香蒲水浸提液对铜绿微囊藻的化感作用

通过液液萃取、薄层层析分离出香蒲叶浸提液中化感活性最高的组分,利用气质联用(GC-MS)技术对其化感物质进行分离与结构鉴定来探究香蒲(Typha orientalis)不同组织部位浸提液对铜绿微囊藻(Microcystis aeruginosa)的化感作用及其作用机理。结果表明:香蒲不同部位对铜绿微囊藻的抑制作用强弱不同,抑制活性为叶浸提液根浸提液蒲黄浸提液,其中叶浸提液最高抑制率72.35%;香蒲叶浸提液对铜绿微囊藻的化感作用表现出低浓度(≤5 g·L-1)促进、高浓度(≥10 g·L-1)抑制现象(低促高抑效应),铜绿微囊藻细胞内叶绿素a含量下降,细胞膜透性增大,SOD活性和MDA含量均呈现先增加后下降的趋势;GC-MS结构表征结果表现为香蒲叶中主要化感物质为酚酸、脂肪酸、酯、酮和酰胺类等物质。该结果为香蒲应用于爆发水华的水生态系统治理提供了一定的理论指导意义。     

苜蓿化感作用的初步研究

利用生物测定法和气相色谱 /质谱 (GC/MS)联用技术对中国北方广为种植的紫花苜蓿的化感作用进行了初步研究。结果表明 ,紫花苜蓿全草的水和 95 %乙醇浸提液对玉米和小麦种子萌发均有显著的抑制作用 ,进一步用柱层析对苜蓿浸提液进行了初步分离 ,分离组份对玉米和小麦种子萌发的影响有所变化。抑制玉米萌发的活性物质主要在弱极性的二氯甲烷组份中 ,而抑制小麦萌发的活性物质不仅存在于二氯甲烷组份中 ,也存在于水中。采用GC/MS技术 ,初步推测苜蓿的化感物质是酚和皂甙两类化合物     

野生露珠杜鹃林不同分解层的土壤化感潜力

探明野生杜鹃群落不同层次土壤浸提物的化学成分及其含量差异,为阐明杜鹃群落天然更新障碍与化感作用之间的关系提供基础数据,从化学生态学角度解释群落天然更新障碍的原因。通过种子发芽试验,比较凋落物层(L层)、腐殖质层(H层)和土壤表层(S层)浸提液对自身种子萌发的化感效应,采用内标法结合气相色谱-质谱(GC/MS)联用等技术鉴定土壤浸提液所含的有机化合物。(1)种子发芽试验显示,露珠杜鹃不同土壤层浸提液的化感效应不同。L层的抑制作用最为强烈,其浸提液显著抑制自身种子的萌发。H层和S层对自身具有一定的抑制作用,与对照相比不显著;(2)鉴定了不同层次土壤浸提液中所含的有机化合物,L层、H层和S层均检测出31种组分,其中相对含量大于5%的组分分别有6、8和8种。L层浸提液主要化感成分为丙三醇和棕榈酸,分别达到总量的19.56%和19.17%;H层主要化感成分为2-羟基丙酸和棕榈酸,分别达到总量的14.05%和12.48%;S层土壤浸提物的主要化感成分为棕榈酸和2-羟基乙酸,分别达到总量的14.91%和12.79%。野生露珠杜鹃不同土壤浸提物的化感物质含量以L层最高,L层作为群落土壤化感物质的主要来源;从化感物质组分来分,长链脂肪酸和有机酸类是H层和S层主要的化感物质种类,长链脂肪酸类和醇类是L层主要的化感物质种类。杜鹃群落林下土壤中存在的化感物质可能是其天然更新障碍的重要原因之一。     

芦苇叶水提物的化感活性分析及潜在化感成分的筛选

采用不同溶剂对芦苇﹝Phragmites australis ( Cav.) Trin. ex Steud.﹞叶片水提物进行萃取,并以小麦( Triticum aestivum Linn.)和萝卜( Raphanus sativus Linn.)种子为实验材料对不同萃取物的化感效应进行检测;采用薄层层析和柱层析对抑制作用最强的正丁醇萃取物进行进一步分离,并采用GC-MS法对生物活性较高的组分进行组成成分分析;在此基础上,选择相对含量高并具有代表性的潜在化感成分进行生物活性检测,以期筛选出芦苇叶中的潜在化感成分。结果显示：随质量浓度(20、100和500 mg·L-1)提高,芦苇叶水提物的石油醚、二氯甲烷、乙酸乙酯、正丁醇和水萃取物对小麦和萝卜种子萌发的抑制作用均逐渐增强,其中正丁醇萃取物的抑制作用最强。在正丁醇萃取物的11个组分中,Fr.5、Fr.6、Fr.7、Fr.9和Fr.10组分均能显著抑制萝卜或小麦幼苗的生长,经质量浓度500 mg·L-1各组分处理液处理后萝卜或小麦幼苗的株高、根长及单株鲜质量均显著低于对照(P<0.05)。采用GC-MS法从Fr.5、Fr.6、Fr.7、Fr.9和Fr.10组分中分别鉴定出11、15、15、12和22种成分,分别占各组分总相对含量的83.02%、91.31%、87.36%、97.92%和94.34%,主要成分包括糖类、醇类、有机酸类、酮类、酰胺类和酯类。对14种潜在化感成分生物活性的检测结果显示这些成分对小麦幼苗生长有明显的抑制作用,其中,经质量浓度20 mg·L-1油酸酰胺、棕榈酸甲酯、亚油酸、2-苯乙胺、2-甲基烯丙醇和4-羟基-3-甲氧基苦杏仁酸处理后,小麦幼苗的株高、根长及单株鲜质量显著低于对照。综合分析结果显示：芦苇叶水提物具有较强的化感活性,其潜在的化感成分为油酸酰胺、棕榈酸甲酯、亚油酸、2-苯乙胺、2-甲基烯丙醇和4-羟基-3-甲氧基苦杏仁酸。     

不同作物秸秆对辣椒的化感效应

采用室内生物测定、室外盆栽的实验方法,研究了不同大田作物秸秆浸提液、腐解液及腐解物对辣椒的化感效应。结果表明:秸秆浸提液和腐解液对辣椒种子萌发的化感效应不同,浸提液抑制辣椒种子萌发,且随着浸提液浓度的增加,辣椒发芽指数、胚根长显著或极显著受到抑制,其中以大豆秸秆浸提液的抑制作用最强;秸秆腐解液对辣椒种子的萌发总体上表现为促进效应,腐解物对辣椒的株高、茎粗、鲜质量及根系活力均具有促进作用,且与腐解 物的添加量呈正比;不同秸秆腐解物的化感效应大小依次为玉米秸秆>水稻秸秆>大豆秸秆。     

水稻化感作用及其生理生化特性的研究

选用具有强化感作用的6个水稻品种为供体,大田稗草为受体,研究了水稻化感作用及生理生化特性,结果表明,提高水稻叶片浸提液浓度,可以相应增强对稗草生长的抑制效果,这种抑制效果与杂草的种植密度呈负相关;化感水稻叶片浸提液能显著抑制物质稗草体内超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的活性,从而影响其生长;苯丙氨酸氨解酶（PAL）的活性大小与酚类物质的含量吴正相关;多种酚类物质的化感作用之间可能是增效的,也可能是拮抗的。     

栗与美国板栗化感作用的比较

以种子发芽率和发芽势为种子萌发参数, 以胚根、胚芽长度和鲜重变化为幼苗生长参数, 研究了栗(Castanea mollissima)叶水提取物对双子叶植物莴苣(Lactuca sativa)、萝卜(Raphanus sativus)、黄瓜(Cucumis sativus), 和单子叶植物洋葱(Allium cepa)、水稻(Oryza sativa)、小麦(Triticum aestivum)的化感作用; 比较了美国板栗(Castanea dentata)与栗叶水提取物及经X-5大孔树脂分离获得的11个洗脱组分间的化感作用强弱; 利用液相色谱-质谱联用技术结合标准物质的反证实验, 解析了美国板栗与栗化感作用最强分离组分的物质结构。研究结果表明, 栗叶化感效应强于美国板栗; 美国板栗与栗化感作用最强分离组分中(5:5洗脱组分)存在绿原酸、对羟基苯甲酸、原儿茶酸和没食子酸等物质。在利用栗基因恢复美国板栗种群时, 有必要考虑美国板栗与栗植物化学生态特性的差异。     

黄瓜种子及其萌发期的化感作用研究

以大白菜、萝卜、番茄和黄瓜种子为受体,采用实验室培养皿种子发芽生物测试法研究了黄瓜种子浸提液、种子萌发、胚根和芽苗分泌物、芽苗腐解物和芽苗浸提液的化感效应。结果表明:(1)黄瓜种子浸提液对大白菜、萝卜、番茄和黄瓜种子萌发均有化感抑制作用,即黄瓜种子内含有某些化感抑制物质。(2)在水浸提过的黄瓜种子萌发过程中,它不仅对其近邻套种的大白菜、萝卜和番茄种子萌发产生化感抑制作用,而且其胚根和芽苗分泌物对后茬播种的4种蔬菜种子发芽也表现出不同程度的化感抑制作用;黄瓜芽苗腐解物和芽苗水浸提液也对各受体蔬菜种子发芽与生长产生不同程度的化感抑制作用,且随着腐解芽苗量的增加或浸提液浓度的升高,各受体蔬菜种子的发芽指标值、化感效应指数值和综合效应值随之降低。(3)黄瓜种子浸提液及芽苗各器官的化感物质对黄瓜种子的萌发与生长产生了自毒作用,且黄瓜芽苗腐解物、芽苗浸提液、胚根及芽苗分泌物对受体黄瓜的自毒作用均为最大。研究发现,黄瓜种子浸提液、种子萌发时期以及芽苗各器官的化感物质主要是通过抑制受体胚根的生长而起化感抑制作用,即受体蔬菜种子胚根对化感效应最为敏感;因黄瓜种子及萌发期释放化感物质的途径有所不同,导致受体大白菜、萝卜、黄瓜和番茄的化感响应也不相同;在黄瓜种子萌发和芽苗生长的早期,化感物质即开始在芽苗体内进行合成与积累,一部分可通过胚根和芽苗分泌途径释放到环境中,另一部分可通过芽苗腐解途径释放化感物质,并对受体蔬菜种子萌发与生长表现出较强的化感抑制作用。     

胜红蓟化感作用研究 Ⅸ.主要化感物质在土壤中的转化

在田间条件下,胜红蓟挥发油能抑制花生的出苗和生长发育,进一步用高效液相色谱（HPLC）研究证实;胜红蓟挥发油中的主要化感物质胜红蓟素在土壤中的转化与土壤的有机质和营养元素水平显著相关,在高有机质和营养元素水平的土壤中,胜红蓟素先聚合形成二聚体,26d后又解聚成胜红蓟素,然后逐步降解成苯甲酸类,2-甲基丙酸和乙酸等小分子,而在低有机质和营养元素水平的土壤中,胜红蓟素不经过二聚化过程,而是直接降解成小分子,经液质（LC/MS）和核磁共振（NMR）等技术分离鉴定了胜红蓟素二聚体的结构,生物测定表明：胜红蓟素二聚体对花生和黑牧草的生长没有抑制活性。     

大豆地上部水浸液的化感作用及化感物质的鉴定

研究了重迎茬大豆花荚期植株地上部水浸液及其提取物对大豆种子萌发及幼苗生长的化感作用 ,并鉴定了有明显化感作用组分的物质。结果表明 :与对照组比 ,0 .1 0 g DW/ ml重茬大豆水浸液显著地抑制了大豆胚根生长 ;3 .75× 1 0 - 2g DW/ ml、7.5 0× 1 0 - 2 g DW/ ml正茬、重茬、迎茬大豆水浸液对砂培大豆苗的生长有显著的化感抑制作用 ;重茬、正茬大豆水浸液乙酸乙酯提取的酸性组分、XAD-4树脂提取组分 ,在 0 .6～ 1 .2 g DW/ ml浓度下 ,对大豆种子萌发初期的萌发率及萌发过程中的胚根生长亦表现出显著的化感抑制作用 ;重茬水浸液乙酸乙酯提取的中性、碱性组分在 1 .2 g DW/ ml浓度下 ,对大豆胚根生长有显著抑制作用。上述各组分化感作用大小随组分浓度增加而增加。在相同浓度处理下水浸液及其提取物的化感抑制作用有重茬大于正茬的趋势 ,统计检验差异均不显著。 GC-MS鉴定结果显示 ,正茬、重茬乙酸乙酯酸性组分、XAD-4树脂提取组分中的有机化合物主要是 :丁二酸、苯甲酸、邻羟基苯甲酸、4-羟基苯甲酸、对羟基肉桂酸、2 -甲基苯酚、2 -甲氧基 -4-乙烯基苯酚、间苯三酚、2 -甲基苯甲醛、3 -甲基苯甲醛、5 ,6,7,7a-四氢 -4,4,7a-三甲基 -2 (4 H) -苯并呋喃酮、2 ,4-戊二烯腈等。     

Studies on the oxidation products from 2,4-diaminotoluene by hydrogen peroxide and their mutagenicities. II

2,4-Diaminotoluene (DAT) was reacted with hydrogen peroxide at room temperature for 2 days, and the resulting red precipitates were separated into 5 fractions on silica gel column chromatography. On the gas chromatographic (GC) study, the first fraction (Fr. 1), which is mutagenic (1425 and 1391 revertants/micrograms in the absence and presence of S9 respectively) in Salmonella typhimurium TA98, contained several peaks. Fr. 1 was further separated into 4 subfractions (Fr. 1-I-Fr. 1-IV) by silica gel column chromatography. The red crystals were separated from Fr. 1-III and the structure of the compound was determined to be 1,8-diamino-2,7-dimethylphenazine from physicochemical and chemical evidence. Further, o-nitro-p-toluidine, p-nitro-o-toluidine, 3,3'-diamino-4,4'-dimethylazobenzene and 3,3'-diamino-4,4'-dimethylazoxybenzene were identified with authentic and synthesized samples by gas chromatography/mass spectrometry. These compounds without nitrotoluidines were mutagenic, and phenazine, azo and azoxy compounds induced 49, 301 and 245 revertants/nmole in Salmonella typhimurium TA98 with 25 microliters S9 per plate, respectively.     

铜藻岩藻黄素提取及纯化工艺研究

为了对岩藻黄素的提取、纯化进行系统研究,进而为高纯度岩藻黄素的工业化生产提供研究基础,筛选了适用于提取铜藻（Sargassum horneri）鲜藻中岩藻黄素的有机溶剂,并通过单因素实验和正交实验确定了最佳的提取溶剂浓度、提取温度、提取时间、料液比等工艺参数。随后采用硅胶柱层析法进行纯化,并通过单因素实验确定了最佳的硅胶柱床高度、上样量和洗脱流速。最后采用制备液相法对经层析纯化的岩藻黄素进一步纯化。结果表明,有机溶剂萃取的最佳工艺条件为：甲醇浓度90%,提取温度50 ℃,提取时间1 h,料液比1∶10,此条件下岩藻黄素提取率达到(0.258 9±0.003 6) mg·g-1鲜重（FW）［(1.078 8±0.015 0) mg·g-1干重（DW）］。硅胶柱层析的最佳工艺条件为：硅胶柱床高度10 cm,上样量6 g,洗脱流速10 mL·min-1,此条件下岩藻黄素得率为0.176 5 mg·g-1FW（0.735 3 mg·g-1 DW）,纯度为87.01%±0.88%。经制备液相进一步纯化后,岩藻黄素得率为0.127 1 mg·g-1 FW（0.529 4 mg·g-1 DW）,纯度为99.27%±0.22%。研究所用工艺简单,岩藻黄素得率高,为高纯度岩藻黄素的制备提供了实验基础。     

马兰化学成分研究 （Ⅱ）

采用冷渗漉提取的方法提取马兰的化学成分。经硅胶柱色谱和Sephadex LH-20进行分离纯化,通过理化方法和波谱数据分析进行结构鉴定。从80%乙醇冷渗漉提取物的水不溶物中分离并鉴定了12个化合物,分别为正十六烷酸(1)、6-羟基-桉烷-4(14)-烯(2)、β-谷甾醇(3)、α-菠菜甾醇(4)、香草醛(5)、β-20(21),24-二烯-达玛烷-3-酮(6)、豆甾醇(7)、木栓酮(8)、羽扇豆酮(9)、α-香树脂醇(10)、表木栓醇(11)、神经酰胺(12)。化合物2、6、9、12均为首次从该种植物中分得。     

Studies on the oxidation products from 2,4-diaminotoluene by hydrogen peroxide and their mutagenicities. II.

2,4-Diaminotoluene (DAT) was reacted with hydrogen peroxide at room temperature for 2 days, and the resulting red precipitates were separated into 5 fractions on silica gel column chromatography. On the gas chromatographic (GC) study, the first fraction (Fr. 1), which is mutagenic (1425 and 1391 revertants/μg in the absence and presence of S9 respectively) in Salmonella typhimurium TA98, contained several peaks. Fr. 1 was further separated into 4 subfractions (Fr. 1-I-Fr. 1-IV) by silica gel column chromatography. The red crystals were separated from Fr. 1-III and the structure of the compound was determined to be 1,8-diamino-2,7-dimethylphenazine from physicochemical and chemical evidence.Further, o-nitro-p-toluidine, p-nitro-o-toluidine, 3,3′-diamino-4,4′-dimethylazobenzene and 3,3′ diamino-4,4′-dimethylazoxybenzene were identified with authentic and synthesized samples by gas chromatography/mass spectrometry. These compounds without nitrotoluidines were mutagenic, and phenazine, azo and azoxy compounds induced 49, 301 and 245 revertants/nmole in Salmonella typhimurium TA98 with 25 μl S9 per plate, respectively.     

南方红豆杉毛状根诱导体系的建立及毛状根中 紫杉醇的分离纯化研究

目的:研究发根农杆菌对南方红豆杉转化的转化体系和毛状根中紫杉醇的提取纯化工艺;方法:采用发根农杆菌对南方红豆杉不同外植体进行诱导,并对影响毛状根转化的因素进行分析。采用超临界CO2流体萃取法、硅胶柱色谱法以及高效液相法对毛状根中的紫杉醇进行分离提纯;结果:成功构建了南方红豆杉毛状根诱导体系。其中农杆菌的种类、外植体类型、预培养和共培养时间、培养基中激素浓度均影响毛状根的转化率。毛状根经高压纸电泳检测显示表达冠瘿碱。毛状根中紫杉醇经超临界CO2法提取及硅胶柱色谱法分离提纯,采用高效液相法分析显示其纯度可达到98%。结论:发根农杆菌诱导南方红豆杉产生的毛状根中可产生紫杉醇,可作为紫杉醇的主要来源。     

Plant secondary substances and insects behaviour

Allelochemicals are storing in different location in plant tissues as inactive form. Number of identified compounds may now exceed 100,000. Environmental factors have an effect on allelochemicals concentration in plants. Many allelochemicals classified as toxins or deterrents for herbivorous insects. Allelochemicals play a major role in feeding or ovipositing stimulants for some specialist insects. Consumption and assimilation of herbivorous insects had affected by the type of allelochemicals in host plants. Allelochemicals have an acute or chronic toxicity on herbivorous insects. Most specialist herbivorous insects rely heavily of ingested plant allelochemicals. Plant allelochemicals may influence an insect's susceptibility to pathogens such as bacteria, fungi and nematode. Specialists herbivorous insect can be using the allelochemicals in their host plants as protection against natural enemies. Some herbivorous insects are synthesising the aggregation, attracting, alarm or mating pheromone from the allelochemicals in their host plants.     

Separation and Qualitative Study of Mentha asiatica Boriss. Essential Oil Components by prep-GC,GC-QTOF-MS,NMR and Prediction of Bioactive Compounds

In order to understand the material basis of wild Mentha asiatica Boris. in Xinjiang, the chemical constituents of essential oil extracted from aerial parts of this plant were studied. A total 52 components were detected and 45 compounds were identified. First of all, the essential oil was separated by silica gel column chromatography, and divided into several parts according to the results of thin layer chromatography. Eight fractions were obtained, and then each fragment was preliminarily screened for antibacterial activity. It was found that all eight fragments had certain antibacterial activity in different level. Then the fractions were subjected to preparative gas chromatography (prep-GC) for further isolation. Ten compounds were identified by ¹³C-NMR, ¹H-NMR and gas chromatography-quadrupole time of flight-Mass spectrometry (GC-QTOF-MS). They are sabinene, limonene and β-caryophyllene, (1R*,3S*,5R*)-sabinyl acetate, piperitone oxide, rotundifolone, thymol, piperitone, 4-hydroxypiperiditone, cedrol. After screened by bioautography, 4-hydroxypiperone and thymol were showed best antibacterial activity. The inhibitory effects of the two isolated compounds on Candida albicans and their related mechanisms were studied. The results showed that, 4-hydroxypiperone and thymol significantly reduced ergosterol content on the surface of Candida albicans cell membrane in a dose-dependent manner. This work has accumulated experience for the development and utilization of Xinjiang characteristic medicinal plant resources and new drug research and development, and provided scientific basis and support for the later research and development of Mentha asiatica Boris.     

Development of an analytical approach for identification and quantification of 5-α-androst-16-en-3-one in human milk

A method was developed for the quantification of 5-α-androst-16-en-3-one in human breast milk based on application of a stable isotope dilution assay using 5α-androst-16-en-3-one-6, 6-d₂. The procedure includes extraction of the human milk by hexane with subsequent clean-up of the obtained extract by gel permeation and silica gel column chromatography. The extracted samples were analyzed by gas chromatography–mass spectrometry. Using this method 5-α-androst-16-en-3-one could be identified and for the first time quantified in a concentration range of 26–155 ng/kg in human milk.     

大豆异黄酮的分离与纯化

天然大豆异黄酮作为健康食品在防治骨质疏松和癌症方面具有一定功效。由于化合物结构相似,异黄酮甙元特别是高纯度黄豆黄素（glycitein）的获得有一定难度,文献报道大多是通过盐酸水解异黄酮甙的方法获得,而这种方法对环境污染和工厂生产设备腐蚀较大。本文报道了用醇溶剂进行固相提取以及硅胶柱色谱方对含有三种大豆异黄酮甙元的混合物产品进行分离。通过低成本和无环境污染的固相提取方法得到纯度为97％的黄豆黄素和纯度超过95％的大豆黄素（daidzein）;95％纯度的另一种大豆甙元金雀异黄素（genistein）则通过硅胶柱色谱分离得到。应用硅胶柱色谱,一次性分离了一种含有两个异黄酮甙：大豆甙（daidzin）和黄豆甙（glycitin）的产品。