青稞β—1，3—葡聚糖酶Ⅱ基因启动子的克隆分析及表达载体的构建

根据本实验室克隆并公布的藏青稞β—1，3—葡聚糖酶Ⅱ(BGH Ⅱ)基因序列，以藏青稞QB09基因组DNA为模板，构建DNA步移文库，并通过步移技术克隆出BGH Ⅱ起始密码子上游调控序列BGHP．鉴定和分析表明，BGHP具备大多数高等植物启动子的保守元件，预测它对BGHⅡ基因的表达具有一定的作用．为鉴定BGHⅡ基因的基本启动子元件，将基因5′侧翼序列做缺失片段分析，利用PCR方法从BGHP中得到3个不同大小两端带有Hind Ⅲ、BamH I酶切位点的片段BGHP1、BGHP2和BGHP3，定向插人载体pMGFP4(pBI221改建，报告基因为GFP)中，取代原有的CaMV35S启动子，构建了由驱动报告基因GFP的植物表达载体BGHPVl、BGHPV2和BGHPV3，用于大麦的遗传转化，为进一步研究其功能奠定了基础．图5表1参15     

菊酯类杀虫剂抗性棉铃虫para型钠离子通道基因的部分序列测定与突变

昆虫神经系统para型钠离子通道是菊酯类杀虫剂的主要靶标，对10多种昆虫的研究表明，钠离子通道基因发生点突变与昆虫对菊酯类杀虫剂的抗性密切相关．通过RT-PCR扩增的方法，我们获得了编码溴氰菊酯抗性和敏感品系棉铃虫钠离子通道域Ⅱ-Ⅳ的cDNA片段．核苷酸序列及推导的氨基酸序列分析结果表明，溴氰菊酯抗性品系棉铃虫钠离子通道基因不存在其它昆虫中报道的kdr和super-kdr抗性突变，但是我们发现了一个甲硫氨酸(M)到亮氨酸(L)的新突变，该突变位于域Ⅱ和Ⅲ之间．由于昆虫钠离子通道高度保守，因此，该突变很可能与棉铃虫对溴氰菊酯的抗性有关．另外，通过PCR扩增的方法，克隆了棉铃虫钠离子通道域Ⅱ-Ⅳ的基因片段，该片段全长7334bp，包含有13个外显子和12个内含子，与果蝇和烟蚜夜蛾相比，内含子在基因中的位置基本一致，进一步表明昆虫钠离子通道基因在进化过程中高度保守．图5表1参22     

模拟酸雨对珍稀濒危植物红豆杉幼苗的影响研究

利用模拟酸雨实验对珍稀濒危植物曼地亚红豆杉和南方红豆杉生长及生理生化指标进行研究。结果表明,pH4.5的模拟酸雨促进了曼地亚红豆杉(Taxus media)和南方红豆杉(T.mairei)的株高生长(与对照相比分别促进了19%和72%),pH3.5的模拟酸雨对其有一定的影响,但不显著。pH4.5和pH3.5的酸雨处理下,曼地亚红豆杉基径比对照分别下降了46%和42%;南方红豆杉的相应值分别是34%和43%。总的来说,模拟酸雨促进了这两种红豆杉叶绿素的质量分数,类胡萝卜素的质量分数变化因种而异。酸雨处理提高了曼地亚红豆杉抗氧化系统酶活性,与对照相比约提高了5%～142%;南方红豆杉的相应值为-16%～35%。曼地亚红豆杉游离脯铵酸和丙二醛(MDA)的质量分数比对照提高了13%～73%;而南方红豆杉的相应质量分数基本等于或低于对照。与南方红豆杉相比,曼地亚红豆杉对酸雨更加敏感。     

CMS高粱保持系叶绿体基因ps1A1和ps1A2片段的克隆与序列比对

以15种包括同质异核和同核异质高粱材料的总：DNA为模板进行RAPD分析，196个随机引物中，引物Y-19扩增得到一个很有规律的差异片段SAY-192300。该片段只出现在4个保持系(B)和3个恢复系(R)中，而不出现在6个不育系(A)和2个杂种F1中，进一步对cms—Al、cms—A2两套8个材料(A／B／R／F1)的总DNA、mtDNA和cpDNA用引物Y-19进行扩增，发现片段SAY-192300仅在cpDNA得到，Southern杂交结果证实了片段SAY-192300的叶绿体来源，同时也表明在cms胞质中该片段所在区未发生插入、缺失等大的结构变异,DNA序列测定结果表明，该片段为叶绿体ps1A1和ps1A2基因的部分序列。图6表2参19。     

邻苯二酚2,3-双加氧酶基因克隆、定位和高效表达

采用特异性引物，以菲、芘降解菌株ZL5的代谢性质粒为模板，扩增出邻苯二酚2，3—双加氧酶(C23O)基因，将该基因和表达载体pET—30a( )连接，转化E.coli JM109(DE3)，获得了高效表达的转化子，SDS—PAGE结果表明，转化子的C23O蛋白不仅在细胞内存在，而且能被分泌到胞外，薄层扫描显示，转化子细胞内和细胞外表达蛋白总量占细胞总蛋白的42％，酶活分析表明，分布在转化子细胞内、外的表达蛋白都具有较高的C23O比活力，Southern杂交将菌株ZL5的C23O基因定位在内生质粒的不同酶切片段上。图5表1参12。     

桃ACC合酶基因组DNA（pACSG01）的序列分析及其表达

以桃基因组DNA为模板，用套式PCR技术扩增并克隆了桃ACC合酶基因片段，将其克隆（定名为pACSG01）并进行序列测定，表明该自然全长1320bp，并富含HindⅢ和EcoRⅠ位点，与其它ACC合酶基因结构序列有一定的相似性，内含有两个内含子，编码的氨基酸序列与已克隆桃ACC合酶cDNA推导的氨基酸序列的同源性分别为57．4％和56．8％，pACSG01与我们已克隆的两个桃ACC合酶cDNA基因表达有所不同，RNA点杂交和RT－PCR结合Southern杂交分析表明，该基因在成熟和乙烯处理的果实均不表达，伤处理、LiCl和生长素处理也不能诱导核基因的表达，在衰老花瓣中也不表达，图3参18。     

微量元素对红豆杉菌根菌生长的影响

微量元素对植物的生长发育有着十分重要的意义。对太行山野生红豆杉(Taxus chinensis var.mairei)菌根菌菌丝在分别加有硫酸锌、钼酸铵以及硫酸亚铁的PDA培养基中的生长发育状况进行了试验研究。所得数据同PDA对照组进行比较分析后,结果表明:Mo元素对红豆杉菌根菌菌丝的生长发育有很好的促进作用;Fe元素明显地促进红豆杉菌根菌菌丝的生长发育;Zn元素在一定范围内对红豆杉菌根菌菌丝的生长发育有一定的抑制作用。实验结果揭示出红豆杉菌根菌生长发育的一些营养因素。     

苏云金芽胞杆菌菌株WB9编码活性因子的基因分析

采用PCR—RFLP及PCR技术对苏云金芽胞杆菌新分离菌株WB9的ICP、VIP、几丁质酶和肠毒素4种活性因子的编码基因进行了分析．结果表明，该菌株含有编码ICP的cry1Aa、cry1Ab、cry1Cb、cry1Fa和cry1GaS个cry1基因型，编码Vip3A蛋白的vip3A基因以及编码3种肠毒素的hblA、bceT和entS基因；不含编码几丁质酶的基因．将vip3A、hblA、bceT和entS基因PCR产物回收纯化后直接测序，经在线的BLAST软件进行同源性分析，前两个基因片段与Bt已公布基因序列的相应片段同源性分别为99％和96％～98％，bceT基因片段与蜡质芽孢杆菌bceT相应片段同源性为97％．     

云南红豆杉的保护和开发利用

云南红豆杉(Taxus yunnanensis W. C. ChengL. K. Fu)是红豆杉属植物中紫杉醇含量最高的树种,同时也是我国红豆杉属中分布最广和资源蕴藏量最丰富的物种.然而,近年来由于人为大量砍伐,云南红豆杉野生资源遭到严重破坏,加上生长缓慢且天然更新困难,目前处于濒危状态.对云南红豆杉天然资源的保护和药用人工林的培育对于云南红豆杉资源的可持续利用具有重要的意义.对云南红豆杉资源分布现状及种群和繁殖生态学等方面的研究进行综述,发现云南红豆杉的濒危是由其自身繁殖机制引起的天然更新困难、对环境的适应能力较低以及人为因素共同导致的.同时从紫杉醇含量的影响因素及获取方式、人工药用林培育方面总结了云南红豆杉的开发利用现状,发现云南红豆杉中紫杉醇的含量受其自身特性及外在因素的综合影响;包含紫杉醇在内的紫杉烷类物质目前主要通过直接提取和半合成两种方式获取;此外在全合成、组织或细胞培养及真菌诱导等方式上也有了新的研究进展.最后,建议从就地保护、迁地保护及引种回归3个方面对云南红豆杉天然资源进行综合保护,同时大力推进紫杉烷工业化合成方式的研究和云南红豆杉人工药用原料林的培育,提高云南红豆杉的资源利用效率,在保护云南红豆杉天然林的同时实现资源的合理利用.(表1参129)     

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     

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.     

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.     

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.     

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.     

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.     

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.     

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.