深圳坝光银叶树湿地园林木有害生物调查及防控

对坝光银叶树Heritiera littoralis 湿地园开展有害生物调查,收集并鉴定有害生物72 种,其中有害杂草5 种、寄生植物2 种、病害20 种、虫害45 种。红火蚁Solenopsis invicta 和薇甘菊Mikania micrantha 属于外来入侵检疫性有害生物,八点广翅蜡蝉Ricania speculum 在公园内危害严重,对主要有害生物应加强预测预报、及时防治。     

Diversity of Glomale Mycorrhizal Fungi in Maize/Sesbania Intercrops and Maize Monocrop Systems in Southern Malawi

The study investigated diversity and frequency of occurrence of glomale (Arbuscular- or Vesicular Arbuscular-) mycorrhizal fungi on three farming systems in a drought prone and nitrogen deficiency site in southern Malawi. The farming systems comprised of two agroforestry systems of Sesbania sesban (L) Merr intercropped with maize and Sesbania macrantha E Phillips & Hutch. intercropped with maize and a maize monocrop systems without fertilizer, with nitrogen, phosphorus and a combination of nitrogen and phosphorus. Species diversity and species frequency of occurrence were examined in soil samples obtained in the dry and wet seasons. Twelve glomale mycorrhizal species were recorded, four species being in the genus Acaulospora, four in Glomus, two in Gigaspora and two in Scutellospora. Species diversity in the two agroforestry systems were not significantly (p ≤ 0.05) different but had lower species diversity than maize monocrop with only Sesbania macrantha intercropped with maize significantly (p ≤ 0.05) lower. Species diversity was significantly increased by the inorganic nitrogen fertilizer. Inorganic phosphorus fertilizer had no effect. The study shows that the occurrence and persistence of glomale species are influenced by agroforestry combinations, and that the spores of most species are tolerant to dry conditions. Only four species responded to fertilizer application with the occurrence of spores of some species high and some low. Management practices have great implication in the persistence of spore propagules of glomale species. The order Glomale was revised in 2001 and upgraded to a phylum Glomeromycota.     

Populations, Community Composition and Biomass of Aquatic Organisms in the Floodwater of Rice Fields and Effects of Field Management

As rice fields are managed for improved rice production, many studies on aquatic organisms in the floodwater have been restricted to agronomically important members such as blue-green algae (BGA) and Azolla , and ecological studies on common aquatic organisms were relatively limited to specific group members. In the present paper, studies on all the aquatic organisms in the floodwater, from protozoa to insects and fishes, were reviewed worldwide from the viewpoint of rice field ecology, paying attention to their populations, seasonal variations, antagonistic relations and biomass, and the effect of field management. Most studies conducted in India and in the Southeast Asian countries dealt with the BGA and green algal populations. In contrast, protozoa and aquatic animals were extensively studied in rice fields in Europe. Japanese studies in the past were intermediate between the previously cited studies. They generally described specific groups depending on the authors' interests, and surprisingly few studies dealt with the community level and prey-predator relationships. Field management such as fertilizer and pesticide application, water management and plowing was found to play a major role in determining the populations of aquatic organisms. Overview of research on aquatic organisms in the floodwater of rice fields suggests several directions for future studies.     

Arbuscular mycorrhizal fungi are directly and indirectly affected by glyphosate application

Glyphosate is a systemic non-selective herbicide, the most widely used in the world. Alongside with its use in agricultural and forestry systems, this herbicide is used in grasslands in late summer with the aim of promoting winter species with the consequent increase in stocking rate. However, its effects on non-target organisms, such as arbuscular mycorrhizal fungi (AMF), are unclear. Arbuscular mycorrhizal fungi (AMF) colonize the root of more than 80% of terrestrial plants, improving their growth and survival, and therefore playing a key role in ecosystem structure and function. The aim of this work was to investigate the possible pathways through which glyphosate application affects AMF spores viability and root colonization in grassland communities. Our hypothesis is that glyphosate application can damage AMF directly (through contact with spores and external hyphae) or indirectly through the changes it generates on host plants. The experiment had a factorial array with three factors: (1) plant species, at two levels (Paspalum dilatatum and Lotus tenuis), (2) doses of glyphosate, at three levels (0 l ha⁻¹, 0.8 l ha⁻¹ and 3 l ha ⁻¹), and (3) application site, at two levels: soil (direct pathway) and plant foliage (indirect pathway). Spore viability was reduced even under the lowest glyphosate rate, but only when it was applied on the soil. Total root colonization for both species was similarly decreased when glyphosate was applied to plant foliage or on soil, with no difference between 0.8 and 3 l ha⁻¹. The number of arbuscules was 20% lower when glyphosate was applied on plant foliage, than when it was applied on the soil. Our findings illustrate that glyphosate application negatively affects AMF functionality in grasslands, due to different causes depending on the herbicide application site. While, under field conditions, the occurrence of direct and/or indirect pathways will depend on the plant cover at the time of glyphosate application, the consequences of this practice on the plant community structure will vary with the mycorrhizal dependence of the species composition regardless of the pathway involved.     

Open geospatial technology standards and their potential in plant pest risk management—GPS-enabled mobile phones utilising open geospatial technology standards Web Feature Service Transactions support the fighting of fire blight in Norway

The threat posed by invasive alien species to global crop production and biodiversity is recognized as one of the major drivers of global change today. Here we report on a successful implementation and operational use of new technology to support risk- and crisis management in the case of fighting an invasive plant disease. The open geospatial standards Web Feature Service Transaction (WFS-T) was implemented in software for GPS-enabled mobile phones communicating with a GeoServer backend in order to track down and eradicate disease outbreaks and susceptible host plants. This new technology offering georeferenced events online from field provides new possibilities for real time action in risk and crisis management.     

双抗体夹心ELISA在转基因生物检测中的应用研究

随着转基因产品种类和数量的不断增加,转基因生物及其产品的安全性也越来越受到关注,各国一直以来也都高度重视转基因生物及产品的管理.作为转基因生物安全评价的第一步,转基因生物的准确检测显得格外重要.该文综述了双抗夹心ELISA方法的原理、技术要点及其在转基因植物及产品检测中的应用,并对该方法在转基因动物检测中的应用前景进行分析,为双抗夹心ELISA方法的合理利用提供依据.     

Are survival and reproduction of Enchytraeus albidus (Annelida: Enchytraeidae) at risk by feeding on Bt-maize litter?

Enchytraeids are saprophagous soil organisms, appearing in high abundances and contributing to ecological processes within the soil. For decades they have been used as model species for biological research. In the framework of research on genetically modified plants, however, they have not been considered to date. Following the ISO/DIS guideline, survival and reproduction of Enchytraeus albidus, fed with diets containing Bt-maize (N4640Bt Cry1Ab, DKC5143Bt Cry3Bb1) leaf material were analysed. For comparison, diets with the corresponding untransformed near-isolines (N4640, DKC5143) were examined. Additionally a high quality control diet (oat flakes) was included. Survival and reproduction showed no significant differences between the Cry3Bb1 treatment and the treatment with the untransformed counterpart. For the Cry1Ab treatment survival was significantly higher than for the treatment with the corresponding near-isoline. In contrast, reproduction was significantly lower for the Cry1Ab treatment compared to that for the isoline. For the Cry3Bb1 treatment, no effect was shown on survival or reproduction. For the Cry1Ab variety and its untransformed counterpart, a contrasting result was detected, which is unlikely to be caused by the Bt-protein but rather by differences in other plant components. Overall survival and reproduction were highest for the control.     

利用微滴数字PCR分析转基因生物外源基因拷贝数

微滴数字PCR(droplet digital PCR,ddPCR)是一种基于泊松分布原理的核酸分子绝对定量技术,在核酸分子的绝对计数/定量领域具有极大的应用潜力。本研究基于ddPCR平台,以转基因水稻(Oryza sativa)T1c-19和转人乳铁蛋白基因基因山羊(Capra hircus)134为例,建立了转基因生物(genetically modified organisms,GMOs)外源基因拷贝数分析方法,并比较了其与传统的实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)和Southern blot方法的准确性。实验数据表明,T1c-19的杀虫晶体蛋白基因(insecticidal crystal protein,Cry1C*)在qRT-PCR和ddPCR测定结果比较一致,约为2拷贝,但已报道的Southern blot分析结果为1拷贝;ddPCR对bar基因的分析结果高于qRT-PCR,分别为2.09拷贝和1.51拷贝。转人乳铁蛋白基因(human lactoferrin,HLF)山羊134在qRT-PCR和ddPCR的分析结果基本一致,均测得含有1拷贝的HLF基因。研究结果表明,微滴数字PCR方法是一种经济、快速和准确的外源基因拷贝数分析新方法,灵敏度和准确性高,将会在拷贝数分析中广泛应用。     

利用QuantStudioTM 3D数字PCR分析转基因玉米MON863含量

QuantStudioTM 3D数字PCR (QuantStudioTM 3D digital PCR,3D-dPCR)是一种基于超高密度亲疏水微孔芯片实现数字PCR分液原理的新型核酸绝对定量平台,在转基因生物定量领域具有极大的应用前景.本研究基于3D-dPCR平台,以转基因玉米(Zea mays)MON863混合样品为例,建立基于单重和双重数字PCR体系的转基因生物(genetically modified organisms,GMOs)含量分析方法.与传统qRT-PCR比较发现,在缺乏样品纯度、纯合度信息的情况下,数字PCR能够较好地排除这些因素的影响,测定准确的量值.研究结果表明,QuantStudioTM 3D数字PCR是一种适用于转基因生物含量分析的精确定量方法,还可反映转基因玉米种子的基因型.本研究基于3D-dPCR建立的转基因玉米MON863单重和双重定量方法为转基因检测提供了新的方法和参考.     

Ecotoxicological evaluation of Harbour sediments using marine organisms from different trophic levels

Background, Aim and Scope  The toxicity of contaminated sediments should be evaluated considering the direct exposure of laboratory organisms to whole sediments and the indirect exposure to elutriates or extracts (Tay et al. 1992, Byrne and Halloran 1999, Nendza 2002). The alga Dunaliella tertiolecta is indicated for the use in toxicity bioassays because it is highly sensitive to several xenobiotics. Harpacticoid copepods have been already used for toxicity testing and Tigriopus fulvus is a promising Mediterranean target-species in ecotoxicology (Todaro et al. 2001, Faraponova et al. 2003, Pane et al. 2005a). In this study, the toxicity of sediments collected in harbour sites of the Northeastern Adriatic Sea was evaluated by growth inhibition test with free living and alginate-immobilized Dunaliella tertiolecta and acute toxicity test with nauplii and adult Tigriopus fulvus with the aim of pointing out the importance to utilize model organisms from different trophic levels in sediment ecotoxicology. Methodology  Elutriates and whole sediments were tested on free living and immobilized (Pane et al. 1998) algal cells, and on laboratory reared copepods. Free-living D. tertiolecta were exposed to diluted elutriates in a static, multi-well plate system. Naalginate immobilized D. tertiolecta were placed in polystyrene inserts fitted with polyester mesh bottoms and exposed to a thin layer (2 mm) of whole sediments in multi-well plates (EPS 1992, Pane and Bertino 1999). Toxicity tests with copepods were carried out on Tigriopus fulvus nauplii (elutriates) and adults (whole sediments and elutriates). Same-aged nauplii useful for toxicity tests were obtained by egg sac detaching and consequent hatching stimulation (Pane et al. 2006). Newborn nauplii (I–II stage) were exposed to elutriates in multi-well plates provided with polystyrene inserts. Adult T. fulvus maintained in polystyrene inserts fitted with polyester mesh bottoms were placed in contact with a thin layer (2 mm) of whole sediment placed on multi-well plate bottoms. All end-points were evaluated after 96 h. Results  In general, the effects increased with the increasing of elutriate concentration up to 50%; the stimulation or inhibition of algal growth was statistically significant in comparison to the control. The inhibiting elutriates induced EC₅₀ variations of algal growth ranging from 66.9% to 74.3%. The mortality of T. fulvus nauplii was always < 25% after treatment with 100% elutriates and < 10% after treatment with 50% dilution; no effect was shown up with 25% dilution; therefore LC₅₀ was not calculable. The effect of elutriates was negligible on adult copepods and LC₅₀ values were never calculable; percent mortality always resulted in < 10% after treatment with whole sediments. Discussion  Both experimental systems gave substantially similar results after exposition to whole sediments and elutriates. During the experiment with algal cells, the immobilization in Na-alginate and the employment of inserts which allowed the contact of organisms with sediments and their easy counting were particularly useful. Likewise, the employment of inserts of adequate mesh size in the tests with copepods allowed the contact of organisms with the sediment and made organism handling and counting easy, as well as the evaluation of mortality. The methodology here described and the utilization of the proposed test-species could have an importance also considering that the current trend in ecotoxicological research is towards finding the most appropriate organism for specific areas of concern by using indigenous species (Mariani et al. 2006) and towards the major significance of chronic and reproductive end-points. Conclusions  Based on the above results, it can be stated that the bioassay with Dunaliella tertiolecta could be a good estimation tool for the ecotoxicological assessment of marine sediments. The immobilization of algae in Na-alginate was seen to be useful to evaluate the toxicity of whole sediments; the employment of polystyrene inserts allowed an improvement of the procedures. T. fulvus nauplii and adults, as other harpacticoids such as Tigriopus japonicus (Yoon et al. 2006), satisfy the basic criteria for the employment of a standard species in marine bioassays. To date only pelagic Acartia tonsa are utilized in the standardized procedure to evaluate the risk assessment of chemicals or wastewaters (ISO 1999). As, on the contrary, the exposure of copepods to solid-phase contaminants it is not yet standardized, the employment of polystyrene inserts improved the procedures for T. fulvus too. So, the rapidity and the possibility to solve practical problems could be the main attractive features of this technique (Pane et al. 2005a) when applied to whole sediments. Recommendations and Perspectives  The methodology here developed being also applicable to long term and reproduction tests should be recommended because it provides relevant information in comparison with other frequently applied, standardized biotests with crustaceans (ISO 1999). The procedure has been shown to be easily applicable to selected marine organisms. ESS-Submission Editor: Prof. Dr. Henner Hollert (henner.hollert@bio5.rwth-aachen.de)