鳗弧菌(Vibrio anguillarum)胞外产物中蛋白酶的纯化及其性质

对鳗弧菌 (Vibrioanguillarum)胞外产物中蛋白酶的纯化方法及其性质进行了研究 .其中 ,蛋白酶的纯化步骤包括 :(1)硫酸铵沉淀 ;(2 )SephadexG 10 0凝胶过滤 ;(3)DEAE Sepharose色谱分离 ;(4)DEAE Cellulose (DE 32 )色谱分离 .经上述纯化步骤得到两种蛋白酶 ,经SDS PAGE分析 ,Mr分别为 37.4× 10 3 和 33.1× 10 3 .以偶氮酪蛋白 (azocasein)作底物测其水解酶活性 ,结果表明二者差别显著 ,前者明显高于后者 .而且 ,Mr37.4× 10 3 的蛋白酶在pH 7～ 10范围内均显示较高活性 ,在 4 0～ 6 0℃范围内稳定性好 .结果表明该蛋白酶是一种金属蛋白酶 .关于该蛋白酶对牙鲆(Paralichtysolivaceus)的毒性作用尚在研究之中 .图 6表 3参 17     

维生素C二步发酵中L-山梨糖脱氢酶的性质及作用

从氧化葡萄糖酸杆菌细胞质中分离纯化出L 山梨糖脱氢酶 (SDH) ,其分子量为 46× 10 3 ,表观分子量为 190× 10 3 ;在测试范围内 ,最适pH是 7.4,温度为 5 5℃ ,最稳定的 pH是 7.0 ,温度为 30℃以下 ;FeCl3 促进酶活 ,CoCl2 抑制酶活 .该酶活力与发酵产物 2 酮基 L 古龙酸的合成呈正相关 ;伴生菌促进产酸菌生长和代谢 ,并使该酶比活力增加 ,从而提高发酵系统中该酶的总活力 .图 11表 5参 9     

镉、汞、锌3种重金属的雌激素样作用（Estrogenic Activity of Cadmium, Mercury, and Zinc）

为探讨镉、汞、锌3种重金属的雌激素样作用,采用双杂交酵母法测定了醋酸镉、氯化汞、醋酸锌单独作用时对重组基因酵母β-半乳糖苷酶诱导活性的EC50值.实验结果表明,醋酸镉浓度为1.0×10-5mol·L-(1实验浓度范围10-7~10-2mol·L-1),氯化汞浓度为5.0×10-7mol·L-(1实验浓度范围10-9~10-5mol·L-1),醋酸锌浓度为1.0×10-4mol·L-(1实验浓度范围10-9~10-2mol·L-1)时对酵母β-半乳糖苷酶诱导活性最大,分别达到1.3、0.95和1.1U.不能求出镉、汞、锌单独作用时对β-半乳糖苷酶诱导活性的EC50值.一味计算EC50值不仅难以评价重金属的雌激素活性,而且限制重组受体基因酵母法的普遍运用.一些重金属通过MCF-7细胞法(E-screen)呈阳性,通过重组hERα酵母法YES检测呈阴性,有可能是重金属通过与ERβ作用而产生效应.若构建出含ERβ基因的双杂交酵母菌株,与现有的方法互补将能完善环境雌激素的体外测评系统.     

酶催化荧光法测定大气水相中过氧化氢

本文报道的酶催化荧光技术测定大气水相中H_2O_2,可以在野外采集样品,经简单前处理后带回中心实验室测定。方法的最低检测限为2×10~(-8)M,在1×10~(-7)M至2×10~(-5)MH_2O_2浓度范围内工作曲线呈线性,在2 × 10~(-6)M和1×10~(-5)M时的变异系数小于3.0％.若用流动注射方式工作,样品量小于1ml,2—3min 可以测定一个样品.     

人参皂苷β-葡萄糖苷酶的分离纯化及其酶学特性

通过硫酸铵分级沉淀、透析和高效液相色谱等技术分别从两株黑曲霉Aspergillusniger 4 8g和A .niger 84 8g中分离纯化出了具有水解人参皂苷葡萄糖苷键的 β 葡萄糖苷酶 ,并对其酶学特性进行了研究 .结果表明 :两株黑曲霉A .niger 4 8g和A .niger 84 8g所产的 β 葡萄糖苷酶的表观分子量不同 ,分别为 34× 10 3 和 31× 10 3 kD ;两株菌所产的 β 葡萄糖苷酶的反应特性基本相同 .图 4表 4参 15     

脆江蓠凝集素的部分性质及细胞凝集作用

脆江蓠藻体的磷酸盐缓冲液浸提液 ,经硫酸铵分级沉淀 ,再经DEAE -Sepharose和SephadexG 10 0层析纯化 ,得到脆江蓠凝集素 (GBL) .经测定该凝集素是一种分子量为 33.1× 10 3 ～ 33.8× 10 3 的糖蛋白 ,分子中含有 11%的中性糖 ,具有很高的耐热性 .GBL的凝血活性可被某些单糖或双糖及卵粘蛋白抑制 ,当浓度为 2 .34 μg/mL时引起兔红细胞凝集 ,浓度为 18.75 μg/mL时凝集绵羊红细胞 ,但对鸡、鸭、鸽子红细胞及人A、O、或B型血细胞没有凝集作用 .GBL能够凝集两种单细胞藻类 ,当浓度为 18.75 μg/mL时能凝集盐生杜氏藻 ,但藻细胞密度减少到 18.3× 10 4 /mL时不能发生凝集作用 ;GBL浓度为 9.37μg/mL时引起蛋白核小球藻凝集 ,但当藻细胞密度减少到 2 8.9× 10 4 /mL也不产生凝集反应 .GBL还能够凝集正常的枯草芽孢杆菌和经过热处理的酿酒酵母细胞 .图 3表 6参 14     

几种化感物质对杉木幼苗生长的影响

采用培养皿滤纸法 ,研究了不同浓度的肉桂酸、苯甲酸、对羟基苯甲酸对杉木 (Cunninghamialanceolata)幼苗生长的影响 .结果表明肉桂酸、苯甲酸、对羟基苯甲酸分别在 1× 10 -5molL-1、1× 10 -3molL-1、1× 10 -4 molL-1浓度时降低了叶绿素含量 (P =0 .0 5 ) ,而在 1× 10 -6molL-1、1× 10 -4 molL-1、1× 10 -5molL-1浓度时抑制了杉木幼苗胚根和胚芽的生长 (P =0 .0 1) .3种酚类物质对胚根生长的抑制作用明显高于对胚芽生长的抑制作用 .在 3种酚类物质中 ,肉桂酸对杉木幼苗生长的抑制作用最强 ,对羟基苯甲酸次之 ,苯甲酸最弱 .这表明酚类物质能在不同程度上抑制杉木幼苗的生长 ,降低其生产力 ,可能是连栽杉木人工林生产力降低的因素之一 .图 2表 1参 14     

白眉蝮蛇毒精氨酸酯酶对CHL细胞增殖抑制和染色体畸变的影响

研究了蝮蛇清栓酶的有效成分精氨酸酯酶体外对中国仓鼠肺成纤维细胞 (CHL)增殖和染色体畸变的影响 .精氨酸酯酶的浓度以酶活单位U表示 .细胞增殖抑制试验和染色体畸变试验都用两种方法 :直接法和代谢活性法 .精氨酸酯酶设 3个浓度 :1.0× 10 -3 U /mL ,0 .5× 10 -3 U /mL和 0 .2 5× 10 -3U/mL .其中最高浓度比临床治疗浓度高约10 0倍 .细胞增殖抑制试验采用细胞贴壁培养 ,乙醇固定 ,结晶紫染色 ,单层细胞密度计测定细胞密度 .染色体畸变试验采用给药后 2 4h和 48h收集中期分裂细胞 ,低渗固定 ,制片 ,Giemsa染色 ,观察 10 0个中期分裂细胞的染色体结构异常及多倍体的出现率 .试验结果表明 ,精氨酸酯酶在上述浓度对CHL细胞的增殖没有抑制作用 ,对CHL细胞的染色体结构也没有影响 .表 2参 10     

嗜热脂肪芽孢杆菌过氧化氢酶基因perA在大肠杆菌中的高效表达

利用PCR技术得到嗜热脂肪芽孢杆菌 (Bacillusstearothermophilus)过氧化氢酶基因 perA ,将该基因与表达载体 pKK2 2 3 3连接构建重组质粒pK perA ,转化大肠杆菌过氧化氢酶HPⅠ和HPⅡ双缺突变株UM 2 ,得到重组大肠杆菌UM 2 1.酶活测定结果表明 ,表达产物具有正常的生物学活性 .SDS PAGE电泳结果显示出明显的特异性表达条带 ,单体Mr =86× 10 3 ,与嗜热脂肪芽孢杆菌所产酶相同 .实验表明 ,重组质粒在宿主UM 2中有较好的稳定性 ,在无选择压力条件下传代 6 0次基本保持稳定 ,传代 10 0次重组质粒保留 80 %以上 .摇瓶实验确定重组菌的最佳表达条件为 :IPTG浓度 ,0 .75mmol/L ;诱导时间 3h ;培养基起始 pH 6 .5 ;诱导温度 37℃ ;装液量 5 0mL/ 2 5 0mL .在优化条件下 ,重组菌产生的过氧化氢酶占菌体总蛋白的 8% ,酶活力可达 35U/mL ,是原始菌株BacillusstearothermophilusIAM110 0 1的 11.7倍 .图 2表 1参 10     

藤茶绿枝扦插技术初探

使用NAA、IAA、IBA、ABT ,浓度分别为 10 0× 10 -6、2 0 0× 10 -6、30 0× 10 -6、5 0 0× 10 -6,基质分别为细炉灰渣、洁净河沙、湿锯末进行藤茶深枝扦插试验 ,试验结果表明 :半木质化枝条经 30 0 - 5 0 0× 10 -6NAA处理 3-5s ,基质用湿锯末 ,在 2 0 - 2 5℃温度下 ,相对湿度 90 %的大棚喷雾环境中 ,成苗率高、质量好     

Rapid,recurring, structured survey versus bioblitz for generating biodiversity data and analysis with a multispecies abundance model

A bioblitz inexpensively and quickly generates biodiversity data, but bioblitzes are often conducted with haphazard, unreplicated sampling. Results tend to be taxonomically, geographically, or temporally biased, lack metadata, and consist of lists of observed taxa that do not enable further analyses or correction for imperfect detection. A rapid, recurring, structured survey (RRSS) uses a structured sampling design and temporal and spatial replication to survey randomly selected sites on a conservation property. We participated in a loosely structured bioblitz and a subsequent RRSS at Big Canoe Creek Nature Preserve in Springville (St. Clair County), Alabama (USA) to compare observed richness derived from the 2 survey approaches. The RRSS data structure enabled us to fit models that accounted for imperfect detection to estimate abundances, occupancy probabilities, and habitat associations. The loosely structured bioblitz data could not be used in such models. We present a new integrated multispecies abundance model that we applied to avian RRSS data. Our model extension enables estimation for the community, employs data augmentation to estimate the number of undetected species, and incorporates covariates. The RRSS generated a more comprehensive and less biased list of observed taxonomic richness than the loosely structured bioblitz (e.g., 73 vs. 45 bird species and 104 vs. 63 insect families from the RRSS vs. loosely structured bioblitz, respectively). Models fit to the RRSS data identified seasonal patterns in avian community composition and allowed for estimation of habitat–occupancy relationships for insect taxa. The RRSS protocol has potential for broad transferability as a standardized, quick, and inexpensive way to inventory biodiversity and estimate ecological parameters while providing an outreach opportunity.     

Compliance with and ecosystem function of biodiversity offsets in North American and European freshwaters

Land-use change via human development is a major driver of biodiversity loss. To reduce these impacts, billions of dollars are spent on biodiversity offsets. However, studies evaluating offset project effectiveness that examine components such as the overall compliance and function of projects remain rare. We reviewed 577 offsetting projects in freshwater ecosystems that included the metrics project size, type of aquatic system (e.g., wetland and creek), offsetting measure (e.g., enhancement, restoration, and creation), and an assessment of the projects’ compliance and functional success. Project information was obtained from scientific and government databases and gray literature. Despite considerable investment in offsetting projects, crucial problems persisted. Although compliance and function were related to each other, a high level of compliance did not guarantee a high degree of function. However, large projects relative to area had better function than small projects. Function improved when projects targeted productivity or specific ecosystem features and when multiple complementary management targets were in place. Restorative measures were more likely to achieve targets than creating entirely new ecosystems. Altogether the relationships we found highlight specific ecological processes that may help improve offsetting outcomes.     

A palaeo-hydrogeological model for arsenic contamination in southern and south-east Asia

An argument is presented in which areas of natural arsenic contamination of modern groundwaters throughout Asia have a common origin. Arsenic originally accumulated in oceanic ferro-manganoan sediments of the eastern Palaeo-Tethys. This was further concentrated through oceanic crustal extinction in what later became the south-east Chinese accreted mineralised terrain. Proto-Himalayan uplift of this area created the palaeo-drainage systems of the Ganges – Brahmaputra, Irrawaddy, Mekong, and Red Rivers, with consequent headwater erosion of arsenic-rich sediments. Their downstream deposition as immature and easily redistributed Neogene sandstones, silts, and iron-rich clays has created secondary and tertiary reservoirs of adsorbed and authigenic arsenic, from which the current arsenic-rich groundwaters have evolved. Considering river basins within the above palaeo-hydrogeological framework provides a basis for assessing the risk of arsenic in groundwater basins of south and south-eastern Asia.     

Cost‐effective conservation of an endangered frog under uncertainty

How should managers choose among conservation options when resources are scarce and there is uncertainty regarding the effectiveness of actions? Well‐developed tools exist for prioritizing areas for one‐time and binary actions (e.g., protect vs. not protect), but methods for prioritizing incremental or ongoing actions (such as habitat creation and maintenance) remain uncommon. We devised an approach that combines metapopulation viability and cost‐effectiveness analyses to select among alternative conservation actions while accounting for uncertainty. In our study, cost‐effectiveness is the ratio between the benefit of an action and its economic cost, where benefit is the change in metapopulation viability. We applied the approach to the case of the endangered growling grass frog (Litoria raniformis), which is threatened by urban development. We extended a Bayesian model to predict metapopulation viability under 9 urbanization and management scenarios and incorporated the full probability distribution of possible outcomes for each scenario into the cost‐effectiveness analysis. This allowed us to discern between cost‐effective alternatives that were robust to uncertainty and those with a relatively high risk of failure. We found a relatively high risk of extinction following urbanization if the only action was reservation of core habitat; habitat creation actions performed better than enhancement actions; and cost‐effectiveness ranking changed depending on the consideration of uncertainty. Our results suggest that creation and maintenance of wetlands dedicated to L. raniformis is the only cost‐effective action likely to result in a sufficiently low risk of extinction. To our knowledge we are the first study to use Bayesian metapopulation viability analysis to explicitly incorporate parametric and demographic uncertainty into a cost‐effective evaluation of conservation actions. The approach offers guidance to decision makers aiming to achieve cost‐effective conservation under uncertainty.     

Parasitic wasps orient to green leaf volatiles

Summary Undamaged plants emit low levels of green leaf volatiles (GLVs), while caterpillar-damaged and artificially damaged plants emit relatively higher levels of certain GLVs. Female braconid parasitoids,Microplitis croceipes, oriented to both damaged plants and to individual GLVs in no-choice tests in a wind tunnel, but seldom oriented to undamaged plants. Female ichneumonid parasitoids,Netelia heroica, also oriented to individual GLVs in a wind tunnel. Males of both wasp species failed to orient to the GLVs. These data show that leaf-feeding caterpillars can cause the release of GLVs, and that parasitic wasps can respond to these odors by flying upwind (chemoanemotactic response), which brings the wasps to their caterpillar hosts. This supports the hypothesis that plants communicate with members of the third trophic level,i.e., plants under herbivore attack emit chemical signals that guide natural enemies of herbivores to sites of plant damage. In this interaction, the GLVs serve as tritrophic plant-to-parasitoid synomones. That parasitoids from two different wasp families oriented to GLVs suggests that the response may be widespread among the Hymenoptera.Mention of a commercial or proprietary product does not constitute an endorsement by the U.S. Department of Agriculture     

Potential for Electropositive Metal to Reduce the Interactions of Atlantic Sturgeon with Fishing Gear

Atlantic sturgeon (Acipenser oxyrhynchus) populations have been declared either endangered or threatened under the U.S. Endangered Species Act. Effective measures to repel sturgeon from fishing gear would be beneficial to both fish and fishers because they could reduce both fishery‐associated mortality and the need for seasonal and area closures of specific fisheries. Some chondrostean fishes (e.g., sturgeons and paddlefishes) can detect weak electric field gradients (possibly as low as 5 Μv/cm) due to arrays of electroreceptors (ampullae of Lorenzini) on their snout and gill covers. Weak electric fields, such as those produced by electropositive metals (typically mixtures of the lanthanide elements), could therefore potentially be used as a deterrent. To test this idea, we recorded the behavioral responses of juvenile Atlantic sturgeon (31–43 cm fork length) to electropositive metal (primarily a mixture of the lanthanide elements neodymium and praseodymium) both in the presence and absence of food stimuli. Trials were conducted in an approximately 2.5 m diameter × 0.3 m deep tank, and fish behaviors were recorded with an overhead digital video camera. Video records were subsequently digitized (x, y coordinate system), the distance between the fish and the electropositive metal calculated, and data summarized by compiling frequency distributions with 5‐cm bins. Juvenile sturgeon showed clear avoidance of electropositive metal but only when food was present. On the basis of our results, we conclude that the electropositive metals, or other sources of weak electric fields, may eventually be used to reduce the interactions of Atlantic sturgeon with fishing gear, but further investigation is needed. El Potencial del Metal Electropositivo para Reducir las Interacciones del Esturión Atlántico con Instrumentos de Pesca Bouyoucos, Bushnell & Brill 13–003     

The role of time and species identities in spatial patterns of species richness and conservation

Many conservation actions are justified on the basis of managing biodiversity. Biodiversity, in terms of species richness, is largely the product of rare species. This is problematic because the intensity of sampling needed to characterize communities and patterns of rarity or to justify the use of surrogates has biased sampling in favor of space over time. However, environmental fluctuations interacting with community dynamics lead to temporal variations in where and when species occur, potentially affecting conservation planning by generating uncertainty about results of species distribution modeling (including range determinations), selection of surrogates for biodiversity, and the proportion of biodiversity composed of rare species. To have confidence in the evidence base for conservation actions, one must consider whether temporal replication is necessary to produce broad inferences. Using approximately 20 years of macrofaunal data from tidal flats in 2 harbors, we explored variation in the identity of rare, common, restricted range, and widespread species over time and space. Over time, rare taxa were more likely to increase in abundance or occurrence than to remain rare or disappear and to exhibit temporal patterns in their occurrence. Space–time congruency in ranges (i.e., spatially widespread taxa were also temporally widespread) was observed only where samples were collected across an environmental gradient. Fifteen percent of the taxa in both harbors changed over time from having spatially restricted ranges to having widespread ranges. Our findings suggest that rare species can provide stability against environmental change, because the majority of species were not random transients, but that selection of biodiversity surrogates requires temporal validation. Rarity needs to be considered both spatially and temporally, as species that occur randomly over time are likely to play a different role in ecosystem functioning than those exhibiting temporal structure (e.g., seasonality). Moreover, temporal structure offers the opportunity to place management and conservation activities within windows of maximum opportunity.     

Main nutrient compounds in food bodies of Mexican <Emphasis Type="Italic">Acacia</Emphasis> ant-plants

Summary. Myrmecophytic plants use obligate ant mutualists as a constitutive indirect defence mechanism. These plants often produce cellular food bodies (FBs) to nourish their resident ants. Lipids, proteins, and even highly specialised compounds such as glycogen have been reported from FBs, but detailed chemical analyses of FB composition have so far been presented only for Southeast Asian Macaranga and Central American Piper myrmecophytes. Here we report the chemical composition of FBs of five myrmecophytic Acacia (Fabaceae) species from Mexico using HPLC (carbohydrates and proteins) and GC-MS (lipids). Feeding experiments revealed no hints on any use of external food sources by the inhabiting Pseudomyrmex ants. These ants obviously rely completely on FBs and extrafloral nectar provided by their hosts. The total content of nutrients in Acacia FBs was 15-25% of FB dry mass, being much lower than in Macaranga or Piper FBs. Proteins were dominating (8-14 % dm) in Acacia FBs and thus were present in higher amounts than in Macaranga FBs, yet in lower amounts than in Piper. Lipids contributed 1-9 % of dry mass, showing a lower proportion than in FBs of Macaranga or Piper. Carbohydrates made up 3-11 % dm, reaching in most Acacia species the same range as observed in Macaranga and in Piper FBs. Water content was 18-24 % of FB fresh mass, and structural tissue obviously made up a much higher proportion in Acacia FBs than in Macaranga or Piper FBs. Both characters might represent an adaptation to producing FBs unprotected at the leaf tips under dry conditions. Acacia FBs contain all amino acids and all fatty acids that are considered essential for insects, and their contents of lipids and proteins are higher than in the leaves from which they are ontogenetically derived. This indicates a putatively adaptive enrichment of nutritionally valuable compounds in structures functioning as ant-food.     

Examining the uncertain origin and management role of martens on Prince of Wales Island,Alaska

Conservation biologists are generally united in efforts to curtail the spread of non‐native species globally. However, the colonization history of a species is not always certain, and whether a species is considered non‐native or native depends on the conservation benchmark. Such ambiguities have led to inconsistent management. Within the Tongass National Forest of Alaska, the status of American marten (Martes americana) on the largest, most biologically diverse and deforested island, Prince of Wales (POW), is unclear. Ten martens were released to POW in the early 1930s, and it was generally believed to be the founding event, although this has been questioned. The uncertainty surrounding when and how martens colonized POW complicates management, especially because martens were selected as a design species for the Tongass. To explore the history of martens of POW we reviewed other plausible routes of colonization; genetically and isotopically analyzed putative marten fossils deposited in the late Pleistocene and early Holocene to verify marten occupancy of POW; and used contemporary genetic data from martens on POW and the mainland in coalescent simulations to identify the probable source of the present‐day marten population on POW. We found evidence for multiple routes of colonization by forest‐associated mammals beginning in the Holocene, which were likely used by American martens to naturally colonize POW. Although we cannot rule out human‐assisted movement of martens by Alaskan Natives or fur trappers, we suggest that martens be managed for persistence on POW. More generally, our findings illustrate the difficulty of labeling species as non‐native or native, even when genetic and paleo‐ecological data are available, and support the notion that community resilience or species invasiveness should be prioritized when making management decisions rather than more subjective and less certain conservation benchmarks.