多糖产生菌T与β-胡萝卜素产生菌C1-B的融合研究

以多糖产生菌Ｔ和胡萝卜素产生菌Ｃ１－Ｂ为双亲,用原生质体融合技术选育出了１株既产多糖又产胡萝卜素的杂交品系,在对这两株菌的原生质体形成、再生的研究基础上,以ＰＥＧ６０００加新生磷酸钙为助融剂,对其进行了原生质体的融合,融合率达３．４８×１０－７,并选择出了遗传性能稳定的融合子Ｆ．该融合子具有双亲优势,得到的发酵液多糖和β胡萝卜素产量分别达８．７ｇ／Ｌ和１３．７ｍｇ／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.     

树状多节孢原生质体的制备和再生

研究了酶系组成、酶解时间、酶解温度、渗透压稳定剂、PH值,培养基成分、培养方式,预处理、保护剂等因素对紫杉醇产生菌树状多孔孢原生质体制备和再生的影响。结果表明：将树状多节孢在PDA液体培养其中静止培养2－3d,离心收集菌体,用PH5．5－6．0的0．7mol/L的NaCl配制成的含有3％纤维素酶、2％蜗牛酶和1％溶菌酶的复合酶,30℃恒温酶解9h,原生质体制备率最高。但是考虑到原生质体的再生,参酶解7h左右为宜,获得的原生质体经过纯化,用0.7mol/LNaCl配制的PDA再生培养基进行双层平板培养法再生,原生质体的再生率最高。本研究为以后通过原生质体诱变、转化、融合构建紫杉醇工程菌株奠定了基础,图3表9参16     

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．     

离子选择性微电极用于外界胁迫下细胞液中钙分布检测的新模式

离子选择性微电极(ISME)是一种电化学传感器。在单细胞检测中可用来测定细胞内各种离子的浓度和胞外空间的离子流。本文提出了一种离子选择性微电极用于单细胞检测的新方法,即用离子选择性微电极测量植物细胞原生质体破裂时形成的离子浓度脉冲信号,进而分析细胞液中离子浓度的分布。并用此方法研究了在低温、纳米氧化铜(CuO NPs)、重金属(铽、镉)和乙醇等不同因素作用下,芦荟(Aloe vera)细胞原生质体的细胞液中Ca~(2+)的浓度分布。实验结果表明,在冷冻、CuO NPs、硝酸铽和氯化镉处理后细胞液中的Ca~(2+)浓度分布发生分层现象,而乙醇处理则不会出现这种现象。这种方法具有抗干扰能力强和操作简便的优点,为更全面地揭示细胞对外界刺激的响应特征提供了新的视角,也可为各种环境和生态毒理学评价提供新的途径。     

冷冻胁迫对芦荟叶肉细胞原生质体中Ca2+、Na+分布的影响

低温和冻害是造成巨大农业损失和植物死亡的主要逆境因子。为揭示冷冻胁迫对植物细胞中离子分布的影响,选择芦荟细胞原生质体为受试材料,使用离子选择性微电极检测了经冷冻处理后的芦荟原生质体在低渗液中破裂时产生的Ca~(2+)浓度脉冲信号,并同时检测Na~+浓度脉冲信号作为对比。研究了冷冻温度、解冻时间和ZnO NPs处理等因素对冷冻胁迫下芦荟原生质体中Ca~(2+)分布的影响。结果表明,与未经冷冻处理的原生质体相对比,经过冷冻处理的原生质体破裂后,其Ca~(2+)脉冲信号前沿处发生明显的"凹陷",这说明,原生质体中Ca~(2+)分布出现分层现象,靠近细胞中心浓度较高而细胞膜附近浓度较低。这一分层现象在温度为-7℃时开始出现,原生质体解冻5 h后仍未消失。经过ZnO NPs预处理后再进行冷冻的原生质体,其Ca~(2+)脉冲凹陷深度明显减小。而当用ZnO NPs处理解冻后的原生质体时,其Ca~(2+)分层现象消失。冷冻胁迫下芦荟原生质体内Ca~(2+)分布发生显著变化,表明原生质体内Ca~(2+)分布变化与其抗寒反应存在一定关系。与Ca~(2+)相反,Na~+的分布几乎不受冷冻因素的影响。ZnO NPs处理对冷冻芦荟原生质体中Ca~(2+)浓度分布分层有明显的缓解作用,表明一定浓度范围的ZnO NPs在缓解冷冻造成的Ca~(2+)流动性下降,维持细胞Ca~(2+)分布的调控能力方面,具有一定的积极影响。     

Zn^2＋对水鳖的毒性作用

研究了不同浓度Zn^2 毒害对水鳖叶片膜脂过氧化作用及其体细胞超微结构的影响。结果表明,随着Zn^2 浓度的增加,叶绿素含量、SOD、CAT、POD活性先后升降,叶绿素a/b值值则逐渐减小,而MDA含量增加,O2^-产生速率也逐渐增大。同功酶分析表明,Zn^2 可以诱导新的POD同功酶,电镜观察发现,低浓度Zn^ 处理下,水鳖叶细胞中核质空泡化,叶绿体膨胀变形,线粒体嵴膨胀,溶酶体、小泡数量增加,且出现了多膜复合体;随着Zn^2 浓度的增加,核质消失,叶绿体和线粒体解体,而根尖分生细胞中则出现了多核仁现象。     

非营养缺陷型原生质体融合选育角蛋白酶生产菌

以两性霉素B抗性作为标记,通过单株灭活,进行栖土曲霉种内非营养缺陷型原生质体融合.在40%的PEG(Mr＝6000)促融下,融合频率为1.62×10-5.连续传接10代后,获得稳定的融合子.对孢子体积、核DNA含量、生长速度进行了测定,并利用RAPD技术分析比较了亲本及融合子的基因组DNA指纹多态性,证明融合子为杂合二倍体,产角蛋白酶活力较亲本显著提高.图3表4参17     

外源降解菌对黄麻根区净化能力的生物强化作用

通过原生质体电融合得到经EGFP标记的外源细菌(蒽高效降解菌An和表面活性剂产生菌P)与黄麻根际优势菌融合的2株融合子Tu-An与融合子Tu-P,然后将两种融合子以及出发菌株分别定殖到播种了黄麻(Corchoruscapsulari)的蒽污染土壤中,对不同污染物浓度下的细菌定殖、植物生长及蒽降解做了初步研究.研究表明,在相同的初始接种量下,融合子的定殖数量明显高于出发菌株;在两种融合子共存的条件下,定殖数量也略高于单一融合子的定殖数量.在接种外源菌的土壤中,植物获得了更好的生长条件,得以良好生长;而没有投加降解菌的组别中,黄麻的生长明显受到了较严重的抑制.投加外源降解菌可以促进生物降解,达到生物强化的目的.实验研究说明,把外源降解菌投加到污染土壤中进行生物强化修复是可行的.图3表2参20     

生物技术在南方果树研究中的应用

综述了微繁、胚培养、胚乳培养、花药培养、原生质体培养与融合、转基因和基因克隆、分子标记等生物技术近年来在柑桔等南方果树上的应用.     

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.     

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.     

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     

Human Influence and Classical Biogeographic Predictors of Rare Species Occurrence

Biogeographic theory predicts that rare species occur more often in larger, less‐isolated habitat patches and suggests that patch size and connectivity are positive predictors of patch quality for conservation. However, in areas substantially modified by humans, rare species may be relegated to the most isolated patches. We used data from plant surveys of 81 meadow patches in the Georgia Basin of Canada and the United States to show that presence of threatened and endangered plants was positively predicted for patches that were isolated on small islands surrounded by ocean and for patches that were isolated by surrounding forest. Neither patch size nor connectivity were positive predictors of rare species occurrence. Thus, in our study area, human influence, presumably due to disturbance or introduction of competitive non‐native species, appears to have overwhelmed classical predictors of rare species distribution, such that greater patch isolation appeared to favor presence of rare species. We suggest conservation planners consider the potential advantages of protecting geographically isolated patches in human‐modified landscapes because such patches may represent the only habitats in which rare species are likely to persist. Influencia Humana y Predictores Biogeográficos Clásicos de la Ocurrencia de Especies Raras     

Seasonal dynamics of macro-algae in the South Lake of Tunis

The macro-algae communities observed in the south lake of Tunis are characterized by the predominance of nitrophilous algae which are in the order of biomass importance:Ulva, Cladophora andEnteromorpha. We have noted seasonal changes of alga distribution. The wind appears to be one of the most important factors influencing this distribution. The total biomass reaches a maximum in the spring. Rapid decomposition of the biomass leads to a severe ecological imbalance, resulting in crises of anoxia and fish death. A restoration project has already started. It aims at removal of contaminated muds and the introduction of a new circulation system. The main objectives of this work were to collect information on the distribution and biomass of the phytobenthic communities as a first step in the constitution of a database for further comparison.     

Patterns and biases of climate change threats in the IUCN Red List

International Union for Conservation of Nature (IUCN) Red List assessments rely on published data and expert inputs, and biases can be introduced where underlying definitions and concepts are ambiguous. Consideration of climate change threat is no exception, and recently numerous approaches to assessing the threat of climate change to species have been developed. We explored IUCN Red List assessments of amphibians and birds to determine whether species listed as threatened by climate change display distinct patterns in terms of habitat occupied and additional nonclimatic threats faced. We compared IUCN Red List data with a published data set of species’ biological and ecological traits believed to infer high vulnerability to climate change and determined whether distributions of climate change‐threatened species on the IUCN Red List concur with those of climate change‐threatened species identified with the trait‐based approach and whether species possessing these traits are more likely to have climate change listed as a threat on the IUCN Red List. Species in some ecosystems (e.g., grassland, shrubland) and subject to particular threats (e.g., invasive species) were more likely to have climate change as a listed threat. Geographical patterns of climate change‐threatened amphibians and birds on the IUCN Red List were incongruent with patterns of global species richness and patterns identified using trait‐based approaches. Certain traits were linked to increases or decreases in the likelihood of a species being threatened by climate change. Broad temperature tolerance of a species was consistently related to an increased likelihood of climate change threat, indicating counterintuitive relationships in IUCN assessments. To improve the robustness of species assessments of the vulnerability or extinction risk associated with climate change, we suggest IUCN adopt a more cohesive approach whereby specific traits highlighted by our results are considered in Red List assessments. To achieve this and to strengthen the climate change‐vulnerability assessments approach, it is necessary to identify and implement logical avenues for further research into traits that make species vulnerable to climate change (including population‐level threats).