Conserved eukaryotic fusogens can fuse viral envelopes to cells

Caenorhabditis elegans proteins AFF-1 and EFF-1 [C. elegans fusion family (CeFF) proteins] are essential for developmental cell-to-cell fusion and can merge insect cells. To study the structure and function of AFF-1, we constructed vesicular stomatitis virus (VSV) displaying AFF-1 on the viral envelope, substituting the native fusogen VSV glycoprotein. Electron microscopy and tomography revealed that AFF-1 formed distinct supercomplexes resembling pentameric and hexameric "flowers" on pseudoviruses. Viruses carrying AFF-1 infected mammalian cells only when CeFFs were on the target cell surface. Furthermore, we identified fusion family (FF) proteins within and beyond nematodes, and divergent members from the human parasitic nematode Trichinella spiralis and the chordate Branchiostoma floridae could also fuse mammalian cells. Thus, FF proteins are part of an ancient family of cellular fusogens that can promote fusion when expressed on a viral particle.     

Accumulation,partitioning, and assimilation of nitrate in Opuntia ficus‐indica

Nitrate levels and nitrate reductase activity (NRA) of the widely cultivated prickly‐pear Opuntia ficus‐indica were measured for 5‐year‐old orchard plants in the field between flowering and fruit ripening (May‐August) and for rooted cladodes (stem segments) in a glasshouse in pots that were supplied with 0.8,4, or 16 mM nitrate during the early growth of new cladodes (6 weeks). Nitrate levels were much higher in the cladodes than in the fruit peels or the roots; in both cladodes and fruit peels, nitrate levels were higher in the inner water‐storing layer (parenchyma) than in the outer green photosynthetic layer (chlorenchyma). NRA was confined to the cladode chlorenchyma and was higher in new cladodes than in the underlying cladodes. The orchard study suggested that the nitrate accumulated in 2‐ and 3‐year‐old cladodes served as a reserve for the growth of new organs. At the beginning of the reproductive season, such older cladodes had high nitrate levels of 7 mg/g DW in the chlorenchyma and 45 mg/g DW in the parenchyma, which decreased by 72% and 43%, respectively, by the end of the season. The glasshouse experiments indicated that the cladodes were more important for nitrate reduction than the roots, particularly under high external nitrate concentrations.     

荒漠区珍稀和野生果树的引种驯化

能在热带荒漠地区进行商业化栽培的作物种类是有限的,园艺作物尤其如是。本研究的目的就是通过引种驯化为热带荒漠区开发新品种。我们以下述两个来源的多年生植物为基础:干旱区和半干旱区人民食用的有营养价值的水果或干果的野生植物种;第二是从当地市场或栽培者那里能收集到的珍稀种类。以色列的内盖夫荒漠气候条件变幅宽广,水土类型多样,是本项研究的理想地点。本研究利用内盖夫的异质特点对向荒漠区引种驯化的不少植物的适宜性进行了预选试验。本文总结了从1984年开始的4年工作,并叙述了总的进展,详细介绍了六种预选植物。     

Physical and visual properties of grape rachis as affected by water vapor pressure deficit

Rachis browning of table grapes after harvest is a significant problem, and water loss is considered the primary factor in browning. The major rachis desiccation and browning occurs during marketing at ambient temperatures and relative humidity (RH) which create high water vapor pressure deficits (WVPD). In this study the effect of WVPD and its components on rachis browning were examined on the two white seedless cultivars ‘Superior’ and ‘Thompson’. The grape clusters were stored at 20 °C or at 10 °C with low (70%) or high (>95%) RH, thus creating 4 WVPD levels. At each WVPD the clusters were held in open punnets, punnets sealed with low density polyethylene film or microperforated polyethylene, and examed every 2 or 3 d for weight loss, berry firmness, rachis dry weight and subjective rachis index. In addition, the rachis were photographed and image analysis employed to identify the level of browning. The results show that image analysis gave very similar patterns to subjective evaluation of rachis browning with correlation coefficients up to 0.90. However, image analysis detected an increase in browning before subjective evaluation. There was poor overall correlation between cluster weight loss and rachis dry weight to browning for ‘Superior’ grapes but a good correlation for ‘Thompson’. Rachis of ‘Superior’ suffered extensive browning at 20 °C even at high RH while rachis of ‘Thompson’ remained relatively green under similar conditions. ‘Thompson’ grape rachis remained green during the entire examination period (11 d) when held at high RH in either 10 °C or 20 °C. At high WVPD, microperforated packaging offered better control of browning in ‘Superior’ grapes than closed packaging, while clusters of ‘Thompson’ retained green rachis after 4 d in open punnets, and after 7 d in covered punnets. In summary, detailed analysis of rachis browning shows that water loss is an important but not the only factor in browning. Quantitative and objective measurement of rachis browning is likely to facilitate better communication of experimental data and higher resolution of processes which lead to browning.     

Induction of Resistance to Penicillium digitatum in Grapefruit by the Yeast Biocontrol Agent Candida oleophila

ABSTRACT The yeast Candida oleophila, the base of the commercial product Aspire, is recommended for the control of postharvest decay in citrus and pome fruit. Its modes of action include nutrient competition, site exclusion, and direct mycoparasitism. In the present study, we showed that application of Candida oleophila to surface wounds or to intact 'Marsh Seedless' grapefruit elicited systemic resistance against Penicillium digitatum, the main postharvest pathogen of citrus fruit. The induction of pathogen resistance in fruit was already pronounced 24 h after elicitation; it was distance, concentration, and time dependent and restricted to the peel tissue closely surrounding the yeast application site. The induction of pathogen resistance required viable yeast cells at concentrations of 10(8) to 10(9) cells ml(-1). Nonviable autoclaved or boiled yeast cells or lower yeast concentrations were ineffective in enhancing fruit disease resistance. Application of Candida oleophila cell suspensions to grapefruit peel tissue increased ethylene biosynthesis, phenylalanine ammonia lyase activity, and phytoalexin accumulation, and increased chitinase and beta-1,3-endoglucanase protein levels, indicated by western immunoblotting analysis. Scanning electron microscope observations revealed that spore germination and germ tube growth of Penicillium digitatum were markedly inhibited in wounds made near the yeast-treated sites. Overall, this study provides evidence that induced resistance against postharvest decay of citrus fruit should be considered an important component of the multiple modes of action of the yeast Candida oleophila.     

The efficacy of ultrasonic fumigation for disinfestation of storage facilities against postharvest pathogens

Inoculum of postharvest pathogens can accumulate inside storage rooms and contaminate new batches of fruit and vegetables, but this chain can be broken by disinfecting storage facilities between storage periods. Quaternary ammonium (QA) has been known for over 50 years as an efficient disinfectant against microorganisms, with wide applications in the food industry. The aim of this study was to determine the efficacy of didecyldimethylammonium chloride (Sporekill, designated here as QA^sk), against development of Botrytis cinerea after direct exposure or by ultrasonic fogging. Following direct exposure to a concentration of QA^sk below 5 mg L^?1, a population of 10⁴ conidia of B. cinerea was inactivated after 2 min; ultrasonic fogging with QA^sk at 500 mg L^?1 took 30 min to achieve consistent inactivation. Fumigation at 20 °C was considerably more effective than fumigation at 5 °C, and similar results were obtained for three other postharvest pathogens, Penicillium expansum, Colletotrichum gloeosporioides and Alternaria alternata. These results show that conidia of B. cinerea are highly sensitive to direct exposure to QA^sk, but that effective sanitation of a storage facility by ultrasonic fumigation requires a QA^sk concentration two orders of magnitude greater.     

Measurement of isotype-specific antibody responses to Aujeszky's disease virus in sera and mucosal secretions of pigs.

Enzyme-linked immunosorbent assays (ELISAs) for the detection of porcine IgM, IgA, IgG1 and IgG2 antibodies directed against Aujeszky's disease virus (ADV) are described. ADV-specific IgA and IgM were detected in an antibody capture assay, and ADV-specific IgG1 and IgG2 were detected in an indirect double antibody sandwich assay. A selected set of samples was tested in the four ELISAs and in a 24 h virus neutralization assay. Comparison of the results showed that the ELISAs were isotype-specific, sensitive, and reproducible. Samples with ADV antibody of one isotype showed that ADV-specific IgG1, IgG2 and IgM were able to neutralize the virus in vitro. In vitro neutralization of virus can be enhanced by complement. ADV-specific IgA neutralized virus only weakly. ADV-infected cells activated complement in the absence of antibody. Specific IgG2 and IgM enhanced complement activation. Analysis of the time course of antibody responses after infection or vaccination revealed that the isotype-specific ELISAs are suitable to study the humoral antibody response of pigs to the virus in mucosal secretions. Wild-type virus (strain NIA-3) and an attenuated vaccine strain (Bartha) administered intranasally induced mucosal IgM and IgA responses to the virus. In contrast, a killed vaccine (Nobivac) administered intramuscularly induced only weak mucosal IgM responses. The attenuated vaccine strain primed for a mucosal IgA memory response evoked upon challenge infection with wild-type virus.     

Mobilization of Antimony and Arsenic in Soil and Sediment Samples – Evaluation of Different Leaching Procedures

This study aims to evaluate the performance of different leaching schemes with respect to the mobilization of antimony and arsenic from polluted samples collected at different sites in Mansfeld District, Germany. Besides the elution by water the leaching by artificial acidic rain and by two different schemes of sequential extraction were employed for estimation of the mobilization of antimony and arsenic. The samples were characterized by X-ray fluorescence analysis for their total concentration of metalloids, metals and main constituents. It was found that both antimony and arsenic show little mobilization with de-ionized water as well as artificial acidic rain in single step batch procedures (≤ 0.13% of the total content). Although the percentage leached is very low, the concentrations in the resulting solutions are of ecotoxicological relevance. BCR procedure indicate a very strong binding of Sb and of As in the samples. Less than 20% of the total content can be leached in sum in all leaching steps, of it most under strongly oxidizing conditions. This scheme seems not suitable for a detailed investigation of possible mobilization processes under environmental conditions for the metalloids under investigation. The four-step extraction procedure by Wenzel et al. gives a more detailed pattern of the binding of antimony and arsenic. This procedure was found to be a suitable scheme for evaluating the possible mobilization processes from the samples contaminated by ore processing waste, especially by change through other ions or under reducing conditions.     

Induction of Chitinase and β-1,3-Endoglucanase Proteins by UV Irradiation and Wounding in Grapefruit Peel Tissue

UV irradiation enhanced the resistance of grapefruit against the development of green mold décay caused byPenicillium digitatum, the main postharvest pathogen of citrus fruit, and significantly inhibited the fungus’ growth at the fruit wound sites. Immunoblotting analysis using specific citrus chitinase and β-1,3-endoglucanase antibodies, showed that UV irradiation, wounding of the fruit, or a combination of these two treatments, induced the accumulation of a 25 kD chitinase protein in the fruit’s peel tissue. On the other hand, UV irradiation or wounding of the fruit alone was unable to induce the accumulation of 39 and 43 kD β-1,3-endoglucanase proteins, but the combination of the two treatments increased these protein levels. It is suggested that both chitinase and β-1,3-endoglucanase may play a role in the UV-induced resistance of grapefruit againstP. digitatum. Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel. No. 403/99. http://www.phytoparasitica.org posting June 3, 1999.     

Fruit cracking in Cereus peruvianus,a new cactus crop

Summary

Fruit of Cereus peruvianus (L.) Miller, a newly developed cactus crop, is subject to cracking. Fruit growth follows a double sigmoid curve, with a significant size increase in the first and third stages and moderate increase in the middle stage. Peel and seed grow in the first two stages, while pulp growth and maturation (e.g. peel colour change and stretching, sugar accumulation, fruit softening) occur in the final stage. Sensitive, moderately sensitive, and tolerant clones are subject to 100, 50–60 and 6–9% cracking, respectively, correlated to relative growth rate of the pulp in the third stage. Fruit cracking, lengthwise splitting of the peel, and exposure of the pulp occurred in fully mature fruits towards the end of their growth. Continuous monitoring of fruit growth revealed expansion and cracking during the day and shrinkage during the night, as expected for CAM plants. Cracking was not correlated with the threshold cracking turgor, which declined during ripening in all clones. GA₃ sprays during colour break reduced cracking in the two most susceptible clones to 20% and 60%, while paclobutrazol increased cracking. Cracking rates were the same in two ecozones differing in salinity, temperatures and relative humidity. It is suggested that breeding and hormone treatments could reduce cracking significantly.