性的研究

【目的】为植物-内生细菌的生态关系研究, 以及植物内生细菌资源的利用提供一定的依据。【方法】采用微生物学传统分离培养的方法从对叶榕果实中分离到内生细菌54株, 通过限制性酶切分析(ARDRA)共有16个操作分类单元(Operational taxonomic units, OTUs), 对16株代表菌株16S rDNA序列系统发育分析。【结果】16个菌株都找到了与其相似性最高的菌株, 相似性达到95%?100%。其中6株为芽孢杆菌Bacillus属, 为对叶榕果实内生细菌优势菌属; 3株为Staphylococcus属, 2株为Pseudomonas属, 1株为Serrata属, 1株为非培养细菌的同源菌, 1株为Kocuria属, 1株为Delftia属, 还有1株为Acinetobacter属。【结论】这16株内生菌在系统发育树中明显聚为两大支; 在参与抑菌试验的14株内生菌中, 有13株对受试菌有不同程度的拮抗作用。尤其是其中的芽孢菌属的Swx15和不动杆菌属的Swx25菌株, 抑菌作用较强, 且有较广的抑菌谱性。     

百里香杀螨活性成分的分离及鉴定

为了探讨百里香的杀螨活性成分,以山楂叶螨(Tetranychus viennensis)为供试对象,采用生物活性示踪法从百里香(Thymus mongolicus)乙醇提取物中分离纯化出5种活性成分,其化学结构经MS、1H-NMR、13C-NMR分析鉴定为百里香酚、香芹酚、松油烯-4醇、豆甾醇和β-谷甾醇.采用玻片浸渍法测试了5种化合物对山楂叶螨的触杀活性,结果表明,百里香酚和香芹酚对山楂叶螨有较强的触杀活性,12 h和24 h的触杀LC50值分别为0.103、0.135 mg·mL-1和0.048、0.096 mg·mL-1;松油烯-4-醇也有一定的杀螨活性,12 h和24 h的触杀LC50值分别为0.320和0.231 mg·mL-1;而两种甾醇类化合物β-谷甾醇和豆甾醇对山楂叶螨没有明显的触杀作用.分析认为百里香酚可能是百里香的主要杀螨活性成分之一.     

肉苁蓉中苯乙醇甙类成分的分离和鉴定

从列当科药用植物肉苁蓉(Cistanche deserticola)中分得四个苯乙醇甙类成分,经光谱分析,确定这四个成分是海胆甙(echinacoside),肉苁蓉甙A(cistanoside A),麦角甾甙(acteoside)和2＇-乙酰麦角甾甙(2＇-acetylacteoside)。以上成分在该植物中属首次分得。     

拟双角斯氏线虫共生细菌的分离与鉴定

从我国东北地区采集的拟双角斯氏线虫(Steinernema ceratophorum)肠道内分离到1株具有较强生物活性功能的致病杆菌菌株CB43。形态特征及生理生化特征测定结果表明,CB43菌株与致病杆菌属(Xe-norhabdus)的基本特性相似;对寄主线虫的产量有明显的促进作用,其代谢物对细菌和真菌均有较强的抑制活性,符合线虫共生菌的基本特性和功能。16S rRNA序列及根据16S rRNA序列构建的系统发育树中,CB43菌株与Xenorhabdus budapestensis序列同源性最高,形成一个类群。但CB43菌株不能产生吲哚,在麦芽糖、海藻糖、D-葡萄糖酸和乙酸利用等生化特征与X.budapestensis存在一定的差异,可能是由于菌株的生态差异造成的。根据形态及生理生化特征,结合16S rRNA序列分析,CB43菌株属于Xenorhabdus budapestensis。     

3-caren-5-one from Kaempferia galanga

The rhizomes of Kaempferia galanga afforded a new monoterpene ketone whose structure was elucidated as 3-caren-5-one by spectroscopic means.     

Biochemical and molecular profiling of micropropagated and conventionally grown Kaempferia galanga

Kaempferia galanga L., family Zingiberaceae, is used extensively in the preparation of both traditional and modern medicines. Buds of rhizomes of K. galanga were incubated on Murashige and Skoog (MS) medium supplemented with 1 mg/l benzyladenine and 0.5 mg/l indole-3-acetic acid (IAA) to induce shoot proliferation. Micropropagated plantlets subjected to random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) marker-based molecular profiling revealed uniform banding patterns similar to those of the mother plants. After 2 years of culture in vitro, plantlets were transplanted to the field, evaluated for different agronomic traits, and their rhizomes were subjected to biochemical profiling using quantitative and qualitative assays of essential oils. Gas chromatography and mass spectroscopy analysis of rhizome oil of micropropagated plants showed the presence of 10 major components which were similar to those detected in the mother plants, and accounted for 95.5% of the total compounds. The compound ethyl-p-methoxy cinnamate accounted for 59.5% of the total compounds detected, followed by ethyl cinnamate, 3-carene, pentadecane, borneol, bornyl acetate, delta-selinene, camphor, alpha-piene and immidazole, 5-carbonylvinyl-4-nitro. Biochemical and molecular profiling of micropropagated clones revealed that an in vitro micropropagation protocol could be effectively used for commercial propagation of true-to-type K. galanga for a stable supply of the medicinal compounds present in this plant species.     

皂荚中一种杀鼠活性成分的分离鉴定

采用70%乙醇渗漉提取,正丁醇萃取,常压硅胶柱反复层析,结合杀鼠活性追踪的方法,从豆科植物皂荚(Gleditsia sinensis Lam.)的果实中分离到一个含有7个糖基的三萜皂苷,通过TLC、mp、IR、MS、1H-NMR和13C-NMR等方法鉴定为3-O-β-D-吡喃木糖基-(1→2)-α-L-吡喃阿拉伯糖基-(1→6)-β-D-吡喃葡萄糖基齐墩果酸28-O-β-D-吡喃木糖基-(1→3)-β-D-吡喃木糖基-(1→4)-α-L-吡喃鼠李糖基-(1→2)-[(6S,2E)-6-羟基-2,6-二甲基-2,7-辛二烯醇-(1→6)]-β-D-吡南葡萄糖酯;对小鼠的毒性测定表明,试鼠经口灌胃的耐受量为200～800 mg/kg,LD50为508.31 mg/kg,95%可信区间为449.40～574.95 mg/kg.该三萜皂苷为皂荚杀鼠的活性成分.     

Isolation and Identification of trans-2- and trans-3-Hydroxy-1,8-cineole Glucosides from Alpinia galanga

Three hydroxy-1,8-cineole glucopyranosides, (1R,2R,4S)- and (1S,2S,4R)-trans-2-hydroxy-1,8-cineole β-D-glucopyranosides, and (1R,3S,4S)-trans-3-hydroxy-1,8-cineole β-D-glucopyranoside, which are possible precursors of acetoxy-1,8-cineoles as unique aroma components, were isolated from the rhizomes of greater galangal (Alpinia galanga W.). Their structures were analyzed by FAB-MS and NMR spectrometry, and the absolute configulation of each aglycone was determined by using a GC-MS analysis with a capillary column coated with a chiral stationary phase. The composition of the diastereomers of (1R,2R,4S)- and (1S,2S,4R)- trans-2-hydroxy-1,8-cineole β-D-glucopyranosides in the rhizomes was determined as 3:7 by a GC-MS analysis after preparing the trifluoroacetate derivatives of the glucosides.     

Sucrose,benzylaminopurine and photoperiod effects on in vitro culture of Kaempferia galanga Linn

Abstract

Kaempferia galanga is a monocotyledonous plant of the Zingiberaceae family, commonly utilized for medicinal purposes. This study evaluates the effect of different concentrations of sucrose, benzylaminopurine (BA) and photoperiod on in vitro propagation of K. galanga. Murashige and Skoog (MS) medium supplemented with 5 mg L^?1 BA and 30 g L^?1 sucrose, and a photoperiod with 4 h of light induced the highest shoot proliferation (7.4 ± 1.0 shoots/explant) and the highest number of roots/shoot (31.3 ± 3.2). On the contrary, the maximum shoot height (4.7 ± 0.7 cm) and the highest number of leaves/shoot (4.7 ± 0.2) were obtained from cultures using MS medium supplemented with 30 g L^?1 sucrose but without BA, and exposed to 16 h of light. Hence MS medium supplemented with 5 mg L^?1 BA and 30 g L^?1 sucrose, and incubated under a 4 h light/20 h dark photoperiod was chosen as the optimal protocol for mass multiplication of K. galanga. This in-vitro technique can facilitate the production of a large number of uniform plants of K. galanga, irrespective of the seasonal factor, and could be used as a tool for conservation of the species.     

海南粗榧内生真菌的分离与初步鉴定

从海南粗榧(Cephalotaxus hainanensis L.)健康的树皮、枝、叶中分离得到72株内生真菌.依据其菌落形态和显微特征,将其中68株归为半知茵类,即产孢的33株鉴定为5个属,未产孢的35株中除1株鉴定为丝核菌属外,其余34株暂归为无孢菌群.还有4株属于担子茵纲.结果表明,海南粗榧内生真菌对宿主具有一定的组织专一性,树皮的真菌在数量、种群及其组成方面与枝、叶的相比存在较大差异.     

Polyoxygenated cyclohexane derivatives and other constituents from Kaempferia rotunda L

Methanol extracts of Kaempferia rotunda L. rhizomes yielded seven compounds including six polyoxygenated cyclohexane derivatives identified as (-)-6-acetylzeylenol (1), four acylated derivatives of 1-benzoyloxymethyl-1,6-epoxycyclohexan-2,3,4,5-tetrol (3-6), a Diels-Alder adduct of 3-benzoyl-1-benzoyloxymethylcyclohexa-4,6-dien-2,3-diol (7), and a triacylated derivative of salicin (9). The cyclohexane diepoxide, crotepoxide (8), was also obtained. Spectroscopic methods were used for structure determination. The methanol extract of the rhizomes of K. rotunda and (-)-2-acetyl-4-benzoyl-1-benzoyloxymethyl-1,6-epoxycyclohexan-2,3,4,5-tetrol (2-acetylrotepoxide B; 6), had antifeedant activity against larvae of Spodoptera littoralis. (-)-Zeylenol (2) also showed antifeedant activity, whereas (-)-6-acetylzeylenol (1) was inactive.     

蕨菜黄酮分离鉴定

采用75%的乙醇提取经乙醚脱脂的蕨菜粉末,提取液回收乙醇后以石油醚、氯仿、乙酸乙酯、正丁醇依次萃取粗分段,硅胶柱层析、乙酸乙酯-甲醇系统对乙酸乙酯部位梯度洗脱,对其乙酸乙酯洗脱部分反复凝胶柱层析获得2个黄酮单体,分别鉴定为紫云英苷(1)、芦丁(2),其中化合物1为首次从蕨菜中分离得到。