红豆杉细胞培养中紫杉醇高产细胞株的筛选及其稳定性分析

对中国红豆杉〔Taxus chinensis(Pilger)Rehd.〕、云南红豆杉(T. yunnanensis Cheng et L.K.Fu)和东北红豆杉(T. cuspidata Sieb.et Zucc.)不同部位的愈伤组织进行培养及分析比较。结果表明,中国红豆杉愈伤组织生长较快,其叶片诱导的愈伤组织紫杉醇含量较高,且紫杉醇含量与培养物的外观特征有明显相关性,颜色浅、块状或颗粒较明显的细胞团紫杉醇含量较高。运用细胞看护培养技术,从中国红豆杉愈伤组织中筛选出生长速率达0.52g.L-1.d-1、紫杉醇含量超过0.01%的细胞优株,经20次继代培养,其生长和紫杉醇含量均较稳定。     

红豆杉育苗栽培及苗木生长动态

紫杉醇是当前倍受关注的抗癌新药,美国国立癌症研究所(NCI)预测,紫杉醇将成为主要的抗癌药物之一[1].其原料植物红豆杉[Taxus chinersis (Pilger) Rehd.]资源的过度开发利用,导致供求矛盾十分突出.     

山西陵川南方红豆杉群落种间联结与生态位特征研究

基于对山西陵川县境内野生南方红豆杉群落的样地调查,采用总体相关性VR检验、Pearson相关系数、Spearman秩相关系数和生态位测度方法,在群落尺度上对南方红豆杉群落的种间联结性和生态位特征进行了研究。结果显示:(1)南方红豆杉(Taxus chinensis(Pilger)Rehd.var.mairei(Lemee et Levl.)Cheng et L.K.Fu)、荆条(Vitex negundo L.var.heterophylla(Franch.)Rehd.)、鹅耳枥(Carpinus turczaninowii Hance)、连翘(Forsythia suspensa(Thunb.)Vahl)和三裂绣线菊(Spiraea trilobata L.)的重要值和生态位宽度较大,为群落中的建群种和优势种;(2)物种间生态位重叠指数总体偏高,种间竞争较强,尤其是南方红豆杉与其他物种之间;(3)群落总体呈显著负联结,大多数种对间呈中性关联(χ23.84,P0.05);(4)群落中物种间的Pearson相关系数、Spearman秩相关系数与生态位重叠指数之间表现为极显著的正相关(P0.01),而生态位宽度与生态位重叠指数之间未表现出线性关系,即物种间的竞争关系随种间关联性的增大而增强,而与生态位宽度没有直接关系。本研究可为保护和利用本区现有野生南方红豆杉资源提供合理的理论支撑。     

寡聚糖诱导悬浮培养南方红豆杉细胞的凋亡

在真菌(Fusarium oxysporum f.vasinfectum (Atkinson) Snyder et Hansen)寡聚糖诱导悬浮培养南方红豆杉(Taxus chinensis (Pilger) Rehd.var.mairei (Lemee et Lévl.) Cheng et L.K.Fu)细胞生产紫杉醇的体系中发现细胞出现凋亡,次生代谢增强.电镜观察到细胞核质和原生质出现凝集现象,液泡内出现大量的高电子致密体.核DNA经琼脂糖凝胶电泳,呈200 bp的整数倍的梯状条带(ladders);而对照组细胞核DNA完整,呈大片段,细胞完整,细胞器发达,但紫杉醇合成速率很低.加入寡聚糖后,细胞防御系统开启,细胞生长停止,次生代谢物酚类物质大量积累且次生壁加厚,多酚氧化酶活性迅速提高,苯丙烷类代谢途径的关键酶苯丙氨酸解氨酶的活性在1 h后急速提高,目的产物紫杉醇在诱导后72 h达到峰值,比对照组提高了6倍,且细胞凋亡的出现与紫杉醇合成的峰值具有时间上的一致性.     

从中国红豆杉细胞培养物中分离鉴定紫杉醇

中国红豆杉(Taxus chinensis(Pilger．)Rehd)细胞在培养过程中合成了紫杉醇,我们利用固液分离、大孔吸附树脂吸附富集、有机溶剂萃取、硅胶柱层析、低压硅胶柱层析、重结晶等方法,从中国红豆杉的细胞培养物中分离纯化了紫杉醇,用多种核磁共振波谱方法(^1H NMR,^13C NMR,DEPT,^1H-^1H-COSY,NOESY,HMQC,HMBC)结合质谱(FABMS),红外光谱、紫外光谱等方法鉴定了它的化学结构,证明与源于天然红豆杉植物材料提取的紫杉醇为同一物质。     

几种促进剂的添加时间对中国红豆杉细胞悬浮培养紫杉醇合成的影响

采用正交实验检测中国红豆杉[Taxus chinensis(Pilger)Rehd.]细胞悬浮培养中水杨酸、硝酸银、氨基酸前体、D-果糖和硫酸镧的添加时间对细胞生长和紫杉醇(taxol)积累的影响.这些促进剂的添加时间对中国红豆杉细胞悬浮培养的生长没有明显的影响,但能明显促进紫杉醇的合成,当在细胞培养的第14 d添加1.67 mg/L硝酸银,第18 d添加0.1 mg/L水杨酸,第21 d添加氨基酸前体,第21 d添加10 g/L D-果糖和2 mg/L硫酸镧时对紫杉醇的促进作用最明显,在此最优组合处理时紫杉醇含量达到10.05 mg/L,相对于最差组合处理时紫杉醇含量仅有1.77 mg/L,紫杉醇含量提高5.7倍,这些因素的添加时间对紫杉醇合成的相互作用达不到显著水平.     

福建省毛竹混交林群落结构特征的比较

通过设置临时标准地对龙栖山、武夷山和天宝岩3个国家级自然保护区的3种毛竹混交林群落类型结构进行调查与统计分析.结果表明,在物种数量上从多到少依次为:武夷山国家级自然保护区的毛竹 少叶黄杞(Phyllostachys pubescences Mazel ex H. Delehaie Engelhardtia fenzelii Merr.)混交林(68科125属230种),龙栖山国家级自然保护区的毛竹 南方红豆杉[Taxus chinensis (Pilger) Rehd. var. mairei (Lemee et Lévl.) Cheng et L. K. Fu]混交林(69科135属194种)和天宝岩国家级自然保护区的毛竹 长苞铁杉(Tsuga longibracteata Cheng)混交林(41科71属105种).3个毛竹混交林群落结构分明,毛竹在混交林群落乔木层中占绝对优势的地位.乔木层、灌木层和草本层层次清晰,还具有层间层.毛竹 南方红豆杉群落和毛竹 长苞铁杉群落乔木层可分为3个亚层,而毛竹 少叶黄杞群落乔木层可分为2个亚层.毛竹 少叶黄杞群落的灌木层物种数和草本层盖度均比其他2个群落高.     

寡聚糖诱导悬浮培养南方红豆衫细胞的凋亡

在真菌（Fusarium oxysporum f.vasinfectum(Atkinson)Snyder et Hansen）寡聚糖诱导悬浮培养南方红豆衫（Taxus chinensis(Pilger)Rehd.var.mairei(Lemee et Levl.）Cheng et L.K.Fu）细胞生产紫杉醇的体系中发现细胞出现凋亡，次生代谢增强，电镜观察到细胞核质和原生质出现凝集现象，液泡内出现大量的高电子致密林。核DNA器发达，但紫杉醇合成速率很低，加入寡聚糖后，细胞防御系统开启，细胞生长停止，次生代谢物酚类物质大量积累且次生壁加厚，多酚氧化酶活性迅速提高，苯丙烷类代谢途径的关键酶苯丙氨酸解氨酶的活性在1h后急速提高，目的产物紫杉醇在诱导后72h达到峰值，比对照组提高了6倍，且细胞凋亡的出现与紫杉醇合成的峰值具有时间上的一致性。     

基于ISSR标记的南方红豆杉野生种群和迁地保护种群的遗传多样性和遗传结构分析

采用ISSR标记方法,对南方红豆杉〔Taxus chinensis(Pilger)Rehd.var.mairei(Lemée et Lévl.)Cheng et L.K.Fu〕3个野生种群(包括江西黄港和黄沙种群以及福建枫溪种群)、2个迁地保护栽培种群及2个迁地保护衍生种群(均位于江苏南京中山植物园和江西庐山植物园)的遗传多样性和遗传结构进行了分析和比较。结果显示:用8个引物从南方红豆杉的基因组总DNA中共扩增出73条带,其中多态性条带62条。2个迁地保护衍生种群的多态性条带百分率(PPB)、Nei’s多样性指数(h)和Shannon信息指数(I)较高,3个野生种群的PPB、h和I值总体上居中,而2个迁地保护栽培种群的PPB、h和I值较低。合并后的迁地保护衍生种群以及野生种群均有较高的遗传多样性,二者的PPB、h和I值相近,分别为78.08%和82.19%、0.207 6和0.205 8、0.322 9和0.325 9;但二者的遗传结构存在差异,迁地保护衍生种群的遗传分化系数(GST)为0.068 9,明显低于野生种群(0.168 5)。合并后的迁地保护栽培种群的遗传多样性相对较低,PPB、h和I值分别为60...     

4种红豆杉属植物遗传多样性和遗传关系的RAPD分析

采用RAPD标记研究了分属于4种红豆杉属(Taxus Linn.)植物的68个单株的遗传多样性,并采用UPGMA方法分析68个单株的遗传关系.结果表明:12条RAPD引物共扩增出109条带,其中多态性条带108条,多态性条带百分率为99.1％;平均每条引物扩增出9.1条带.南方红豆杉[T.wallichiana var.mairei (Lemée et Lévl)L.K.Fu et Nan Li.]种内的多态性条带百分率和观察等位基因数均最高;欧洲红豆杉(T.baccata Linn.)种内的有效等位基因数、Nei's基因多样度和Shannon's信息指数均最高;须弥红豆杉(T.wallichiana Zucc.)种内的各项遗传多样性指数均最低.供试4种植物的种内遗传多样度、种间遗传多样度、基因流和种间遗传分化系数分别为0.1745、0.358 6、0.401 7和0.554 5,表明55.45％的遗传变异发生在种间.南方红豆杉和须弥红豆杉遗传距离最近;曼地亚红豆杉(Taxus×media Rehd.)和须弥红豆杉的遗传距离最远.通过聚类分析可将68个单株分为3组,欧洲红豆杉的18个单株和曼地亚红豆杉的18个单株分别各自聚为1组;须弥红豆杉的16个单株和南方红豆杉的16个单株聚为1组,其中须弥红豆杉的16个单株和南方红豆杉的16个单株又各自聚为1个亚组,且南方红豆杉的雌、雄单株也分别聚在同一分支上,表明须弥红豆杉和南方红豆杉遗传关系较近,而欧洲红豆杉与其他3种植物的遗传关系较远.     

中国假尾孢属的研究VI.

本文报道中国假尾孢属的16个种,其中有3个新种:藤山柳假尾孢(Pseudocercospora clematoclethrae),仙茅假尾孢(P. curculiginis,),豆付柴假尾孢(P. premnicola), 8个新组合:金粟兰假尾孢(P. chloranthe), 榅桲假尾孢(P. cydoniae),福岗假尾孢(P. fukuokaensis),土连翘假尾孢(P. hymenodictyonis),南五味子假尾孢(P. kadsurae), 野岛假尾孢(P. nojimae).绣线菊假尾孢(P. spiraeicola),球形假尾孢(P. sphaeriiformis)和3个中国新纪录。文中对新种进行了描述并绘图。研究的标本保存在中国科学院微生物研究所真菌标本室(HMAS)。     

中国假尾孢属的研究V

本文报导中国假尾孢属的30个种,其中有2个新种:透骨草假尾孢(Pseudocercospora phrymae),槐假尾孢(P. sophorae), 6个新组合:无花果假尾孢(P. fici),爵床假尾孢(P. justiciae),木犀生假尾孢(P. osmanthicola),海桐花假尾孢(P. pittospori),色柱假尾孢(P. polygonorum),花椒假尾孢(P. xanthoxyli)和5个中国新记录。文中对新种和新组合进行了描述并绘图,其他种作了简要描述或讨论。研究的标本保存在中国科学院微生物研究所真菌标本室(HMAS)。     

假尾孢属四个新种

本文报道中国假尾孢属的4个新种:扁担秆生假尾孢(Pseudocercospora grewiigena Y. L. Guo> sp. nov.),忍冬假尾孢(Pseudocercospora lonicerae Y. L. Guo, sp. nov.),红豆树假尾孢(Pseudocercospora ormosiae Y. L. Guo & Y. R. Lin, sp. nov.)欧洲荚蒾假尾孢(Pseudocercosporatinea Y. L. Guo & W. H. Hsieh> sp. nov.)。文中对每个种都进行了描述并附图。研究的标本保存在中国科学院微生物研究所真菌标本室(HMAS)。     

Acht neue Arten der GattungAnthemis (Compositae) aus Persien

The following species are described as new:Anthemis mazandaranica in N. and NW. Iran is allied toA. coelopoda; A. moghanica in NW. Iran is close toA. candidissima andA. sintenisii; A. atropatana also in NW. Iran is similar toA. hyalina; A. gracilis in W. Iran is close toA. plebeia; A. bushehrica in SW. Iran is similar toA. susiana; andA. rhodocentra in S. and E. Iran and in Pakistan is akin toA. austro-iranica, A. gayana, andA. kandaharica.

Anschrift des Herausgebers: Hofrat Univ.-Prof. Dr.Karl Heinz Rechinger, Beckgasse 22, A-1130 Wien, Österreich.     

Occurrence and diversity of mesophilic Shewanella strains isolated from the North-West Pacific Ocean

Although bacteria of the genus Shewanella belong to one of the readily cultivable groups of "Gammaproteobacteria", little is known about the occurrence and abundance of these microorganisms in the marine ecosystem. Studies revealed that of 654 isolates obtained from marine invertebrates (ophiuroid Amphiopholis kochii, sipuncula Phascolosoma japonicum, and holothurian Apostichopus japonicus, Cucumaria japonica), seawater and sediments of the North-West Pacific Ocean (i.e. the Sea of Japan and Iturup Is, Kurile Islands), 10.7% belonged to the genus Shewanella. The proportion of viable Shewanella species varied from 4% to 20% depending on the source of isolation. From the isolation study, representative strains of different phenotypes (from seventy presumptive Shewanella strains) were selected for detailed characterization using phenotypic, chemotaxonomic, and phylogenetic testing. 16S rDNA sequence-based phylogenetic analysis confirmed the results of tentative identification and placed the majority of these strains within only a few species of the genus Shewanella with 98-99% of 16S rDNA sequences identity mainly with S. japonica and S. colwelliana, suggesting that the strains studied might belong to these species. Numerically dominant strains of S. japonica were metabolically active and produced proteinases (gelatinases, caseinases), lipases, amylases, agarases, and alginases. Shewanella strains studied demonstrated weak antimicrobial and antifungal activities that might be an indication of their passive role in the colonization on living and non-living surfaces.     

In vitro culture of adult and juvenile bud explants of Passiflora species

Cultivar E23, an F1 hybrid of P. edulis and P. edulis f. flavicarpa is usually propagated by shoot-tip grafting. Various media were tested to evaluate the potential of E23 for in vitro propagation. Adult tissue was difficult to culture and did not respond to media containing low (<10 µM) concentrations of growth regulators. Growth of adult buds on intact stem sections was promoted by 1 week of dark incubation on MS basal medium plus 150 µM 2iP, 200 µM adenine sulphate and 17.1 µM IAA (3 mg l^–1), and further developed into shoots on MS medium plus 4.9 µM 2iP (1 mg l^–1) and 5.7 µM IAA (1 mg l^–1). By contrast, juvenile shoots of E23, and Passiflora species: edulis f. flavicarpa, edulis, alata, caerulea, mollissima, coccinea, herbertiana and suberosa grew rapidly on MS medium plus 10 µM kinetin and 5 µM IAA. Rapid multiplication was achieved on MS plus 20 µM BA, 10 µM kinetin, 5 µM IAA, and roots initiated on MS plus 5 µM IAA.Abbreviations IAA indole-3-acetic acid - 2iP N6-iso pentenyl adenine - BA N6-benzyl adenine     

竹亚科刚竹属植物的修订(Ⅳ)——Phyllostachys hispida的恢复及其近缘种的分类

在模式标本考证、居群调查和引种栽培的基础上,确认毛壳竹(Phyllostachys hispida S.C.Li,S.H.Wu et S.Y.Chen)与乌竹(P.varioauriculata S.C.Li et S.H.Wu)在新秆性状、秆箨颜色、箨舌高度及颜色、叶鞘毛被上有较大的区别,应作为独立的竹种看待。小叶光壳竹(P.varioauriculata var.glabrata G.H.Lai)也因新秆下部一段呈扁圆形、秆箨基部具长柔毛,叶片很小等特征而明显不同于乌竹,应提升为一个独立的竹种,但由于存在晚出同名问题而给出新名称P.microphylla G.H.Lai。同时还描述了毛壳竹的一新变种,即光壳竹(P.hispida var.glabrivagina G.H.Lai)。日本所产的姬淡竹与毛壳竹属于同一种系,因其学名P.humilis Muroi为一裸名,应予废弃。     

New taxa ofRubiaceae-Rubieae from Iran

Asperula sect.Oppositifoliae is validated. 2 new species ofAsperula and 7 new species and 1 variety ofGalium from Iran are described, discussed in respect to their affinities and illustrated. 2 subspecies are raised to species rank.     

Pollen Aerobiological Monitoring with the personal volumetric air sampler (PVAS). Correlation with a fixed Hirst type sampling station

Summary Monitorings both with a fixed Hirst type volumetric sampler, placed on the balcony of the Medicine Division in Lanciano City Hospital and with a portable volumetric sampler PVAS type by Burkard, placed outside the dwelling-houses of the examined patients have been carried out simulaneously. This has given us further information about the quantity of pollen that single patients affected by pollinosis inhale. Preliminary controls on the capture efficiency of both samplers have been conducted. The results of this survey have shown that the symptom score cards of the eight patients enrolled for the study, have always been well correlated with the average daily pollen concentration of the fixed station (Pearson correlation), and the data obtained by the two samplers have pointed out remarkable differences in the pollen concentration. According to the PVAS, placed near patients, the concentrations were twofold to sixfold as compared to those of the fixed station. This was probably due to the different height of the samplers. However, owing to the short sampling time (30 min.), we aren't able to draw any conclusions. Nevertheless we do hope that more and more patients will undergo this test, so that with several daily surveys over longer periods, we will have more information on the real pollen exposure of single patients. We will verify the threshold of symptoms onset, which is only theoretical nowadays, in the monosensitized patients. Such a program will allow us to carry on useful research both in the respiratory physiopathological and in the allergological fields.