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．     

固定CO2基因工程菌的构建

构建了含有ＲｕｂｉｓＣＯＦｏｒｍ ＩＩ基因的可转化大肠杆菌紫色非硫杆菌穿梭质粒（ＰＭＰＢ２）,将其转化紫色非硫杆菌野生型Ｒｈｏｄｏｐｓｅｕｄｏｍｏｎａｓｐａｌｕｓｔｒｉｓ Ｎｏ．７ 和紫色非硫杆菌ＲｕｂｉｓＣＯＦｏｒｍ Ｉ缺陷型ＲｈｏｄｏｐｓｅｕｄｏｍｏｎａｓｐａｌｕｓｔｒｉｓＮｏ．７ＤＦＩ,获得２ 株ＲｕｂｉｓＣＯ基因工程菌ＭＧ１１ 和ＭＧ１４,二者的ＲｕｂｉｓＣＯ 酶活性比对照菌分别提高７６ ．７％ 和８３．３％ ．连续传４０ 代表明,重组质粒在受体菌中是较稳定的     

苏云金芽孢杆菌无质粒突变株BMB171的转化和表达性能

研究了苏云金芽孢杆菌无质粒突变株ＢＭＢ１７１的转化性能和表达ｃｒｙ１Ａｃ和ｃｒｙ１Ｃａ基因的性能．用ｐＨＴ３１０１、ｐＢＭＢ３０５、ｐＢＭＢ１７３６、ｐＢＭＢ６７１、ｐＢＴＬ１和ｐＨＶ１２４９等６种外源质粒电转化无质粒突变株ＢＭＢ１７１,其转化频率分别是Ｂｔ４Ｑ７、Ｂｔ４Ｄ１０和Ｂｔｉ．ＩＰＳ．７８／１１等３种常用受体菌相应最高转化频率的１０００、６．７、１２．５、６６．７、３５００和２倍,其每μｇＤＮＡ的最高转化子数达１０７．导入ＢＭＢ１７１的外源质粒的稳定性与其复制子类型和质粒大小有关．无质粒突变株ＢＭＢ１７１表达ｃｒｙ１Ａｃ基因的表达量高于对照受体菌Ｂｔ４Ｑ７,略低于Ｂｔ４Ｄ１０和Ｂｔｉ．ＩＰＳ．７８／１１,而ＢＭＢ１７１表达ｃｒｙ１Ｃａ基因的表达量高于这３种受体菌;对小菜蛾３龄幼虫和甜菜夜蛾初孵幼虫的毒力测定结果表明,ＢＭＢ１７１表达ｃｒｙ１Ａｃ基因的杀虫毒力高于Ｂｔ４Ｑ７和Ｂｔ４Ｄ１０,略低于Ｂｔｉ．ＩＰＳ．７８／１;而表达ｃｒｙ１Ｃａ基因的毒力高于这３种受体菌     

微生物降解蔬菜残留农药研究

从富集培养物中筛选到以甲胺磷为唯一碳源生长的菌株ＮＭＪ５和以乐果为唯一碳源生长的菌株ＮＭＬ３,经鉴定分别为芽孢杆菌属（Ｂａｃｉｌｕｓｓｐ．）和假单胞菌属（Ｐｓｅｕｄｏｍｏｎａｓｓｐ．）．它们利用甲胺磷、乐果生长的最适条件及在无机盐培养基中农药最大耐受浓度分别为ｐＨ７．５,ｔ＝３５℃,ρ＝１０００ｍｇ／Ｌ和ｐＨ７．５,ｔ＝３０℃,ρ＝２０００ｍｇ／Ｌ．在无碳基础培养基内单菌株培养８ｄ,５２４ｍｇ／Ｌ的甲胺磷好气降解４２．５％,厌气降解３５．９％,２５０ｍｇ／Ｌ的乐果好气降解５０．２％,厌气降解１６．４％．小区试验表明,ＮＭＪ５、ＮＭＬ３的菌液制剂对普通白菜的变种南农矮脚黄（Ｂｒａｓｉｃａｃａｍｐｅｓｔｒｉｓｓｐ．ｃｈｉｎｅｎｓｉｓＬ．ｖａｒ．ｃｏｍｍｕｎｉｓＴｓｅｎｅｔＬｅｅ）中残留的甲胺磷、乐果有明显的去除作用．     

汉沽生物稳定塘净化效果研究

汉沽生物稳定塘建于１９９３年．它由１０级池塘组成,ＡＴ７５ｈｍ２,用于处理城市污水（主要是化工废水）．根据２年试运行期间化学和生物学资料,分析了稳定塘的净化效果．结果表明,该稳定塘净化效能较好．主要污染物ＢＯＤ５、ＣＯＤＣｒ、ＮＨ３Ｎ和ＰＯ４Ｐ的平均去除率分别为８６．２％、７０．１％、６０．３％和６７．５％;平均年去除ＢＯＤ５４２９７．８ｔ、ＣＯＤＣｒ６１７９．８ｔ、ＮＨ３Ｎ１５４．６ｔ和ＰＯ４Ｐ８１．９ｔ．但由于有机物负荷量过高,使出水水质较差．文中提出若干改善出水水质的措施和建议     

全期表达外源基因的重组杆状病毒

构建了含早期、晚期、极晚期启动子、全期表达ＨＢｓＡｇ 基因的重组杆状病毒ｖＴｎＮＰＶＰ３５ＨＢｓＯＣＣ＋ ．结果ＨＢｓＡｇ 表达时间由感染后１２ ｈ 提前到４ ｈ,而其表达量,特别是病毒感染早期的表达水平亦显著提高,其中２４ ｈ 和９６ ｈ 的表达量分别比对照提高１８０ ％ 和６０ ％ ． 转录起始位点分析结果显示,该重组毒株可在病毒感染的早、晚期形成多个转录本     

紫云英根瘤分离菌株质粒多样性的研究

从广东、广西、湖南、湖北、江西、浙江、江苏７省紫云英（ＡｓｔｒａｇａｌｕｓｓｉｎｉｃｕｓＬ．）主要分布区４６个采集点的紫云英根瘤中分离到１０１个菌株．检测发现有１６种不同质粒类型,所含质粒１～４个不等,含两个质粒的菌株占绝大部分,质粒Ｍｒ为３６×１０６～３６４×１０６．用克隆Ｍ．ｈｕａｋｕｉ７６５３Ｒ的ｎｏｄＤＢＣ４．２ｋｂ片段为探针（α３２ｐｄＣＴＰ）进行的分子杂交结果表明,所有这些菌株均含有共生质粒,通常为第一大质粒,但Ｃ、Ｌ、Ｍ、Ｎ和Ｏ型为第二大质粒,共生质粒Ｍｒ在１３１×１０６～３６４×１０６之间．比较质粒类型与１６Ｓ２３ＳｒＤＮＡＩＧＳ（ｐＨｒ,ｐ２３ＳＲ０１）ＰＣＲＲＦＬＰ分析结果表明,ＩＧＳ类型不同,即使在同一植株分离所得的根瘤其质粒类型常不同,但在同一植株上分离菌株的质粒类型相同,则其ＩＧＳ常表现相同．表明质粒的类型是细菌染色体背景和细菌外环境（如共生条件等）共同作用的结果     

邻苯二酚1,2—双加氧酶（C120）基因的定位,克隆和表达

放射性同位素标记ｔｆｄＣ基因片段作探针,Ｓｏｕｔｈｅｒｍｂｌｏｔ杂交定位Ｌ１菌株的邻苯二酚１,２－双加氧酶基因位于ＰｓｔＩ的Ｉ片段和ＢａｍＨＩ的Ｍ和Ｎ片段。     

人G—CSF转基因鼠系与正常鼠系比较研究

利用人ＧＣＳＦ基因组基因和ｃＤＮＡ基因作为目的片段,将其受控于２．６ｋｂ的小鼠乳清酸蛋白（ＷＡＰ）基因的调控区下,通过显微注射法获得了两只整合有人ＧＣＳＦ转基因小鼠,通过繁殖建立了稳定的转基因系．一些表型参数测定表明,转基因鼠与正常鼠无明显差别,为对转基因动物进行安全性评价提供了依据     

BACILLUS SP. BT-7木聚糖酶基因克隆与鉴定

报道了从一株有较高半纤维素酶活性的Ｂａｃｉｌｕｓｓｐ．ＢＴ７克隆内切木聚糖酶基因的结果．以大肠杆菌ＸＬ１ｂｌｕｅ为宿主菌,采用水解圈检测法,在含有ＲＢＢ木聚糖（４氧甲基Ｄ葡萄糖苷Ｄ木聚糖Ｒｅｍａｚｏｌ亮蓝Ｒ）的平板上分离到能水解ＲＢＢ木聚糖的阳性克隆３个,对它们进行的限制酶作图和Ｓｏｕｔｈｅｒｎ杂交分析表明,它们属于３种不同的内切木聚糖酶基因     

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.     

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     

Potential for Electropositive Metal to Reduce the Interactions of Atlantic Sturgeon with Fishing Gear

Atlantic sturgeon (Acipenser oxyrhynchus) populations have been declared either endangered or threatened under the U.S. Endangered Species Act. Effective measures to repel sturgeon from fishing gear would be beneficial to both fish and fishers because they could reduce both fishery‐associated mortality and the need for seasonal and area closures of specific fisheries. Some chondrostean fishes (e.g., sturgeons and paddlefishes) can detect weak electric field gradients (possibly as low as 5 Μv/cm) due to arrays of electroreceptors (ampullae of Lorenzini) on their snout and gill covers. Weak electric fields, such as those produced by electropositive metals (typically mixtures of the lanthanide elements), could therefore potentially be used as a deterrent. To test this idea, we recorded the behavioral responses of juvenile Atlantic sturgeon (31–43 cm fork length) to electropositive metal (primarily a mixture of the lanthanide elements neodymium and praseodymium) both in the presence and absence of food stimuli. Trials were conducted in an approximately 2.5 m diameter × 0.3 m deep tank, and fish behaviors were recorded with an overhead digital video camera. Video records were subsequently digitized (x, y coordinate system), the distance between the fish and the electropositive metal calculated, and data summarized by compiling frequency distributions with 5‐cm bins. Juvenile sturgeon showed clear avoidance of electropositive metal but only when food was present. On the basis of our results, we conclude that the electropositive metals, or other sources of weak electric fields, may eventually be used to reduce the interactions of Atlantic sturgeon with fishing gear, but further investigation is needed. El Potencial del Metal Electropositivo para Reducir las Interacciones del Esturión Atlántico con Instrumentos de Pesca Bouyoucos, Bushnell & Brill 13–003     

The role of time and species identities in spatial patterns of species richness and conservation

Many conservation actions are justified on the basis of managing biodiversity. Biodiversity, in terms of species richness, is largely the product of rare species. This is problematic because the intensity of sampling needed to characterize communities and patterns of rarity or to justify the use of surrogates has biased sampling in favor of space over time. However, environmental fluctuations interacting with community dynamics lead to temporal variations in where and when species occur, potentially affecting conservation planning by generating uncertainty about results of species distribution modeling (including range determinations), selection of surrogates for biodiversity, and the proportion of biodiversity composed of rare species. To have confidence in the evidence base for conservation actions, one must consider whether temporal replication is necessary to produce broad inferences. Using approximately 20 years of macrofaunal data from tidal flats in 2 harbors, we explored variation in the identity of rare, common, restricted range, and widespread species over time and space. Over time, rare taxa were more likely to increase in abundance or occurrence than to remain rare or disappear and to exhibit temporal patterns in their occurrence. Space–time congruency in ranges (i.e., spatially widespread taxa were also temporally widespread) was observed only where samples were collected across an environmental gradient. Fifteen percent of the taxa in both harbors changed over time from having spatially restricted ranges to having widespread ranges. Our findings suggest that rare species can provide stability against environmental change, because the majority of species were not random transients, but that selection of biodiversity surrogates requires temporal validation. Rarity needs to be considered both spatially and temporally, as species that occur randomly over time are likely to play a different role in ecosystem functioning than those exhibiting temporal structure (e.g., seasonality). Moreover, temporal structure offers the opportunity to place management and conservation activities within windows of maximum opportunity.     

Main nutrient compounds in food bodies of Mexican <Emphasis Type="Italic">Acacia</Emphasis> ant-plants

Summary. Myrmecophytic plants use obligate ant mutualists as a constitutive indirect defence mechanism. These plants often produce cellular food bodies (FBs) to nourish their resident ants. Lipids, proteins, and even highly specialised compounds such as glycogen have been reported from FBs, but detailed chemical analyses of FB composition have so far been presented only for Southeast Asian Macaranga and Central American Piper myrmecophytes. Here we report the chemical composition of FBs of five myrmecophytic Acacia (Fabaceae) species from Mexico using HPLC (carbohydrates and proteins) and GC-MS (lipids). Feeding experiments revealed no hints on any use of external food sources by the inhabiting Pseudomyrmex ants. These ants obviously rely completely on FBs and extrafloral nectar provided by their hosts. The total content of nutrients in Acacia FBs was 15-25% of FB dry mass, being much lower than in Macaranga or Piper FBs. Proteins were dominating (8-14 % dm) in Acacia FBs and thus were present in higher amounts than in Macaranga FBs, yet in lower amounts than in Piper. Lipids contributed 1-9 % of dry mass, showing a lower proportion than in FBs of Macaranga or Piper. Carbohydrates made up 3-11 % dm, reaching in most Acacia species the same range as observed in Macaranga and in Piper FBs. Water content was 18-24 % of FB fresh mass, and structural tissue obviously made up a much higher proportion in Acacia FBs than in Macaranga or Piper FBs. Both characters might represent an adaptation to producing FBs unprotected at the leaf tips under dry conditions. Acacia FBs contain all amino acids and all fatty acids that are considered essential for insects, and their contents of lipids and proteins are higher than in the leaves from which they are ontogenetically derived. This indicates a putatively adaptive enrichment of nutritionally valuable compounds in structures functioning as ant-food.     

Examining the uncertain origin and management role of martens on Prince of Wales Island,Alaska

Conservation biologists are generally united in efforts to curtail the spread of non‐native species globally. However, the colonization history of a species is not always certain, and whether a species is considered non‐native or native depends on the conservation benchmark. Such ambiguities have led to inconsistent management. Within the Tongass National Forest of Alaska, the status of American marten (Martes americana) on the largest, most biologically diverse and deforested island, Prince of Wales (POW), is unclear. Ten martens were released to POW in the early 1930s, and it was generally believed to be the founding event, although this has been questioned. The uncertainty surrounding when and how martens colonized POW complicates management, especially because martens were selected as a design species for the Tongass. To explore the history of martens of POW we reviewed other plausible routes of colonization; genetically and isotopically analyzed putative marten fossils deposited in the late Pleistocene and early Holocene to verify marten occupancy of POW; and used contemporary genetic data from martens on POW and the mainland in coalescent simulations to identify the probable source of the present‐day marten population on POW. We found evidence for multiple routes of colonization by forest‐associated mammals beginning in the Holocene, which were likely used by American martens to naturally colonize POW. Although we cannot rule out human‐assisted movement of martens by Alaskan Natives or fur trappers, we suggest that martens be managed for persistence on POW. More generally, our findings illustrate the difficulty of labeling species as non‐native or native, even when genetic and paleo‐ecological data are available, and support the notion that community resilience or species invasiveness should be prioritized when making management decisions rather than more subjective and less certain conservation benchmarks.