EoNPV的限制性酶切图谱及polyhedrin基因和egt基因的定位

用８种限制性内切酶酶解茶尺蠖Ｅｃｔｒｏｐｉｓｏｂｌｉｑｕａ核型多角体病毒（ＥｏＮＰＶ）基因组ＤＮＡ，获得大于０．８８ｋｂ的片段数分别为７、３８、１９、１７、１６、９、３２和１４．由此统计，基因组平均大小大于１１８．５ｋｂ，即Ｍｒ约大于７８．６×１０６．用ＢｍＮＰＶ的多角体蛋白（ｐｈ）基因和ＡｃＭＮＰＶ的ｅｇｔ基因为探针，应用Ｓｏｕｔｈｅｒｎ杂交方法，将ＥｏＮＰＶ的ｐｈ基因定位于ＢａｍＨＩＡ、ＣｌａＩＤ、ＥｃｏＲＩＭ、ＥｃｏＲＶＬ、ＨｉｎｄⅢＦ、ＰｓｔＩＡ、ＳａｌＩＨ和ＸｈｏＩＢ片段上，将ｅｇｔ基因定位于ＢａｍＨＩＡ、ＣｌａＩｌ、ＥｃｏＲＩＫ、ＥｃｏＲＶＡ、ＨｉｎｄⅢＨ、ＰｓｔＩＡ、ＳａｌＩＢ、ＸｈｏＩＩ片段上．通过克隆和酶切鉴定ＥｃｏＲＩＭ和ＥｃｏＲＶＬ片段，测定ｐｈ基因部分序列，并以ＥｃｏＲＩＭ为探针对基因组酶切印迹进行杂交，制作了ＥｏＮＰＶｐｈ基因和ｅｇｔ基因区的酶切图谱，推定ｅｇｔ基因保守区位于ｐｈ基因上游约４．５５ｋｂ处．     

粘虫颗粒体病毒及其增效因子对粘虫核型多角体病毒的增效作用

生物测定的结果表明,粘虫颗粒体病毒ＰｕＧＶ－Ｐｓ及其增效因子对粘虫核型多角体病毒ＰｓＮＰＶ均有明显的增效作用．粘虫颗粒体病毒不仅能提高粘虫感染ＰｓＮＰＶ的死亡率,而且ＮＰＶ、ＧＶ两种病毒混合感染使粘虫幼虫代谢发育受到抑制,表现为生长缓慢、体重减少等．增效因子的增效作用表现在能显著提高粘虫幼虫的死亡率,降低核型多角体病毒的半致死浓度ＬＣ５０,并缩短半致死时间ＬＴ５０．     

黄地老虎NPV增效作用的研究

对黄地老虎核型多角体病毒（ＡｓＮＰＶ）的多角体蛋白和病毒粒子组分进行分离纯化，分别与棉铃虫核型多角体病毒（ＨａＮＰＶ）混合感染三龄棉铃虫幼虫，发现此二种组分均能够提高棉铃虫幼虫的死亡率，缩短半致死时间，降低半致死浓度。使用酶联免疫吸附测定（ＥＬＩＳＡ）技术检验ＡｓＮＰＶ多角体蛋白和病毒粒子与粘虫颗粒体病毒增效因子（ＰｓＧＶ－ＳＦ）和ＨａＮＰＶ多角体蛋白的同源性，发现ＡｓＮＰＶ多角体蛋白和病毒粒子与     

HaNPV多角体蛋白在宿主不同组织中的时相性表达

应用亲和凝胶过滤层析对多角体蛋白多克隆抗血清进行纯化,辣根过氧化物酶进行标记,用于ＥＬＩＳＡ法检测中肠、血淋巴、脂肪体中的多角体蛋白的含量变化．结果：中肠在感染２４ｈ含量最高,４８ｈ最低,７２ｈ较４８ｈ略有升高,９６ｈ和１２０ｈ与７２ｈ基本保持一致;血淋巴在感染４８ｈ含量最高,然后下降至９６ｈ,１２０ｈ又稍有升高;脂肪体在感染７２ｈ开始增加,直至１２０ｈ．三种组织中多角体蛋白含量变化并不同步．从表达的时相性看,与ＮＰＶ的感染过程基本一致,即从中肠至血腔再到脂肪体．免疫电镜观察表明,感染７２ｈ的中肠,在胞浆中合成的多角体蛋白此时已转运到细胞核中     

柞蚕多角体病毒的纯化和鉴定

对柞蚕多角体病毒(ApNPV)的多角体颗粒进行了分离和纯化,并在显微镜下进行了形态观察．提取和纯化了柞蚕多角体病毒的基因组DNA,对DNA进行了部分限制性内切酶图谱鉴定．结果表明,在ApNPV基因组中只含少量的EcoRI位点,与家蚕核型多角体病毒差别很大,并提示这两种病毒的复制机制可能不同．图3参5。     

马尾松毛虫质型多角体病毒结构蛋白的抗体制备及免疫性

质型多角体病毒的病毒粒子结构蛋白是由衣壳蛋白VP1(Capsid protein,CP)、塔状蛋白VP3(Turret protein,TP)和大突起蛋白VP5(Large protrusion protein,LPP)组成,这3个蛋白分别由病毒基因组片段S1、S4和S7编码.为了解质型多角体病毒结构蛋白的免疫定位情况,本研究从马尾松毛虫质型多角体病毒湖南株中提取病毒核酸,回收、纯化S1、S4和S7片段,经RT-PCR扩增得到对应片段的ORF全长或抗原区的片段,克隆于原核表达载体上,在大肠杆菌BL21菌株中进行了表达,免疫家兔制备了多克隆抗体.Western blot结果显示DpCPV基因组的S1、S4和S7片段分别编码其结构蛋白VP1、VP3和VP5,与推测的结果一致.免疫胶体金的方法也证明S1、S4和S7编码的蛋白定位于病毒粒子衣壳的外表面.本研究首次通过免疫学方法确定了质型多角体病毒结构蛋白在病毒粒子上的定位,为质型多角体病毒的结构蛋白功能和侵染机制研究提供了免疫学方面的基础,对质型多角体病毒的研究有较重要的意义.     

荧光增白剂对重组苜蓿银纹夜蛾核型多角体病毒的增效作用

以β-半乳糖苷酶基因为标志基因的重组苜蓿银纹夜蛾核型多角体(AcMNPV-hsp70／lacz)研究光增白剂对该病毒的增效作用及其作用方式．通过病毒毒力的生物测定表明：光增白剂FB-28能提高甜菜夜蛾幼虫的死亡率，降低半致死浓度，缩短半致死时间；光增白剂的增效作用与感染虫龄有关．此外，应用组织切片技术对中肠组织病理以及血淋巴细胞感染观察表明：在幼虫的病毒接种物中添加合适浓度的荧光增白剂不会改变病毒在幼虫中肠组织中的感染部位，却可以提高幼虫对病毒的敏感性，产生更多的病灶．图版1图4表3参19     

转化血型用蕃茄α－D－半乳糖苷酶的分离、纯化及理化性质

成熟蕃茄匀浆后，经硫酸铵盐析，ＤＥＡＥ－ＳｅｐｈａｄｅｘＡ－５０离子交换层析，ＳｅｐａｄｅｘＧ－１００凝胶过滤和Ｍｅｌｉｂｉｏｓｅ－Ａｇａｒｏｓｅ亲和层析，获得了α－Ｄ－半乳糖苷酶（Ｃ．Ｅ．３．２．１．１１）。酶制剂经ＰＡＧＥ检测为一条带；ＳＤＳ－Ｇ－ＰＡＧＥ测得酶Ｍｒ为３４０００；比活力５２．９Ｕ／ｍｇ·；提纯倍数为５２９０１产率为４５％．酶专－催化以α－Ｄ－半乳糖为末端ａ－（１，３）连接的糖苷键，以ＰＮＰＧ（对硝基苯－α－Ｄ－半乳糖昔）为废物，酶催化反应的Ｋｍ＝０．１１ｍｍｏｌ／Ｌ，Ｖｍａｘ为６７μｍｏｌ·ｍｇ１－·ｍｉｎ－１．ｔ稳定范是０～３５℃；ＰＨ稳定范围是４．０～７．０．最适ｐＨ为５．１．半乳糖是酶的竞争性抑制剂；Ｃｕ２＋、Ｚｎ２＋、Ｍｎ２＋、Ｆｅ３＋、Ａｇ＋和ＥＤＴＡ对酶活性无影响．纯酶制剂可作为Ｂ型血向Ｏ型血转化的工具酶液．     

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.     

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.     

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.     

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.     

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     

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.