适用于流式细胞术的三角褐指藻核DNA染色方法

为制备可用于配置有488 nm激发器的流式细胞仪检测核DNA含量的三角褐指藻细胞,取对数生长期的三角褐指藻细胞,分别采用70%乙醇、1%戊二醛、1%甲醛溶液进行细胞固定,经RNase A消化后的样品用碘化丙啶(PI)染色,用SYBR-green I对未经RNase A消化的细胞染色,在荧光显微镜下观察细胞染色情况,用流式细胞仪检测细胞核DNA含量.结果显示:在荧光显微镜下观察到,3种不同固定液固定的细胞均表现为SYBR-green I只对藻细胞核染色,PI对于藻的细胞核与细胞质均有着色.流式结果的DNA直方图显示,SYBR-green I染色的细胞的DNA直方图有明显的G1与G2/M峰,拟合度(RCS)在3左右,变异系数(CV)在8.6%左右.而PI染色细胞的DNA直方图表现为变异系数值偏大,拟合度较低,没有明显的G1与G2/M峰.本研究表明,SYBR-green I是一种无需对细胞进行RNase A处理,可应用于配置有488 nm激光器流式细胞仪的核酸染料,可为研究在整个细胞周期的调控机制奠定基础,同时也为其他浮游植物细胞核的染色提供参考.     

白玉兰落叶水浸出液抑制蓝藻生长和叶绿素荧光特性分析

利用浮游植物荧光仪对暴露于不同浓度白玉兰落叶水浸出液下微囊藻生长、最大光合作用效率(Fv/Fm)、实际光合作用效率[Y(Ⅱ)]、光能利用效率(alpha)和最大相对电子传递速率(r ETRmax)进行为期15 d的检测,分析白玉兰落叶浸出液对微囊藻的抑制效应和叶绿素荧光特性影响.结果发现,白玉兰落叶浸出液能有效抑制微囊藻的生长,呈明显浓度抑制型变化,抑藻能力随时间的延长而下降.低浓度(0.4、0.8、1.2、1.6 g·L-1)浸出液胁迫下,对微囊藻叶绿素荧光参数无显著影响;高浓度(2.0 g·L-1)浸出液胁迫下,在早期(4 d内)对荧光参数有极显著抑制作用.三维荧光图谱表明,在投量为2.0 g·L-1时,第15天色氨酸及酪氨酸荧光峰强度约为1.2 g·L-1投量情况下的1/3,同时腐殖酸的荧光峰强度减弱.第7—15天,藻细胞生长的半抑制浓度EC50值最小约为0.5—0.7 g·L-1.     

水稻根系生长发育对CO_2浓度升高的响应及其品种间的差异

Minirhizontrons是一种非破坏性、定点、可直接观测和研究植物根系的新方法。利用微根管Minirhizotrons在试验田的温室大棚内研究CO2浓度升高作用下的水稻根系生长发育,试验采用完全随机处理,探讨CO2浓度升高(800μmol·mol-1)对水稻(Oryza sativa L.)生物量和根系形态的变化差异。结果表明,与CO2对照相比,CO2浓度升高显著增加4个水稻品种(2种杂交籼稻和2种常规籼稻)的地上部生物量,增幅为8.58%~12.66%,平均增加10.61%。CO2浓度升高条件下,根的生物量分别增加了3.16%~12.13%,平均增加8.64%。高CO2浓度对根系形态的影响表明,4种水稻根系对CO2浓度升高都有积极的响应。CO2浓度升高条件下,各根系指标在水稻不同生育期都有显著增加,根长密度、表面积、体积和根数的平均增幅分别为10%~27%、21%~24%、20%~58%和4%~18%。但在水稻生长发育过程中,品种间也存在着差异。CO2浓度升高和对照处理,籼型杂交稻威优644(V644)和金优207(JY207)的根长密度和根数表现出相似的变化趋势;高CO2浓度处理时其根长密度平均都增加了10%,根数平均增加4%和8%。CO2浓度升高和对照处理,2种籼型杂交水稻的根体积和表面积表现出较快的增长幅度,都呈现出近线性的生长趋势;CO2浓度升高处理下其根体积平均增加40%和25%,表面积平均都增加了24%。CO2浓度升高和对照处理,籼型常规稻湘晚12号(XW12)和丰华占(FHZ)的生长变化趋势表现一致,生长发育后期达到一个近似饱和的拐点。CO2浓度升高条件下其根长密度、根数和根体积分别平均增加27%和24%、18%和11%、58%和20%,根表面积平均都增加了21%。     

固定化多环芳烃降解酶——谷胱甘肽S-转移酶的研究

以自制壳聚糖为载体,戊二醛为交联剂固定谷胱甘肽S-转移酶,研究了交联剂浓度、酶用量、pH值、温度及时间等因素对固定化酶活性的影响.结果表明,戊二醛浓度0.5%,pH7.0,温度20℃,时间12h以及液态酶与壳聚糖凝胶1:1配比是谷胱甘肽S-转移酶最佳固定化条件,固定化酶催化菲降解的最佳反应时间是9h.另外,固定化酶与游离酶相比,稳定性和可操作性都有较大的提高.     

海洋赤潮藻球形棕囊藻在氮磷富营养下的细胞增殖

利用常见海洋赤潮微藻球形棕囊藻(Phaeocystis globosa)为试验研究材料,以舵海洋微藻营养液为对照(1P1N:磷质量浓度为5×10-3g·L-1.氮质量浓度为75×10-3g·L-1),设置3组富磷和富氮营养处理(3P1N:磷质量浓度为15×10-3 g·L-1,氮质量浓度为75×10-3 g·L-1;1P3N:磷质量浓度为5××10-3 g·L-1,氮质量浓度为225×10-3 g·L-1;3P3N:磷质量浓度为15××10-3 g·L-1,氮质量浓度为225×10-3 g·L-1),利用细胞记数和叶绿素荧光测定等方法研究了藻细胞在不同富磷和富氮条件的增殖情况.结果显示,不同浓度磷和氮营养下的藻体荧光值变化在试验周期内均呈现"S"型曲线,表明藻细胞的生长经历缓慢期,快速期和平缓期3个阶段;同时,不同的富磷和富氮营养条件对球形棕囊藻的叶绿素荧光值有一定的影响,其中在对照1P1N下的藻体荧光值最低,在试验结束时(第10天)只有850 μg·L-1,而在3P1N,1P3N和3P3N条件下的藻体荧光值均达到900 μg·L-1以上,显著高于1P1N下的藻体荧光值,表明富磷和富氮营养可以促进藻细胞的生长增殖,但在试验设置的不同富磷和富氮营养下的藻体荧光值之间没有显著的差异.就不同磷和氮营养条件下的藻最大比生长速率而言,3P3N和3P1N条件下的最大,均达到0.77 d-1,明显高于1P1N和1P3N条件下的藻最大比生长速率(分别只有0.70 d-1和0.69 d-1).此外,试验结束时细胞密度的变化趋势与藻体荧光值相似,富磷和富氮营养条件下的细胞密度显著高于1P1N下的细胞密度,而富磷和富氮营养条件下的细胞密度间也不存在显著的差异.研究结果揭示,水体中的高磷和高氮营养浓度是导致藻细胞大量快速增殖的一个主要因素,而利用叶绿素荧光来测定藻细胞增殖是一种快速、简便,灵敏和可靠的方法,可在今后赤潮监测过程中多加利用,以能及时、准确地预测预报赤潮爆发.从而减少其对环境和经济的影响.     

KMnO4/H2SO4引发制备淀粉改性絮凝剂的研究

以高锰酸钾为引发剂,制备淀粉-丙烯酰胺接枝共聚物.研究酸浓度、引发剂浓度、单体浓度、温度和加料方式对接枝反应的影响.结果表明,少量KMnO4(0.45mmol·l-1)和H2SO4(0.8mmol·l-1)可有效引发淀粉-丙烯酰胺接枝共聚反应,接枝效率达到90%以上,分子量大大提高,对高岭土的絮凝效果较好.     

微波辅助提取-氢化物发生原子荧光光谱法测定土壤中As（Ⅲ）和As（Ⅴ）

建立了微波辅助提取,采用氢化物发生原子荧光光谱法不进行价态分离,直接测定土壤中As（Ⅲ）和As（Ⅴ）的方法。通过加入抗坏血酸作为抗氧化剂,以磷酸溶液、L-半胱氨酸分别为提取剂和还原剂,在不还原与还原两种情况下分别进行测量,根据荧光强度与As（Ⅲ）和As（Ⅴ）的质量浓度关系方程式计算提取液中它们的质量浓度。对实验条件进行优化后,As（Ⅲ）、As（Ⅴ）的方法检出限均可达到0.05μg.g-1,实验精密度（RSD,n=11）分别为1.3%和2.1%,样品加标平均回收率分别为As（Ⅲ）为93.2%~110.0%,As（Ⅴ）为92.6%~104.0%。该方法不需要高效液相色谱等大型设备、简单可靠、易于操作、分析速度快、精密度高,具有在各分析测试实验室推广应用价值。方法已用于实际土壤样品中不同价态无机砷质量分数的测定。     

宁波市某人工泻湖浮游生物变化特征及影响因素分析

以宁波市北仑区梅山水道形成的人工泻湖为研究对象,在不同季节进行水质及浮游生物调查,分析其浮游生物时空分布特征与水质的关系。4个采样点共检出浮游植物66种,以硅藻为绝对优势种,检出浮游动物25种,主要为桡足类、少量轮虫及网纹虫;拦坝后水道内浮游生物密度有了数量级增长,各项生物评价指数降低,但各采样点仍处于中污染水平。监测理化参数表明,研究水域在拦坝后盐度下降、悬浮物浓度下降,氮磷含量无明显变化;水域大部分点位处于中度富营养化水平。结合浮游生物分布与理化参数进行分析,发现堤坝合龙后,水道内侧海水淡化、悬浮物含量下降,导致浮游生物密度上升、生物多样性下降、出现淡水优势种群;营养盐含量不是浮游生物生长的限制因子,对浮游生物分布无显著影响。     

基于美拉德反应产物作为荧光和共振瑞利散射的传感平台快速检测阿斯巴甜

源于葡萄糖和L-精氨酸(glucose and L-Arginine,GLA)的美拉德反应的产物与Cu^2+的螯合作用形成具有良好的光学活性和水溶性的GLA-Cu^2+复合物,建立荧光(fluorescence,FL)和共振瑞利散射(resonance rayleigh scattering,RRS)的新型传感平台,可用来检测阿斯巴甜(Aspartame,APM)。研究表明,由于GLA与Cu^2+螯合,GLA的蓝色荧光将被Cu^2+淬灭,同时出现了一个增强的RRS特征峰。当加入一定量的APM后,系统的荧光恢复,RRS峰强度降低。因此,GLA-Cu^2+-APM体系形成荧光“关-开”模式和RRS“开-关”模式的传感平台,这个传感平台展现出很高的灵敏度和良好的选择性,可用于样品溶液中痕量APM的快速检测。其线性范围分别为0.3~300.0μmol/L(FL)和0.4~800.0μmol/L(RRS),检测限(LOD)为26.0 nmol/L(FL)和39.0 nmol/L(RRS)。应用此传感平台的FL法测定水样中的APM取得了令人满意的结果。     

高效液相色谱法测定水体中的阿特拉津

为建立检测水体中阿特拉津的高效液相色谱法,以ODSC18(250mm×4.6mm×5μm)柱为色谱柱,以甲醇∶水=5∶1为流动相,流速0.6mL·min-1,采用紫外检测器,用外标法测定水体中阿特拉津含量。结果表明,方法线性范围为0.052~13.0mg·L-1,线性相关系数r=0.9998;对浓度低于线性的样品,取100mL样品提取后测定,方法的检出限为0.0002mg·L-1,对含1.95μg·L-1、32.5μg·L-1、72.8μg·L-1阿特拉津水质样品进行测定,其相对标准偏差为1.61%-6.85%,加标回收率为84.6%~96.9%。采用净化方法时的加标回率为74.9%~92.9%。对阿特拉津含量在线性范围内的水样可直接过0.45μm膜后测定,加标回收率为97.0%~99.6%。该方法适用于水体中痕量和常量阿特拉津的测定。     

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.     

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     

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).