不同品种酿酒葡萄表皮微生物群落多样性分析

葡萄酒酿造是多种微生物参与代谢的过程,其中的微生物在没有外源接种的情况下通常认为源自酿酒葡萄本身。应用高通量测序技术,分析沙城地区不同品种酿酒葡萄表皮的微生物群落,旨在从源头上了解葡萄酒酿造原料的品质,为研究酿酒葡萄微生物对酿酒品质的影响提供理论依据。结果显示,无论细菌还是真菌丰富度最大的均为雷司令,细菌主要分布在变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、厚壁菌门(Firmicutes)等8个门,优势细菌主要是欧文氏菌属(Erwinia)、假单胞菌属(Pseudomonas)、芽孢杆菌属(Bacillus)等6个属。真菌仅分布在子囊菌门(Ascomycota)、担子菌门(Basidiomycota)和接合菌门(Zygomycota)3个门,链格孢菌属(Alternaria)、枝孢属(Cladosporium)、茎点霉属(Phoma)、镰孢属(Fusarium)等9个主要的属。细菌中心OTU最相似菌种分别为节杆菌(Arthrobacter sp.)、假单胞菌(Pseudomonas sp.)等,真菌为枝孢菌(Cladosporiumsp.)、茎点霉(Phoma sp.)、链格孢菌(Alternaria sp.)以及大量未知种属的OTU。研究表明,葡萄品种是影响微生物群落的最重要的因素,推测酿酒葡萄上的一些微生物会对葡萄植株的健康、葡萄果实的品质以及葡萄酒酿造产生有益或有害的影响。     

山东莴苣亚族植物的修订

对山东现有的莴苣亚族的植物标本进行了研究,确定山东莴苣亚族植物有24种,分属于12个属,即：苦苣菜属(Sonchus L.)、乳苣属(mulgedium Cass.)、山柳菊属(Hieracium L.)、黄鹌菜属(Youngia Cass.)、福王草属(Prenanthes L.)、翅果菊属(Pterocypsela Shih)、莴苣属(Lactuca L.)、苦荬菜属(Ixeris Cass.)、小苦荬属(Ixeridium (A.Gray) Tzvel.)、沙苦荬属(Chorisis DC.)、黄瓜菜属(Paraixeris Nakai)和蒲公英属(Taraxacum L.)。发现1个新记录种：华蒲公英(Taraxacum borealisinense Kitam.)。     

南方红豆杉产紫杉醇内生真菌的分离鉴定

从南方红豆杉(Taxus chinensisvar.mairei)植株分离到内生真菌63株,其中24株经鉴定分属于19个种(属).真子囊菌纲的2个属2株,半知菌纲的14个属22株.真子囊菌纲的无毛毛壳属(Achaetomiumsp.)的1株为新发现种.半知菌纲的葡萄孢属(Botrytissp.)1株和无孢菌群(Mycelia sterlia)2株的培养液,经浓缩萃取,薄层层析检测,Rf值分别为0.404、0.404、0.401,与紫杉醇对照品Rf值一致,为南方红豆杉产紫杉醇内生真菌.     

俞藤属—葡萄科一新属

作者在研究中国葡萄科植物过程中发现分布于中国亚热带、尼泊尔和印度卡西山区一些植物种自成一类,与葡萄科现有属的定义不符,命名为一新属-愈藤属（YuaC.L.Li)。文中讨论了它与爬山虎属（Parthenocissus Planch.)、崖爬藤属（TetrastigmaPlanch.)、乌蔹莓属（CayratiaJuss.)和蛇葡萄属（Ampelopsis Michx.)的区别,并在野外和栽培条件下观察了某些重要属级鉴定性状在个体发育过程中的表现,以证明其可靠。为了正确鉴定这个属和种类,给出了包括这些属在内的国产葡萄科植物分属检索表和该属的分种检索表,最后,讨论了俞藤属的地理分布及其意义。     

内生真菌对草坪植物病原真菌抑制作用的比较

分别从野生牧草羽茅(Achnatherum sibiricum (L.) Keng)、栽培种高羊茅(Festuca arundinacea Schreb.)(品种Millennium)、栽培种黑麦草(Lolium perenne L.) (品种Justus)中分离出内生真菌Neotyphodium sp.、N. coenophialum和N. lolli,通过体外培养法比较了这3种内生真菌对草坪植物病原真菌的抑制作用.结果表明,从羽茅中分离的内生真菌Neotyphodium sp.在两菌相交前对所有供试的病原真菌都有一定的抑制作用,其中对枝孢霉属(Cladosporium sp.)、弯孢霉属(Curvularia sp.)和拟茎点属(Phomopsis sp.)病原真菌的抑制效果尤为显著,对峙培养3d后的抑菌率分别达70.1%、52.3%和30.9%,营养竞争作用、重寄生作用是其主要的拮抗机制;从高羊茅中分离的内生真菌N. coenophialum对枝孢霉属病原真菌存在一定的抑制作用;而从黑麦草中分离的内生真菌N. lolli与病原真菌对峙培养时,病原真菌菌落慢慢侵占整个营养空间,内生真菌停止生长并逐渐褐变死亡.体外培养结果说明Neotyphodium sp.对供试病原真菌的拮抗效果优于N. coenophialum和N. lolli,由此推测Neotyphodium sp.与宿主植物羽茅的共生可能有利于宿主植物抵抗病原真菌的侵扰.     

三种水生植物内生真菌多样性及其抗真菌活性

从华凤仙(Impatiens chinensis L.)、问荆(Equisetum arvense L.)和轮叶狐尾藻(Myriophyllum verticillatum L.) 3种植物中共分离到内生真菌155株。经鉴定, 它们分属于26个不同的分类单元。以杨桃炭疽菌(Colletotrichum gloeosporioides)、香蕉疫霉(Phytophthora nicotianae)和腐皮镰刀菌(Fusarium solani)等6种植物病原真菌为指示菌, 对3种水生植物内生真菌代谢产物进行抗菌活性研究。结果表明, 有37株内生真菌(占分离菌株总数的23.9%)显示出对一种或多种病原菌的抑菌活性。来自华凤仙、问荆和轮叶狐尾藻的抗菌活性菌株比例分别为29.2%、13.9%及37.1%, 都远低于已报道的陆生植物内生真菌中活性菌株的比例。抗菌活性菌株主要分布于枝孢属(Cladosporium)、木霉属(Trichoderma)和地霉属(Geotrichum)等5个属中。     

浙江葡萄属(Vitis L.)植物新资料

该文报道了在浙江葡萄属(Vitis L.)分类研究中的新发现:(1)描述了开化葡萄(V. kaihuaica Z.H. Chen,F. Chen et W. Y. Xie)、秀丽葡萄(V. amoena Z. H. Chen,F. Chen et W. Y. Xie) 2新种和腺枝龙泉葡萄(V. longquanensis var. glandulosa Z. H. Chen,F. Chen et W. Y. Xie) 1新变种;(2)将V. adenoclada Hand.-Mazz.作为毛葡萄(V. heyneana Roem. et Schult.)的变种处理,即腺枝毛葡萄[V. heyneana Roem. et Schult. var. adenoclada (Hand.-Mazz.) Z. H. Chen,F. Chen et W. Y. Xie];(3)报道了蓝果刺葡萄[V. davidii(Roman. Du Caill.) F9ex var. cyanocarpa (Gagnep.) Sarg.]在浙江的分布新记录。     

葡萄属野生资源分类研究进展

葡萄属野生资源在整个葡萄属(Vitis L.)的系统演化中占有十分重要的地位。葡萄属野生资源分布广泛,种间杂交容易,形态变异较为复杂,造成分类工作难度较大。长期以来,部分野生种的分类一直存在争议。本文从世界葡萄属植物的分类历史出发,对葡萄属野生种分类研究进展进行了综述。传统的分类方法不能够很好地解决不同种之间的亲缘关系,近十几年来,多种分子标记被应用于葡萄属的系统发育研究,并构建了葡萄属系统发育的基本框架。但是,分子系统学研究工作开展的远远不够,而且多数研究仍然受到资源的限制。未来建议加强葡萄属野生资源的收集和保护,并开展更全面的分类学修订工作。同时,充分利用分子系统学的相关技术手段,进一步阐明各野生种的分类地位。     

葡萄属植物花粉形态的研究

利用扫描电子显微镜对葡萄属Vitis L．37个类群(包括32种、1亚种和4变种)以及其近缘属麝香 葡萄属Muscadinia(Planch．)Small一种植物的花粉进行了观察,结果表明,葡萄属植物的花粉形态特征 在区别并鉴定属下的种类时有较大价值,但对种以上的分类以及研究系统演化无多大意义。葡萄属植 物的花粉形态特征可以分为4类：(1)花粉粒扁球形或圆球形(P/E值为O.97-1.08),沟较宽,可见被有 较密颗粒状突起的沟膜,属于该类的有紫葛V．coignetiae Pull．ex Planch．;(2)花粉粒长球形至近长圆球 形(P/E值为1.25～1.76),沟较宽,可见有较疏颗粒状突起的沟膜,该类型包括了庐山葡萄V．hui Cheng 等共10种1亚种3变种;(3)花粉粒长圆球形(P/E值为1.87～2.1),沟边缘紧靠合呈直线状,但在赤道 轴萌发孔处沟边缘分开呈菱形,该类型有秋葡萄V．romanetii Roman du Caill．ex Planch．和锈毛刺葡萄V． davidii(Roman du Caill．)Foex．var．ferruginea Merr．et Chun等1种1变种;(4)花粉粒长球形或超长球形 (P/E值为1.30～2.0),沟边缘紧靠成直线状,麝香葡萄Muscadinia rotundifolia(Michx .)Small等21种的花粉属于此类型。     

40种杂草的丙酮提取物对3种植物病原真菌的抑菌活性

在离体条件下研究了40种杂草的丙酮提取物对辣椒疫霉(Phytophthora capsici Leonian)、尖孢镰刀菌(Fusarium oxysporium Schl.)和灰葡萄孢(Botrytis cinerea Pets.)3种植物病原真菌的抑菌活性.结果表明,有7种杂草的丙酮提取物至少对1种供试病原菌的抑制率在60%以上.其中,黄花蒿(Artemisia annua L.)、苍耳(Xanthiumsibiricum Patrin)和荔枝草(Salvia plebeia R. Br.)的丙酮提取物对3种病原菌的抑制率都在60%以上;鳢肠(Ecliptaprostrata L.)的丙酮提取物对辣椒疫霉和尖孢镰刀霉的抑制率在60%以上;车前(Plantago asiatica L.)、夏至草[Lagopsis supina(Steph.)IK.-Gal.]和泽漆(Euphorbia helioscopia L.)的丙酮提取物对灰葡萄孢的抑菌效果较好.     

岩黄连引种栽培研究

岩黄连 ( Corydalis saxicola Bunting)为石山特有的多年生草本药用植物 ,局限分布裸露石山 ,生于石缝、石穴 ,资源稀少且濒于枯竭。通过近四年的引种栽培研究 ,表明其对丘陵区酸性土壤亦有较强的适应性 ,能正常生长发育 ,种子发芽率可达 4 0 %～ 50 % ,药材产量 (干品 ) 90 0～ 12 0 0 kg/hm2 。报道了其生物学特性和主要的栽培技术。     

Comparative analysis of the chemical profile of wild and cultivated populations of Corydalis saxicola by high-performance liquid chromatography

Studies on the simultaneous determination and chemical fingerprinting of alkaloids in Corydalis saxicola Bunting. (Yanhuanglian) were performed for authentication purposes. Ninety samples prepared from different parts of C. saxicola, including whole plants, roots, stems, leaves and flowers, from wild and cultivated populations, were submitted to quantitative determination and fingerprint analysis. Five major alkaloids, namely, tetradehydroscoulerine, dehydroapocavidine, dehydroisoapocavidine, coptisine and dehydrocavidine, were quantitatively analysed by reversed-phase HPLC with acceptable recoveries (>98.2%). Chemical fingerprinting of C. saxicola was established and involved 11 markers. The results indicated that there were no obvious differences between the chemical profiles of wild and of cultivated C. saxicola populations, and that the mean alkaloid contents of the five marker compounds in cultivated populations were significantly higher than those of the wild plants. The highest content of total alkaloids (up to 28.8 mg/g) was found in roots of C. saxicola. The total alkaloids of the leaves were approximately 50% of those of roots, suggesting that the leaves may be employed as an alternative source of alkaloids. Chemical fingerprints and quantitative HPLC analysis will have a positive impact on the conservation and cultivation of this medicinal plant.     

Simultaneous determination of eight bioactive alkaloids in Corydalis saxicola by high-performance liquid chromatography coupled with diode array detection

Corydalis saxicola Bunting (Papaveraceae), a traditional folk medicine, has been used to treat hepatic diseases for a long time. Owing to its signicant clinical effectiveness against hepatitis, cirrhosis and hepatoma, C. saxicola and its preparation are widely applied. In this study, eight alkaloids, namely isocorydine, scoulerine, dehydrocheilanthifoline, dehydrodiscretamine, dehydroisoapocavidine, dehydrocavidine, palmatine and berberine, which have been previously proven to possess potential antitumour activity, were selected as the chemical markers of C. saxicola. To evaluate the quality of C. saxicola, a simple, accurate and reliable HPLC-DAD method was developed for the simultaneous determination of the above eight compounds. Separation was achieved on a Gemini C(18) column (5 microm, 250 x 4.6 mm i.d., Phenomenex Inc., CA, USA) with a gradient solvent system of 20 mM aqueous ammonium acetate-acetonitrile, at a flow-rate of 1.0 mL/min and detected at 270 and 280 nm. All eight calibration curves showed good linearity (R(2) > 0.9992). The method was reproducible with intra- and inter-day variations of less than 5%. The recovery was in the range of 96.09-102.80%. This assay was successfully utilised to quantify the eight alkaloids in C. saxicola from different locations. The results demonstrated that this method is simple, reliable and suitable for the quality control of this medicinal herb.     

岩黄连规范化种植标准操作规程(SOP)

该操作规程明确规定了岩黄连的种质特征、产地环境、种子标准、育苗技术、栽培技术措施、采收加工、外观品质、成分含量、农药残留以及包装、储运方法等。基地生产应用表明:生长一年采收,平均每株鲜重0.2~0.3kg平均每株产品约0.02~0.03kg;产品以干燥品计岩黄连总碱含量茎叶部不低于0.1%、根部不低于0.2%;产品各项卫生质量均达到国家相关规定。     

放大培养对岩黄连细胞悬浮培养的影响

为了优化岩黄连细胞悬浮培养的条件,研究了在放大培养过程中,岩黄连细胞生长与主要营养成分的代谢动力学,以及生物碱的产生情况。结果表明,在不同培养体系下,细胞生长曲线均呈现"S"型。随着培养体积从50、100 mL放大到500 mL和1 L,培养液中碳源、氮源和磷源的消耗减慢,细胞生物量减少,生物碱产量降低。其中100 mL培养体系所获生物量最高,达到15.2 g/L,生物碱产量也最高,脱氢卡维丁为8.35 mg/mL,小檗碱为4.58 mg/mL。根据本文结果,提出了岩黄连细胞培养条件的优化和大规模细胞培养生产岩黄连生物碱应采取的策略。     

DNA topoisomerase I inhibitory alkaloids from Corydalis saxicola

Chemical studies of the Chinese herb Corydalis saxicola Bunting led to the isolation and identification of 14 alkaloids, 1-14. Seven of these compounds, 4-9 and 11, were obtained from this plant for the first time. Feruloylagmatine (7) is the first guanidine-type alkaloid to be identified in the family Papaveraceae and in dicotyledonous plants. All of the isolated compounds were assayed for inhibitory activity against human DNA topoisomerase I. A DNA cleavage assay demonstrated that these alkaloids specifically inhibit topoisomerase through stabilization of the enzyme-DNA complex. Among the isolated alkaloids, (-)-pallidine (8) and (-)-scoulerine (11) showed strong inhibitory activities toward topoisomerase I that were comparable to camptothecin, a typical topoisomerase I inhibitor. A preliminary structure-activity relationship study suggested that the quaternary ammonium ion might play an important role in topoisomerase I inhibition by the isoquinoline alkaloids. These data indicated that DNA topoisomerase I inhibition represents probably one of the anticarcinogenic mechanisms of C. saxicola.     

Alkaloids from Corydalis saxicola and their anti-hepatitis B virus activity

Eight protoberberine-type alkaloids and two indole alkaloids were isolated from the MeOH extracts of the herb Corydalis saxicola Bunting (Papaveraceae). Their structures were identified as dehydrocavidine (1), dehydroapocavidine (2), dehydroisoapocavidine (3), berberine (4), dehydroisocorypalmine (5), coptisine (6), tetradehydroscoulerine (7), berbinium (8), 1-formyl-5-methoxy-6-methylindoline (9), and 1-formyl-2-hydroxy-5-methoxy-6-methylindoline (10). Compounds 3, 9, and 10 are new alkaloids. All compounds were tested for anti-HBV activity against the 2.2.15 cell line in vitro. Dehydrocavidine (1), dehydroapocavidine (2), and dehydroisoapocavidine (3) exhibited inhibitory activity against HBsAg and HBeAg, but no cytotoxicity against the 2.2.15 cell line.     

岩黄连光合与蒸腾特性及其对光照强度和CO2浓度的响应

采用LI-6400便携式光合测定系统(Li-CorInc.,USA)对岩黄连叶片的气体交换进行了测定。结果表明(1)岩黄连叶片的光饱和点(LSP)为329.18μmol.m-2.s-1左右,光补偿点(LCP)为12.76μmol.m-2.s-1,最大净光合速率为2.96μmol.m-2.s-1,暗呼吸速率(Rd)为0.17μmol.m-2.s-1。光饱和点和光补偿点都比效低,表明岩黄连对光照的要求不高,属于阴生植物。(2)4月份,岩黄连Pn随CO2浓度升高而逐渐增大。当CO2浓度由50μmol.mol-1增加到600μmol.mol-1,Pn几乎呈直线上升,600~1000μmol.mol-1范围内逐渐缓和,到1000μmol.mol-1以后Pn变化平稳。由曲线估算CO2饱和点(CSP)大约在1000μmol.mol-1左右。CO2的补偿点为68.80μmol.mol-1。羧化效率为0.0308μmol.m-2.s-1。(3)岩黄连叶片水分利用率(WUE)随有效光辐射强度(PAR)的增强呈抛物线状变化,PAR在200μmol.m-2.s-1内呈直线上升,到200μmol.m-2.s-1时WUE达最大值,大于200μmol.m-2.s-1后WUE呈逐渐下降趋势。     

岩黄连细胞生长与营养物质消耗的动态学研究

在岩黄连细胞悬浮培养过程中,对培养液pH值,碳源、氮源和磷酸盐含量,以及细胞生物量和生物碱含量进行测定,分析其动态变化过程。结果显示培养液pH值在培养初期降低,后逐渐升高;碳源在培养过程中逐渐被利用,磷酸盐和氮源在培养中期几乎耗尽,其中磷酸盐的消耗速率最快;悬浮细胞的生长周期为20 d左右,第18天细胞鲜重和干重达最大,而第21天脱氢卡维丁和小檗碱的含量最高,分别为8.22mg/L和4.31mg/L。结果表明营养物质(碳、氮和磷)的吸收与细胞生长以及生物碱的合成密切相关,营养元素的相对消耗速率为磷>氮>碳,推测氮和磷是影响岩黄连细胞培养的主要因素。     

广西栽培与野生岩黄连的质量比较研究

通过对广西栽培与野生岩黄连(Corydalis saxicola Bunting)全草的性状、组织构造、化学反应、薄层层析、紫外吸收的比较,表明栽培岩黄连与当地野生品质量基本一致,为广西发展岩黄连资源提供科学依据。