番茄果实特异表达启动子ESp1的克隆及表达

E8启动子是番茄果实成熟乙烯响应性基因E8的启动子区。这个区域中存在着一些响应乙烯以及在果实发育和成熟过程中起调节作用的顺式作用元件，调节E8基因在果实成熟过程中表达（Deikmaa et al．，1998）。本研究根据GenBank中E8基因的启动子区序列设计引物，从番茄基因组中克隆了该启动子，构建了番茄果实特异表达载体，并通过RT-PCR和gus基因检测了该表达载体的果实特异表达活性。     

嗜酸乳杆菌真空喷雾干燥微胶囊化技术的研究

以阿拉伯胶、β-环糊精、可溶性淀粉、明胶、脱脂奶粉为壁材，根据正交试验确定微生态菌株嗜酸乳杆菌细胞微胶囊化的最佳真空喷雾干燥进风温度、壁材浓度和芯材浓度，并对其粒子直径进行分析。结果表明，β-环糊精与阿拉伯胶以9∶1配比为理想壁材，喷雾干燥进风温度170℃、壁材浓度20%和芯材浓度6%为最佳组合，活菌数达10⁸ cfu/g以上，微胶囊粒子直径平均为20 μm。     

Bioassay-directed Fractionation of Organic Extracts of Marine Surface Sediments from the North and Baltic Sea - Part II: Results of the biotest battery and development of a biotest index (8 pp)

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Part I: Determination and identification of organic pollutants Part II: Results of the biotest battery and development of a biotest index

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Preamble. This series of two papers presents the results of an interdisciplinary research project (ISIS) dealing with bioassay-directed fractionation of marine sediment extracts. Part I presents the extraction and fractionation procedure as well as the results of chemical analysis, including non-target analysis of sediments. Part II describes the results of the biotest battery in relation to chemicals possibly causing parts of the observed effects. A biotest index is used to compare the toxicities of the samples.

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AUTHORS / AFFILIATIONS Ninja Reineke (3), Werner Wosniok (4), Dirk Danischewski (1), Heinrich Hühnerfuss (3), Angelika Kinder (5), Arne Sierts-Herrmann (5), Norbert Theobald (2), Hans-Heinrich Vahl (6), Michael Vobach (1), Johannes Westendorf (6) and Hans Steinhart (5).

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(1) Federal Research Centre for Fisheries, Institute for Fishery Ecology, Palmaille 9, 22767 Hamburg, Germany (2) Federal Maritime and Hydrographic Agency, Bernhard-Nochtstr. 78, 20359 Hamburg, Germany (3) University of Hamburg, Institute for Organic Chemistry, Martin-Luther-King-Platz 6, 20146 Hamburg, Germany (4) University of Bremen, Institute of Statistics, Bibliothekstr. 1, 28334 Bremen, Germany (5) University of Hamburg, Institute for Food Chemistry, Grindelallee 117, 20146 Hamburg, Germany (6) University of Hamburg, University Hospital Hamburg-Eppendorf, Department for Toxicology, Vogt-Kölln-Str. 30, 22527 Hamburg, Germany (7) Eurofins Wiertz-Eggert-Jörissen, Stenzelring 14b, 21107 Hamburg, Germany

Goal, Scope and Background

The ecological relevance of contaminants in mixtures is difficult to assess, because of possible interactions and due to lacking toxicity data for many substances present in environmental samples. Marine sediment extracts, which contain a mixture of environmental contaminants in low concentrations, were the object of this study. The extracts were investigated with a set of different biotests in order to identify the compound or the substance class responsible for the toxicity. For this goal, a combination of biotests, biotest-directed fractionation and chemical analysis has been applied. Further on, a strategy for the development of a biotest index to describe the toxicity of the fractions without a prior ranking of the test results is proposed. This article (Part II) focuses on the biological results of the approach.

Methods

The toxicological potential of organic extracts of sediments from the North Sea and the Baltic Sea was analyzed in a bioassay-directed fractionation procedure with a set of biotests: luciferase reporter gene assays on hormone receptor and Ah receptor, arabinose resistance test, fish embryo test (Danio rerio), comet assay, acetylcholinesterase inhibition test, heat-shock protein 70 induction, oxidative stress and luminescence inhibition test (Vibrio fischeri). The test results provided the basis for the calculation of a biotest index by factor analysis to compare the toxicity of the samples and fractions.

Results and Discussion

Results of 11 biotests on different fractionation levels of the samples were described and discussed with regard to the occurrence of contaminants and their toxic potentials. Polychlorinated biphenyls, polycyclic aromatic hydrocarbons, quinones, brominated indoles and brominated phenols were in the focus of interest. A biotest index was constructed to compare the toxic responses in the samples and to group the biotest results.

Conclusion

The procedure presented in this study is well suited for bioassay-directed fractionation of marine sediment extracts. However, in relatively low contaminated samples, high enrichment factors and sufficient fractionation is necessary to allow identification of low concentrations of contaminants which is required to link effects and possible causes. In the present case, the relation between substances and effects was difficult to uncover due to relatively low concentrations of pollutants compared to the biogenic matrix and to the remaining complexity of the fractions. The results, with respect to the brominated phenols and indoles in the samples, highlight the successful use of bioassay directed fractionation in the case of high concentrations and high toxicity.

Recommendation and Outlook

In general, it has been shown that a marine risk assessment requires focusing on the input of diffuse sources and taking into account the fact of mixture toxicity. Effects resulting from biogenic substances will make the assessment of the influence of anthropogenic substances even more difficult.     

拮抗细菌C27活性物质对小麦白粉病菌发育的影响

采用有机溶剂萃取、温室盆栽试验以及离体叶组织病理学染色技术,测定拮抗细菌C27发酵液不同组分对小麦白粉病菌发育的影响.结果表明,桔抗细菌C27产生的拮抗物质对小麦白粉病的防效达69.89%.对小麦白粉病菌的孢子萌发、附着胞的形成及吸器原体的形成均无抑制作用,但可明显提高附着胞的畸形率.抑制指状吸器的发育,降低入侵率.分析认为拮抗细菌C27严生的拈抗物质在降低小麦白粉痛的初始菌源数量和控制小麦白粉病的流行危害中将会发挥重要的作用.     

响应面法优化海洋沉积物微生物总DNA提取方法

海洋沉积物中的微生物是自然界生态系统中不可或缺的组分,在维持环境与海洋生态平衡方面发挥着重要作用.在海洋沉积物微生物的研究中,细胞裂解率低、片段不够完整等问题导致无法高效地提取微生物的总DNA片段.本实验采用响应面分析法对CTAB-SDS-冻融DNA提取法进行优化.根据单因素的实验结果,选取溶菌酶含量、65℃水浴时间、蛋白酶K含量作为影响因子,利用Box-Behnken进行实验设计,将DNA浓度作为响应值,进行DNA提取方法的响应面优化实验.结果发现:当溶菌酶含量为1.13 mg/mL、65℃水浴时间为100 min、蛋白酶K含量为0.21 mg/mL时,DNA提取效果达到最好.验证实验表明:优化后提取DNA的实际值与理论值相近,具有可行性,优化后提取的DNA片段在纯度、浓度方面高于优化之前.     

Protective efficacy of vaccination with NcMIC3 and NcMIC8 against Neospora caninum infection in mice

Microneme proteins (MICs) are important for Apicomplexan parasite invasion due to their adhesion to host cells. Several studies have indicated that Neospora caninum MIC3 and MIC8 are important adhesion factors and potential vaccine candidates against neosporosis. In this study, we evaluated the protective efficacy of recombinant proteins and DNA vaccines of NcMIC3 and NcMIC8. BALB/c mice were immunized with rNcMIC3, rNcMIC8, pcDNA3.1-NcMIC3 and pcDNA3.1-NcMIC8 respectively, and challenged with N. caninum tachyzoites. The immune responses were evaluated through cytokine, antibody measurements and the parasite burden in the mice brain tissues. Serological analysis showed that recombinant protein vaccines induced higher levels of immunoglobulin G (IgG) than other groups. The percentage of IgG1 and IgG2a in the recombinant protein groups was higher than the other groups, and with a predominance of IgG1 over IgG2a, suggesting that recombinant protein vaccines elicited a Th2-type immune response, while DNA vaccines mainly produce a Th1-type immune response. In addition, mice immunized with rNcMIC3 and rNcMIC8 a had lower parasite burden in brain tissue compared with the other groups. These results demonstrate that rNcMIC3 and rNcMIC8 could induce humoral and Th2-type immune response, leading to a considerable level of resistance against neosporosis.     

基于模拟Landsat-8 OLI数据的小麦秸秆覆盖度估算

田间秸秆作为农业生产过程中的重要物质,其覆盖度的遥感估算具有十分重要的意义。Landsat-8 OLI影像作为Landsat系列影像的最新数据产品,具有更精细的光谱特征,明确其在秸秆覆盖度估算中的表现具有重要的现实意义。该研究使用ASD Field Spec 4 Hi-Res地物光谱仪,以实测田间小麦秸秆光谱反射率为数据源,模拟Landsat-8 OLI、Landsat-5TM、Aster、Hyperion影像波段反射率,构建光谱指数,并建立小麦秸秆覆盖度估算模型,通过对比分析,评估Landsat-8OLI数据的估算能力。结果表明,基于Landsat-8 OLI1和OLI2波段构建的NDIOLI21指数模型估算结果最优,决定系数(coefficient of determination,R2)为0.60,均方根误差(root mean square error,RMSE)为9.56%,平均相对误差(mean relative error,MRE)为9.83%,优于Landsat-5 TM构建的光谱指数,且仅次于Aster构建的木质素-纤维素吸收指数(lignin cellulose absorption,LCA)和短波红外归一化差异秸秆指数(shortwave infrared normalized difference residue index,SINDRI)以及Hyperion构建的纤维素吸收指数(cellulose absorption index,CAI)。因此,波段更多、波段划分更加精细的Landsat-8OLI构建的光谱指数在小麦秸秆覆盖度估算方面达到了一定精度,具有良好的应用前景。     

混菌发酵苹果渣生产饲料蛋白的研究

试验利用太空诱变获得的优良菌种黑曲霉ZM-8和酿酒酵母、热带假丝酵母以及白地霉复配制成复合菌剂固体发酵苹果渣。通过正交试验获得了最适的培养基配方和复合菌剂的最佳配方。试验确定的最适培养基配方为硫铵7g、白糖4g、尿素2g、食盐4g,产物中粗蛋白的含量达26.4%,提高了312.50%。复合菌剂的最佳配方为黑曲霉突变株ZM-80.5g、酿酒酵母1.5g、白地霉1.0g、热带假丝酵母1.5g,产物中粗蛋白的含量达27.9%,提高了335.94%。     

海洋氧化短杆菌15E产碱性蛋白酶的发酵条件

对氧化短杆菌15E菌株(Brevibacterium oxydans)产碱性蛋白酶的发酵条件进行研究,结果表明,培养基初始pH值9.0、培养温度28℃、摇床转速200.rmin-1条件下培养40 h,菌体生长最适且总酶活力达到最大;菌株在添加5 g.L-1牛肉膏、1 g.L-1葡萄糖的酪蛋白培养基中生长和产酶均达到最佳;人工海水中的K+、Ca2+和Mg2+浓度较为适合15E菌株生长和产酶,其中K+对菌株产酶起关键作用.     

宁康霉素对植物病原真菌的抑制作用

宁康霉素是由海洋细菌BAC-9912发酵获得的生物农药,对多种植物病原真菌有良好的抑制作用。本研究发现宁康霉素强烈抑制玉米纹枯病菌和苹果轮纹病菌的菌丝,可使玉米纹枯病菌菌丝枯萎和产生畸形。比较宁康霉素与农利灵和速克灵对玉米纹枯病菌的抑菌效果,结果显示宁康霉素的抑菌效果优于两种化学农药。