中药甘松中的白藜芦醇低聚体类成分

采用硅胶、凝胶等柱色谱以及高效液相制备色谱等现代分离方法,从甘松(Nardostachys chinensis Batal.)根及根茎70%乙醇冷浸渗漉提取液的乙酸乙酯萃取部位中分离得到5个白藜芦醇低聚体类化合物,通过物理化学性质和波谱学方法鉴定其化学结构分别为:-α-viniferin(1)、kobophenol A(2)、蛇葡萄素A(ampelopsin A,3)、isohopeaphenol(4)、grandiphenol A(5)。化合物1~5均为首次从该属植物中分离得到。     

甘松属植物化学和药理学研究进展

对甘松属植物的化学和药理学研究进展进行了综述。其主要成分为倍半萜类;主要药理作用有镇静、平滑肌解痉、降压、抗心律失常、增强耐缺氧能力、抗心肌缺血、抑菌等。     

大花甘松(Nardostachys grandiflora DC.)和中华甘松(N.chinensis Batalin)的精油成分及其在香料上的应用研究

本文报道滇产大花甘松和蜀产中华甘松精油的化学成分。利用气相色谱、光谱数据、化学降解和衍生物的制备,从大花甘松精油中鉴定了五种成分:甘松醇(Ⅰ)(69.4％),△~(9,10)-马兜铃烯(Ⅶ)(0.2％),△~(1,10)-马兜铃烯(Ⅷ)(2％),β-橄榄烯(Ⅸ)(2.9％)和1,2,9,10-四去氢马兜铃烯(Ⅳ)(2.9％)。中华甘松精油中上述五种成分的含量分别为:Ⅰ(8％),Ⅶ(2.1％),Ⅷ(8.6％),Ⅸ(32.8％),Ⅳ(13.1％)。根据植物的形态、地理分布和精油中马兜铃烷型倍半萜化合物的含量和转换关系看来,甘松香属几种植物中似乎可认为大花甘松为其原始种,并由大花甘松进化到中华甘松和匙叶甘松。 甘松酯(Ⅵ)具浓郁持久的木香香气,甘松酮(Ⅲ)具独特的檀香香气和能与玫瑰、铃兰协调的特点。该二化合物特别适于调制檀香型、玫瑰檀香型和馥奇型等香精,并具有良好的定香作用,是一类有价值的香料。     

甘松油生产技术

一、甘松与甘松油简介甘松属败酱科甘松香属多年生草本植物。分为中华甘松(Nardostachys Chinensis Batalin)和长匙甘松(Nardostachys jtamansi),是一种珍贵的经济植物,其主要分布我国四川、云南、甘肃、青海、西藏等省(区),以及印度、尼泊尔等国家。生长于阴湿山坡及湿     

中国半扇毛螨属两新种记述(蜱螨亚纲,肉食螨科)

记述肉食螨科Cheyletidae,半扇毛螨属Hemicheyletia 的2新种:采自贵阳储藏中药材连翘内的多毛半扇毛螨Hemicheyletiapolyseta sp.nov.,采自贵阳储藏中药材甘松内的贵阳半扇毛螨Hemicheyletia guiyangensis sp.nov..标本保存在贵州大学昆虫研究所.     

甘产野生资源植物—中华甘松

中华甘松是一种野生植物,是甘肃、四川、青海三省交界处特有珍贵资源植物。它的根茎自古以来为民间驱虫、熏室之物,具有理气止痛,开郁醒脾,健胃之功,是我国常用中药。还能提取挥发油——甘松油,是香料工业主要原料之一,在皂用、化妆香精中均可使用,其蒸馏残渣又可做农业杀虫剂和良好的饲料,以及制香的添加剂。已往,仅四川省生产甘松油,用于香料工业,我省资源尚未利用,仍处于自生自灭。为充分开发利用甘肃优势资源植物,我们对甘产中华甘松的形态、分布地区,生态条件以及对精油进行测试研究,现报告如下:     

浙贝母鲨烯合酶核心功能区基因克隆与序列分析

目的:克隆浙贝母次级代谢途径的关键酶——鲨烯合酶(SQS)的基因,有助于对次生代谢产物的合成进行调控。方法:根据NCBI上公布的8种植物SQS蛋白序列,分析高度保守区并设计简并引物。从浙贝母幼根中提取总RNA,利用RT-PCR技术成功扩增出高度保守区基因片段。再利用GSP引物和RACE技术中克隆出3’末端片段。结果:获得浙贝母SQS 1 122bp核苷酸序列,编码374个氨基酸。Protein-Blast比对发现与印度甘松、柴胡、马铃薯、拟南芥、紫杉五种植物的同源性分别达到76%、74%、75%、74%、73%,包含了底物结合位点、Mg2+结合位点、催化活性位点等。该序列在Genbank中的登录号为:KF551097。结论:通过蛋白质特征区、保守区以及进化树分析,初步确定该基因为SQS基因。这是首次从药用植物浙贝母中克隆得到甲羟戊酸途径关键酶SQS的基因核心功能片段,为浙贝母次生代谢工程研究打下良好的基础。     

浙贝母角鲨烯合酶全长cDNA的克隆及功能分析

鲨烯是甾醇和其他三萜类化合物的关键代谢中间体,其生物合成由鲨烯合酶（squalene synthase,SQS）催化,该酶将2分子法呢基焦磷酸转化为鲨烯。浙贝母异甾体生物碱的生物合成途径与三萜类化合物类似。在本研究中,基于cDNA末端的快速扩增（RACE）技术克隆了浙贝母鲨烯合酶 (FtSQS）基因的全长cDNA,GenBank登录号为KF551097.2。通过生物信息学方法对FtSQS进行详细表征,包括保守区检测、序列同源分析、二级和三级结构预测及系统发育树分析。结果表明,其开放阅读框（ORF）为1 230 bp,编码409个氨基酸,FtSQS氨基酸序列与印度甘松、截形苜蓿、紫衫、马铃薯、柴胡、金铁锁和拟南芥的SQS氨基酸同源性分别达到73.84%、73.23%、72.24%、70.66%、70.66%、69.44%和68.14%。启动子分析表明,FtSQS的5′上游区域具有与生理和环境因素相关的各种潜在因素。为了获得可溶性FtSQS表达,从羧基末端截断24个疏水氨基酸,构建了原核表达载体pGEX-2T-FtSQSΔTM,并在大肠杆菌BL21(DE3)中表达。SDS-PAGE检测到约66 kD的重组FtSQSΔTM蛋白。体外酶促反应证明,FtSQS可以催化FPP转化成鲨烯。qRT-PCR分析FtSQS mRNA在叶中的表达量最高,茎、根次之,而在鳞茎中表达水平最低。这提示,叶子是浙贝母碱生物合成的主要活性器官。FtSQS的鉴定及功能研究为浙贝母次生代谢产物的研究提供了重要依据。     

外源多胺对铜胁迫下水鳖叶片多胺代谢、抗氧化系统和矿质营养元素的影响

采用营养液水培的方法,研究了外源亚精胺(Spd)和精胺(Spm)对Cu胁迫下水鳖叶片3种形态多胺(PAs)、抗氧化系统及营养元素的影响。结果表明:(1)Cu胁迫使水鳖叶片腐胺(Put)急剧积累,Spd和Spm明显下降,从而使(Spd+Spm)/Put比值也随之下降。外源Spd和Spm显著或极显著逆转Cu诱导的PAs变化,抑制Put的积累,缓解Spd和Spm的下降,从而提高了(Spd+Spm)/Put比值。(2)外源Spd和Spm抑制了Cu胁迫诱导的多胺氧化酶(PAO)的增加,缓解了二胺氧化酶(DAO)的下降。(3)与单一Cu胁迫相比,Spd和Spm显著或极显著提高了超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)活性和抗坏血酸(AsA)、谷胱甘肽(GSH)、游离脯氨酸(Pro)含量,从而降低了超氧阴离子(O2.-)产生速率和过氧化氢(H2O2)含量,极显著降低了丙二醛(MDA)含量,缓解了Cu诱导的氧化胁迫。(4)外源Spd和Spm显著或极显著缓解了Cu胁迫下矿质营养元素吸收平衡的紊乱。以上结果均说明了外施Spd和Spm可增加水鳖对Cu胁迫的耐受性。     

中国兰科植物新资料

报道了中国兰科2个新记录属及5个新记录种:固唇兰属(Plocoglottis)、栖林兰属(Drymoda)、滇南固唇兰(Plocoglottis bokorensis)、栖林兰(Drymoda siamensis)、泰国牛角兰(Ceratostylis siamensis)、景洪白蝶兰(Pecteilis hawkesiana)、三脉贝母兰(Coelogyne trinervis)、浮萍毛兰(Eria spirodela).对2个属及5个种进行了特征描述,并对比了近似种.     

Flower-inducing and -inhibiting activities of Phloem Exudate from Cotyledons of Pharbitis Seedlings

The flower-inducing and -inhibiting activities of phloem exudate (PE) prepared from cotyledons of Pharbitis seedlings were examined, using apex cultures in vitro from Pharbitis as a bioassay system.The PE was prepared from photoperiodically-induced cotyledons (SD-PE). The SD-PE was subjected to the following fractionations: When the SD-PE was extracted with CHCl₃ and then ethyl acetate, the inducing activity was located in the final aqueous fraction. The activity was localized in the diffusate when the aqueous fraction was dialyzed (molecular weight cut off was 10,000). The diffusate was fractionated by ion exchange chromatography, and flower-inducing activity was found in the fraction adsorbed onto anion exchange resin. When the fraction was applied to a Sep-Pak C18 cartridge, the activity eluted with 25% MeOH. As a result of the above fractionation, activity was increased about 30-fold.The nature of the flower-inhibiting activity of the PE taken from cotyledons exposed to continuous-light conditions was examined (CL-PE). The inhibiting activity was decreased as the cotyledons were exposed to longer dark periods; it appeared to be heat-stable. The CL-PE also inhibited flowering in Lemna. The CL-PE was subjected to the following fractionations: When the CL-PE was extracted with CHCl₃ and ethyl acetate, activity was located in the final aqueous fraction. Activity was localized in the diffusate when the aqueous fraction was dialyzed (molecular weight cut off was 10,000). When the diffusate was fractionated by ion exchange chromatography, the activity was found in the flow-through fraction. When the fraction was applied to a hydroxyapatite cartridge, the activity eluted with 25 mM sodium phosphate buffer. When the fraction was re-dialyzed (molecular weight cut off was 1,000), the diffusate contained the activity. As a result of the above fractionation, activity was increased about 10-fold.     

产碱性蛋白酶芽孢杆菌的鉴定

通过测量比较在碱性蛋白平板上产生的蛋白水解圈直径,从土壤中筛选到一株高产蛋白酶菌株Bacillus sp.HFBL0079,根据生理生化特性、16S rDNA序列,鉴定为B.amyloliquefaciens。其最适培养温度为35°C-37°C,最适生长pH 8.0,在特定培养条件下16 h达到稳定期,菌体生长和蛋白酶合成同步进行。以大豆分离蛋白为氮源时发酵液具有最高酶活。发酵液在pH 10时具有最高酶活,表明为碱性蛋白酶。该菌株产生的碱性蛋白酶可水解多种天然蛋白质,对胶原蛋白水解度高于其他蛋白质,对羽毛角蛋白也有一定水解能力,提示该酶具有一定新颖性。     

一株产乙酰酯酶细菌筛选、鉴定及酶学性质研究

【目的】利用筛选培养基,从肉牛瘤胃液中分离筛选产乙酰酯酶的细菌菌株,并研究菌株的产酶特征。【方法】利用厌氧培养技术,以木质素为唯一碳源,筛选并驯化所得菌株。根据菌株16S rDNA序列分析、革兰氏染色、伊红美蓝培养基培养、甲基红试验和柠檬酸盐利用试验,鉴定菌株。采用对-硝基苯乙酯测定酶活力。【结果】筛选得到产乙酰酯酶活力较高细菌菌株RB1,初步鉴定为Escherichia coli。菌株RB1的生长曲线表明,0 42 h为菌株的延迟期,42 60 h为菌株的对数期,60 66 h为菌株的稳定期,66 86 h为菌株的衰亡期。菌株所产乙酰酯酶最适温度为40°C,最适pH为8.0,在最适温度与pH条件下,培养基中添加玉米秸秆粉,乙酰酯酶最高酶活力达到0.52 U/mL。【结论】筛选获得产乙酰酯酶的细菌菌株RB1,其乙酰酯酶活力高于已报道的菌株,是一株具有研究和应用潜力的产乙酰酯酶的菌株。     

BrdU抗血清制备和应用的研究——Ⅰ.高度特异的BrdU抗血清的制备与特性

半抗原BrU通过与BSA偶联制备了完全抗原,经过光吸收、SDS聚丙烯酰胺凝胶电泳和琼脂糖凝胶电泳的测定表明,核苷-蛋白质复合物符合制备的要求,每个BSA上估计大约平均有10.3个BrU。用常规免疫的方法获得兔抗BrdU的抗血清,与BrU-EA的双向扩散效价高达32。抗血清稀释128万倍时仍可见明显的ELISA阳性反应。与以前所报道的BrdU抗血清不同,该抗血清具有高水平的识别能力,已达到BrdU单克隆抗体的识别水平,无须纯化即可用于染色体及核酸的研究。     

濒危植物秦岭冷杉补光育苗技术研究

秦岭冷杉种子生活力差、成苗率低(2.2%)是制约种群恢复的薄弱环节。通过对播种后苗期2年连续补光处理,幼苗表现出良好的生长效应,苗高10.5 cm(对照为7.3 cm),地径4.5 mm(对照为2.7 mm),保存率88.6%(对照66.4%),延长了幼苗生长期,增加了苗木生物量积累,加快了苗木培育进程。     

辣根过氧化物酶同工酶C基因在巴斯德毕赤酵母中的克隆与鉴定

目的：克隆辣根过氧化物酶同工酶C基因,为此基因的表达作准备。方法：用PCR方法从辣根的总DNA中扩增得到一种辣根过氧化物酶同工酶C基因HRPC2,通过PCR的方法去除内含子后连接到pMD18-T载体上,测序证明正确后,用限制性内切酶切下目的基因,插入到巴斯德毕赤酵母表达载体pPIC9K中,构建成重组质粒pC2EX9K。再将辣根过氧化物酶同工酶C基因在毕赤酵母中进行克隆、鉴定。结果：重组质粒pC2EX9K转化毕赤酵母后,经PCR鉴定,证明形成了目的基因的克隆。结论：应用毕赤酵母作为受体菌,pPIC9K为载体,成功克隆了HRPC2。     

Effect of phenolic compounds on cellulose degradation by some white rot basidiomycetes

Abstract Cellulose degradation by several white rot fungi was investigated. In most fungi cellulase production was stimulated by lignin-related phenolics. Detailed investigation of Tremetes versicolor showed that this stimulation was not directly effected by phenols but was due to an indirect induction. The phenol was oxidized by laccase to quinone. The quinone was then reduced by the enzyme cellobiose: quinone-oxidoreductase while cellobiono-lactone was formed from cellobiose. The cellobiono-lactone was responsible for the increased cellulase production in submerged cultures with cellulose as the sole carbon source.     

白腐菌紫外诱变选育高产漆酶突变株及其产酶条件研究

以白腐菌为出发菌株,利用紫外线(UV)进行诱变,筛选高产漆酶突变菌株。通过测定致死率绘制出发菌株的致死曲线,采用PDA-RBBR平板变色法进行初筛,ABTS检测酶活对突变株进行摇瓶复筛。结果表明:利用15 w紫外灯在照射距离为30 cm,照射时间为120 s,致死率为72.1%的条件下进行诱变处理,获得一株高产菌株,其酶活提高79.54%,经过5代传代培养,未见酶活下降,具有较好的遗传稳定性,进一步研究了初始pH值,接种量和培养基装液量等对诱变菌株产酶的影响,结果表明在最佳的培养条件pH值6.0,15%的装液量于28℃下,酶活达214.9 U/L。     

Marginal bone loss around axial and straight implants supported with prefabricated SFI-Bar with mandibular overdentures

To assess the role of prefabricated SFI-Bar in peri-implant bone loss around immediately axially loaded and straight implants. This study comprised of 40 complete denture wearer patients who received two axially parallel implants connected by SFI-Bars in group I and two 15° mesially tilted implants connected by SFI-Bars in group II. Peri- implant bone loss (PiBL) was measured at 1 year, 2 years and 3 years. The mean PiBL at 1 year in group I was 0.21 mm and I group II was 0.22, at 2 years in group I was 0.26 mm and in group II was 0.23 mm and at 3 years, in group I was 0.29 mm and in group II was 0.34 mm. The difference was significant at 3 years (P< 0.05). The mean mesial PIBL at 1 year in group I was 0.18 mm, in group II was 0.20 mm, at 2 years in group I was 0.19 mm and in group II was 0.07 mm and at 3 years, in group I was 0.25 mm and in group II was 0.29 mm. The difference found to be significant in each time duration in both groups (P< 0.05).The mean distal PIBL at 1 year in group I was 0.23 mm, in group II was 0.22 mm, at 2 years in group I was 0.33 mm and in group II was 0.39 mm and at 3 years, in group I was 0.34 mm and in group II was 0.39 mm. The difference found to be significant at 2 and 3 years in both groups (P< 0.05). Authors found that mandibular overdentures retained with Prefabricated SFI-Bar with axial and straight inserted implants may be useful in patients with reduced bone height.     

生长激素释放激素和人血清白蛋白融合蛋白的克隆表达

目的:通过与人血清白蛋白(HSA)融合,延长生长激素释放激素(GHRH)在体内的半衰期。方法:根据毕赤酵母偏爱密码子重新设计GHRH的核酸序列,并通过化学合成和重叠PCR法将GHRH的N端与HSA的C端通过一个11肽的接头连接,获得GHRH和HSA融合的全长基因序列。构建pPIC9-HSA-GHRH表达载体,电击转化毕赤酵母GS115感受态细胞,通过表型筛选和诱导表达实验得到蛋白表达工程菌,对表达产物进行分离纯化和生物学活性分析。结果:克隆了HSA-GHRH融合基因,构建了pPIC9-HSA-GHRH融合表达载体;电击转化后通过表型筛选和诱导表达实验得到蛋白表达工程菌;经分离纯化后,对表达产物的生物学活性分析显示其在体内有促进生长的作用。结论:与人血清白蛋白的融合有效地提高了GHRH的表达水平,并延长了GHRH的半衰期。