新的端粒酶活性抑制蛋白LPTS-L的制备

LPTS基因是利用定位候选克隆策略克隆的一个新的肝相关候选肿瘤抑制基因。LPTS基因编码一个全长为328氨基酸的蛋白质(LPTS-L),该蛋白具有抑制细胞端粒酶活性的功能。为了进一步研究LPTS-L蛋白的结构与功能,利用DNA重组技术,将LPTS-L的cDNA克隆到表达载体pET-24a中构建重组克隆pET-24-LPTS,并在大肠杆菌BL-21中进行融合表达,获得可溶形式的LPTS-L融合蛋白。采用Ni Sepharose4B柱亲和层析,可以获得纯度较高的蛋白,但不适合大量制备。通过设计引物去掉了pET-24a载体上的6×His tag将LPTS-L基因进行了非融合表达,然后采用磷酸纤维素P11阳离子交换层析纯化LPTS-L蛋白,纯度可达到55%。再经Sephadex G-100凝胶过滤,LPTS-L蛋白的纯度可达到80%。Western blot实验显示经纯化后的LPTS-L蛋白可与兔抗GST-LPTS-L的多抗发生特异性结合。采用TRAP法测定蛋白质活性,结果显示纯化得到的LPTS-L蛋白可抑制端粒酶的活性,与采用Ni Sepharose4B纯化获得的LPTS-L融合蛋白比较,其抑制效率基本一致。因此,所建立的技术可以有效地制备LPTS-L蛋白。     

Programmed cell death during development of cowpea (Vigna unguiculata (L.) Walp.) seed coat

The seed coat develops primarily from maternal tissues and comprises multiple cell layers at maturity, providing a metabolically dynamic interface between the developing embryo and the environment during embryogenesis, dormancy and germination of seeds. Seed coat development involves dramatic cellular changes, and the aim of this research was to investigate the role of programmed cell death (PCD) events during the development of seed coats of cowpea [Vigna unguiculata (L.) Walp.]. We demonstrate that cells of the developing cowpea seed coats undergo a programme of autolytic cell death, detected as cellular morphological changes in nuclei, mitochondria, chloroplasts and vacuoles, DNA fragmentation and oligonucleosome accumulation in the cytoplasm, and loss of membrane viability. We show for the first time that classes 6 and 8 caspase‐like enzymes are active during seed coat development, and that these activities may be compartmentalized by translocation between vacuoles and cytoplasm during PCD events.     

Quantifying the role of steric constraints in nucleosome positioning

Statistical positioning, the localization of nucleosomes packed against a fixed barrier, is conjectured to explain the array of well-positioned nucleosomes at the 5′ end of genes, but the extent and precise implications of statistical positioning in vivo are unclear. We examine this hypothesis quantitatively and generalize the idea to include moving barriers as well as nucleosomes actively packed against a barrier. Early experiments noted a similarity between the nucleosome profile aligned and averaged across genes and that predicted by statistical positioning; however, we demonstrate that aligning random nucleosomes also generates the same profile, calling the previous interpretation into question. New rigorous results reformulate statistical positioning as predictions on the variance structure of nucleosome locations in individual genes. In particular, a quantity termed the variance gradient, describing the change in variance between adjacent nucleosomes, is tested against recent high-throughput nucleosome sequencing data. Constant variance gradients provide support for generalized statistical positioning in ∼50% of long genes. Genes that deviate from predictions have high nucleosome turnover and cell-to-cell gene expression variability. The observed variance gradient suggests an effective nucleosome size of 158 bp, instead of the commonly perceived 147 bp. Our analyses thus clarify the role of statistical positioning in vivo.     

拉萨河流域亏组山植物物种丰富度和群落特征研究

为研究草地植物群落、物种丰富度及其分布格局与影响因子之间的关系,该文以拉萨河流域林周县卡孜乡亏组山为研究地点,对山体垂直样带(3900~5100 m)植物群落特征、植物物种丰富度与各影响因子之间的关系进行了研究。研究区域共设置了13个样带(每隔100 m设一个样带),每样带设置0.5 m×0.5 m的5个样方进行植被调查,运用主成分分析(PCA)和双向指示种分析法(TWINSPAN)对植物群落进行排序和分类,运用冗余分析法(RDA)对群落及其分布格局与影响因子之间的关系进行分析,对植物物种丰富度与各因子之间的关系进行了回归分析。结果表明:该山体植物可分为3个群落类型,含7个群丛;影响区域植物群落物种组成和分布格局的主要环境因子为海拔,其次是坡度;物种丰富度与海拔、分种盖度呈单峰关系,与总盖度、坡度、地上生物量呈正相关。该研究结果为区域植物物种多样性和山地植物资源的保护和可持续利用提供了科学依据。     

Effects of the geomorphometric characteristics of the local terrain on floristic composition in the central Brazilian Amazon

The objective of this study was to identify the effects of local geomorphometry on the abundance, richness and floristic composition of tree species in the central Brazilian Amazon. Forty‐six 0.25‐ha plots in different phyto‐ecologic sites were sampled, and their trees were inventoried. Geomorphometric data (elevation, slope, aspect, plan and profile curvatures) were derived from Shuttle Radar Topography Mission data. A detrended correspondence analysis (DCA) was used to examine the floristic distribution patterns among plots. In addition, geomorphometric variables were submitted to multiple regression analysis to identify the variables influencing floristic composition (represented by the first DCA component), abundance and species richness. Correlation analyses between the number of individuals from each species and the first DCA component were performed to evaluate the contribution of each species. Analysis of the results could not confirm an effect of geomorphometry alone on species richness and abundance, although floristic composition was significantly influenced by profile curvature and elevation. Despite the relatively low variation in altitude at the study site, species were found to be sensitive to terrain peculiarities such as elevation and profile curvature, which can constrain particular ecologic niches and contribute to the spatial distribution patterns of species.     

Phylogenetic and phenotypic originality and abundance in a cerrado plant community

The originality of a species is how much that species contributes to the rarity of traits in a community. Here we tested the relation between abundance and both phylogenetic and phenotypic originality. We measured nine traits associated with defence against herbivory, as well as phylogenetic information and abundance for woody plant species in a woodland cerrado in southeastern Brazil. About 90% of the species accounted for about 50% of the phylogenetic and phenotypic originality: most woody species had low originality. Abundance was related to tougher leaves, lower specific leaf area and lower originality based on nutritional quality. Our results suggest that herbivory may reduce the abundance of species with low resistance to herbivory and with different nutritional quality. Nevertheless, abundance was not related to either phylogenetic or phenotypic originality, so extinction of rare species may not endanger overall community function as long as more abundant species are retained. We argue that this is a consequence of the low complementarity of a large number of woody species.     

Floral biology of Eriope blanchetii (Lamiaceae) in coastal sand dunes of NE Brazil

Abstract The floral biology of the sand dune shrub Eriope blanchetii (Benth) Harley was investigated in a wild population located within an environment protection area near Salvador, NE Brazil. Inflorescence and flower development were monitored, and the reproductive biology was assessed. Inflorescences are terminal panicles, each bearing from one to 38 small, zygomorphic flowers. The duration of an inflorescence is about 2 months, with few new flowers each day. The rewards to visitors are pollen and nectar, the latter being produced in small quantities at the corolla base, near the stylopodium. Three sequential phases of flower development were identified, based on colour changes, pollen and nectar availability, and stigmatic receptivity. Anthesis takes place throughout the day within each plant, and flowers are strongly protandrous. Flowers are short‐lived, with individual variations of 7–30 h until corolla abscission. This variation is probably associated to a post‐pollinating response, which reduces the length of the female phase and could be advantageous for the economy of nectar production and for increased efficient pollination of the remaining flowers on a plant. The results of the pollination tests (apomixis, spontaneous and manual autogamy, manual cross‐pollination, and control) reinforced the role of pollinators for the reproductive success of E. blanchetii, given that the production of fruits from spontaneous pollination is unlikely to be resulting from protandry and given the morphological floral features. Taken together, the morphological and physiological aspects of E. blanchetii floral biology favour cross‐pollination, and it is likely that most of the fruits in nature are produced by outcrossing.     

浙江天童国家森林公园常绿阔叶林主要组成种的种群结构及更新类型

 以浙江天童国家森林公园常绿阔叶林为对象,应用永久样方法和每木调查法调查了群落种类组成和结构,并对主要组成种的种群结构进行了分析。依据胸径级频率分布的形状,将各树种的种群结构归纳为5种类型,并结合生物学、生态学特征,分别讨论了其更新类型,同时对调查群落所处的演替阶段进行了诊断。结果表明：1）群落为6种共优势群落,出现木本植物69种。2）种群结构为单峰型的马尾松（Pinus massoniana）、枫香（Liquidambar formosana）和檫木（Sassafras tzumu）等为阳性高大乔木,无正常更新能力,更新类型是先锋群落优势种或是顶极性先锋种;间歇型的木荷（Schima superba）、黑山山矾（Symplocos heishanensis）和红楠（Machilus thunbergii）等为具有耐荫性的常绿阔叶树种,更新具有波动性和机会性,是演替系列或顶极群落优势种;逆-J字型的米槠（Castanopsis carlesii）、栲树（Castanopsis fargesii）和长叶石栎（Lithocarpus harlandii）等为强耐荫性的常绿阔叶树种,通过幼苗库和根萌生枝进行更新,是顶极群落的优势种;浙江新木姜子（Neolitsea aurata var. chekiangensis）、厚皮香（Ternstroemia gymnanthera）等为L字型;连蕊茶（Camellia fraternna）、老鼠矢（Symplocos stellaris）等为单柱型,此二型是群落亚乔木层和灌木层的主要组成种。3）调查群落已进入演替的顶极阶段,但仍处于其前期,将进一步向以米槠、栲树为主要优势种的方向发展。     

Distribution of Epitopes of Trypanosoma cruzi Amastigotes During the Intracellular Life Cycle within Mammalian Cells

ABSTRACT. In this study we have examined the distribution of epitopes defined by monoclonal antibodies raised against Trypanosoma cruzi amastigotes during the intraceullar life cycle of the parasite. We have raised monoclonal antibodies towards amastigote forms and performed preliminary immunochemical characterization of their reactivities. MAB 1D9, 3G8, 2B7, 3B9, and 4B9, and 4B9 react with carbohydrate epitopes of the parasite major surface glycoprotein—Ssp-4 defined by MAB 2C2 [5]: MAB 4B5 reacts with a noncarbohydrate epitope in all developmental stages of the parasite, and MAB 3B2 also detects a noncarbohydrate epitope preferentially in T. cruzi flagellared forms. Vero cells infected with tissue culture-derived trypomastigotes of clone D11 (G strain) were fixed at different times during the intraceullular proliferation of parasites, and processed for immjno-electron microscopy and confocal immunoflurescence with the different monoclonal antibodies. We observed that while the surface distribution of MAB 2C2 and 4B9 epitopes was uniform throughout the cycle, MAB 1D9, 3G8, and 2B7 reacted with cytoplasmic membrance-bound compartments of the amastigotes. MAB 3B9 displayed a unique surface dentate pattern in some amastigotes. MAB 4B5 recognized a curved-shaped structure at the flagellar pocket region in some intracellular amastigotes and localized to the membrane in dividing forms. In intracellular trypomastigotes, MAB 4B5 also displayed a punctate pattern near the flagellar pocket.     

上海乡土水生植物资源及其在水生态恢复与水景观建设中的应用潜力

水生植物作为水生生态系统的主体, 对发挥水生生态系统的自维持、自循环功能有重要作用。研究通过相关资料的查阅, 建立上海地区乡土水生植物名录, 并对其科属组成、区系特征、生活型、生长型等进行统计分析。结果表明上海地区乡土水生植物共计35 科83 属160 种(含变种), 单属科、单种属的比例较高, 均达65%以上; 植物区系组成丰富、成分复杂, 以热带成分占优势, 达64.6%; 生活型以挺水植物为主, 沉水植物次之, 浮水植物最少; 生长型类型丰富, 以草本型、禾草型居多, 20 种生长型可进一步归为表征相似生态学特征和功能地位的6 个生长型组。在水生态恢复与水景观建设中, 仅有68.8%的景观水体有水生植物应用,且应用种类在2 种以下的占79.2%。乡土水生植物应用不足, 一半以上为观赏性强的外来物种, 应用频率较高的为挺水植物, 对具有良好净化效果的沉水植物重视不够。因此, 在水生态恢复与水景观建设中, 建议加强乡土水生植物资源的繁育栽培, 在充分利用乡土水生植物资源配置群落的基础上, 根据水质的富营养及基底状况, 通过不同生长型组水生植物的应用, 构建沉水-浮水-挺水植物群落复合体, 并通过近自然型护岸的营造, 形成水生-湿生复合生态系统。运用植被工程学的原理和方法, 采用生态浮岛、生态沉岛等技术营造水生植被, 将强人工化的水景观建成具生命的水生生态系统。