Catalog of Micro-Tom tomato responses to common fungal, bacterial, and viral pathogens

Lycopersicon esculentum cultivar Micro-Tom is a miniature tomato with many advantages for studies of the molecular biology and physiology of plants. To evaluate the suitability of Micro-Tom as a host plant for the study of pathogenesis, Micro-Tom plants were inoculated with 16 well-known fungal, bacterial, and viral pathogens of tomato. Athelia rolfsii, Botryotinia fuckeliana, Oidium sp., Phytophthora infestans, and Sclerotinia sclerotiorum caused typical symptoms and sporulated abundantly on Micro-Tom. Micro-Tom was resistant to Alternaria alternata, Corynespora cassiicola, and Fusarium oxysporum. When Micro-Tom was inoculated with 17 isolates of Ralstonia solanacearum, many isolates induced wilt symptoms. Agrobacterium tumefaciens also was pathogenic, causing crown galls on stem tissue after needle prick inoculation. In Micro-Tom sprayed with Pseudomonas syringae pv. tomato, P. s. pv. tabaci, or P. s. pv. glycinea, bacterial populations did not increase, and yellow lesions appeared only on leaves sprayed with P. s. pv. tomato. Tomato mosaic virus, Tomato aspermy virus, and Cucumber mosaic virus systemically infected Micro-Tom, which developed symptoms characteristic of other cultivars of tomato after infection with the respective virus. These results indicated that Micro-Tom was generally susceptible to most of the important tomato pathogens and developed typical symptoms, whereas certain pathogens were restricted by either hypersensitive resistance or nonhost resistance on Micro-Tom. Therefore, an assortment of Micro-Tom–pathogen systems should provide excellent models for studying the mechanism of susceptible and resistant interactions between plants and pathogens.     

Interaction between protein, phytate, and microbial phytase. In vitro studies

The interaction between protein and phytate was investigated in vitro using proteins extracted from five common feedstuffs and from casein. The appearance of naturally present soluble protein-phytate complexes in the feedstuffs, the formation of complexes at different pHs, and the degradation of these complexes by pepsin and/or phytase were studied. Complexes of soluble proteins and phytate in the extracts appeared in small amounts only, with the possible exception of rice pollards. Most proteins dissolved almost completely at pH 2, but not after addition of phytate. Phytase prevented precipitation of protein with phytate. Pepsin could release protein from a precipitate, but the rate of release was increased by phytase. Protein was released faster from a protein-phytate complex when phytase was added, but phytase did not hydrolyze protein. Protein was released from the complex and degraded when both pepsin and phytase were added. It appears that protein-phytate complexes are mainly formed at low pH, as occurs in the stomach of animals. Phytase prevented the formation of the complexes and aided in dissolving them at a faster rate. This might positively affect protein digestibility in animals.     

Classification of Dutch and German avian reoviruses by sequencing the sigma C protein

We have amplified, cloned and sequenced (part of) the open reading frame of the S1 segment encoding the sigma C protein of avian reoviruses isolated from chickens with different disease conditions in Germany and The Netherlands during 1980 up to 2000. These avian reoviruses were analysed phylogenetically and compared with sequences of avian reoviruses in the Genbank database. The avian reoviruses could be grouped in 5 different genotyping clusters and this classification was identical when the sequences were compared of the 5' end, the 3' end or the whole open reading frame of the sigma C protein. Therefore sequencing of either part of the gene encoding the sigma C protein seems to be reliable for classification. We were unable to identify a correlation between sigma C sequences of the avian reoviruses and the disease condition they were isolated from. The sequences found in The Netherlands and in Germany are, like those in Taiwan, more dispersed than the known avian reovirus sigma C sequences in the USA and Australia. We did not establish temporal or geographic differences in the avian reoviruses studied.     

PCR-based differentiation of Fusarium oxysporum ff. sp. lycopersici and radicis-lycopersici and races of F. oxysporum f. sp. lycopersici

The pathogenic type (form and race) of Fusarium oxysporum, which generates wilt symptoms on tomato, was rapidly identified with a polymerase chain reaction (PCR)-based technique. We compared the partial nucleotide sequences of endo polygalacturonase (pg1) and exo polygalacturonase (pgx4) genes from isolates of F. oxysporum ff. sp. lycopersici (FOL) and radicis-lycopersici (FORL) from Japan and designed specific primer sets (uni, sp13, sp23, and sprl) based on the nucleotide differences that appeared among the pathogenic types. PCR with the uni primer set amplified a 670∼672-bp fragment from all isolates of FOL and FORL. With the sp13 primer set, an amplicon of 445 bp was obtained only from isolates of FOL race 1 and 3. With the sp23 primer set, a 518-bp fragment was obtained from isolates of FOL race 2 and 3. The sprl primer set yielded a 947-bp fragment from isolates of FORL, but not from FOL. A combination of amplifications with these primer sets effectively differentiated the pathogenic types of F. oxysporum in tomato.     

细胞分裂素:杨树气孔运动的调节剂

通过在三种杨树无性系，Ｉ－２１４（Ｐｏｐｕｌｕｓ×ｅｕｒａｎｅｒｉｃａｎａｃｖ．Ｉ－２１４）（ｌｔａｌｉｃａ）、中东杨（Ｐ．ｂｅｒｏｌｉｎｅｎｓｉｓ）（Ｂｅｒｏｌｉｎｅｎｓｉｓ）和群众杨（Ｐ．ｐｏｐｕｌａｒｉｓ３５－４４）（Ｐｏｐｕｌａｒｉｓ）一年生盆栽插条苗的木质部导入ＡＢＡ和细胞分裂素，研究了这两类激素在气孔调控中的作用。尽管不同无性系的气孔在对ＡＢＡ的敏感性上存在显著差异，但ＡＢＡ仍可导致气孔的关闭，然而在蒸腾流中的细胞分裂素（与ＡＢＡ共导入或分别导入）可以明显地抑制ＡＢＡ的作用。并且玉米素还能推迟土壤干旱所诱导的气孔关闭，在水分胁迫条件下，内源细胞分裂素浓度下降而同时ＡＢＡ上升．据此提出了复合胁迫信号的概念，即在根冠通讯中，是ＡＢＡ和细胞分裂素共同调控气孔的运动。另外还研究了玉米素、激动素、６－ＢＡ等不同细胞分裂素与ＡＢＡ的相互作用，结果发现６－ＢＡ与玉米素和激动素的作用相反，它不能抑制ＡＢＡ的作用，反而促进其对气孔的关闭     

Trend analysis of Trichinella in a red fox population from a low endemic area using a validated artificial digestion and sequential sieving technique

Freezing of fox carcasses to minimize professional hazard of infection with Echinococcus multilocularis is recommended in endemic areas, but this could influence the detection of Trichinella larvae in the same host species. A method based on artificial digestion of frozen fox muscle, combined with larva isolation by a sequential sieving method (SSM), was validated using naturally infected foxes from Latvia. The validated SSM was used to detect dead Trichinella muscle larvae (ML) in frozen muscle samples of 369 red foxes from the Netherlands, of which one fox was positive (0.067 larvae per gram). This result was compared with historical Trichinella findings in Dutch red foxes. Molecular analysis using 5S PCR showed that both T. britovi and T. nativa were present in the Latvian foxes, without mixed infections. Of 96 non-frozen T. britovi ML, 94% was successfully sequenced, whereas this was the case for only 8.3% of 72 frozen T. britovi ML. The single Trichinella sp. larva that was recovered from the positive Dutch fox did not yield PCR product, probably due to severe freeze-damage. In conclusion, the SSM presented in this study is a fast and effective method to detect dead Trichinella larvae in frozen meat. We showed that the Trichinella prevalence in Dutch red fox was 0.27% (95% CI 0.065-1.5%), in contrast to 3.9% in the same study area fifteen years ago. Moreover, this study demonstrated that the efficacy of 5S PCR for identification of Trichinella britovi single larvae from frozen meat is not more than 8.3%.     

Fish larvae: zooplankton relationships in microcosm simulations of earthen nursery ponds. II. brackish water system

A simulation of the effects of predation intensity on zooplankton composition in brackish water nursery ponds was carried out in order to address the problem that commercial fish nurseries encounter in obtaining enough zooplankton of adequate species composition and size when fish larvae start to feed. The experimental system consisted of twelve 130 l containers with treatments of four densities (0, 1, 2, or 4 larvae l⁻¹) of common carp (Cyprinus carpio L.), stocked on the 6th day after filling the containers. Zooplankton-environment relationships were explored using factor analysis. Factor analysis allowed identifying several groups of zooplankters that responded in different ways to fish larvae predation pressure. The first factor represented a general measurement of rotifer abundance, and the second identified the direct effect of size-selective fish predation. Since no rotifers were present in the filling water, all these species were autochthonous populations that hatched from resting forms in the sediment and reproduced. In the absence of fish predation, this led to a steep rotifer increase. Fish predation started when the rotifer concentration was just starting to increase and their direct predation reduced and delayed the rotifer abundance peak. This effect increased with the increase in fish larvae density. Estimations of rotifer consumption by fish larvae in this experiment were higher than similar calculations from data of the literature, which led us to test the hypothesis that factors other than direct predation were affecting rotifer population dynamics. The mechanisms involved in rotifer population regulation are discussed. It was concluded that in commercial nurseries, increased larvae production can be achieved by keeping the larvae density at an intermediate level and stocking fish to match the increasing phase of the rotifer peak. Under reasonable larvae density (up to 2 l⁻¹) it seems that the direct predation effect of fish larvae on rotifer dynamics is minor, compared to fish induced self regulation.     

Cartilage acidic protein-1B (LOTUS), an endogenous Nogo receptor antagonist for axon tract formation

Neural circuitry formation depends on the molecular control of axonal projection during development. By screening with fluorophore-assisted light inactivation in the developing mouse brain, we identified cartilage acidic protein-1B as a key molecule for lateral olfactory tract (LOT) formation and named it LOT usher substance (LOTUS). We further identified Nogo receptor-1 (NgR1) as a LOTUS-binding protein. NgR1 is a receptor of myelin-derived axon growth inhibitors, such as Nogo, which prevent neural regeneration in the adult. LOTUS suppressed Nogo-NgR1 binding and Nogo-induced growth cone collapse. A defasciculated LOT was present in lotus-deficient mice but not in mice lacking both lotus- and ngr1. These findings suggest that endogenous antagonism of NgR1 by LOTUS is crucial for normal LOT formation.     

Role of histone H3 lysine 27 methylation in X inactivation

The Polycomb group (PcG) protein Eed is implicated in regulation of imprinted X-chromosome inactivation in extraembryonic cells but not of random X inactivation in embryonic cells. The Drosophila homolog of the Eed-Ezh2 PcG protein complex achieves gene silencing through methylation of histone H3 on lysine 27 (H3-K27), which suggests a role for H3-K27 methylation in imprinted X inactivation. Here we demonstrate that transient recruitment of the Eed-Ezh2 complex to the inactive X chromosome (Xi) occurs during initiation of X inactivation in both extraembryonic and embryonic cells and is accompanied by H3-K27 methylation. Recruitment of the complex and methylation on the Xi depend on Xist RNA but are independent of its silencing function. Together, our results suggest a role for Eed-Ezh2-mediated H3-K27 methylation during initiation of both imprinted and random X inactivation and demonstrate that H3-K27 methylation is not sufficient for silencing of the Xi.     

Virus, bacteriophages and water purification

Water can be a vector of viral disease, but direct virological analysis of water has logistic and practical limitations. Viruses of major importance for water hygiene (e.g. hepatitis and gastro-enteritis viruses) cannot yet be grown in tissue culture. Therefore, as in bacteriological quality procedures, model organisms are required for the evaluation of virological quality of water and the effectiveness of virus removal by water treatment processes. On the basis of published information, the F specific RNA (FRNA) phages have been chosen for this purpose. For the enumeration of the phages a particular Salmonella typhimurium strain with an artificially introduced F plasmid was developed as a host strain and was found to give accurate and reliable results. FRNA phages were found in very high numbers (10(2)-10(5) pfu/ml) in all types of waste water investigated. FRNA phages are seldom found in non-faecally contaminated waste water. Surprisingly low numbers are found in faeces. FRNA phages in waste water effluent were found to be highly resistant to chloramines and relatively resistant to UV inactivation. The FRNA phages can thus effectively be used as indicator organisms for human pathogenic viruses in the evaluation of disinfection processes for water treatment plants.