Mutants in the lipopolysaccharide of Brucella ovis are attenuated and protect against B. ovis infection in mice

Brucella spp. are Gram-negative bacteria that behave as facultative intracellular parasites of a variety of mammals. This genus includes smooth (S) and rough (R) species that carry S and R lipopolysaccharides (LPS), respectively. S-LPS is a virulence factor, and mutants affected in the S-LPS O-polysaccharide (R mutants), core oligosaccharide or both show attenuation. However, B. ovis is naturally R and is virulent in sheep. We studied the role of B. ovis LPS in virulence by mutating the orthologues of wadA, wadB and wadC, three genes known to encode LPS core glycosyltransferases in S brucellae. When mapped with antibodies to outer membrane proteins (Omps) and R-LPS, wadB and wadC mutants displayed defects in LPS structure and outer membrane topology but inactivation of wadA had little or no effect. Consistent with these observations, the wadB and wadC but not the wadA mutants were attenuated in mice. When tested as vaccines, the wadB and wadC mutants protected mice against B. ovis challenge. The results demonstrate that the LPS core is a structure essential for survival in vivo not only of S brucellae but also of a naturally R Brucella pathogenic species, and they confirm our previous hypothesis that the Brucella LPS core is a target for vaccine development. Since vaccine B. melitensis Rev 1 is S and thus interferes in serological testing for S brucellae, wadB mutant represents a candidate vaccine to be evaluated against B. ovis infection of sheep suitable for areas free of B. melitensis.     

Can the structure of dormant cambium and the widths of phloem and xylem increments be used as indicators for tree vitality?

We investigated the structure and width of the dormant cambium and of the increments of phloem and xylem of Quercus robur to estimate their potential as indicators for tree vitality. The samples were taken from three woodlands, two in Slovenia [Krakovo forest (KRA) and Murska Suma (MUS)] and one in Croatia [Kobiljak (KOB)], with reported tree decline. The number of dormant cells seems to reflect the initial capacity of the cambium to accomplish cell division. With the exception of two trees at KRA, cell production was always higher on the xylem side than on the phloem side. The annual phloem increments were narrower, less variable among trees and with clear lower and upper limits. With increased cambial cell productivity, the share of the xylem in the total annual radial increment increased following a curvilinear function. In trees with an annual radial increment >3.5 mm, the xylem size represented more than 90 % of the total radial growth. The anatomical variables analyzed show that the most limiting environmental conditions seem to prevail at KRA, whereas the conditions at MUS seem to be most favorable in terms of radial growth. Analysis of the width and structure of xylem and phloem increments, the number of dormant cambial cells and their inter-relationships can provide additional information on the vitality of oaks.     

杜鹃红山茶在海南中部地区扦插育苗技术研究

为掌握杜鹃红山茶引种到海南中部地区的扦插育苗技术,通过扦插实验实验表明:杜鹃红山茶在11月的扦插生根效果最好,9月与3月的生根率也达到80%以上,故较好的扦插时节在中秋后至入夏之前;使用浓度为500mg/L的NAA浸泡枝条下部,生根率最高;半木质化的插穗生根效果最好;基质则以红壤土体积:河沙体积=1:1的混合基质效果最理想。     

用地高辛标记的PCR－ELISA技术快速检测转基因鱼

取含有小鼠重金属螯合蛋白（mMT)基因启动子及人生长激素（hGH)基因的鱼样品，以Qiagene核酸纯化技术对鱼组织DNA进行纯化。常规PCR检测显示，mMT启动基因和hGH基因的PCR产物分别为240bp和130bp，与设计相符。PCR产物的核酸测序结果证实其扩增产物具特异性。地高辛-PCR（Dig-PCR)标记反应显示，2个基因均产生1条Dig标记的特异产物带。Dig-PCR-ELISA检测敏感性试验显示，对mMT启动子和hGH基因的检测敏感性可达10^-3，与常规PCR结合琼脂糖胶电泳检测方法相比，检测敏感性可提高至100-1000倍。试验证明，针对mMT和hGH基因所建立的Dig-PCR-ELISA技术特异可靠。     

聚合物包膜控释肥料膜壳累积、降解及对土壤质量影响的研究进展

聚合物包膜控释肥料是实现化肥提质增效的新型肥料之一,其控释效果依赖于包膜材料。控释肥料养分释放后的残留膜壳是农业土地上微塑料的重要来源,膜壳及其降解产物是否对土壤质量安全造成影响尚不清楚。本文综合了国内外膜壳残留及其降解的研究进展,从聚合物包膜控释肥料残留膜壳在农田中的累积现状、降解性能研究及其对土壤物理–化学–生物学效应等土壤质量参数的影响进行了阐述,包括:1)现有文献指出,控释肥料膜壳在农田中的累积量与施用量基本相同,其在不同类型农田土壤及不同土层中的赋存、分布和累积现状不同;2)不同类型控释肥料膜壳在土壤中的降解过程主要与聚合物材质相关,降解率可通过聚合物材料改性来提高,其降解过程、产物及机制值得探索;3)控释肥料膜壳降解产物对土壤物理、化学、生物学性状及作物生长的影响程度取决于聚合物材料类型、累积量及土壤类型。短期内低浓度的膜壳累积对土壤理化性质、土壤生物及作物生长无不良影响,长期累积或高浓度下膜壳影响土壤质量变化的阈值亟需明确。据此,进一步提出,1)应广泛开展不同种类控释肥料膜壳在农田中的赋存、分布和累积的现场监测评估工作;2)开发控释肥料膜壳及其次级降解物分离提取、鉴定和定量表征的分析技术和方法,建立健全标准化方法;3)了解控释肥料膜壳及其降解产物在土壤中的去向和迁移过程及其对不同类型土壤及不同作物品种的影响,探寻膜壳残留量与效应之间的关系及其影响机制;4)研制开发天然、生物及可降解控释肥料膜材,研究膜壳降解与养分释放之间的相互关系,构建新型可生物降解控释肥料膜壳降解与养分释放关系数据模型,以期为聚合物包膜控释肥料的应用、开发及标准的制定提供科学依据。     

黄芪ISSR-PCR反应体系的建立及优化

为获得最优黄芪ISSR-PCR反应体系,以山西省浑源县官儿乡蒙古黄芪为试验材料,利用单因素试验法对反应体系中的4个因素(模板DNA,dNTPs,Taq DNA聚合酶,ISSR引物)分别设置浓度梯度进行体系优化。结果表明,20μL黄芪ISSR-PCR最优体系为:模板DNA质量浓度6.0 ng/μL,dNTPs浓度0.03 mmol/L,Taq DNA聚合酶浓度0.05 U/μL,引物浓度0.80 mmol/L。最后用不同产地的黄芪检测该体系,结果发现,PCR产物多态性良好。优化体系的确立为今后黄芪种质资源的分子评价奠定了技术基础。     

Emissions of CO2, CH4 and N2O from Southern European peatlands

Peatlands play an important role in emissions of the greenhouse gases CO₂, CH₄ and N₂O, which are produced during mineralization of the peat organic matter. To examine the influence of soil type (fen, bog soil) and environmental factors (temperature, groundwater level), emission of CO₂, CH₄ and N₂O and soil temperature and groundwater level were measured weekly or biweekly in loco over a one-year period at four sites located in Ljubljana Marsh, Slovenia using the static chamber technique. The study involved two fen and two bog soils differing in organic carbon and nitrogen content, pH, bulk density, water holding capacity and groundwater level. The lowest CO₂ fluxes occurred during the winter, fluxes of N₂O were highest during summer and early spring (February, March) and fluxes of CH₄ were highest during autumn. The temporal variation in CO₂ fluxes could be explained by seasonal temperature variations, whereas CH₄ and N₂O fluxes could be correlated to groundwater level and soil carbon content. The experimental sites were net sources of measured greenhouse gases except for the drained bog site, which was a net sink of CH₄. The mean fluxes of CO₂ ranged between 139 mg m⁻² h⁻¹ in the undrained bog and 206 mg m⁻² h⁻¹ in the drained fen; mean fluxes of CH₄ were between −0.04 mg m⁻² h⁻¹ in the drained bog and 0.05 mg m⁻² h⁻¹ in the drained fen; and mean fluxes of N₂O were between 0.43 mg m⁻² h⁻¹ in the drained fen and 1.03 mg m⁻² h⁻¹ in the drained bog. These results indicate that the examined peatlands emit similar amounts of CO₂ and CH₄ to peatlands in Central and Northern Europe and significantly higher amounts of N₂O.     

Bioaccumulation and Effects of Heavy Metals in Crayfish: A Review

Metal pollution is a global problem which represents a growing threat to the environment. Because of bioaccumulation and negative effects of heavy metals, their bioavailability needs to be monitored. Many studies showed accumulation of metals in crayfish tissues as dose- and time-dependent without significant differences in tissue concentration levels comparing males and females. Muscles and exoskeleton were considered as specific for accumulation of mercury and nickel, respectively. Cadmium, zinc, copper, lead, and chromium accumulated mainly in hepatopancreas. By analyzing these specific tissues, it is possible to deduce the bioavailability and, by presumption, the level of environmental pollution by specific metals. However, in the case of zinc and copper, their utility is limited to assessing bioavailability because rapid depuration of these metals renders them less useful for long-term environmental monitoring programs. The literature reporting heavy metal impacts on freshwater crayfish, with reference to accumulation levels, is reviewed and summarized with respect to their suitability as bioindicators. Summarized published data from unpolluted or control localities can be used as referential values in crayfish, and consequently help with evaluation of monitored sites.     

Phytotoxic Effect of the Uranium on the Growing Up and Development the Plant of Corn

The aim of this article is the determination of uranium accumulation in plants tissue in shoots and roots of corn—maize (Zea mays), grown on two types of soils, pseudogley and chernozem, together with its phytotoxic effect on the plant growth and development. The soils was contaminated with different rates (10 to 1,000 mg U(VI) kg^?1) of uranyl nitrate (UO₂(NO₃)₂·6H₂O). Vegetative tests performed with maize indicated uranium phytotoxic effect on plant height, yield, and germination of seeds. This effect was stronger on the plants grown on pseudogley in comparison with those grown on chernozem. Soil properties determined the tolerance and accumulation of U in plants. A linear dependence between the content of uranium in soil and in plants tissue, including maximal content of 1,000 mg U?kg^?1, indicates that maize could be used for phytoremediation of uranium-contaminated soils.