Differential resistance of oilseed rape cultivars (Brassica napus ssp. oleifera) to Verticillium longisporum infection is affected by rhizosphere colonisation with antagonistic bacteria, Serratia plymuthica and Pseudomonas chlororaphis

The effect of a seed treatment with the antagonistic bacteria Serratia plymuthica (strain HRO-C48) and/or Pseudomonas chlororaphis (strain MA 342) on the infection of oilseed rape with Verticillium longisporum was assessed with ten different cultivars. Soil was inoculated with microsclerotia and mycelium of a V. longisporum culture. Seeds were treated with rifampicin-resistant antagonistic bacteria at a rate of log₁₀ 6–7 cells per seed. Resistance against V. longisporum infection did not differ between cultivars and was generally low. A significant disease reduction recorded as area under disease progress curve (AUDPC) was obtained with both antagonistic rhizobacteria with no significant difference between the treatments. Percent of healthy plants was approximately 70% in all bacterial treatments. Significant differences were observed between the cultivars ranging from 46.5% (cultivar Titan) to 72.6% (Trabant). The combined use of both bacteria could not provide additional control effects. The bacterial density in the rhizosphere was not related to the control effect, but increased by log₁₀ 2 on infection with V. longisporum. Growth promotion effects were also not related to the control effect. At present, neither the application of chemical fungicides nor breeding for resistance against V. longisporum in oilseed rape can provide a solution for this increasingly problematic plant pathogen. The present results now open perspectives to control V. longisporum in oilseed rape by making use of cultivars, which express resistance against this pathogen on interaction with the antagonistic rhizobacteria S. plymuthica or P. chlororaphis.     

Impact of formulation procedures on the effect of the biocontrol agent Serratia plymuthica HRO-C48 on Verticillium wilt in oilseed rape

Verticillium wilt is an important disease in oilseed rape with an increasing importance worldwide. Currently, there are no methods available to suppress the pathogen. A biological protection strategy on the basis of the plant-beneficial bacterium Serratia plymuthica HRO-C48 to control Verticillium dahliae in oilseed rape was developed. Three different techniques to apply the biocontrol agent to seeds, namely pelleting, film coating and bio-priming, were evaluated considering the influence on the control activity, cell stability during storage and practical feasibility. Neither the treatment nor the inoculum density was found to influence the abundances of HRO-C48 in the rhizosphere after 30 days. Serratia treatment using bio-priming and pelleting resulted in a statistically significant biocontrol in comparison to the non-bacterized controls. Additionally, survival of HRO-C48 differed between treatments, and was the highest using bio-priming at 20°C, and pelleting at 4°C. In conclusion, the procedure of bio-priming, which was developed in line with this study, resulted in a stable and efficient formulation of S. plymuthica on rape seed. This technology opens a possibility to develop a commercial Serratia formulation to protect oilseed against V. dahliae.     

Inoculation methods using Rhodococcus erythropolis strain P30 affects bacterial assisted phytoextraction capacity of Nicotiana tabacum

In this study different bacterial inoculation methods were tested for tobacco plants growing in a mine-soil contaminated with Pb, Zn, and Cd. The inoculation methods evaluated were: seed inoculation, soil inoculation, dual soil inoculation event, and seed+soil inoculation. Each inoculum was added at two bacterial densities (10⁶ CFUs mL^?1 and 10⁸ CFUs mL^?1). The objectives were to evaluate whether or not the mode of inoculation or the number of applied microorganisms influences plant response. The most pronounced bacterial-induced effect was found for biomass production, and the soil inoculation treatment (using 10⁶ CFUs mL^?1) led to the highest increase in shoot dry weight yield (up to 45%). Bacterial-induced effects on shoot metal concentrations were less pronounced; although a positive effect was found on shoot Pb concentration when using 10⁸ CFUs mL^?1 in the soil inoculation (29% increase) and in the seed+soil inoculation (34% increase). Also shoot Zn concentration increased by 24% after seed inoculation with 10⁶ CFUs mL^?1. The best effects on the total metal yield were not correlated with an increasing number of inoculated bacteria. In fact the best results were found after a single soil inoculation using the lower cellular density of 10⁶ CFUs mL^?1.     

Quantitative and specific detection of the biocontrol agent, Serratia plymuthica, in plant extracts using a real-time TaqMan? assay

A Serratia plymuthica-specific TaqMan? assay was designed based on the consensus nucleotide sequence from the 3??- end of the luxS gene present in all S. plymuthica strains tested. The specificity of the assay was demonstrated by testing 21 Serratia spp. strains and 30 isolates belonging to various species that can potentially coexist with S. plymuthica in the same environment. Positive reactions in the TaqMan? assay were observed only for S. plymuthica isolates and not for other bacteria. The TaqMan? assay could detect down to 1.95 ng of S. plymuthica DNA, down to 5 bacterial cells per reaction (100?cfu ml^?1) in vitro, down to 50 bacterial cells per reaction (1,000?cfu ml^?1) in spiked potato root extracts and down to 5 bacterial cells per reaction (100?cfu ml^?1) in spiked potato haulm extracts. We used this assay to quantify S. plymuthica A30 cells in potato and tomato haulms and roots grown from S. plymuthica A30-inoculated potato seed tubers and tomato seeds. The results were comparable with the spread-plating of plant extracts on a newly developed S. plymuthica A30 selective medium (CVTR2Arif). The TaqMan? assay can be used to quantify S. plymuthica isolates in different ecosystems and in complex substrates.     

Beneficial microorganism survival on seed, roots and in rhizosphere soil following application to seed during drum priming

Priming is a technique used to improve seedling establishment of direct-seeded crops such as onion and carrot, resulting in a quick and uniform emergence. This work investigated the application of four selected beneficial microorganisms (Pseudomonas chlororaphis MA342, Pseudomonas fluorescens CHA0, Clonostachys rosea IK726d11 and Trichoderma harzianum T22) to onion and carrot seed during drum priming, and their subsequent survival and establishment in the rhizosphere once the seed was planted. Different application rates of fungi (7 log₁₀ cfu g⁻¹ dry seed) and bacteria (6 log₁₀ cfu g⁻¹ dry seed) were required on onion to achieve the end target of 5 log₁₀ cfu g⁻¹ dry seed, whereas a lower rate (5 log₁₀ cfu g⁻¹ dry seed for both bacteria and fungi) was successful on carrot. Microorganism-treated seed was planted in soil in the glasshouse and root and rhizosphere soil samples were taken at 2, 4 and 8 weeks post-planting. All seed-applied microorganisms were recovered throughout the experiment, although differences in the survival patterns were seen. The bacterial isolates declined in number over time, with P. fluorescens CHA0 showing better overall survival than P. chlororaphis MA342, particularly on the roots and in the rhizosphere soil of carrot. In contrast to the bacteria, the fungal isolate C. rosea IK726d11 showed good survival on both onion and carrot, and increased significantly in number throughout the 8-week period. Trichoderma harzianum T22 remained relatively constant in number throughout the experiment, but showed better survival on carrot than onion roots. Similar results were found in three different soil-types.     

Microbial population dynamics on seeds during drum and steeping priming

In the UK, seeds are primed commercially via drum or steeping priming processes to improve seed germination and seedling establishment but the microbial population dynamics occurring during these processes are unknown. Consequently, changes in culturable bacterial and fungal populations that occurred during laboratory and commercial scale drum priming of carrot, leek and parsnip seed and during laboratory scale and commercial scale steeping priming of carrot and sugar beet seed were investigated. Populations of bacteria and pseudomonads increased by 2 and 4 log₁₀ cfu g^–1 seed during priming to reach between 7 and 10 log₁₀ cfu g^–1 seed with less than 1 log₁₀ cfu g^–1 seed decrease after redrying, irrespective of seed type or priming process. Pseudomonads represented approximately 10% of the culturable bacterial population. Fungal populations also increased by between 1 and 3 log₁₀ cfu g^–1 seed during priming of carrot, leek and parsnip seed and were little affected by redrying. Addition of a tefluthrin coating (a standard commercial insecticide treatment) during the drying stage of commercial scale drum priming had little effect on microbial populations. During steeping priming, populations of bacteria and pseudomonads on sugar beet also increased by 2 and 4 log₁₀ cfu g^–1 seed with less than 1 log₁₀ cfu g^–1 seed decrease after redrying. However, fungi did not increase in the same manner on sugar beet, suggesting that the sugar beet seed itself may have an inhibitory effect on fungal growth. The presence of thiram in the steeping liquid (a standard commercial fungicide treatment) inhibited fungal proliferation on carrot and sugar beet seed. Spore forming bacteria generally occurred at low levels on all seeds (between 2 and 5 log₁₀ cfu g^–1seed) and did not exhibit any characteristic population changes during priming or redrying. Clearly some culturable populations of microorganisms can increase reproducibly during these priming processes and survive the redrying procedure, thereby demonstrating the potential for a novel method of introducing microbial inoculants onto seed.     

Microbial Diversity Inside Pumpkins: Microhabitat-Specific Communities Display a High Antagonistic Potential Against Phytopathogens

Recent and substantial yield losses of Styrian oil pumpkin (Cucurbita pepo L. subsp. pepo var. styriaca Greb.) are primarily caused by the ascomycetous fungus Didymella bryoniae but bacterial pathogens are frequently involved as well. The diversity of endophytic microbial communities from seeds (spermosphere), roots (endorhiza), flowers (anthosphere), and fruits (carposphere) of three different pumpkin cultivars was studied to develop a biocontrol strategy. A multiphasic approach combining molecular, microscopic, and cultivation techniques was applied to select a consortium of endophytes for biocontrol. Specific community structures for Pseudomonas and Bacillus, two important plant-associated genera, were found for each microenvironment by fingerprinting of 16S ribosomal RNA genes. All microenvironments were dominated by bacteria; fungi were less abundant. Of the 2,320 microbial isolates analyzed in dual culture assays, 165 (7%) were tested positively for in vitro antagonism against D. bryoniae. Out of these, 43 isolates inhibited the growth of bacterial pumpkin pathogens (Pectobacterium carotovorum, Pseudomonas viridiflava, Xanthomonas cucurbitae); here only bacteria were selected. Microenvironment-specific antagonists were found, and the spermosphere and anthosphere were revealed as underexplored reservoirs for antagonists. In the latter, a potential role of pollen grains as bacterial vectors between flowers was recognized. Six broad spectrum antagonists selected according to their activity, genotypic diversity, and occurrence were evaluated under greenhouse conditions. Disease severity on pumpkins of D. bryoniae was significantly reduced by Pseudomonas chlororaphis treatment and by a combined treatment of strains (Lysobacter gummosus, P. chlororaphis, Paenibacillus polymyxa, and Serratia plymuthica). This result provides a promising prospect to biologically control pumpkin diseases.     

Inhibition of Vancomycin-Resistant <Emphasis Type="Italic">Enterococcus</Emphasis> by Continuous-Flow Cultures of Human Stool Microflora With and Without Anaerobic Gas Supplementation

A continuous-flow competitive exclusion (CFCE) culture model of human stool microflora was used to examine whether supplemental anaerobic gas is necessary for maintenance of anaerobes and inhibition of vancomycin-resistant Enterococcus (VRE). CFCE cultures of human stool microflora were maintained with supplemental nitrogen, without supplemental nitrogen, or with percolated room air. Cultures with or without supplemental nitrogen maintained >9 log₁₀ CFU mL^–1 of obligate anaerobes and eliminated 10⁶ CFU mL^–1 of VRE. When room air was percolated into the culture, anaerobes were detected at 2 log₁₀ CFU mL^–1, and the same VRE inoculum was not eliminated (P < 0.001). These data demonstrate that human stool CFCE cultures maintain high levels of obligate anaerobes and inhibit VRE without the addition of supplemental anaerobic gas.     

Latitude, seed predation and seed mass

Aim We set out to test the hypothesis that rates of pre‐ and post‐dispersal seed predation would be higher towards the tropics, across a broad range of species from around the world. We also aimed to quantify the slope and predictive power of the relationship between seed mass and latitude both within and across species. Methods Seed mass, pre‐dispersal seed predation and post‐dispersal seed removal data were compiled from the literature. Wherever possible, these data were combined with information regarding the latitude at which the data were collected. Analyses were performed using both cross‐species and phylogenetic regressions. Results Contrary to expectations, we found no significant relationship between seed predation and latitude (log₁₀ proportion of seeds surviving predispersal seed predation vs. latitude, P = 0.63; R² = 0.02; n = 122 species: log₁₀ proportion of seeds remaining after postdispersal seed removal vs. latitude, P = 0.54; R² = 0.02; n = 205 species). These relationships remained non‐significant after variation because of seed mass was accounted for. We also found a very substantial (R² = 0.21) relationship between seed mass and latitude across 2706 species, with seed mass being significantly higher towards the tropics. Within‐species seed mass decline with latitude was significant, but only about two‐sevenths, as rapid as the cross‐species decline with latitude. Results of phylogenetic analyses were very similar to cross‐species analyses. We also demonstrated a positive relationship between seed mass and development time across ten species from dry sclerophyll woodland in Sydney (P < 0.001; R² = 0.77; Standardized Major Axis slope = 0.14). These data lend support to the hypothesis that growing period might affect the maximum attainable seed mass in a given environment. Main conclusions There was no evidence that seed predation is higher towards the tropics. The strong relationship between seed mass and latitude shown here had been observed in previous studies, but had not previously been quantified at a global scale. There was a tenfold reduction in mean seed mass for every c. 23° moved towards the poles, despite a wide range of seed mass within each latitude.     

树鼩细菌感染模型的建立及抗菌药物的治疗效果评价(英文)

在抗微生物感染药物开发过程中, 动物模型是必不可少的。虽然目前已经用啮齿类动物建立了一些细菌感染动物模型, 但在小型灵长类动物中还很少见。这里首次报道两个树鼩细菌感染动物模型。第一种是在三度烫伤后的皮肤表面接种 5×106 CFU 的金黄色葡萄球菌构建的皮肤烫伤感染模型。这个数量的金黄色葡萄球菌可以造成 7 d 持续性感染, 并且在第 4天可以看到明显的炎症反应。第二种是用绿脓杆菌构建的涤纶补片感染模型, 接种 2×106 CFU 的绿脓杆菌同样可以引起持续 6 d 感染, 并在第三天在伤口处观察到大量的脓液。进一步用这两种模型评价头孢哌酮钠和左氧氟沙星的治疗效果。左氧氟沙星和头孢哌酮钠在皮肤烫伤感染模型中能分别将 100 mg 皮肤组织中的细菌降低到 4log10 和 5log10 CFU, 并且在涤纶补片植入感染模型中这两种抗生素都能显著地将感染的细菌降低了 4log10 CFU (P<0.05)。结果表明用金黄色葡萄球菌和绿脓杆菌成功构建了两个细菌感染的树鼩模型。此外, 树鼩对金黄色葡萄球菌和绿脓杆菌很敏感, 适合用于构建细菌感染动物模型和评价新的抗细菌感染药物的效果。     

Seed moisture content affects afterripening and smoke responsiveness in three sympatric Australian native species from fire‐prone environments

Germination of freshly collected seeds of three sympatric herbaceous species native to fire‐prone environments in south‐western Australia was significantly improved through the application of novel combinations of dry heat, gibberellic acid, smoke water and dry afterripening. For fresh seeds, combinations of dry heat, gibberellic acid and/or smoke water resulted in >80% germination in Austrostipa elegantissima (Poaceae) and Stylidium affine (Stylidaceae) seeds and >60% germination in Conostylis candicans (Haemodoraceae) seeds, compared with <10% germination of control seeds. For fresh seeds, two broad germination patterns were observed in response to smoke water: nil – low germination for both control and smoke water‐treated seeds (A. elegantissima and S. affine); and a significant smoke response (35%) compared with control seeds (1%) (C. candicans). During afterripening, high germination for A. elegantissima seeds was achieved following 3 months storage of seeds at equilibrium relative humidities of 23–75%, but seeds stored at 5–13% equilibrium relative humidities took 6–36 months to achieve similar levels of germination. Germination of C. candicans seeds also increased after 3 months storage, to >60% at each equilibrium relative humidity and further increases over time were slight. For S. affine seeds >60% germination was achieved only after 36 months storage at 50% equilibrium relative humidity. Seeds from all three species were smoke‐responsive at some point, but the interaction/effects of afterripening on the smoke response varied significantly between species. This study highlights an apparent effect of seed dormancy status on response to smoke and a surprisingly high level of ecological variation in pre‐germination requirements (cues) for these co‐occurring species that may relate to variation(s) in microsite selection forces operating on the soil seed bank of the different species.     

Sodium trimetaphosphate and hexametaphosphate impregnated with silver nanoparticles: characteristics and antimicrobial efficacy

This study aimed to synthesize and characterize materials containing silver nanoparticles (AgNP) with polyphosphates (sodium trimetaphosphate (TMP) or sodium hexametaphosphate (HMP), and evaluate their effect against Candida albicans and Streptococcus mutans. The minimum inhibitory concentration (MIC) was determined, which was followed by the quantification of the biofilm by counting colony-forming units (CFUs), the amount of metabolic activity, and the total biomass. The MICs revealed greater effectiveness of composites containing 10% Ag (TMP + Ag10% (T10) and HMP + Ag10% (H10)) against both microorganisms. It was observed that T10 and H10 reduced the formation of biofilms by 56–76% for C. albicans and by 52–94% for S. mutans for total biomass and metabolic activity. These composites promoted significant log reductions in the number of CFUs, between 0.45–1.43 log₁₀ for C. albicans and 2.88–3.71 log₁₀ for S. mutans (p < .001). These composites demonstrated significant antimicrobial activity, especially against S. mutans, and may be considered a potential alternative for new dental materials.     

The roles of inorganic nitrogen salts in maintaining phytochrome- and gibberellin A3-mediated germination control in skotodormant lettuce seeds

Skotodormant seeds of Lactuca sativa Grand Rapids imbibed in darkness for 10 days (10-day DS) germinated poorly upon terminal treatment with red light (R) or gibberellin A₃ (GA₃). Inorganic nitrogen salts in the imbibition solutions reduced seed skotodormancy. Ten-day DS seeds, imbibed in 25 mm salt solutions followed by terminal R, germinated 99% if imbibed in NH₄NO₃, 70% if imbibed in KNO₃ or NH₄Cl, and 55% if imbibed in NaNO₃. Seeds imbibed in higher salt concentrations germinated fully upon terminal R treatment. Seeds imbibed in 25 mm NH₄Cl or in 50 mm NH₄NO₃ germinated completely upon GA₃ treatment. Osmotic effects of imbibition media accounted for only part of the effect, since seeds imbibed in 50 mm CaCl₂ or NaCl germinated poorly following R or GA₃ treatment. Seeds imbibed in 500 mm polyethylene glycol (PEG) 1000 or mannitol solutions for 10 days still exhibited skotodormancy. Treatments of R or GA₃ did not stimulate germination in seeds imbibed in mannitol, but germination was complete if seeds were given 1-h acid immersion plus a water rinse before the terminal R or GA₃ treatment. Seeds imbibed in 50–500 mm PEG during 10-day DS germinated significantly better in response to terminal R. Terminal GA₃ significantly improved germination only in seeds imbibed at 500 mm PEG. P_fr appeared to function in mannitol-imbibed seed only after an acid treatment. Seed exposure to inorganic nitrogen salts during the 10-day DS maintained seed sensitivity to terminal R or GA₃ treatment. The depth of seed skotodormancy was related to the availability of inorganic nitrogen and also involved the levels of P_fr or endogenous GA₃.Abbreviations FR far red - DS dark storage - R red - GA₃ gibberellin A₃ - PEG polyethylene glycol - SHAM salicylhydroxamic acid - ANOVA analysis of variance - GLM general linear model - LSD least squares difference - P_fr far-red absorbing form of phytochrome     

烟草种子内生细菌群落结构与多样性

【目的】为了解烟草种子内生细菌的群落结构和多样性。【方法】分别对3个品种(K326、云烟85、云烟87)烟草种子内生细菌的16S rRNA基因V3–V4区进行扩增,采用Illumina MiSeq测序技术对扩增片段进行高通量测序,并对3个品种烟草种子内生细菌群落结构和多样性进行分析。【结果】3个品种种子共获得的V3–V4区高质量序列片段128558条,Shannon指数计算为2.03–3.73,K326和云烟85内生菌多样性指数高于云烟87。3品种烟草种子内生细菌的优势门均为变形菌门Proteobacteria、放线菌门Actinobacteria、厚壁菌门Firmicute和拟杆菌门Bacteroidete。3个品种烟草种子内生细菌共有菌属共有27个,K326和云烟85的最优势菌属为假单胞菌属(Pseudomonas),云烟87的最优势菌属为大肠杆菌志贺菌属(Escherichia-shigella)。16S功能预测显示各种子中产生了丰度较高的蛋白质、核苷酸、糖类、辅酶及代谢产物合成的有益功能信息。【结论】烟草种子内生细菌多样性丰富,不同品种种子细菌群落组成基本相似,其丰度存在一定差异性。种子中存在的潜在有益细菌包括假单胞菌、类芽孢杆菌、根瘤菌、马赛菌、藤黄单胞菌、萨勒河菌、Lelliottia菌等,具有大量代谢相关的有益功能。研究结果为今后烟草种子内生菌的功能研究和利用以及种子病害生物防控提供参考信息。     

Colonization Pattern of the Biocontrol Strain Pseudomonas chlororaphis MA 342 on Barley Seeds Visualized by Using Green Fluorescent Protein

Pseudomonas chlororaphis MA 342 is a potent biocontrol agent that can be used against several seed-borne diseases of cereal crops, including net blotch of barley caused by the fungus Drechslera teres. In this study, strain MA 342 was tagged with the gfp gene (encoding the green fluorescent protein) in order to study the fate of cells after seed inoculation. The gfp-tagged strain, MA 342G2, had the same biocontrol efficacy as the wild type when it was applied at high cell concentrations to seeds but was less effective at lower cell concentrations. By comparing cell counts determined by microscopy to the number of CFU, we found that the number of culturable cells was significantly lower than the total number of bacteria on seeds which were inoculated and dried for 20 h. Confocal microscopy and epifluorescence stereomicroscopy were used to determine the pattern of MA 342G2 colonization and cell aggregation on barley seeds. Immediately after inoculation of seeds, bacteria were found mainly under the seed glume, and there was no particular aggregation pattern. However, after the seeds were sown, irregularly distributed areas of bacterial aggregation were found, which reflected epiphytic colonization of glume cells. There was a trend towards bacterial aggregation near the embryo but never within the embryo. Bacterial aggregates were regularly found in the groove of each seed formed by the base of the coleoptile and the scutellum. Based on these results, we suggest that MA 342 colocalizes with the pathogen D. teres, which facilitates the action of the fungistatic compound(s) produced by this strain.     

Reduction of Salmonella on alfalfa seeds using peroxyacetic acid and a commercial seed washer is as effective as treatment with 20 000 ppm of Ca(OCl)2

Aims: The efficacy of a commercial seed washer and 1 and 3% peroxyacetic acid or 20 000 ppm calcium hypochlorite for reducing Salmonella on alfalfa seeds was investigated. Methods and Results: Alfalfa seeds were inoculated with Salmonella Stanley to achieve c. 5 log CFU g^?1. Seeds were then treated with 1 or 3% peroxyacetic acid or 20 000 ppm calcium hypochlorite for 15 min in a commercial seed washer that uses air to enhance contact of the sanitizer with the seed. Experiments were also conducted using industry and laboratory methods. An c. 1‐log reduction in number of Salm. Stanley was demonstrated regardless of the chemical treatment or method of treatment. Although this 1‐log reduction was significant (P < 0·05), differences among the treatments were not significant. Treating the seed with 1 and 3% peroxyacetic acid resulted in similar Salm. Stanley reductions of 1·77 and 1·34 log, respectively, not being statistically significant (P > 0·05). Conclusions: These results suggest that under conditions tested, 1 or 3% peroxyacetic acid solutions are equally effective as 20 000 ppm of Ca(OCl)₂ in the reduction of Salm. Stanley on alfalfa seed when used in conjunction with a commercial seed washer. Significance and Impact of the Study: A 1% peroxyacetic acid solution could potentially be used in place of 20 000 ppm of Ca(OCl)₂ for treatment of seeds used for sprouting_. The commercial seed washer did not enhance removal of Salm. Stanley from alfalfa seeds, but did facilitate removal of excess soil from seeds.     

High synergistic antibacterial,antibiofilm, antidiabetic and antimetabolic activity of Withania somnifera leaf extract-assisted zinc oxide nanoparticle

Nanotechnology is currently gaining immense attention to combat food borne bacteria, and biofilm. Diabetes is a common metabolic disease affecting majority of people. A better therapy relies on phytomediated nanoparticle synthesis. In this study, W. somnifera leaf extract-assisted ZnO NPs (Ws-ZnO NPs) was synthesized and characterized. From HR-TEM analysis, it has been found that the hexagonal wurtzite particle is 15.6 nm in size and − 12.14 mV of zeta potential. A greater antibacterial effect of Ws-ZnO NPs was noticed against E. faecalis and S. aureus at 100 µg mL⁻¹. Also, the biofilm of E. faecalis and S. aureus was greatly inhibited at 100 µg mL⁻¹ compared to E. coli and P. aeruginosa. The activity of α-amylase and α-glucosidase enzyme was inhibited at 100 µg mL⁻¹ demonstrating its antidiabetic potential. The larval and pupal development was delayed at 25 µg mL⁻¹ of Ws-ZnO NPs. A complete mortality (100%) was recorded at 25 µg mL⁻¹. Ws-ZnO NPs showed least LC₅₀ value (9.65 µg mL⁻¹) compared to the uncoated ZnO NPs (38.8 µg mL⁻¹) and leaf extract (13.06 µg mL⁻¹). Therefore, it is concluded that Ws-ZnO NPs are promising to be used as effective antimicrobials, antidiabetic and insecticides to combat storage pests.

     

Seed production and seed quality in a calcareous grassland in elevated CO2

In diverse plant communities the relative contribution of species to community biomass may change considerably in response to elevated CO₂. Along with species‐specific biomass responses, reproduction is likely to change as well with increasing CO₂ and might further accelerate shifts in species composition. Here, we ask if, after 5 years of CO₂ exposure, seed production and seed quality in natural nutrient‐poor calcareous grassland are affected by elevated CO₂ (650 μ L L^?1 vs 360 μ L L^?1) and how this might affect long‐term community dynamics. The effect of elevated CO₂ on the number of flowering shoots (+ 24%, P < 0.01) and seeds (+ 29%, P = 0.06) at the community level was similar to above ground biomass responses in this year, suggesting that the overall allocation to sexual reproduction remained unchanged. Compared among functional groups of species we found a 42% increase in seed number (P < 0.01) of graminoids, a 33% increase (P = 0.07) in forbs, and no significant change in legumes (? 38%, n.s.) under elevated CO₂. Large responses particularly of two graminoid species and smaller responses of many forb species summed up to the significant or marginally significant increase in seed number of graminoids and forbs, respectively. In several species the increase in seed number resulted both from an increase in flowering shoots and an increase in inflorescence size. In most species, seeds tended to be heavier (+ 12%, P < 0.01), and N‐concentration of seeds was significantly reduced in eight out of 13 species. The fraction of germinating seeds did not differ between seeds produced in ambient and elevated CO₂, but time to germination was significantly shortened in two species and prolonged in one species when seeds had been produced in elevated CO₂. Results suggest that species specific increases in seed number and changes in seed quality will exert substantial cumulative effects on community composition in the long run.     

莫莫格扁秆藨草恢复湿地土壤种子库对长芒稗入侵的响应

湿地恢复过程中,时常有外来种或本地杂草入侵。土壤种子库作为未来植被的潜在种源,对湿地恢复效果具有重要的指示意义。在莫莫格国家级自然保护区,以恢复白鹤栖息地(扁秆藨草(Scirpus planiculmis)沼泽)为目的,进行了退化湿地的水文恢复;但退化湿地恢复2a后,一年生杂草长芒稗(Echinochloa caudata)在大部分区域成为建群种。以长芒稗入侵湿地和扁秆藨草自然湿地为研究对象,对比分析了长芒稗和扁秆藨草的土壤种子库及生长结实特征。结果表明,在自然湿地扁秆藨草种子库规模是长芒稗的18.42倍,而在恢复湿地长芒稗种子库大小是扁秆藨草的5.04倍。与自然湿地相比,扁秆藨草种子库密度在入侵湿地明显减少,但仍保留了一定量具有活力的种子(664.32±105.98)粒/m~2,这与研究区扁秆藨草较高的种子生产力(9210.4±1513.4)粒/m~2及种子较强的浮力(FP50=39.7d)有关,说明扁秆藨草具备通过种子库或水传播恢复的潜力。长芒稗土壤种子库密度在入侵湿地高达(3345.9±520.3)粒/m~2,明显高于自然湿地种子库规模(P0.01),说明恢复湿地受长芒稗入侵影响严重,这与长芒稗较高的种子生产力(7621.4±376.25)粒/m~2及较弱的种子浮力(FP0=5d)有关,同时也表明长芒稗通过水传播扩散的能力较弱。另外,研究区长芒稗平均高度超过1m,且盖度较大,不仅阻碍扁秆藨草种子的水播,也降低了到达地表的光照水平,从而抑制扁秆藨草更新。因此,在莫莫格受长芒稗入侵湿地,于开花结实前收获长芒稗地上植物体及凋落物应是限制长芒稗扩展、同时促进扁秆藨草恢复的有效措施。     

Improved delivery of biocontrolPseudomonas and their antifungal metabolites using alginate polymers

Alginate polymer was evaluated as a carrier for seed inoculation with a genetically modified strainPseudomonas fluorescens F113LacZY, which protects sugar-beet againstPythium-mediated damping-off. F113LacZY survived in alginate beads at 5 log₁₀ CFU/ bead or higher counts for 8 weeks of storage, regardless of the conditions of incubation. In plant inoculation experiments, colonisation of the growing area of the root by F113LacZY, derived from alginate beads placed in the soil next to the seed or from an alginate coating around the seeds, was improved compared with application of just free cells of the strain. F113LacZY trapped in alginate beads was an effective producer of antifungal phloroglucinols as indicated by direct HPLC quantification of phloroglucinols and in vitro inhibition of both the indicator bacteriumBacillus subtilis A1 and the pathogenic fungusPythium ultimum. Alginate polymer represents a promising carrier for the delivery of biocontrol inoculants for root colonisation and production of antifungal metabolites.