唐鱼卵黄脂磷蛋白的纯化鉴定与免疫原性分析

采用Native-PAGE和SDS-PAGE方法从性成熟雌性唐鱼(Tanichthys albonubes)卵巢组织提纯了卵黄脂磷蛋白(Lv).已确定被纯化的唐鱼Lv在Native-PAGE(4%～7.5%)电泳中分子量(Mr)为314 ×103.用纯化的唐鱼Lv免疫大白鼠获得鼠源多克隆抗血清.以Native-PAGE和SDS-PAGE制备的唐鱼Lv及裂解后的组分作为抗原所制备的抗血清,与经17β-雌二醇(E2)诱导的雄性唐鱼、去除卵巢的雌性唐鱼以及唐鱼卵巢的匀浆液能发生免疫反应,并且印迹位置与雌性特异蛋白卵黄蛋白原(Vtg)的位置相当.但是两种血清和未经E2诱导的雄鱼整体匀浆液并无反应,显示Lv及其大分子亚基的抗血清与Vtg和Lv两种蛋白是特异的.结果表明,唐鱼Lv裂解组分的抗血清可用于检测唐鱼的Vtg.图3参19     

弱阴离子交换-超滤两级纯化浓缩真鲷卵黄蛋白原

利用弱阴离子交换色谱,从雌激素17β-雌二醇诱导的真鲷鱼血清中分离纯化出卵黄蛋白原粗组分,以超滤法进一步除去杂质蛋白,并脱盐浓缩,获得纯度较高的卵黄蛋白原(Vtg),两级纯化得到的Vtg经常规聚丙烯酰胺凝胶电泳(Native-PAGE)分析,证实真鲷鱼Vtg有三种形式,分子量分别为665kDa,635kDa和355kDa;变性聚丙烯酰胺凝胶电泳(SDS-PAGE)分析结果表明,它们由185kDa左右的蛋白亚基组成.     

唐鱼卵黄蛋白原的诱导、纯化与鉴定

为探讨利用雄性唐鱼(Tanichthys albonubes)卵黄蛋白原(Vitellogenin,Vtg)作为生物标志物检测环境雌激素的可行性,采用浸浴法用50μg·L-117β-雌二醇(E2)对雄性唐鱼进行染毒,30d后将鱼体整体匀浆进行常规聚丙烯酰胺凝胶电泳(Native-PAGE)以分析Vtg的产生;并采用Mg2+-EDTA选择性沉淀和Q Sepharose阴离子交换层析对Vtg进行分离纯化.结果表明,E2诱导下,雄性唐鱼产生了雌性特异蛋白Vtg,并且可在体内积累;利用Mg2+-EDTA选择性沉淀和Q Sepharose阴离子交换层析的两步纯化方法可分离纯化唐鱼整体匀浆Vtg;经Native-PAGE鉴定,确定唐鱼Vtg的分子量为440kDa.以上结果提示,雄性唐鱼Vtg可以作为环境雌激素监测的有效生物学标记物。     

印楝素对亚洲玉米螟的生物活性及蛋白质合成的影响

室内生物测定表明,印楝素对玉米螟幼虫具有明显的毒杀作用和抑制生长发育的活性.浓度3×10-6 ~15×10-6印楝素处理幼虫,均出现活动性降低、化蛹延迟、蛹出现畸形的现象,甚至形成永久性幼虫,且与处理浓度有一定的相关性.同时发现,经印楝素处理后,亚洲玉米螟48h蛹蛋白质的含量和组分与对照相比出现明显的差异,含量与处理浓度有较好的相关性.SDS-PAGE结果显示, 经3×10-6 ~15×10-6印楝素处理,昆虫的蛹均出现分子量Mr55×103、50×103两种新蛋白组分,减少了21. 6×103、20×103两种蛋白组分;另外10×10-6和15×10-6印楝素处理还减少了22×103蛋白组分, 15×10-6处理还出现9×103新蛋白组分; 这6种差异蛋白可能是印楝素分子作用的相关蛋白. 图2表1参20     

17β-雌二醇与1-萘酚对雄性罗非鱼（GIFT Oreochromis niloticus）雌激素效应的比较

在实验室条件下,研究了暴露于不同质量浓度1-萘酚（0.064、0.8、2 mg.L-1）与17β-雌二醇（1、10、100μg.L-1）下,对雄性罗非鱼（GIFT Oreochromis niloticus）性腺系数、血清雌二醇质量浓度的影响及血清中卵黄蛋白原的诱导效应,以期对它们的环境雌激素效应有所了解。研究结果显示：暴露于1-萘酚环境下30 d后,雄性罗非鱼的性腺系数有所降低,当质量浓度为2 mg.L-1时,与对照组相比,差异显著（p〈0.05）,雄性罗非鱼血清中雌二醇的量,此时亦达到最大值,为209.3 pg.mL-1,与对照组相比,差异显著（p〈0.05）;但是,血清中卵黄蛋白原并无变化。暴露于17β-雌二醇环境中的雄性罗非鱼,30 d后,其性腺系数有所降低,当质量浓度为10、100μg.L-1时,与对照组相比,差异显著（p〈0.05）;血清中的雌二醇也随着暴露质量浓度的增多,逐渐增多,当质量浓度为100μg.L-1时,差异极显著（p〈0.01）;血清中卵黄蛋白原的质量浓度明显高于对照组,与暴露质量浓度之间呈剂量效应关系。结果显示,17β-雌二醇具有很强的雌激素效应,会使罗非鱼性腺系数下降,诱导血清中雌二醇、卵黄蛋白原的生成;而1-萘酚会使罗非鱼性腺系数下降,诱导血清雌二醇的生成,但是并不诱导卵黄蛋白原的生成,所以推测其雌激素效应可能较弱,需要进一步研究。     

多氯联苯对孔雀鱼卵黄蛋白原的诱导及检测

为探讨利用孔雀鱼(Poecilia reticulata)卵黄蛋白原(Vitellogenin,Vtg)作为生物标志物检测环境雌激素的可行性,采用浸浴法分别使用17β-雌二醇(E2)和多氯联苯(PCBs)对雄性孔雀鱼成鱼进行染毒,30d后测定其性腺系数(GSI)和肝指数(LSI),并将鱼体整体匀浆进行常规聚丙烯酰胺凝胶电泳(Native-PAGE)及磷、脂和糖蛋白特异染色分析.结果表明,PCBs对雄性孔雀鱼具有雌激素效应,和E2均可以诱导雄性孔雀鱼体内产生Vtg,但诱导组雄鱼GSI及LSI与对照组比较均无显著性差异(p>0.05).Native-PAGE及磷、脂、糖蛋白特异性分析表明,孔雀鱼Vtg是一种富含磷、脂、糖的蛋白,具有VtgⅠ、VtgⅡ和VtgⅢ等3种形式,其分子量分别为642kDa、541kDa和441kDa.雄性孔雀鱼Vtg可作为环境雌激素监测的有效生物标记物.     

近平滑假丝酵母NAD(H)依赖型次级醇脱氢酶的分离纯化及酶学性质

从近平滑假丝酵母(Candida parapsilosis CCTCC 203011)中分离得到了NAD(H)依赖型的次级醇脱氢酶.粗酶经乙醇沉淀、Blue Sepharose Fast Flow亲和层析、DEAE Sepharose离子交换层析后在SDS-PAGE上显示为单一条带,其酶蛋白的亚基分子量(Mr)为37.5×103.该酶还原反应的最适pH为6.0,最适温度为45℃,是一个含Zn2 金属酶.该脱氢酶具有较高的底物的选择性,对2-酮具有较高的还原活力;同时,也能作用于醇类发生氧化反应,对次级醇的氧化活力最高.以β-羟基苯乙酮为底物,用纯化得到的酶催化反应后,还原产物为(R)-苯基乙二醇,因此该酶为(R)-专一性次级醇脱氢酶,是一种生产(R)-苯基乙二醇的有效的生物催化剂.经LC-MASS-MASS分析得到了酶蛋白中3个肽段的氨基酸序列,通过比对发现该酶与NCBI编号为BAA24528的次级醇脱氢酶具有较高的同源性.图4表3参17     

一种嗜热细菌来源角质酶的分离纯化及酶学性质

通过跟踪发酵液中pNPB水解酶活性,对角质诱导的Thermobifida fusca 口发酵液进行分离纯化.采用活性炭脱色、硫铵沉淀、Phenyl HP疏水色谱、DEAE sephamse阴离子交换色谱等方法,分离纯化得到电泳纯PNPB水解酶.该酶水解角质可得到角质单体,是一种角质酶.SDS-PAGE电泳结果显示,角质酶表观分子量约为29×10~3.该酶的最适温度为60℃.在40℃和60℃下均具有良好的热稳定性.最适pH为8.0,pH稳定范围为6.0～9.0.该角质酶的生化性质适合在纺织工业中应用.图8表2参17     

海栖热袍菌耐高温β-甘露糖苷酶基因的克隆及表达

以海栖热袍菌(Thermotoga maritima)MSB8基因组DNA为模板,克隆得到β-甘露糖苷酶基因(man2).测序分析表明,该基因全序列为2358bp,编码785个氨基酸,分子量约为92×103.根据蛋白质氨基酸的同源性分析,该β-甘露糖苷酶与新阿波罗栖热袍菌(Thermotoga neapolitana)Man2(accession No.AAK52304.1)的同源性最大,达到80%.将此基因连接至表达载体pET-28a( )并转化到大肠杆菌BL21细胞中,经IPTG诱导,β-甘露糖苷酶活力达5.96U/mL.粗酶的温度稳定性分析表明,该酶的热稳定性好,90℃处理10min,活力回收率65%,具有重要的工业应用前景.图3表1参17     

生孢噬纤维菌荚膜多糖的分离纯化及其性质

生孢噬纤维菌(Sporocytophaga)是能降解纤维素的滑动细菌,它可将滤纸和棉花纤维素完全降解.本文对生孢噬纤维菌产生的荚膜多糖的分离纯化以及性质进行了研究.以滤纸平板法培养生孢噬纤维菌,加入1%苯酚后,荚膜多糖溶解在水相中,再经氯化十六烷基吡啶(CPC)沉淀,分离出荚膜多糖提取物.利用Sephadex G-75柱层析法、胰蛋白酶和Sevag法对荚膜多糖提取物进行纯化;并利用比旋光度检查、红外光谱、液相色谱以及气相色谱等分析手段对精制后的荚膜多糖进行分析.结果表明,分离纯化得到了纯度较高的生孢噬纤维菌荚膜多糖.Sephadex G-75柱层析分析表明,荚膜多糖提取物中含有结合蛋白,除去结合蛋白后,多糖分子量约为6.5×103;气相色谱分析表明,该荚膜多糖主要由Gal、Glc、GlcA以及Man等单糖组成,其比值约为3.513.33.61;同时该荚膜多糖的红外光谱显示,该多糖所含的甲基、亚甲基等基团的量比正常多糖多;红外光谱图中881cm-1处的峰位显示,该多糖含有不典型β糖苷键特征吸收峰.图5参8     

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.     

Threats to intact tropical peatlands and opportunities for their conservation

Large, intact areas of tropical peatland are highly threatened at a global scale by the expansion of commercial agriculture and other forms of economic development. Conserving peatlands on a landscape scale, with their hydrology intact, is of international conservation importance to preserve their distinctive biodiversity and ecosystem services and maintain their resilience to future environmental change. We explored threats to and opportunities for conserving remaining intact tropical peatlands; thus, we excluded peatlands of Indonesia and Malaysia, where extensive deforestation, drainage, and conversion to plantations means conservation in this region can protect only small fragments of the original ecosystem. We focused on a case study, the Pastaza‐Marañón Foreland Basin (PMFB) in Peru, which is among the largest known intact tropical peatland landscapes in the world and is representative of peatland vulnerability. Maintenance of the hydrological conditions critical for carbon storage and ecosystem function of peatlands is, in the PMFB, primarily threatened by expansion of commercial agriculture linked to new transport infrastructure that is facilitating access to remote areas. There remain opportunities in the PMFB and elsewhere to develop alternative, more sustainable land‐use practices. Although some of the peatlands in the PMFB fall within existing legally protected areas, this protection does not include the most carbon‐dense (domed pole forest) areas. New carbon‐based conservation instruments (e.g., REDD+, Green Climate Fund), developing markets for sustainable peatland products, transferring land title to local communities, and expanding protected areas offer pathways to increased protection for intact tropical peatlands in Amazonia and elsewhere, such as those in New Guinea and Central Africa which remain, for the moment, broadly beyond the frontier of commercial development.     

Human Influence and Classical Biogeographic Predictors of Rare Species Occurrence

Biogeographic theory predicts that rare species occur more often in larger, less‐isolated habitat patches and suggests that patch size and connectivity are positive predictors of patch quality for conservation. However, in areas substantially modified by humans, rare species may be relegated to the most isolated patches. We used data from plant surveys of 81 meadow patches in the Georgia Basin of Canada and the United States to show that presence of threatened and endangered plants was positively predicted for patches that were isolated on small islands surrounded by ocean and for patches that were isolated by surrounding forest. Neither patch size nor connectivity were positive predictors of rare species occurrence. Thus, in our study area, human influence, presumably due to disturbance or introduction of competitive non‐native species, appears to have overwhelmed classical predictors of rare species distribution, such that greater patch isolation appeared to favor presence of rare species. We suggest conservation planners consider the potential advantages of protecting geographically isolated patches in human‐modified landscapes because such patches may represent the only habitats in which rare species are likely to persist. Influencia Humana y Predictores Biogeográficos Clásicos de la Ocurrencia de Especies Raras     

Seasonal dynamics of macro-algae in the South Lake of Tunis

The macro-algae communities observed in the south lake of Tunis are characterized by the predominance of nitrophilous algae which are in the order of biomass importance:Ulva, Cladophora andEnteromorpha. We have noted seasonal changes of alga distribution. The wind appears to be one of the most important factors influencing this distribution. The total biomass reaches a maximum in the spring. Rapid decomposition of the biomass leads to a severe ecological imbalance, resulting in crises of anoxia and fish death. A restoration project has already started. It aims at removal of contaminated muds and the introduction of a new circulation system. The main objectives of this work were to collect information on the distribution and biomass of the phytobenthic communities as a first step in the constitution of a database for further comparison.     

Patterns and biases of climate change threats in the IUCN Red List

International Union for Conservation of Nature (IUCN) Red List assessments rely on published data and expert inputs, and biases can be introduced where underlying definitions and concepts are ambiguous. Consideration of climate change threat is no exception, and recently numerous approaches to assessing the threat of climate change to species have been developed. We explored IUCN Red List assessments of amphibians and birds to determine whether species listed as threatened by climate change display distinct patterns in terms of habitat occupied and additional nonclimatic threats faced. We compared IUCN Red List data with a published data set of species’ biological and ecological traits believed to infer high vulnerability to climate change and determined whether distributions of climate change‐threatened species on the IUCN Red List concur with those of climate change‐threatened species identified with the trait‐based approach and whether species possessing these traits are more likely to have climate change listed as a threat on the IUCN Red List. Species in some ecosystems (e.g., grassland, shrubland) and subject to particular threats (e.g., invasive species) were more likely to have climate change as a listed threat. Geographical patterns of climate change‐threatened amphibians and birds on the IUCN Red List were incongruent with patterns of global species richness and patterns identified using trait‐based approaches. Certain traits were linked to increases or decreases in the likelihood of a species being threatened by climate change. Broad temperature tolerance of a species was consistently related to an increased likelihood of climate change threat, indicating counterintuitive relationships in IUCN assessments. To improve the robustness of species assessments of the vulnerability or extinction risk associated with climate change, we suggest IUCN adopt a more cohesive approach whereby specific traits highlighted by our results are considered in Red List assessments. To achieve this and to strengthen the climate change‐vulnerability assessments approach, it is necessary to identify and implement logical avenues for further research into traits that make species vulnerable to climate change (including population‐level threats).     

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