利用原生质体诱变技术筛选脱落酸高产菌株

以葡萄孢属TB-3菌株为出发菌株制备其原生质体．在纤维素酶浓度为20g／L，蜗牛酶浓度为3g／L的酶解系统中，25℃酶解3h，其原生质体制备数可达67×105mL-1，在KYM上的再生率为46．1％，在KPDA上的再生率为33．6％，经过3轮原生质体紫外线诱变后回复再生，及对大量再生突变株进行发酵筛选，获得高产稳定株TB－3H8，其发酵液中ABA的质量浓度可达1．4g／L     

产碱性果胶酶的枯草芽孢杆菌ZGL14原生质体制备与再生条件

原生质体融合技术是一种重要的微生物育种方法.以能产碱性果胶酶的枯草芽孢杆菌ZGL14为出发菌株,探讨影响其原生质体制备和再生的主要因素.通过研究菌龄、溶菌酶浓度、酶解时间、酶解温度、渗透压稳定剂的p H值、再生培养基类型和培养方式等对枯草芽孢杆菌ZGL14原生质体制备与再生的影响,确定其最优化条件.结果显示:枯草芽孢杆菌ZGL14原生质体制备的最优条件为菌龄9 h,溶菌酶浓度0.2 mg/m L,酶解时间10 min,酶解温度32℃,渗透压稳定剂的p H值7.0.在此条件下,枯草芽孢杆菌ZGL14原生质体的形成率为98%,再生率达30%.枯草芽孢杆菌ZGL14原生质体再生培养基以琥珀酸钠作为渗透压稳定剂并结合Ca~(2+)再生效果较好,再生方式以单层培养较好.本研究获得了枯草芽孢杆菌ZGL14原生质体制备和再生的最佳条件,可为实现其融合育种提供技术支持.     

平菇原生质体的高效制备及再生优化条件

以杂优1号平菇为出发菌株,对其原生质体制备、再生的各关键因子及条件进行比较研究,结果表明,其原生质体制备的优化条件为:以0.6 mol/L MgSO4为稳渗剂配制浓度为15 mg/mL新鲜溶壁酶液,菌丝与酶液比w/V=1mg:8μL,取培养6 d的菌丝200 mg,自然pH下28℃小平板酶解6 h,原生质体得率可达2.80×107个/mL.原生质体再生实验结果表明,50 U/mL链霉素能有效控制平菇原生质体再生的杂菌污染.孟加拉红对平菇原生质体再生具强抑制作用,抑制率达93%.进一步研究证实,0.6 mol/L的MsSO4、KCI等无机盐稳渗剂对原生质体制备的得率较高,而0.6 mol/L的蔗糖作为稳渗剂的平菇原生质体再生率较高.图4表3参23     

红豆杉内生真菌发酵培养基和原生质体制备酶系统的筛选

在对红豆杉内生真菌(Ozoniumsp.)适生碳源、氮源和生长情况研究的基础上,通过正交试验筛选了其发酵培养基和原生质体制备的酶系统;用L9(34)安排了四因素三水平并考虑交互作用的正交试验,对实验结果进行了分析.结果表明,最优的发酵培养基为果糖1%、蔗糖1%、蛋白胨0.2%、酵母粉0.5%、KH2PO40.5%、MgSO4·7H2O0.3%、VB10.001%;分离原生质体的最优酶系统为1.5%溶壁酶 0.5%蜗牛酶 1.5%纤维素酶 1.0%溶菌酶;用此酶系统在30℃条件下酶解3h,原生质体的产量达6.55×107个/mL酶液;经荧光素二醋酸酯(FDA)染色评估原生质体活力,表明该条件下分离的原生质体活力较高,原生质体的再生率为2.56%.该研究为利用生物技术手段改良紫杉醇生产菌奠定了基础.图6表4参32     

GUS基因在诸葛菜子叶原生质体中的瞬间表达

本文以诸葛菜无菌苗子叶组织为材料，采用原生质体培养获得成功，建立了原生质体培养高频再生体系，植板率３％，植株再生频率１００％。在此基础上，本文首次研究了ＰＥＧ介导转化诸葛菜原生质体的影响因素，系统地试验了ＰＥＧ法转化子叶原生质体的过程，发现：最适质粒量为２５－３０μｇ；ＰＥＧ终浓度为１５％，ｐＨ８．０；另外，表达效率还与质粒的大小有关，较小的质粒具有相对较高的转化效率；而ＣＴ－ＤＮＡ以及热击处理未     

诸葛菜下胚轴和子叶原生质体培养

以诸葛菜无菌苗的下胚轴和子叶组织为材料，分离原生质体，ＦＷ产量分别为５×１０８／ｇ和１×１０７／ｇ．下胚轴原生质体经纯化后在原生质体培养基中作液体浅层暗培养，培养密度为５×１０４／ｍＬ，培养第二天即出现第一次分裂，７～９ｄ后分裂频率约５０％，两周后出现大量细胞团；转入扩增培养基中，即扩增出大量小愈伤，植板率约为５％；愈伤转入分化培养基后即可诱导分化成苗，分化率达１００％，随后转到生根培养基中，即形成完整植株．子叶原生质体的培养方式与下胚轴原生质体培养基本相同，结果也类似．     

氢化可的松高转化菌株的选育及其发酵条件

利用酮康唑抗性突变选育氢化可的松高转化菌株，即将经过紫外线诱变处理的新月弯孢霉原生质体倾注于含有酮康唑最小抑制浓度(10μmoL／L)的培养基平板上，获得了136株酮康唑抗性突变株，氢化可的松转化率高于出发菌株的有14株．其中抗性突变株KA-91，其氢化可的松转化率为出发菌株的1．42倍，达到68．05％．研究了菌株的发酵性能，优化了培养基组成和发酵工艺条件，氢化可的松转化率达到73．6％．图7表3参5     

CO2浓度和光合光量子通量密度对叶用甘薯组培苗光合自养和过氧化物酶活力的影响

以新兴蔬菜叶用甘薯组织培养苗的茎节段作为外植体，用MS低糖、无激素培养基进行生根培养，设置不同的CO2浓度(Ci)和光合光量子通量密度(PPFD)对组培植株进行环境调控处理，获得小植株净光合速率最高的处理组合(Ci，PPFD)=(8720μmol mol^-1，250μmol m^-2 s^-1)，并在此组合下继续培养21d。结果表明：该调控系统可以有效地调节控制组培苗生长环境中的Ci和PPFD，提高组培苗的光合自养能力，实现低糖、无激素培养，处理组植株生长健壮，叶片数、株高、根鲜重和单株鲜重分别高出CK组57．7％，103．3％，131．6％和225．35％，叶片和根系过氧化物酶活力分别高出CK组26．3％和24．0％。图3表1参18。     

黄孢原毛平革菌对染料和印染废水的降解

白腐真菌黄孢原毛平革菌在合成木素过氧化物酶的限碳培养条件下，可以降解酶性、直接、活性、阳离子等多种类型的印染工业染料、在培养的d5木素过氧化物酶活力最高时，分别加入酸性染料卡布龙红和弱酸大红，质量浓度（ρ/mgL^-1)分别为25、50、100和12．5、25．50，48h后培养液基本脱色，较高浓度下菌膜上有残余染料吸附，5d后染料质量降解率分别是100％、88％、92％和58％、58％、65％、38％。以含有上述两种染料的印染废水置换培养液，并加入葡萄糖1g/L，黄孢原毛平革菌可以直接使废水脱色，菌丝可以重复培养脱色废水至少5批，每批废水的脱色率均大于90％，5批废水总的染料质量降解率约为80％，在重复培养脱色废水的过程中，测不到木素过氧化物酶的活力，说明废水中的染料分子是在细胞表面或进入胞内被降解的，若加入的葡萄糖浓度降低一半以上，菌丝脱色废水的效果将有所下降，图5表5参11     

鞭式节丛孢(Arthrobotrys flagrans CBS583.91)对氯化钠和氯化镉的耐受性

首次报道了鞭式节丛孢的氯化钠和氯化镉耐受性,在查氏平板上培养39d，鞭式节丛孢对氯化钠的最高耐受浓度为0．6mol L^-1，对氯化镉的最高耐受浓度为3．0mmol L^-1；在LCMA平板上，0．18mol L^-1氯化钠抑制三维菌网的形成，0．30mol L^-1氯化钠抑制厚垣孢子的萌发，0．05mmol L^-1氯化镉抑制三维菌网形成，0．07mmol L^-1氯化镉抑制厚垣孢子萌发．图1表3参8     

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.     

Threats to intact tropical peatlands and opportunities for their conservation

Large, intact areas of tropical peatland are highly threatened at a global scale by the expansion of commercial agriculture and other forms of economic development. Conserving peatlands on a landscape scale, with their hydrology intact, is of international conservation importance to preserve their distinctive biodiversity and ecosystem services and maintain their resilience to future environmental change. We explored threats to and opportunities for conserving remaining intact tropical peatlands; thus, we excluded peatlands of Indonesia and Malaysia, where extensive deforestation, drainage, and conversion to plantations means conservation in this region can protect only small fragments of the original ecosystem. We focused on a case study, the Pastaza‐Marañón Foreland Basin (PMFB) in Peru, which is among the largest known intact tropical peatland landscapes in the world and is representative of peatland vulnerability. Maintenance of the hydrological conditions critical for carbon storage and ecosystem function of peatlands is, in the PMFB, primarily threatened by expansion of commercial agriculture linked to new transport infrastructure that is facilitating access to remote areas. There remain opportunities in the PMFB and elsewhere to develop alternative, more sustainable land‐use practices. Although some of the peatlands in the PMFB fall within existing legally protected areas, this protection does not include the most carbon‐dense (domed pole forest) areas. New carbon‐based conservation instruments (e.g., REDD+, Green Climate Fund), developing markets for sustainable peatland products, transferring land title to local communities, and expanding protected areas offer pathways to increased protection for intact tropical peatlands in Amazonia and elsewhere, such as those in New Guinea and Central Africa which remain, for the moment, broadly beyond the frontier of commercial development.     

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.