苦苣苔科植物的组织培养和离体保存

苦苣苔科(Gesneriaceae)植物种类繁多, 全世界约150属3700余种,我国有58属470余种，大部分具有极高的观赏价值，许多是传统的民间草药。虽然我国苦苣苔科植物资源丰富，然而很多种类分布区域狭窄，种群数量稀少，加之生境受到破坏，许多已经面临灭绝的危险。本研究拟通过组织培养、玻璃化超低温保存以及快速繁殖，达到保护和扩繁珍稀濒危苦苣苔科植物的目的。 以药用唇柱苣苔（Chirita medica D. Fang ex W. T. Wang）和粉绿异裂苣苔（Pseudochirita guangxiensis W.T.Wang var. glauca Y. G. Wei et Y. Liu）为材料，取幼嫩叶片为外植体，通过组织培养实验得到最佳诱导不定芽培养基：MS培养基附加30 g l-1蔗糖，7.5 g l-1琼脂，药用唇柱苣苔附加0.10 mg l-1 BA ，0.10 mg l-1 NAA，粉绿异裂苣苔附加0.05 mg l-1IAA，1.00 mg l-1BA。最高不定芽诱导率分别为：90.3%和85.0%。最佳生根培养基：1/2MS培养基附加30 g l-1蔗糖，5 g l-1活性炭，7 g l-1琼脂，生根率为100%，诱导产生6.11条根，根长为18.8mm（药用唇柱苣苔）；1/2MS培养基附加10-20g l-1蔗糖，1 g l-1活性炭，7 g l-1琼脂，诱导生成6.8-7.4条根，均长17.7-22.0mm（粉绿异裂苣苔）。 在组织培养的基础上进行了苦苣苔科植物的玻璃化超低温冷冻保存研究。以烟叶唇柱苣苔（C. heterotricha Merr.）和濒危植物药用唇柱苣苔叶片外植体为材料，经过自然干燥、装载液处理、玻璃化溶液处理、液氮冷冻保存，成功实现了玻璃化超低温冷冻保存，经过液氮冷冻保存后的材料可以继续分化、生长。适当时间的玻璃化试剂处理对于材料无致死作用，不经液氮冷冻，可以达到100%存活。-20 oC 、-40 oC、液氮保存后，存活率随温度下降而下降，表明冷冻致死的原因在于冰晶形成；提高冷冻后成活率的关键是控制干燥脱水，经过适当的自然干燥，材料存活率分别达到50.0%和27.8%。 以叶片为外植体材料，通过组织培养和快速繁殖可以大规模扩繁苦苣苔科植物。主要步骤为：外植体叶片消毒→不定芽诱导培养→生根诱导培养→继代保存或炼苗移栽，经过3-4个月时间可获得大量栽培植株。已成功保存并培养了40余种苦苣苔科植物，包括濒危苦苣苔及高观赏价值苦苣苔。

ROLE OF PHOSPHORUS CYCLING IN ALGAL METABOLISM AND ALGAL SUCCESSION IN LAKE DONGHU, CHINA

The role of phosphorus cycling in algal metabolism was studied in a shallow lake, Donghu, in Wuhan using the methods of measuring cell quota C, N and P, and calculating nutrients uptake rate by algal photosynthesis. The mean daily phosphorus uptake rate of phytoplankton varied between 0.04-0.11 and 0.027-0.053 g/m2/d in station I and station II respectively. The turnover time of phosphorus in phytoplankton metabolism ranged from 0.75-5.0 days during 1979-1986. The available P was 0.176 (+/- 0.156) g/m3 (mean +/- SD) in 1982 and 0.591 (+/- 0.24) g/m3 in 1986. The relationship between P/B ratio (Y) and TP (X: mg/l) was described by the following regression equation Y = 1.163 + 0.512logX (r = 0.731, P < 0.001). The dynamics of algal biomass and algal species succession were monitored as the indicators of environmental enrichment. The small-sized algae have replaced the blue-green algae as the dominant species during 1979-1986. The small-sized algae include Merismopedia glauca, Cryptomonas ovata, Cryptomonas erosa, several species Cyclotella. There has been drastic decrease in algal biomass and an obvious increase in P/B ratio. A nutrient competition hypothesis is proposed to explain the reason of the disappearance of blue-green algae bloom. The drastic change in algal size and the results in high P/B ratio (reaching a maximum mean daily ratio of 1.09 in 1986) may suggest a transition of algal species from K-selection to r-selection in Lake Donghu.

濒危植物毛柄小勾儿茶生存群落的数量分类

应用除趋势对应分析(DCA)与双向指示种分析(TWINSPAN)对濒危植物毛柄小勾儿茶(Berchemiella wilsoniivar.pubipetiolata)69块样地进行了排序与分类,共划分9个群落类型。结果表明,毛柄小勾儿茶生境特异性高。毛柄小勾儿茶在枫香(Liquidambar formosana)林、马尾松(Pinusmassoniana)+杉木(Cunninghamia lanceolata)+青冈栎(Quercus glauca)针阔混交林、杂木林和青榨槭(Acer davidii)+大果山胡椒(Lindera praecox)林中生长较好。这4个群落林下层优势种具有相似性:草本层的优势种中均有求米草(Oplismenus undulatifolius),常有三脉紫菀(Aster ageratoides);灌木层优势种常有大果山胡椒。毛柄小勾儿茶分布于海拔500~900 m的低山区,多分布于湿润、排水良好的沟谷边,生境坡向多为阴坡。

Decomposition of emergent macrophyte roots and rhizomes in a northern prairie marsh

Dry mass, nitrogen and phosphorus content in belowground litter of four emergent macrophytes (Typha glauca Godr., Phragmites australis (Cav.) Trin., Scolochloa festucacea (Willd.) Link and Scirpus lacustris L.) were followed for 1.2 years in a series of experimental marshes, Delta Marsh, Manitoba. Litter bags containing roots and rhizome materials of each species were buried in unflooded soil, or soil flooded at three water depths (1–30, 31–60, > 60 cm). There were few differences in dry mass loss in unflooded or flooded soils, and depth of flooding also had little effect on decomposition rates. In the flooded sites, Scolochloa and Phragmites roots lost more mass (48.9–63.8% and 59.2–85.5%, respectively) after 112 days than Typha and Scirpus (36.3–43.6 and 37.0–47.2%, respectively). These differences continued through to the end of the study, except in the shallow sites where Scirpus roots lost more mass and had comparable mass remaining as Scolochloa and Phragmites. In the unflooded sites, there was little difference between species. All litters lost nitrogen (22.9–90.0%) and phosphorus (46.3–92.7%) during the first 112 days, then levels tended to remain constant. Decay rates for our belowground root and rhizome litters were comparable to published literature values for aboveground shoot litter of the same species, except for Phragmites roots and rhizomes which decomposed at a faster rate (−k = 0.0014−0.0032) than shoots (−k = 0.0003−0.0007, [van der Valk, A.G., Rhymer, J.M., Murkin, H.R., 1991. Flooding and the decomposition of litter of four emergent plant species in a prairie wetland. Wetlands 11, 1–16]).

A novel polypeptide from shark cartilage with potent anti-angiogenic activity

Using guanidine-HCl extraction, acetone precipitation, ultra-filtration and chromatography, a novel polypeptide with potent anti-angiogenic activity was purified from cartilage of the shark, Prionace glauca. N-terminal amino acid sequence analysis and SDS-PAGE revealed that the substance is a novel polypeptide with MW 15500 (PG155). The anti-angiogenic effects of PG155 were evaluated using zebrafish embryos model in vivo. Treatment of the embryos with 20 mu g/ml PG155 resulted in a significant reduction in the growth of subintestinal vessels (SIVs). A higher dose resulted in almost complete inhibition of SIV growth, as observed by endogenous alkaline phosphatase (EAP) staining assay. An in vitro transwell experiment revealed that the polypeptide inhibited vascular endothelial growth factor (VEGF) induced migration and tubulogenesis of human umbilical vein endothelial cells (HUVECs). Exposure of HUVECs in 20 mu g/ml PG155 significantly decreased the density of migrated cells. Almost complete inhibition of cell migration was found when HUVECs were treated with 40-80 mu g/ml PG155. PG155 (20 mu g/ml) markedly inhibited the tube formation of HUVECs and a dose-dependent effect was also found when treatment of HUVECs with PG155 at the concentration from 20 to 160 mu g/ml.

鲨鱼软骨抗血管生成多肽的分离纯化及其生物活性

以血管生成为靶点的抗肿瘤策略是抗肿瘤领域的研究热点，目前已经发现许多天然和化学合成的抗血管生成药物。鲨鱼软骨作为抗新生血管生成因子的重要来源的研究已有20多年的历史，很多研究显示鲨鱼软骨提取物有抗血管生成活性。但鲨鱼软骨活性多肽的完整分子结构一直未见报道；鲨鱼软骨活性多肽干扰血管生成通路的信号途径尚不明确。 本文应用盐酸胍抽提、丙酮分级沉淀、超滤、凝胶层析等分离技术，从青鲨（Prionace glauca）软骨中分离纯化并鉴定了一种新的具有抗新生血管生成活性的多肽。经SDS-PAGE和N-末端氨基酸序列分析显示，该多肽分子量为15500 Da，采用蛋白数据库分析表明该多肽是一种新发现的鲨鱼软骨多肽（Polypeptide from Prionace glauca，PG155）。 体外实验显示，PG155抑制内皮细胞生长因子（vascular endothelial growth factor，VEGF）介导的人脐静脉内皮细胞（human umbilical vein endothelial cell ，HUVEC）迁移和管腔形成，并呈剂量依赖关系。200 μg/ml PG155对牛主动脉内皮细胞（Bovine Aortic Endothelial Cells，BAECs）和HUVECs及以下癌细胞，包括人肝癌细胞（human hepatoma Bel-7402 cells，Bel-7402）、 口腔上皮癌细胞（human oral epidermoid carcinoma KB cells ，KB）、人结肠癌细胞（human colon cancer HCT-18 cells，HCT-18）和人乳腺癌细胞（human breast MCF7 cancer cells ，MCF7）的增殖均无抑制作用，说明PG155无细胞毒作用。20 μg/ml PG155显著抑制HUVEC的迁移和管腔形成；40-80 μg/ml PG155 对VEGF 介导的HUVEC的迁移和管腔形成几乎完全抑制。 体内实验显示，PG155显著抑制斑马鱼胚胎模型新生血管生成，并呈剂量依赖关系。形态学观察表明PG155显著抑制斑马鱼胚胎肠下静脉（subintestinal vessels， SIVs）的生长，随着浓度的升高SIVs的生长可受到完全抑制。碱性磷酸酶染色分析显示，在一定浓度范围内，PG155随着浓度的升高对斑马鱼胚胎整体血管生成抑制作用依次增强。160 μg/ml PG155会引起斑马鱼胚胎心脏功能障碍。 由海洋生物中发现新的肿瘤新生血管生成抑制剂国内外的报道较少，我们的工作表明鲨鱼软骨可作为血管生成抑制剂的重要来源，鲨鱼软骨活性多肽PG155由于具有极低的细胞毒作用，并能抑制VEGF介导的血管生成过程，有希望成为一类新型抗肿瘤药物。

Caracterização das capturas de tartaruga careta (Caretta caretta) e influência de parâmetros ambientais e pesqueiros, na pesca dirigida ao espadarte (Xiphias gladius) nos Açores

Dissertação de mest., Estudos Marinhos e Costeiros, Faculdade de Ciências do Mar e Ambiente, Univ. do Algarve, 2005