活性污泥总DNA不同提取方法的比较

为了快速、简便和经济地获得活性污泥的总DNA,以用于分子生物学研究,采用5种不同的DNA提取方法提取活性污泥的总DNA,并通过提取的核酸总量、纯度、是否含有聚合酶链反应抑制剂等指标综合分析不同的提取方法对活性污泥总DNA提取效果的影响.结果表明:蛋白酶K-SDS-酚氯仿法和柱提取DNA试剂盒法提取活性污泥总DNA效果最好,提取DNA总量多,纯度高,可不经纯化直接进行PCR扩增反应。     

快速提取用于PCR分析的几种蔬菜DNA的方法

利用两种不同的简便方法提取大白菜、大葱和厚皮酣瓜发芽种子的DNA,均能产生具有重现性的RAPD结果这两种方法提取DNA的PCR扩增结果与多次利用苯酚／氯仿提取DNA的结果一致.RNase处理与否对提取DNA的PCR扩增结果没有影响.方法1不用氯仿,但残留蛋白质等容易造成DNA溶解困难;方法2提取的DNA纯度高对于含蛋白质较少的植物器官或组织,可以选用方法1,而对于含蛋白质较多的植物器官或组织,宜选用方法2.两种方法提取的DNA量与用苯酚／氯仿提取DNA的量相似样品之间提取DNA量的多少主要受研磨破碎程度高低所决定.方法1每人每天提取约120个样品,方法2提取约100个样品,比先前的方法快1倍多.所提取的每个样品DNA约能用于100次PCR反应,两种DNA提取方法简便、迅速,为快速进行大量样品的PCR分析提供了条件.     

电化学DNA生物传感器在检测环境有机污染物中的应用

电化学DNA生物传感器是一种以DNA为生物识别元件并通过电化学信号转换器将目标物的存在转变为可检测的电信号的传感装置.因具有灵敏性高、选择性好、制备成本低、操作简单、携带方便、抗干扰性强等优点而被广泛用于环境分析领域.本文简要阐述了电化学DNA生物传感器的检测原理及DNA在电极表面的固定化方法,重点讨论了电化学DNA生物传感器在检测环境有机污染物中的应用,最后对电化学DNA生物传感器的发展方向进行了展望.     

真菌核酸的一种快速提取方法

在总结多种真菌DNA和RNA提取方法的基础上，发展了一种提取真菌核酸新方法，与其它方法相比，该方法具有快速、操作简便等特点，而且提取DNA和RNA步骤基本一致，用该方法提取的DNA完整、纯度高，可用于PR、限制性内切酶酶切和Southern杂交等分子生物学操作；提取RNA产量高，RNA无降解，纯度高，适用Northern和cDNA合成等分子生物学操作。     

海洋微藻活体及乙醇固定状态下基因组DNA的微量提取方法

探讨了海洋微藻活体及固定状态下基因组DNA的提取方法,即作者改进的CTAB法.结果表明,CTAB改进法提取的基因组DNA蛋白质、酚、盐和小分子的污染较少,多糖成份也得到了有效去除.用5%乙醇固定样本提取DNA的效果最好,对原生动物抑制效果也较明显;10%以上的乙醇可杀死原生动物,但对DNA的降解率较大;30%以上的乙醇对细菌抑制较明显,但DNA都有较大程度的降解.本法提取的DNA可用于正常的酶切和PCR.用甲醛、甲醇/冰醋酸等其它固定液固定样品,DNA的得率低或者质量和纯度低.图7表1参27     

识别黄山药、穿龙薯蓣和盾叶薯蓣的分子标记建立

在对黄山药、穿龙薯蓣和盾叶薯蓣的psbA-trnH片段序列分析的基础上,根据3种薯蓣在该片段上的特征序列位点设计了用于识别3种薯蓣的寡核苷酸片段,并作为PCR反应的引物,将3种薯蓣的特征标记引物与psbAf-trn-Hr配合使用建立了识别3种薯蓣的显性和共显性两种检测方式,辅以NaOH碱法快速提取薯蓣干燥根茎总DNA,为3种薯蓣相互间快速、准确的鉴别创造了条件.图3表2参9     

浮游植物叶绿素a测定方法比较

选取规范方法、超声波法、反复冻融法、延时提取法、热丙酮法、丙酮加热法、热乙醇法、混合溶剂法8种常见的浮游植物叶绿素a的测定方法,以实验室培养的微囊藻(Microcystis aeruginosa)水华样品为研究对象进行叶绿素a含量的测定.从叶绿素a提取的完全性、测定数据的稳定性、试验操作的简便性等方面对8种方法进行评价,并对叶绿素a结果测量的单色法和三色法进行分析讨论.试验结果表明,选取的8种测定方法中,包括国家标准方法在内的大多数方法,存在操作耗时长、过程复杂、叶绿素提取不完全的缺点;而丙酮加热法测量叶绿素a的提取效率高、数据稳定性好,操作耗时短、简便,可推荐作为一种常用的浮游植物叶绿素a的测量方法;三色法和单色法测定方法的选取应综合考虑.     

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。     

白菜叶绿体DNA快速提取及分析

采用SDS-蛋白酶法提取了白菜的叶绿体DNA.琼脂糖电泳及紫外分光光度分析表明,所提取的叶绿体DNA质量高,除cpDNA主环外,在胞质雄性不育新种质及原不育系中还存在约1375bp和831bp的两条质粒分子.用所提取的叶绿体DNA作为模板进行RAPD扩增,获得了清晰的扩增图谱.该方法与传统的提取方法相比省时、高效、无污染.图2表1参14     

中孔硅镁胶对水溶液中磷酸根的吸附性能

化肥、农药、含磷洗涤剂以及粪便,是废水中磷的主要来源,而磷浓度超标是引起水体中富营养化的主要原因之一.目前,国内外除磷的方法中主要有化学沉淀法、物理吸附法、离子交换法和微生物降解法等,吸附法因其工艺简单、运行可靠、操作灵活和无二次污染等特点备受关注.硅酸镁为多孔结构,属两性化合物,具有酸碱两种吸附性能,在聚醚精制过程中用于脱酸、脱臭、脱色及脱钾,在煎炸油处理和生物柴油加工过程中用于降低酸价;此外,硅酸镁还广泛地应用于脱除废水中的金属离子和染料.目前,国内外对三硅酸镁和六硅酸镁的报道较多,对其它镁硅配比的硅酸镁报道较少.本课题组系统地研究了不同镁硅物质的量之比的     

Achieving Open Access to Conservation Science

Conservation science is a crisis discipline in which the results of scientific enquiry must be made available quickly to those implementing management. We assessed the extent to which scientific research published since the year 2000 in 20 conservation science journals is publicly available. Of the 19,207 papers published, 1,667 (8.68%) are freely downloadable from an official repository. Moreover, only 938 papers (4.88%) meet the standard definition of open access in which material can be freely reused providing attribution to the authors is given. This compares poorly with a comparable set of 20 evolutionary biology journals, where 31.93% of papers are freely downloadable and 7.49% are open access. Seventeen of the 20 conservation journals offer an open access option, but fewer than 5% of the papers are available through open access. The cost of accessing the full body of conservation science runs into tens of thousands of dollars per year for institutional subscribers, and many conservation practitioners cannot access pay‐per‐view science through their workplace. However, important initiatives such as Research4Life are making science available to organizations in developing countries. We urge authors of conservation science to pay for open access on a per‐article basis or to choose publication in open access journals, taking care to ensure the license allows reuse for any purpose providing attribution is given. Currently, it would cost $51 million to make all conservation science published since 2000 freely available by paying the open access fees currently levied to authors. Publishers of conservation journals might consider more cost effective models for open access and conservation‐oriented organizations running journals could consider a broader range of options for open access to nonmembers such as sponsorship of open access via membership fees. Obtención de Acceso Abierto a la Ciencia de la Conservación     

Some polyphagous Homoptera gain protection from their natural enemies by feeding on the toxic plantsSpartium junceum andErythrina corallodendrum (Leguminosae)

Summary Interactions were studied among alkaloid-containing legumes (Erythrina corallodendrum andSpartium junceum) and non-toxic plants (Citrus sinensis, Cucurbita moschata andEuphorbia tirucalli), several polyphagous homopterans,Aphis craccivora (Aphididae),Icerya purchasi, I. aegyptiaca (Margarodidae),Lepidosaphes ulmi (Diaspididae) andPlanococcus citri (Pseudococcidae), and some major natural enemies of these homopterans. Significant reductions in survival due to negative effects of alkaloid containing as compared with non-alkaloidal plants were recorded for the predatorsRodolia cardinalis andChilocorus bipustulatus, but not forCryptolaemus montrouzieri (Coleoptera: Coccinellidae),Chrysoperla carnea (Neuroptera: Chrysopidae) andSympherobius sanctus (Neuroptera: Sympherobiidae). The development time of the larvae or pupae ofR. cardinalis, C. carnea andS. sanctus was longer on the toxic plants than on the non-toxic ones. The percentage of parasitism ofA. craccivora collected from the non-alkaloidal plantsVicia palaestina andMelilotus albus was much higher than that onS. junceum. The parasitoid complexes ofA. craccivora differed between both plant groups. The nutritive value of honeydew ofI. purchasi andA. craccivora, as expressed by the life span ofEncyrtus infelix (Hymenoptera: Encyrtidae) adults, was also investigated. Life spans were significantly longer when the wasps fed on honeydew produced on non-alkaloidal plants (C. sinensis andPittosporum tobira) than on alkaloid containing plants whenI. purchasi — but notA. craccivora — was the producer. It is suggested that the chemical defense ofE. corallodendrum andS. junceum is exploited by polyphagous phytophages to reduce predation. In nature, population growth and density of four of the investigated homopterans are conspicuously high when they developed on the alkaloid containing plant species, and very low on non-alkaloid plants. The efficiency of their natural enemies may be reduced by sequestration of alkaloids (or other toxic plant compounds) or their transfer into excreted honeydew. Therefore it is assumed that a generalist phytophagous homopteran may be protected from its natural enemies, although at different rates of efficiency, if it can safely sequester the host allelochemical when it develops on toxic species within its host range.     

Sequestration of cucurbitacin analogs by New and Old World chrysomelid leaf beetles in the tribe Luperini

Summary Two South American polyphagous leaf beetles,Diabrotica speciosa andCerotoma arcuata, selectively accumulated the bitter tasting compound 23,24-dihydrocucurbitacin D in their body after ingesting root tissues of cucurbit plants. Similarly, three Asian Cucurbitaceae-feeding specialists in the genusAulacophora were found to sequester the same compound. Cucurbitacin analogs were shown to deter feeding by a bird predator, indicating an allomonal role for these compounds in cucurbitacin-associated chrysomelid leaf beetles both of New and Old Worlds. The strong affinity to cucurbitacins, selective sequestration of the analogs and consequent protection from predators suggested an ecological adaptation mechanism developed in common among these two geographically isolated subtribes in the Luperini.     

Effects of Controlled Fire and Livestock Grazing on Bird Communities in East African Savannas

Abstract: In East Africa fire and grazing by wild and domestic ungulates maintain savannas, and pastoralists historically set fires and herded livestock through the use of temporary corrals called bomas. In recent decades traditional pastoral practices have declined, and this may be affecting biodiversity. We investigated the effects of prescribed fires and bomas on savanna bird communities in East Africa during the first and second dry seasons of the year (respectively before and after the rains that mark the onset of breeding for most birds). We compared abundance, richness, and community composition on 9‐ha burned plots, recently abandoned bomas, and control plots in the undisturbed matrix habitat over a 3‐year period. Generally, recently burned areas and abandoned bomas attracted greater densities of birds and had different community assemblages than the surrounding matrix. The effects of disturbances were influenced by interactions between primary productivity, represented by the normalized difference vegetation index, and time. Bird densities were highest and a greater proportion of species was observed on burned plots in the months following the fires. Drought conditions equalized bird densities across treatments within 1 year, and individuals from a greater proportion of species were more commonly observed on abandoned bomas. Yearly fluctuations in abundance were less pronounced on bomas than on burns, which indicate that although fire may benefit birds in the short term, bomas may have a more‐lasting positive effect and provide resources during droughts. Several Palearctic migrants were attracted to burned plots regardless of rainfall, which indicates continued fire suppression may threaten their already‐declining populations. Most notably, the paucity of birds observed on the controls suggests that the current structure of the matrix developed as a result of fire suppression. Traditional pastoralism appears critical to the maintenance of avian diversity in these savannas.     

What Every Conservation Biologist Should Know about Economic Theory

Abstract: The last century has seen the ascendance of a core economic model, which we will refer to as Walrasian economics. This model is driven by the psychological assumptions that humans act only in a self‐referential and narrowly rational way and that production can be described as a self‐contained circular flow between firms and households. These assumptions have critical implications for the way economics is used to inform conservation biology. Yet the Walrasian model is inconsistent with a large body of empirical evidence about actual human behavior, and it violates a number of basic physical laws. Research in behavioral science and neuroscience shows that humans are uniquely social animals and not self‐centered rational economic beings. Economic production is subject to physical laws including the laws of thermodynamics and mass balance. In addition, some contemporary economic theory, spurred by exciting new research in human behavior and a wealth of data about the negative global impact of the human economy on natural systems, is moving toward a world view that places consumption and production squarely in its behavioral and biophysical context. We argue that abandoning the straightjacket of the Walrasian core is essential to further progress in understanding the complex, coupled interactions between the human economy and the natural world. We call for a new framework for economic theory and policy that is consistent with observed human behavior, recognizes the complex and frequently irreversible interaction between human and natural systems, and directly confronts the cumulative negative effects of the human economy on the Earth's life support systems. Biophysical economics and ecological economics are two emerging economic frameworks in this movement.     

Effects of Coffee Management on Deforestation Rates and Forest Integrity

Knowledge about how forest margins are utilized can be crucial for a general understanding of changes in forest cover, forest structure, and biodiversity across landscapes. We studied forest‐agriculture transitions in southwestern Ethiopia and hypothesized that the presence of coffee (Coffea arabica)decreases deforestation rates because of coffee's importance to local economies and its widespread occurrence in forests and forest margins. Using satellite images and elevation data, we compared changes in forest cover over 37 years (1973–2010) across elevations in 2 forest‐agriculture mosaic landscapes (1100 km² around Bonga and 3000 km² in Goma‐Gera). In the field in the Bonga area, we determined coffee cover and forest structure in 40 forest margins that differed in time since deforestation. Both the absolute and relative deforestation rates were lower at coffee‐growing elevations compared with at higher elevations (?10/20% vs. ?40/50% comparing relative rates at 1800 m asl and 2300–2500 m asl, respectively). Within the coffee‐growing elevation, the proportion of sites with high coffee cover (>20%) was significantly higher in stable margins (42% of sites that had been in the same location for the entire period) than in recently changed margins (0% of sites where expansion of annual crops had changed the margin). Disturbance level and forest structure did not differ between sites with 30% or 3% coffee. However, a growing body of literature on gradients of coffee management in Ethiopia reports coffee's negative effects on abundances of forest‐specialist species. Even if the presence of coffee slows down the conversion of forest to annual‐crop agriculture, there is a risk that an intensification of coffee management will still threaten forest biodiversity, including the genetic diversity of wild coffee. Conservation policy for Ethiopian forests thus needs to develop strategies that acknowledge that forests without coffee production may have higher deforestation risks than forests with coffee production and that forests with coffee production often have lower biodiversity value. Efectos de la Administración Cafetalera sobre las Tasas de Deforestación y la Integridad de los Bosques     

Globalization of the Cashmere Market and the Decline of Large Mammals in Central Asia

As drivers of terrestrial ecosystems, humans have replaced large carnivores in most areas, and human influence not only exerts striking ecological pressures on biodiversity at local scales but also has indirect effects in distant corners of the world. We suggest that the multibillion dollar cashmere industry creates economic motivations that link western fashion preferences for cashmere to land use in Central Asia. This penchant for stylish clothing, in turn, encourages herders to increase livestock production which affects persistence of over 6 endangered large mammals in these remote, arid ecosystems. We hypothesized that global trade in cashmere has strong negative effects on native large mammals of deserts and grassland where cashmere‐producing goats are raised. We used time series data, ecological snapshots of the biomass of native and domestic ungulates, and ecologically and behaviorally based fieldwork to test our hypothesis. In Mongolia increases in domestic goat production were associated with a 3‐fold increase in local profits for herders coexisting with endangered saiga (Saiga tatarica).That increasing domestic grazing pressure carries fitness consequences was inferred on the basis of an approximately 4‐fold difference in juvenile recruitment among blue sheep (Pseudois nayaur)in trans‐Himalayan India. Across 7 study areas in Mongolia, India, and China's Tibetan Plateau, native ungulate biomass is now <5% that of domestic species. Such trends suggest ecosystem degradation and decreased capacity for the persistence of native species, including at least 8 Asian endemic species: saiga, chiru (Pantholops hodgsoni), Bactrian camel (Camelus bactrianus), snow leopard(Panthera uncia), khulan(Equus hemionus), kiang (E. kiang), takhi (E. przewalski), and wild yak (Bos mutus). Our results suggest striking yet indirect and unintended actions that link trophic‐level effects to markets induced by the trade for cashmere. Globalización del Mercado de Cachemira y la Declinación de Mamíferos Mayores en Asia Central     

The lichen-rich coastal heath vegetation on the isle of Anholt, Denmark — conservation and management

The high conservational value of the lichen-rich vegetation and landscape of the marine foreland Ørkenen on the isle of Anholt is treated from a Danish as well as a European perspective. The sensitivity of the lichen-rich vegetation to physical disturbance is emphasized. The impact of invasive species such asPinus mugo as well as the effect of atmospheric deposition of nutrients on the heaths at Anholt is described. Considerations related to the development of a management plan for Ørkenen are presented.     

Behavior and ecology of Old World Luperini beetles of the genusAulacophora (Coleoptera: Chrysomelidae)

Summary The attraction of Old World leaf beetles in the genusAulacophora to kairomones and parakairomones which are effective lures for New WorldDiabrotica andAcalymma were investigated. Beetles captured on sticky traps baited with single and multicomponent lures were no different from the control traps for two species ofAulacophora. Yellow colored traps and squash blossoms are attractive toAulacophora beetles which detect sub-microgram quantities of cucurbitacins on silica gel. Leaf feeding behavior and flight activity data are correlated with varietal preference of threeAulacophora species. The common response byDiabrotica andAulacophora to cucurbitacins reinforces the two groups' coevolutionary association with the Cucurbitaceae. The apparent lack of a common response toCucurbita blossom volatiles suggests recent evolutionary pathways are substantially different for these two groups of beetles.     

Use of stochastic patch occupancy models in the California red-legged frog for Bayesian inference regarding past events and future persistence

Assessing causes of population decline is critically important to management of threatened species. Stochastic patch occupancy models (SPOMs) are popular tools for examining spatial and temporal dynamics of populations when presence–absence data in multiple habitat patches are available. We developed a Bayesian Markov chain method that extends existing SPOMs by focusing on past environmental changes that may have altered occupancy patterns prior to the beginning of data collection. Using occupancy data from 3 creeks, we applied the method to assess 2 hypothesized causes of population decline—in situ die-off and residual impact of past source population loss—in the California red-legged frog. Despite having no data for the 20–30 years between the hypothetical event leading to population decline and the first data collected, we were able to discriminate among hypotheses, finding evidence that in situ die-off increased in 2 of the creeks. Although the creeks had comparable numbers of occupied segments, owing to different extinction–colonization dynamics, our model predicted an 8-fold difference in persistence probabilities of their populations to 2030. Adding a source population led to a greater predicted persistence probability than did decreasing the in situ die-off, emphasizing that reversing the deleterious impacts of a disturbance may not be the most efficient management strategy. We expect our method will be useful for studying dynamics and evaluating management strategies of many species.