碱性β-聚糖酶产生菌选育及产酶条件优化

从碱性土样中分离到数十株产碱性β聚糖酶类的细菌,经摇瓶反复筛选后,得到一株碱性β聚糖酶产量较高的耐碱性细菌．经初步鉴定,属短小芽孢杆菌（Ｂａｃｉｌｕｓｐｕｍｉｌｕｓ）．其纤维素酶作用的最适ｐＨ为７．６,最适θ为６０℃,木聚糖酶的作用最适ｐＨ为９．０,最适θ为５５℃．该菌的最适生长ｐＨ为８．０,最适产酶θ为２８～３２℃．木聚糖与山梨糖分别是木聚糖酶和纤维素酶的良好诱导物．以麸皮为碳源,产酶的最适浓度为５％．添加尿素和（ＮＨ４）２ＳＯ４为氮源可提高纤维素酶酶活２倍,木聚糖酶酶活１倍．发酵周期为６０ｈ,纤维素酶酶活最高可达１．２１ＩＵｍＬ－１,木聚糖酶酶活可达４３ＩＵｍＬ－１．     

一株产木聚糖酶嗜热真菌樟绒枝霉的鉴定及其产纤维质降解酶系分析

对分离得到的一株产耐热木聚糖酶的真菌CAU521进行鉴定,并对其产纤维质降解酶系进行研究.通过菌落形态、显微镜产孢结构以及18S rDNA序列同源性比对等分析,鉴定该菌为樟绒枝霉(Malbranchea cinnamomea),其最适生长温度为45℃,为一株嗜热真菌.该菌能以农业废弃物玉米芯为碳源液体发酵产耐热木聚糖酶,50℃下培养7 d,木聚糖酶的最高酶活力达到173 U/mL.SDS-PAGE和酶谱分析表明该菌株能同时分泌多种纤维质降解酶:4种木聚糖酶、2种纤维素酶、3种葡聚糖酶和1种甘露聚糖酶.结果表明樟绒枝霉CAU521在降解和利用纤维质材料方面具有潜在的应用价值.     

碱性β—聚糖酶产生菌选育及产酶条件优化

从碱性土样中分离到数十株产碱性β－聚糖酶类的细菌经摇瓶反复筛选后,得到一株碱性β－聚产产量较高的耐碱细菌经初步鉴定,属短小芽孢杆菌,其纤维素酶作用的最适ＰＨ为７．６,最适θ为６０℃,木聚糖酶的作用最适ＰＨＪ为９．０,最适θ为５５℃,该菌的最适生长ＰＨ为８．０,最适产酶θ为２８－３２℃,木聚糖与山梨糖分别是木聚糖酶和纤维素酶的良好诱导物,以麸皮为碳源,产酶的最适浓度为５％,添加尿素和（ＮＨ４）２ＳＯ     

耐碱性木聚糖酶产生菌的筛选及发酵条件研究

利用刚果红透明圈法,从造纸厂碱性土壤中筛选到一株木聚糖酶生产菌株X24-14,经培养特性研究及16S rDNA序列分析,初步认为该菌属于纤维菌属(Cellulosimicrobium).经发酵条件优化,该菌在麸皮60 g/L,蛋白胨10 g/L,K2HPO4 7.0 g/L, pH 8.5,接种量为5%, 37 ℃, 200 r/min的条件下发酵培养108 h,可达到最大活力,为2 204 u/mL. 该酶最适反应温度为60 ℃;具有较宽的pH作用范围,在pH 4.2～9.4范围内能保持较高的酶活力,在pH 9.4条件下,仍具有80%的酶活力; pH稳定性较好,在4 ℃、pH 11.0的条件下处理24 h仍能保持75%的酶活力.图3表1参12     

黑曲霉耐热木聚糖酶XYNB在毕赤酵母中的高效表达

高比活木聚糖酶的高效表达是进一步提高木聚糖酶发酵效价、降低生产成本的有效途径.将黑曲霉木聚糖酶基因XynB(不含信号肽)克隆到分泌型表达载体pPIC9K上,线性化后电击转化巴斯德毕赤酵母GS115,G418和PCR鉴定的阳性转化子经0.5%甲醇、在28℃诱导表达.SDS-PAGE分析表明,该蛋白相对分子质量为20×103左右.优化的诱导表达条件为,每隔12 h添加0.5%的甲醇,发酵5 d后,比活达4 757 U/mg;其最适温度为55℃,最适pH为5.0,80℃处理30min后仍有74%的残余酶活.     

一株能分解纤维素的高温耐碱放线菌

对一株分离于堆肥的分解纤维素的放线菌的形态学和生理生化特征进行了研究．结果表明：基丝由分枝发达的不断裂的菌丝组成，气丝发达；孢子单个长在气丝上，孢子表面光滑，细胞壁含meso-DAP；该菌能分解纤维素，液化明胶、水解酪素和淀粉，在35-60℃生长，pH范围为6．0—12．0．按《伯杰氏系统细菌学鉴定手册》进行了鉴定，初步将此菌归为高温单孢菌属(Thermomonospora sp.)的一个新种．研究了产羧甲基纤维素酶(CMCase)和滤纸酶(FPA)的条件．CMCase最适产酶时间为5d，FPA为8d，最适产酶温度为50℃，产酶最适起始pH为9．5—10．5．最适氮源为黄豆粉，最佳碳源为棉花纤维．图6表2参10     

应用假单胞菌进行防蜡作用的研究

假单胞菌L－11菌体与其胞外产物可以作为一种很好的防蜡剂，对L－11的发酵条件进行了优化，确定了最适发酵培养基的配方，发酵培养基以葡萄糖、蛋白胨为其最适碳、氮源，防蜡剂生产的发酵条件为：前16h的发酵温度为35℃，后期将温度降至20℃，发酵液的pH=4．5为发酵终点，此时发酵液在内条件下的防蜡率为78％，将上述所得菌液在大庆油田做了防蜡实验，结果表明，注入L－11的油井洗井周期有所延长，实验证明了L－11的发酵液可以作为一种经济、有效的蜡剂。     

纤维素酶产生菌HS-F9的筛选鉴定和产酶条件优化

为了选育高产纤维素酶菌株,通过刚果红鉴别培养基以及滤纸条崩解实验测定,从牛粪堆肥中筛选到一株产纤维素酶的真菌HS-F9,根据菌株形态特性和18SrRNA基因序列分析,初步鉴定该菌为绿色木霉(Trichoderma viride).利用液体发酵培养产生纤维素酶,研究了碳源、氮源、培养时间、培养温度、培养基起始pH、接种量对菌株HS-F9产酶的影响.结果表明:产EG、CBH和FPA的最适碳源均为CMC-Na;EG和CBH在以蛋白胨为唯一氮源时酶活最高,FPase则在以黄豆粉为唯一氮源时酶活最高;产生EG、CBH和FPA的最适温度分别为30℃、30℃、33℃;最适起始pH为3.0、3.0和4.0;EG和FPase的最适接种量为2%,CBH最适接种量达到了8%;培养时间均以5～6d为宜.在最适条件下培养,该菌株EG、CBH和FPase的酶活分别达到了5275.3U/mL、8502.1U/mL和3619.1U/mL,是未优化前的1.42、1.35和1.32倍.图6表2参23     

嗜热子囊菌光孢变种内切葡聚糖酶Ⅰ基因在毕赤酵母中的表达及部分酶学性质

嗜热子囊菌是一种嗜热真菌,可以产生具有很高工业价值的内切葡聚糖酶.本研究成功表达了嗜热子囊菌内切葡聚糖酶Ⅰ基凶,并获得热稳定的重组内切葡聚糖酶.提取嗜热子囊菌光孢变种(Thermoascus aurantiacus var.levisporus)总RNA,通过RT-PCR方法克隆出内切-β-葡聚糖酶eg1基因的成熟肽编码序列.采用基因重组的方法构建该基因的巴斯德毕赤酵母Pichia pastoris分泌型表达载体pPIC9K-eg1,经线性化后采用电穿孔法将其导入毕赤酵母GS115中,大量筛选后获得高效表达内切葡聚糖酶Ⅰ的毕赤酵母工程菌株GpN24.该菌株采用甲醇诱导120 h后,内切葡聚糖酶Ⅰ的活力可达570.7 U/mL,最适温度为55℃,在90℃的条件下保温30 min后仍具有60%的酶活力;最适pH为5.0,在pH 3.0～5.0的条件下酶活力保持稳定.图6表2参16     

一株嗜酸性产纤维素酶真菌的特性及产酶条件优化

从四川海螺沟原始森林腐土中分离到一株嗜酸性产纤维素酶的真菌X-13,其主要特点是产纤维素酶的最适pH及其纤维素酶最适反应pH均为2.0.在PDA培养基上培养时菌落呈浅黄色至肉桂色,反面呈黄色至棕褐色,产黄色色素;菌丝体透明有隔膜,分生孢子呈球形或近球形.根据菌株的形态特征以及ITS序列同源性和系统发育分析结果,鉴定该菌株为土曲霉(Aspergillus terreus Thom).该菌最佳产酶培养时间为8～10 d;最适产酶温度为30℃,纤维素酶最适反应温度为50℃;最佳碳源、氮源分别为纤维素粉和硫酸铵.通过响应面法对菌株产纤维素酶条件进行优化,使菌株X-13纤维素酶活从1.39 IU/mL提高到2.94 IU/mL,提高了111.5%.     

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     

Representation of the world's biophysical conditions by the global protected area network

Protected areas (PAs) are often implemented without consideration of already existing PAs, which is likely to cause an overrepresentation of certain biophysical conditions. We assessed the representativeness of the current PA network with regard to the world's biophysical conditions to highlight which conditions are underprotected and where these conditions are located. We overlaid terrestrial and marine PAs with information on biophysical conditions (e.g., temperature, precipitation, and elevation) and then quantified the percentage of area covered by the PA network. For 1 variable at a time in the terrestrial realm, high temperature, low precipitation, and medium and very high elevation were underrepresented. For the marine realm, low and medium sea surface temperature (SST), medium and high sea surface salinity (SSS), and the deep sea were underrepresented. Overall, protection was evenly distributed for elevation across the terrestrial realm and SST across the marine realm. For 2 variables at a time, cold and very dry terrestrial environments had mostly low protection, which was also the case for low SST and low and medium SSS across most depths for marine environments. Low protection occurred mostly in the Sahara and the Arabian Peninsula for the terrestrial realm and along the Tropic of Capricorn and toward the poles for the marine realm. Although biodiversity measures are of prime importance for the design of PA networks, highlighting biophysical gaps in current PAs adds a frequently overlooked perspective. These gaps may weaken the potential of PAs to conserve biodiversity. Thus, our results may provide useful insights for researchers, practitioners, and policy makers to establish a more comprehensive global PA network.     

Quantitative composition of the defensive secretion ofBledius species (Coleoptera: Staphylinidae: Oxytelinae) is adapted to naturally occurring predators

Summary The adaptation of defensive secretions to their target organisms was examined for the abdominal gland secretions ofBledius furcatus, B. spectabilis andB. arenarius. Therefore the target organisms of the secretion of theseBledius species (i.e. their predators) had to be identified. At the collection sites examined these were the earwigLabidura riparia, the antCataglyphis bicolor, the flyLispe candicans, different carabids of the generaPogonus, Dichirotrichus, Dyschirius, Bembidion andCalathus and the wading birdsHaematopus ostralegus andCalidris alba. The secretion of the abdominal glands contains the toxin ptoluquinone dissolved in either-dodecalactone and 1-undecene (B. furcatus andB. spectabilis) or in octanoic acid and octyloctanoate (B. arenarius). The ratio of these solvents is species-specific. Application experiments using some of the natural insect predators (L. riparia, C. bicolor, Pogonus, Di. gustavii, Dyschirius) revealed that these solvent ratios provided a more effective deterrent than other possible ratios. Thus by combining the solvents in certain ratios, the capability of cuticular penetration and therefore the effectiveness of the defensive secretions are adapted to their natural targets.     

Democracy versus development: New urban debates

The politics of development includes subtexts of choice, as underpinned by community consensus (or lack of it), against the larger backdrop of a liberal democracy, with its intricate power structures that influence the choice of concepts and often lead to complicity rather than consent. A dilemma exists between the understanding of field-level practitioners and that of academics, whose understandings of progress differ. Progress can (and often does) imply long-term change across generations whereas development is often more a matter of crisis management and delivering tangible results to end-users. This makes it a bone of contention across political systems, irrespective of local or global dynamics. Development can emerge as a tool to be wielded for power and further political mileage at the expense of progress. Progress cannot be studied as a counterpoint to development as the sense of history is often missing in development interventions. Development tends to be piecemeal as its very logic often stems from the rationale of power. This is not to suggest, however, that development is only about politics; it is definitely more than the politics of everyday life and involves all of civil society that requires its critical and immediate attention. In this sense, it has become a priority for politicians with an underlying agenda designed to out-manouevre dissent and all statements of protest.     

A framework to measure the wildness of managed large vertebrate populations

As landscapes continue to fall under human influence through habitat loss and fragmentation, fencing is increasingly being used to mitigate anthropogenic threats and enhance the commercial value of wildlife. Subsequent intensification of management potentially erodes wildness by disembodying populations from landscape-level processes, thereby disconnecting species from natural selection. Tools are needed to measure the degree to which populations of large vertebrate species in formally protected and privately owned wildlife areas are self-sustaining and free to adapt. We devised a framework to measure such wildness based on 6 attributes relating to the evolutionary and ecological dynamics of vertebrates (space, disease and parasite resistance, exposure to predation, exposure to limitations and fluctuations of food and water supply, and reproduction). For each attribute, we set empirical, species-specific thresholds between 5 wildness states based on quantifiable management interventions. We analysed data from 205 private wildlife properties with management objectives spanning ecotourism to consumptive utilization to test the framework on 6 herbivore species representing a range of conservation statuses and commercial values. Wildness scores among species differed significantly, and the proportion of populations identified as wild ranged from 12% to 84%, which indicates the tool detected site-scale differences both among populations of different species and populations of the same species under different management regimes. By quantifying wildness, this framework provides practitioners with standardized measurement units that link biodiversity with the sustainable use of wildlife. Applications include informing species management plans at local scales; standardizing the inclusion of managed populations in red-list assessments; and providing a platform for certification and regulation of wildlife-based economies. Applying this framework may help embed wildness as a normative value in policy and mitigate the shifting baseline of what it means to truly conserve a species.     

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.     

Sexual Predators,Energy Development,and Conservation in Greater Yellowstone

Abstract: In the United States, as elsewhere, a growing debate pits national energy policy and homeland security against biological conservation. In rural communities the extraction of fossil fuels is often encouraged because of the employment opportunities it offers, although the concomitant itinerant workforce is often associated with increased wildlife poaching. We explored possible positive and negative factors associated with energy extraction in the Greater Yellowstone Ecosystem (GYE), an area known for its national parks, intact biological diversity, and some of the New World's longest terrestrial migrations. Specifically, we asked whether counties with different economies—recreation (ski), agrarian (ranching or farming), and energy extractive (petroleum)—differed in healthcare (gauged by the abundance of hospital beds) and in the frequency of sexual predators. The absolute and relative frequency of registered sex offenders grew approximately two to three times faster in areas reliant on energy extraction. Healthcare among counties did not differ. The strong conflation of community dishevel, as reflected by in‐migrant sexual predators, and ecological decay in Greater Yellowstone is consistent with patterns seen in similar systems from Ecuador to northern Canada, where social and environmental disarray exist around energy boomtowns. In our case, that groups (albeit with different aims) mobilized campaigns to help maintain the quality of rural livelihoods by protecting open space is a positive sign that conservation can matter, especially in the face of rampant and poorly executed energy extraction projects. Our findings further suggest that the public and industry need stronger regulatory action to instill greater vigilance when and where social factors and land conversion impact biological systems.     

Comparison of the insecticidal effects of water extracted and intact aromatic plants onAcanthoscelides obtectus,a bruchid beetle pest of kidney beans

Summary Acanthoscelides obtectus (Say) is one of the most damaging pests of kidney beans,Phaseolus vulgaris L. worldwide. However, aromatic plants from the families Lamiaceae, Lauraceae, Myrtaceae and Poaceae can protect kidney beans by direct or delayed insecticidal effect, through increased adult mortality and inhibition of beetle reproduction (both oviposition and adult emergence). The efficiency of hydrodistillated and intact plants fromThymus vulgaris andT. serpyllum, Mentha piperata, Rosmarinus officinalis, Satureia hortensis, Eucalyptus globulus, Laurus nobilis, Origanum vulgare, andCymbopogon nardus was compared. For both extracts,Origanum vulgare had the best effect. The insecticidal effect was induced by more than the essential oils because no significant difference was noticed between distilled and intact plants extract. Inhibition of reproduction was particularly important. These results suggest that lipidic as well as non lipidic, allelochemicals, such as phenolics, or non-protein amino-acids, or flavonoids may be involved in the toxicity of aromatic plants to this beetle.     

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.     

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.