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.     

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     

棉铃虫核多角体病毒基因组限制酶酶切分析及其多角体基因的定位

应用１１种限制性内酶ＢａｍＨⅠ、ＢｓｔＥⅡ、ＥｃｏＲⅠ、ＥｃｏＲⅤ、ＨｉｎｄⅢ、ＫｐｎⅠ、ＰｓｔⅠ、ＳａｌⅠ、ＳｓｔⅠ、ＸｂａⅠａⅠ和ＸⅠⅠ和ＸｈｏⅠ分别将中国棉铃虫核多角体病毒（单粒包埋ＨａＳＮＰＶ）湖北株基因组Ｄ组ＤＮＡ酶切为１０、１２、２２、２１、１３、６、６、４０、６、２１、６个片段，并求得基因组大小平均为１什什Ｍｒ≈７９．１×１０６．以家委核多角体病毒ＢｍＮＰＶ多角体基因为探针，利用Ｓｏｕｔｈｅｒｎ杂交技术将病毒多角体基因定位在ＳａｌⅠ４．２×１０３ｂ左右的片段上．与棉铃虫核多角体病毒其它株系酶切图谱比较结果表明，本株病毒与上海等株系及美洲棉铃虫核多角体病毒ＨａＳＮＰＶＥｌｋａｒ株系酶切图谱相似，它们之间的条带数和大小差异较小，而与已发现的所有多粒包埋型病毒ＨａＳＮＰＶ酶切图主谱差异较大．据此认为ＨａＳＮＰＶ和ＨａＭＮＰＶ属于基因型不同的两类病毒，而ＨａＳＮＰＶ不同分离株的同源性很高     

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．     

湖流域土壤微生物基因组总DNA分离纯化及其质粒文库的初步构建

采用研磨／冻融和SDV蛋白酶K热处理以及CTAB处理等理化方法，直接从太湖流域土壤样品中抽提得到微生物总基因组DNA，所得粗DNA用透析袋法以及Nucleaotrap suspension DNA纯化试剂盒进行了纯化，纯化过的DNA能够满是常用限制性内切酶酶切、细菌16S rDNA通用引物和随机引物进行PCR扩增的要求，将所得纯品DNA经过EcoRI部分酶切之后与酶切并脱磷酸的pSK^ 空载体连接，再以大肠杆菌JM109为宿主细胞初步构建了土壤微生物基因组文库，图6参7     

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     

杆状病毒包涵体蛋白的共同抗原性

在比对已公布的37种杆状病毒包涵体蛋白氨基酸序列的基础上,选定棉铃虫核型多角体病毒(HaNPV)多角体蛋白的两个高度保守多肽(54-113aa和206-245aa)制备多克隆抗体,用Western blot法检测这两个高度保守多肽与多种杆状病毒包涵体蛋白之间的血清学关系.结果表明,HaNPV多角体蛋白的两个多肽的抗体与14种核型多角体病毒的多角体蛋白和5种颗粒体病毒的颗粒体蛋白均有明显的杂交信号,表明杆状病毒的包涵体蛋白之间具有共同的抗原决定簇.根据杆状病毒包涵体蛋白之间的这种血清学关系,进一步讨论了利用免疫金试纸技术检测病毒杀虫剂中包涵体含量的可行性.     

温度和pH值对重组杆状病毒(AcMNPV-BmKIT-Chi)的毒力影响

研究了温度、酸碱度对重组杆状病毒(AcMNPV-BmKIT-Chi)毒力的影响.结果显示,多角体病毒在温度4~50℃,pH6.0~8.0范围内可以保持较高的毒力稳定性;不同虫龄棉铃虫幼虫对重组病毒的毒力敏感性差异显著,低虫龄对重组核型多角体病毒更为敏感;病毒对幼虫存在亚致死作用,饲毒组与对照组幼虫体重差异显著.图3表3参13     

昆虫病毒增效蛋白的研究进展

杆状病毒是专化性很强的一类双链ＤＮＡ病毒,主要感染无脊椎动物,特别是鳞翅目昆虫．杆状病毒科又分为３个亚组：核型多角体病毒ＮＰＶ（Ｎｕｃｌｅａｒｐｏｌｙｈｅｄｒｏｓｉｓｖｉｒｕｓ）、颗粒体病毒ＧＶ（Ｇｒａｎｕｌｏｓｉｓｖｉｒｕｓ）和非包涵体型杆状病毒（...     

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．     

Importance of greater interdisciplinarity and geographic scope when tackling the driving forces behind biological invasions

Invasive non-native species are important drivers of ecosystem change, yet the driving forces of biological invasions themselves are poorly understood. Such information is essential to ensure policies focus on the most relevant drivers, and that future scenarios capture the full range of potential outcomes for invasive non-native species. I carried out a bibliometric analysis of articles published from 2000 to 2020 that address either invasive non-native species or biodiversity and ecosystem services and that also mention 1 or more drivers of ecosystem change. I examined 5 indirect drivers (demographic, economic, governance, sociocultural, and technological) and 6 direct drivers (climate change, invasive non-native species, land-use or sea-use change, natural hazards, pollution, and resource extraction). Using the Web of Science core collection of citation indexes, I undertook searches of article titles and keywords and retrieved 27,462 articles addressing invasive non-native species and 110,087 articles dealing with biodiversity or ecosystem services. Most research to date on biological invasions as well as on biodiversity and ecosystem services has focused on anthropogenic direct drivers of ecosystem change rather than indirect drivers. Yet currently, less than 18% of articles addressing biological invasions examined drivers of ecosystem change, a similar level to that found over 20 years ago for biodiversity or ecosystem services. Knowledge of the drivers of biological invasions is limited, emphasizes tractable drivers over those that require an interdisciplinary approach, and is biased toward developed economies. Drivers generally deemed important for biological invasions, such as governance and resource extraction, accounted for less than 2% of research effort. The absence of a systematic understanding of the forces that drive invasive non-native species and how they interact means that attempts to mitigate or forecast biological invasions are likely to fail. To address biological invasions requires a much better orientation of national and international research on drivers in relation to both their actual importance as well as their policy relevance.     

Mitigating the effects of climate change on the nests of sea turtles with artificial irrigation

For sea turtles, like many oviparous species, increasing temperatures during development threaten to increase embryonic mortality, alter offspring quality, and potentially create suboptimal primary sex ratios. Various methods are being implemented to mitigate the effects of climate change on reproductive success, but these methods, such as breeding programs, translocations, and shading, are often invasive and expensive. Irrigation is an alternative strategy for cooling nests that, depending on location, can be implemented relatively quickly and cheaply. However, multiple factors, including ambient conditions, nest substrate, and species characteristics, can influence irrigation success. Additionally, irrigation can vary in duration, frequency, and the volume of water applied to nests, which influences the cooling achieved and embryonic survival. Thus, it is critical to understand how to maximize cooling and manage risks before implementing irrigation as a nest-cooling strategy. We reviewed the literature on nest irrigation to examine whether artificial irrigation is feasible as a population management tool. Key factors that affected cooling were the volume of water applied and the frequency of applications. Embryonic responses varied with species, ambient conditions, and the timing of irrigation during development. Nest inundation was the key risk to a successful irrigation regime. Future irrigation regimes must identify clear targets, either primary or adult sex ratios, that maximize population viability. Monitoring population responses and adjusting the irrigation regime in response to population characteristics will be critical. Most studies reported on the manipulation of only one or two variables, further research is required to understand how altering multiple factors in an irrigation regime influences the cooling achieved and embryonic responses.     

Positive and Negative Effects of a Threatened Parrotfish on Reef Ecosystems

Species that are strong interactors play disproportionately important roles in the dynamics of natural ecosystems. It has been proposed that their presence is necessary for positively shaping the structure and functioning of ecosystems. We evaluated this hypothesis using the case of the world's largest parrotfish (Bolbometopon muricatum), a globally imperiled species. We used direct observation, animal tracking, and computer simulations to examine the diverse routes through which B. muricatum affects the diversity, dispersal, relative abundance, and survival of the corals that comprise the foundation of reef ecosystems. Our results suggest that this species can influence reef building corals in both positive and negative ways. Field observation and simulation outputs indicated that B. muricatum reduced the abundance of macroalgae that can outcompete corals, but they also feed directly on corals, decreasing coral abundance, diversity, and colony size. B. muricatum appeared to facilitate coral advancement by mechanically dispersing coral fragments and opening up bare space for coral settlement, but they also damaged adult corals and remobilized a large volume of potentially stressful carbonate sediment. The impacts this species has on reefs appears to be regulated in part by its abundance—the effects of B. muricatum were more intense in simulation scenarios populated with high densities of these fish. Observations conducted in regions with high and low predator (e.g., sharks) abundance generated results that are consistent with the hypothesis that these predators of B. muricatum may play a role in governing their abundance; thus, predation may modulate the intensity of the effects they have on reef dynamics. Overall our results illustrate that functionally unique and threatened species may not have universally positive impacts on ecosystems and that it may be necessary for environmental managers to consider the diverse effects of such species and the forces that mediate the strength of their influence. Efectos Positivos y Negativos de un Pez Loro Amenazado Sobre Ecosistemas Arrecifales     

Form-specific fragances fromOphrys insectifera L. (Orchidaceae) attract species of different pollinator genera. Evidence of sympatric speciation?

Two closely related forms ofOphrys insectifera were observed in the field to attract different pollinator species selectively.O. i. ssp.insectifera attracted males of two species ofArgogorytes (Sphecidae, Hymenoptera Aculeata) andO. i. ssp.aymoninii attractedAndrena combinata males (Andrenidae, Apoidea, Hymenoptera Aculeata). A third form,O. aff.i. ssp.insectifera, attracted none of these three species. Volatile compounds from flowers and inflorescences of the three forms (originating from Öland, Sweden, and Aveyron, France) were collected, using entrainment, enfleurage, and solvent extraction techniques, and identified by gas chromatography — mass spectrometry. Scent differences between the three forms were confirmed in the amounts of aliphatic hydrocarbons (C₁₁–C₁₉), methyl esters (C₁₄–C₁₈), short chain aliphatic 1-alcohols (C₆–C₁₂), and monoterpene alcohols (C₁₀).     

Estimation of the surface stress from the streamwise pressure gradient: The Kármán Integral Momentum Equation revisited

A method is developed to estimate the stress at the surface in a portable wind tunnel for wind erosion studies. The boundary layer height and the pressure gradient are used in a simple expression from the Kármán Integral Momentum Equation. Values of friction velocity u _* are within 10% of experimental values obtained through correlation techniques, including measurements of differential pressures with the Murdoch Turbulence Probe MTP and the X-wire, hot-wire anemometer XWA. Wind velocity and stress profiles reveal logarithmic trends and a constant stress layer near the surface in the DAWA portable wind tunnel. Realignment of the statistics with the mean wind is essential.     

Using decision science to evaluate global biodiversity indices

Global biodiversity indices are used to measure environmental change and progress toward conservation goals, yet few indices have been evaluated comprehensively for their capacity to detect trends of interest, such as declines in threatened species or ecosystem function. Using a structured approach based on decision science, we qualitatively evaluated 9 indices commonly used to track biodiversity at global and regional scales against 5 criteria relating to objectives, design, behavior, incorporation of uncertainty, and constraints (e.g., costs and data availability). Evaluation was based on reference literature for indices available at the time of assessment. We identified 4 key gaps in indices assessed: pathways to achieving goals (means objectives) were not always clear or relevant to desired outcomes (fundamental objectives); index testing and understanding of expected behavior was often lacking; uncertainty was seldom acknowledged or accounted for; and costs of implementation were seldom considered. These gaps may render indices inadequate in certain decision-making contexts and are problematic for indices linked with biodiversity targets and sustainability goals. Ensuring that index objectives are clear and their design is underpinned by a model of relevant processes are crucial in addressing the gaps identified by our assessment. Uptake and productive use of indices will be improved if index performance is tested rigorously and assumptions and uncertainties are clearly communicated to end users. This will increase index accuracy and value in tracking biodiversity change and supporting national and global policy decisions, such as the post-2020 global biodiversity framework of the Convention on Biological Diversity.     

Assessing the extent and nature of wildlife trade on the dark web

Use of the internet as a trade platform has resulted in a shift in the illegal wildlife trade. Increased scrutiny of illegal wildlife trade has led to concerns that online trade of wildlife will move onto the dark web. To provide a baseline of illegal wildlife trade on the dark web, we downloaded and archived 9852 items (individual posts) from the dark web, then searched these based on a list of 121 keywords associated with illegal online wildlife trade, including 30 keywords associated with illegally traded elephant ivory on the surface web. Results were compared with items known to be illegally traded on the dark web, specifically cannabis, cocaine, and heroin, to compare the extent of the trade. Of these 121 keywords, 4 resulted in hits, of which only one was potentially linked to illegal wildlife trade. This sole case was the sale and discussion of Echinopsis pachanoi (San Pedro cactus), which has hallucinogenic properties. This negligible level of activity related to the illegal trade of wildlife on the dark web relative to the open and increasing trade on the surface web may indicate a lack of successful enforcement against illegal wildlife trade on the surface web.     

Estimating freshwater turtle mortality rates and population declines following hook ingestion

Freshwater turtle populations are susceptible to declines following small increases in the mortality of adults, making it essential to identify and understand potential threats. Freshwater turtles ingest fish hooks associated with recreational angling, and this is likely a problem because hook ingestion is a source of additive mortality for sea turtles. We used a Bayesian‐modeling framework, observed rates of hook ingestion by freshwater turtles, and mortality of sea turtles from hook ingestion to examine the probability that a freshwater turtle in a given population ingests a hook and subsequently dies from it. We used the results of these analyses and previously published life‐history data to simulate the effects of hook ingestion on population growth for 3 species of freshwater turtle. In our simulation, the probability that an individual turtle ingests a hook and dies as a result was 1.2–11%. Our simulation results suggest that this rate of mortality from hook ingestion is sufficient to cause population declines. We believe we have identified fish‐hook ingestion as a serious yet generally overlooked threat to the viability of freshwater turtle populations.     

Content analysis of newspaper coverage of the Florida panther

Populations of large carnivores are declining globally, and analysis of public discourse about carnivores is useful for understanding public opinion and influences on management and policy. Portrayal of carnivores in the media affects public perceptions and support for their conservation. We conducted a content analysis of 513 articles about Florida panthers (Puma concolor coryi) published from 2003 to 2006 in newspapers with local circulation in core panther habitat in southwest Florida and papers with statewide circulation to compare the differences in the amount of coverage and portrayals of panther risks to people and property on the basis of proximity of human communities to panthers. Local papers published significantly more news articles and significantly longer news articles primarily about panthers. Articles in local and statewide papers used both episodic frames, which focus on specific occurrences (e.g., a panther sighting or predation) and thematic frames, which focus on general trends (e.g., abundance of panthers over time). Local articles more often emphasized risks that panthers might harm people, pets, or livestock than statewide papers. Our results are consistent with theory that proximity to human-carnivore conflict influences perceptions and salience of risks posed by large carnivores. Most articles mentioned panthers as a secondary topic, which we believe was a result of the relevance an endangered carnivore has in discussions of public land management, development, and regulations in Florida. Claims made by sources quoted in each article had a neutral to positive depiction of panthers, and most quotations were from federal and state agency scientists. We suggest continued use by the media of agency sources provides the opportunity for clear, concordant messages about panther management. Content analysis provides a way to monitor media portrayal of carnivores for consistency with agency outreach goals.