胡子鲶败血症病原菌的研究

由患细菌性败血症的胡子鲶苗种分离到6个菌株，经菌株致病力测试、生化鉴定和药物敏感试验，结果表明嗜水气单胞菌是胡子鲶苗种败血症的病原菌，该病原菌对卡那霉素、恶喹酸、呋喃唑酮、丁胺卡那霉素等敏感，而对磺胺等9种药物不敏感。     

Suppressiveness of 18 composts against 7 pathosystems: Variability in pathogen response

Compost is often reported as a substrate that is able to suppress soilborne plant pathogens, but suppression varies according to the type of compost and pathosystem. Reports often deal with a single pathogen while in reality crops are attacked by multiple plant pathogens. The goal of the present study was to evaluate the disease suppression ability of a wide range of composts for a range of plant pathogens. This study was conducted by a consortium of researchers from several European countries. Composts originated from different countries and source materials including green and yard waste, straw, bark, biowaste and municipal sewage. Suppressiveness of compost-amended (20% vol./vol.) peat-based potting soil was determined against Verticillium dahliae on eggplant, Rhizoctonia solani on cauliflower, Phytophthora nicotianae on tomato, Phytophthora cinnamomi on lupin and Cylindrocladium spathiphylli on Spathiphyllum sp., and of compost-amended loamy soil (20% vol./vol.) against R. solani on Pinus sylvestris and Fusarium oxysporum f. sp. lini on flax. From the 120 bioassays involving 18 composts and 7 pathosystems, significant disease suppression was found in 54% of the cases while only 3% of the cases showed significant disease enhancement. Pathogens were affected differently by the composts. In general, prediction of disease suppression was better when parameters derived from the compost mixes were used rather than those derived from the pure composts. Regression analyses of disease suppression of the individual pathogens with parameters of compost-amended peat-based mixes revealed the following groupings: (1) competition-sensitive: F. oxysporum and R. solani/cauliflower; (2) rhizosphere-affected: V. dahliae; (3) pH-related: P. nicotianae; and (4) specific/unknown: R. solani/pine, P. cinnamomi and C. spathiphylli. It was concluded that application of compost has in general a positive or no effect on disease suppression, and only rarely a disease stimulating effect.     

Activity of antifungal proteins against mold in sorghum caryopses in the field

Sorghums were stressed with pathogenic fungi and sprinkling to determine relationships between changes in chitinase and sormatin in caryopses and grain mold resistance. Panicles of 10 cultivars differing in mold resistance and accumulation of antifungal proteins (AFPs) were inoculated at anthesis with Fusarium moniliforme and Curvularia lunata spores. Panicles were sampled at 30 and 50 days after anthesis, and caryopses were evaluated for chitinase and sormatin using western blots. Sprinkling panicles (to mimic rainfall) decreased sormatin and chitinase in most cultivars. Inoculation decreased AFPs in susceptible cultivars, but resistant cultivars maintained or increased AFPs in caryopses. Grain mold resistance corresponded to induction of AFP synthesis in response to sprinkling, fungal stress, and/or adverse field conditions. Sormatin and chitinase appear to be an active part of the defense mechanism of the caryopsis against grain mold.     

番茄青枯病拮抗菌X10的鉴定和田间应用

对番茄青枯病拮抗菌X10进行了形态学特征、生理生化特征测定,结果表明它与侧孢短芽孢杆菌种的特征一致。测定了菌株的16S rDNA序列并根据其构建了系统发育树,在系统发育树中,X10与侧孢短芽孢杆菌形成一个类群,序列同源性为99%。因此将X10鉴定为侧孢短芽孢杆菌。制备了侧孢短芽孢杆菌X10的固体菌剂,在山东肥城科技示范园的蔬菜大棚里进行了小区试验,对田间应用效果进行了测定。结果表明,固体菌剂的两种接种量对番茄青枯病的防治效果分别达到了58.42%和68.68%,同时还能够促进番茄植株生长和增加产量。     

Isolation and identification of locally isolated bacterial strains effective against whitefly Bemisia tabaci

Potent bacterial strains effective against the whitefly, Bemisia tabaci, nymphs (second instar), were isolated from tomato cultivated fields at Fayoum governorate, Giza, Egypt. Of 72 isolates, 12 with the most morphologically distinct-looking bacterial colonies were selected and named A1, A2, A3, A6, A7, A9, A12, A13, A107, B37, B45 and B100. All isolates were preliminarily identified as members of the genus Bacillus based on morphological, physiological and biochemical characteristics. When tested for their pathogenicity against Bemisia tabaci, the 12 isolates revealed varying efficiencies with isolates A1 and A9 being superior, exhibiting maximum mortality of 92.2 and 90.8% on day 10, respectively. Isolate A7 recorded the lowest percentage at 18.3%. Further genetic characterization of the 12 isolates was performed using inter simple sequence repeat (ISSR), randomly amplified polymorphic DNA (RAPD) and 16S rDNA gene sequencing analysis. RAPD and ISSR results confirmed each other. The combined ISSR and RAPD phylogenetic tree showed two major clusters. With 16S rRNA gene analysis, isolate A1 and A12 sequences recorded 100% identity with Bacillus thuringiensis, while isolates A7 and B100 showed 95.7% and 95.6% identity with Bacillus cereus and Bacillus sphaericus, respectively.     

Detection of traces of tetracyclines from fish with a bioluminescent sensor strain incorporating bacterial luciferase reporter genes

Bioluminescent Escherichia coli K-12 strain for the specific detection of the tetracycline family of antimicrobial agents was optimized to work with fish samples. The biosensing strain contains a plasmid incorporating the bacterial luciferase operon of Photorhabdus luminescens under the control of the tetracycline responsive element from transposon Tn10 (Korpela et al. Anal. Chem. 1998, 70, 4457-4462). The extraction procedure of oxytetracycline from rainbow trout (Oncorhynchus mykiss) tissue was optimized. There was neither need for centrifugation of homogenized tissue nor use of organic solvents. The lowest levels of detection of tetracycline and oxytetracycline from spiked fish tissue were 20 and 50 microg/kg, respectively, in a 2-h assay. The optimized assay protocol was tested with fish that were given a single oral dose of high and low concentrations of oxytetracycline. The assay was able to detect oxytetracycline residues below the European Union maximum residue limits, and the results correlated well with those obtained by conventional HPLC (R = 0.81).     

Activity of bacteria strains originaited from sewage sludge against some soil fungi

Abstract

The potential biological activity of three bacterial strains against the four phytopathogens was examined. The bacterial strains were originated from sewage sludge water extract and from the humic acid solution obtained from the sewage sludge. The fresh bacteria cultures and filtered culture broth were used. The noncoated sugar beet seeds were coated by soaking for 15 min in the liquid culture of bacteria, then they were sown in the sterilized soil inoculated by one of the following types of soil fungus: Aphanomyces cochlioides L3, Phoma betas PH8, Pythium irregulare PIR4, Rhizoctonia solani Z10-2. The test proved that Bacillus amytoliquefaciens O1 / O3 and Flaviomonas oryzihabitans K1 / K2 showed antifungal activity on the artificial medium, but they were not able to effectively suppress fungal pathogens in the soil. The bacteria did not injure sugar beet roots, so they can be regarded as microorganisms that are not harmful to plants. It may be concluded that these bacteria have the potential to support antifungal activity of the humis acid and the sewage sludge extracts.     

Activity of 1,4-benzoquinones against formosan subterranean termites (Coptotermes formosanus)

A large number of naturally occurring and synthetic benzoquinones were evaluated for activity against the Formosan subterranean termite, Coptotermes formosanus, with potential use in termite control. Among these bioactive naturally occurring benzoquinones are 2-methyl-5-isopropyl-1,4-benzoquinone, 2-methoxy-6-pentyl-1,4-benzoquinone, 2,3-dimethoxy-5-methyl-6-(3-methyl-2-butenyl)-1,4-benzoquinone, 2,3-dimethoxy-5,6-dimethyl-1,4-benzoquinone, and 2,3-dichloro-5,6-dimethyl-1,4-benzoquinone. All five of these compounds demonstrated 100% mortality against C. formosanus by day 11 at a concentration of 1% (wt/wt) or less. In general, benzoquinones with one or two hydrophobic groups on the 5 and/or 6 positions of the quinone ring along with one or two group(s) on the opposite side of the ring, at the 2 and/or 3 position, led to high rates of mortality against C. formosanus. Quantitative structure-activity relationship (QSAR) studies showed no correlation between lipophilicity (calculated log P) and mortality for the entire group of nonhalogenated benzoquinones. A correlation was observed between C-6 chain length and day 3 percent mortality for 2,3-dimethoxy-5-methyl-6-substituted aliphatic benzoquinones where short chain lengths resulted in higher mortality.     

Bacillus pumilus WP8 exhibits biocontrol efficacy against tomato bacterial wilt via attenuation of the virulence of the pathogenic bacterium

Bacillus pumilus WP8 is a plant growth-promoting rhizobacterium (PGPR) with good biocontrol efficacy against tomato bacterial wilt caused by Ralstonia solanacearum Rs1115. Biocontrol, however, is not due to antagonism of the pathogenic bacterium. Thus, we hypothesised that the biocontrol efficacy of WP8 was achieved by attenuation of Rs1115 virulence. Here, pot experiments for comparison of Rs1115 in different plant parts were conducted to investigate the ability of WP8 to prevent entry of Rs1115 into the regions of the plant above ground. Primary and secondary metabolite contents of WP8 and their inhibitory effects on twitching and swarming motilities of Rs1115 were determined by microscopic examination and crystal violet staining. The effects of WP8 metabolites on the expression of typical virulence genes in Rs1115 were established by quantitative PCR. Rs1115 abundance in the rhizosphere increased with time after inoculation. However, the shoots treated with WP8 were pathogen-free on days 3 and 6 after inoculation. In the WP8 + Rs1115 treatment group, the abundance of Rs1115 in shoots was 1.5?lg units higher on day 9 post-inoculation than that in the Rs1115 treatment group, while less Rs1115 was observed in the leaves. This indicated that WP8 prevented Rs1115 from spreading to the regions of the plant above ground. Furthermore, some heat-resistant secondary metabolites of WP8 (e.g. lipopeptides) inhibited the twitching and swarming motility of Rs1115. Moreover, the metabolites decreased the expression of typical virulence genes in Rs1115. Therefore, WP8 was shown to attenuate Rs1115 virulence, possibly through pumilacidin secretion.     

鱼稻共生条件下陶粒载体水稻浮床提高细菌活性及多样性

以轻质陶粒为主要原料开发出一种新型生物浮床载体,并通过种植水稻构建了陶粒载体水稻浮床对池塘水环境进行修复。研究了陶粒载体水稻浮床对精养池塘的水质净化作用,并采用Biolog-ECO技术分析了陶粒载体水稻浮床对池塘浮游细菌碳源代谢模式和多样性的影响。结果显示,试验组池塘水中总氮(TN)、总磷(TP)和亚硝态氮(NO2–-N)等水质指标在8月份显著低于对照组,高锰酸盐指数(IMn)在8月份和10月份显著低于对照组。试验组池塘浮游细菌群落代谢活性(AWCD)、Shannon指数和丰富度指数在8月份显著高于对照塘,Shannon均匀度指数在10月份显著高于对照塘。浮床种植水稻产量达到5 900 kg/hm~2。结果表明陶粒载体水稻浮床在具备一定生产性能基础上,降低了精养池塘水中氮磷营养水平,改变了精养池塘浮游细菌群落碳源利用模式,并提高了池塘浮游细菌群落代谢活性和功能多样性。结果可为应用陶粒载体水稻浮床调控池塘水环境提供理论指导。     

Effect of calcium concentration in nutrient solution on development of bacterial wilt and population of its pathogen Ralstonia solanacearum in grafted tomato seedlings

We examined the effect of the calcium (Ca) concentration in the nutrient solution on the development of bacterial wilt and the population of its pathogen Ralstonia solanacearum in tomato (Lycopersicon esculentum Mill. cv. Momotarou) seedlings grafted onto the rootstock of a highly resistant cultivar (cv. Hawaii 7998). The grafted seedlings were cultured in a nutrient solution containing Ca at concentrations of 0.4, 4.4, and 20.4 mm, and inoculated with the pathogen by stem puncture at the base of the stem of the rootstock, and the disease incidence was recorded for a period of 21 d. In another experiment, xylem exudates were collected from decapitated scions of the Ca-treated seedlings 5 d after inoculation, and the population of the pathogen in the exudates was counted by plating on a selective medium. The grafted tomato seedlings were highly resistant to bacterial wilt, when cultured in the nutrient solution containing Ca at 20.4 mm. The population of the pathogen in the xylem exudates decreased with increasing concentration of Ca in the solution. However, even in the presence of Ca at a high concentration, infection with the virulent pathogen was observed in the xylem of the scion.     

施硅对水稻白叶枯病抗性及叶片抗氧化酶活性的影响

【目的】水稻白叶枯病是一种细菌性枯萎病害,是限制水稻生产的重要生物因素之一。通过田间接种白叶枯病菌,研究施硅对水稻叶片中丙二醛含量及抗氧化系统酶活性的影响及其抗白叶枯病的机理,为安全有效的防治病害提供理论依据。【方法】以唐粳2号水稻品种为材料,2013年在河北省秦皇岛市进行田间试验,试验在两个施氮水平 [N 180 kg/hm2(正常供氮,N180), 450 kg/hm2(高量供氮,N450)]下设3个硅处理[不施硅(-Si),施硅酸钠(Si1, 以SiO2计,70 kg/hm2), 施硅钙肥(Si2, 以SiO2计,70 kg/hm2)],在水稻孕穗期采用剪叶法接种白叶枯病菌,研究硅对接种后30 d水稻病情指数和第1 d、 3 d、 5 d、 7 d和10 d水稻叶片中丙二醛(MDA)含量、 超氧化物歧化酶(SOD)活性、 过氧化氢酶(CAT)活性、 抗坏血酸过氧化物酶(APX)活性的影响。【结果】接种白叶枯病菌后,正常供氮水平,施硅处理的病情指数比不施硅处理平均降低17.8%(P0.05); 高量供氮水平,施硅钙肥的病情指数比不施硅降低15.1%(P0.05),而施硅酸钠的病情指数差异不显著。接种白叶枯病菌后,施硅处理的水稻叶片MDA均低于不施硅处理,且在正常供氮水平第7 d和高量供氮水平第3 d、 第7 d差异达显著水平。接种白叶枯病菌后,正常供氮水平第1 d、 第7 d和高量供氮水平第1 d、 第5 d,施硅处理的水稻叶片中SOD活性均显著高于不施硅处理,且第1 d施硅钙肥的叶片SOD显著高于施硅酸钠处理; 接种白叶枯病菌后,施硅处理的水稻叶片中CAT活性均高于不施硅处理,但未达显著水平; 高量供氮水平第1 d、 第7 d和第10 d施硅处理的水稻叶片中APX活性均显著高于不施硅处理。【结论】施硅能提高感病水稻叶片中SOD、 CAT和APX的活性,降低水稻叶片中MDA含量,有效清除植物体内活性氧(ROS),从而增强了水稻抗白叶枯病的能力; 在高量供氮水平下,硅钙肥抵御白叶枯病效果好于硅酸钠。     

Activity of nematodes and enchytraeids, bacterial community composition, and functional redundancy in coniferous forest soil

Soil faunal activity stimulates terrestrial carbon and nutrient cycling and maintains soil physical structure. Interestingly, the effects of faunal activity on soil microbial community composition and activity is less clear, although the microbial community evidently is the key element of terrestrial decomposition processes and nutrient cycling. We manipulated the composition of soil fauna in mesh bags with various mesh sizes (5, 45, and 1,000 µm) to exclude faunal groups based on body size. Our experiment indicated that feeding and foraging activity of nematodes and enchytraeids modified the physical–chemical properties of the soil and had long-term effects on soil bacterial community composition (measured by denaturing gel electrophoresis, colony-forming unit counts, and phospholipid fatty acid pattern). However, microbial enzyme (phosphomonoesterase, butyrate-esterase, ß-glucosidase, N-acetyl-glucosamidase, and ß-cellobiosidase) activities (measured by fluorogenic model substrates), decomposition rate of woody and cellulose material, or C and N status of the soil were not altered by the faunal treatment. Instead, enzyme activity was reduced by soil drying. Our results give support to ideas that functional redundancy is a common characteristic of the soil organisms, but no support was found for the idea that the community composition of soil decomposers had a strong link to the regulation of C and N cycles in the soil.     

Activity of meadowfoam (Limnanthes alba) seed meal glucolimnanthin degradation products against soilborne pathogens

Meadowfoam (Limnanthes alba L.) is a herbaceous winter-spring annual grown as a commercial oilseed crop. The meal remaining after oil extraction from the seed contains up to 4% of the glucosinolate glucolimnanthin. Degradation of glucolimnanthin yields toxic breakdown products, and therefore the meal may have potential in the management of soilborne pathogens. To maximize the pest-suppressive potential of meadowfoam seed meal, it would be beneficial to know the toxicity of individual glucolimnanthin degradation products against specific soilborne pathogens. Meloidogyne hapla second-stage juveniles (J2) and Pythium irregulare and Verticillium dahliae mycelial cultures were exposed to glucolimnanthin as well as its degradation products. Glucolimnanthin and its degradation product, 2-(3-methoxyphenyl)acetamide, were not toxic to any of the soilborne pathogens at concentrations up to 1.0 mg/mL. Two other degradation products, 2-(3-methoxymethyl)ethanethioamide and 3-methoxyphenylacetonitrile, were toxic to M. hapla and P. irregulare but not V. dahliae. The predominant enzyme degradation product, 3-methoxybenzyl isothiocyanate, was the most toxic compound against all of the soilborne pathogens, with M. hapla being the most sensitive with EC(50) values (0.0025 ± 0.0001 to 0.0027 ± 0.0001 mg/mL) 20-40 times lower than estimated EC(50) mortality values generated for P. irregulare and V. dahliae (0.05 and 0.1 mg/mL, respectively). The potential exists to manipulate meadowfoam seed meal to promote the production of specific degradation products. The conversion of glucolimnanthin into its corresponding isothiocyanate should optimize the biopesticidal properties of meadowfoam seed meal against M. hapla, P. irregulare, and V. dahliae.     

The bioprotective effect of AM root colonization against the soil-borne fungal pathogen Gaeumannomyces graminis var. tritici in barley depends on the barley variety

The systemic effect of root colonization by the arbuscular mycorrhizal fungus (AMF) Glomus mosseae on the susceptibility of old and modern barley varieties to the soil-borne fungal pathogen Gaeumannomyces graminis var. tritici (Ggt) was studied in a split-root system. Plants were precolonized on one side of the split-root system with the AMF and thereafter the other side of the split-root system was inoculated with the pathogen. At the end of the experiment the level of bioprotection was estimated by quantifying lesioned roots and the determination of the root fresh weight. AM root colonization provided protection in some of the barley genotypes tested, but not in others. This protective effect seemed to vary in the oldest and the most modern barley variety tested.     

Synthesis of phenoxyphenyl pyridine and pyrazine carboxamides. Activity against Cydia pomonella (L.) eggs

Several N-(4-phenoxyphenyl)pyridinecarboxamides and N-(4-phenoxyphenyl)pyrazinecarboxamides were synthesized from commercially available material, and their ovicidal activities against Cydia pomonella (L.) were tested. Some of the tested products showed a moderate activity when <24-h-old eggs were sprayed using a Potter tower. A significant increase in the length of the development period of the eggs was also observed in many cases. A clear correlation between both effects was noticed: the products that produced higher mortality also produced higher increase of the length of the development. These results seem to confirm our hypothesis that these compounds could be defined as a juvenile hormone analogues.     

过鱼设施进口区域水温对集诱鱼效果的影响

库区水温分层与过鱼通道隔断是水利水电工程建设随之产生的生态环境影响之一。过鱼效果是评估过鱼设施建设成功与否的关键指标。对于存在水温分层的高坝工程而言,过鱼设施进口附近存在的温差效应是否会对进口区域的集诱鱼效果产生影响是当前应该解决的首要问题之一。该研究通过建立三维水温水动力数学模型与包含过鱼设施进口段和明渠河流段的物理模型,分别开展了水温水动力预测及实鱼过鱼试验研究工作,研究结果表明：温差效应对进口附近水动力条件影响甚微,影响区域主要取决于鱼道流量,而岸边水温量值则取决于温差的大小;增大明渠流量将增加上溯鱼类发现鱼道进口的难度;沿冷水区域上溯样本量占总样本的39%,沿温水区域上溯样本量占总样本的61%,由此可知,温水区域对洄游鱼类更具有吸引力;温差效应在一定程度上有利于洄游鱼类发现鱼道进口并在进口区域聚集,与冷水区域相比,温水有效区域鱼进入鱼道进口的尝试率提高了17个百分点。研究成果可为相关人员尝试采用改变水温调整洄游鱼类上溯路线,改善鱼道进口集诱鱼,进而提升鱼道过鱼效果提供思路。     

Activity of (-)-epigallocatechin 3-O-gallate against oxidative stress in rats with adenine-induced renal failure

Methylguanidine (MG) is widely recognized as a strong uremic toxin. The hydroxyl radical (*OH) specifically plays an important role in the pathway of MG production from creatinine (Cr). In this study, we investigated whether oral administration of (-)-epigallocatechin 3-O-gallate (EGCg) suppresses MG production in rats with chronic renal failure after intraperitoneal Cr injection. MG production from Cr was significantly increased in rats with adenine-induced renal failure, which was more vulnerable to oxidative stress, compared with that in normal rats. However, oral administration of EGCg 30 min before and after Cr injection effectively inhibited MG production. Our findings suggest that EGCg, an excellent antioxidant from green tea, exerts protective activity in rats with chronic renal failure, resulting in suppression of Cr oxidation influenced by *OH.     

玉米内生芽胞杆菌的抗菌活性物质及其拮抗玉米大斑病菌机理的初步研究

为了研究生防菌株对玉米大斑病菌的抑菌作用,深化对生防菌抗菌机制的认识,本研究从玉米(Zea mays)植株体内分离拮抗玉米大斑病菌(Setosphaeria turcica)的内生细菌,对其抗菌物质及其抑菌机理进行初步研究。结果表明,所分离的内生菌株YY1经形态学观察、生理生化测定及16SrDNA序列分析,鉴定为枯草芽胞杆菌(Bacillus subtilis)。菌株YY1发酵液的硫酸铵沉淀物具有抑菌活性,且在硫酸铵50%饱和度时抑菌活性最强,说明YY1菌株产生的抗菌活性物质可能是蛋白类物质。该菌株及其蛋白粗提液均对禾谷镰刀菌(Fusarium graminearum)、苹果轮纹病菌(Botryosphaeria dothidea)、灰霉病菌(Botrytis cinerea)、玉米弯孢霉叶斑病菌(Curvularia lunata)等7种植物病原真菌有较强的拮抗作用。用蛋白粗提液处理菌丝、分生孢子、原生质体后经显微观察发现,大斑病菌的基内菌丝由丝状畸变为串珠状,当蛋白粗提液浓度为0.78μg/μL时,可完全抑制分生孢子萌发,并导致原生质体裂解。通过抑制孢子萌发过程中信号途径相关基因的半定量RT-PCR分析和玉米大斑病菌不同信号途径相关基因突变体的抑制率统计,初步判定该抑菌过程主要通过cAMP信号转导途径发挥作用。本研究为寻找玉米大斑病菌新的防治方法和途径提供基础资料。     

Stimulation of benzyladenine-induced in vitro shoot organogenesis and endogenous proline in melon (Cucumis melo L.) by fish protein hydrolysates in combination with proline analogues

A previous study demonstrated that proline is beneficial for improving melon in vitro shoot organogenesis. A natural source of proline and proline precursors can be obtained from fish protein hydrolysates (FPH), a byproduct of the fishery industry. Proline analogues azetidine-2-carboxylate and hydroxyproline in combination with standardized FPH were used to stimulate proline synthesis and benzyladenine-induced shoot organogenesis by exploiting the proposed proline-linked pentose phosphate pathway (PPP). In the presence of elevated levels of endogenous proline, potential stimulation of cytokinins and auxin may occur via the PPP and shikimate pathways, respectively. Treatments with FPH singly and in combination with the above proline analogues significantly increased the endogenous proline content and the extent of differentiation, suggesting that in vitro organogenesis is closely linked to proline synthesis, strengthening the hypothesis that purine metabolism via the proline-linked PPP may be important for organogenesis. Thioproline addition resulted in increased proline levels but without corresponding stimulation of organogenesis. This study also provides potential use of fishery waste for value-added application in plant micropropagation industry.