Hiseq Base Molecular Characterization of Soil Microbial Community, Diversity Structure, and Predictive Functional Profiling in Continuous Cucumber Planted Soil Affected by Diverse Cropping Systems in an Intensive Greenhouse Region of Northern China.

Cover crops are key determinants of the ecological stability and sustainability of continuous cropping soils. However, their agro-ecological role in differentially reshaping the microbiome structure and functioning under a degraded agroecosystem remains poorly investigated. Therefore, structural and metabolic changes in soil bacterial community composition in response to diverse plant species were assessed. Winter catch leafy vegetables crops were introduced as cover plants in a cucumber-fallow period. The results indicate that cover crop diversification promoted beneficial changes in soil chemical and biological attributes, which increased crop yields in a cucumber double-cropping system. Illumina high-throughput sequencing of 16S rRNA genes indicated that the bacterial community composition and diversity changed through changes in the soil properties. Principal component analysis (PCA) coupled with non-metric multidimensional scaling (NMDS) analysis reveals that the cover planting shaped the soil microbiome more than the fallow planting (FC). Among different cropping systems, spinach-cucumber (SC) and non-heading Chinese cabbage-cucumber (NCCC) planting systems greatly induced higher soil nutrient function, biological activity, and bacterial diversity, thus resulting in higher cucumber yield. Quantitative analysis of linear discriminant analysis effect size (LEfSe) indicated that Proteobacteria, Actinobacteria, Bacteroidetes, and Acidobacteria were the potentially functional and active soil microbial taxa. Rhizospheres of NCCC, leaf lettuce-cucumber (LLC), coriander-cucumber (CC), and SC planting systems created hotspots for metabolic capabilities of abundant functional genes, compared to FC. In addition, the predictive metabolic characteristics (metabolism and detoxification) associated with host-plant symbiosis could be an important ecological signal that provides direct evidence of mediation of soil structure stability. Interestingly, the plant density of non-heading Chinese cabbage and spinach species was capable of reducing the adverse effect of arsenic (As) accumulation by increasing the function of the arsenate reductase pathway. Redundancy analysis (RDA) indicated that the relative abundance of the core microbiome can be directly and indirectly influenced by certain environmental determinants. These short-term findings stress the importance of studying cover cropping systems as an efficient biological tool to protect the ecological environment. Therefore, we can speculate that leafy crop diversification is socially acceptable, economically justifiable, and ecologically adaptable to meet the urgent demand for intensive cropping systems to promote positive feedback between crop-soil sustainable intensification.

Complete resistance to powdery mildew and partial resistance to downy mildew in a Cucumis hystrix introgression line of cucumber were controlled by a co-localized locus.

Key message A single recessive gene for complete resistance to powdery mildew and a major-effect QTL for partial resistance to downy mildew were co-localized in a Cucumis hystrix introgression line of cucumber. Downy mildew (DM) and powdery mildew (PM) are two major foliar diseases in cucumber. DM resistance (DMR) and PM resistance (PMR) may share common components; however, the genetic relationship between them remains unclear. IL52, a Cucumis hystrix introgression line of cucumber which has been reported to possess DMR, was recently identified to exhibit PMR as well. In this study, a single recessive gene pm for PMR was mapped to an approximately 468-kb region on chromosome 5 with 155 recombinant inbred lines (RILs) and 193 F2 plants derived from the cross between a susceptible line 'changchunmici' and IL52. Interestingly, pm was co-localized with the major-effect DMR QTL dm5.2 confirmed by combining linkage analysis and BSA-seq, which was consistent with the observed linkage of DMR and PMR in IL52. Further, phenotype-genotype correlation analysis of DMR and PMR in the RILs indicated that the co-localized locus pm/dm5.2 confers complete resistance to PM and partial resistance to DM. Seven candidate genes for DMR were identified within dm5.2 by BSA-seq analysis, of which Csa5M622800.1, Csa5M622830.1 and Csa5M623490.1 were also the same candidate genes for PMR. A single nucleotide polymorphism that is present in the 3' untranslated region (3'UTR) of Csa5M622830.1 co-segregated perfectly with PMR. The GATA transcriptional factor gene Csa5M622830.1 may be a likely candidate gene for DMR and PMR. This study has provided a clear evidence for the relationship between DMR and PMR in IL52 and sheds new light on the potential value of IL52 for cucumber DMR and PMR breeding program.

Identification of Chemotaxis Compounds in Root Exudates and Their Sensing Chemoreceptors in Plant-Growth-Promoting Rhizobacteria Bacillus amyloliquefaciens SQR9.

Chemotaxis-mediated response to root exudates, initiated by sensing-specific ligands through methyl-accepting chemotaxis proteins (MCP), is very important for root colonization and beneficial functions of plant-growth-promoting rhizobacteria (PGPR). Systematic identification of chemoattractants in complex root exudates and their sensing chemoreceptors in PGPR is helpful for enhancing their recruitment and colonization. In this study, 39 chemoattractants and 5 chemorepellents, including amino acids, organic acids, and sugars, were identified from 98 tested components of root exudates for the well-studied PGPR strain Bacillus amyloliquefaciens SQR9. Interestingly, mutant stain SQR9Δ8mcp, with all eight putative chemoreceptors completely deleted, lost the chemotactic responses to those 44 compounds. Gene complementation, chemotaxis assay, and isothermal titration calorimetry analysis revealed that McpA was mainly responsible for sensing organic acids and amino acids, while McpC was mostly for amino acids. These two chemoreceptors may play important roles in the rhizosphere chemotaxis of SQR9. In contrast, the B. amyloliquefaciens-unique chemoreceptor McpR was specifically responsible for arginine, and residues Tyr-78, Thr-131, and Asp-162 were critical for arginine binding. This study not only deepened our insights into PGPR-root interaction but also provided useful information to enhance the rhizosphere chemotaxis mobility and colonization of PGPR, which will promote their application in agricultural production.

Detection of NEO in muskmelon fruits inoculated with Fusarium sulphureum and its control by postharvest ozone treatment.

Fusarium rot of muskmelon, caused by Fusarium spp., is one of the most important postharvest decays, that not only causes economic losses but leads to trichothecenes contamination. A rapid and sensitive method was developed for neosolaniol (NEO) analysis in muskmelon inoculated with F. sulphureum, utilizing acetonitrile/water (84:16, v/v) extraction and PriboFast M270 columns purification and UPLC-MS/MS detection. Method validation was evaluated by linearity (R ≥ 0.9990), recovery (88.1-136.9%), precision (RSD ≤ 3.97%) and sensitivity (LOD, 0.5 µg/kg; LOQ, 1.5 µg/kg). The effect of ozone treatment on Fusarium rot development and NEO accumulation in inoculated muskmelon was also evaluated. The results showed that UPLC-MS/MS method was suitable for analyzing NEO in inoculated muskmelon, and 1.10 mg/l ozone treatment for 120 min significantly controlled Fusarium rot development and NEO accumulation in fruits after 5, 8 and 11 days. In vivo tests showed that ozone at 1.10 mg/l effectively degraded NEO in acetonitrile.

Molecular mapping and candidate gene analysis for resistance to powdery mildew in Cucumis sativus stem.

Powdery mildew (PM) of cucumber (Cucumis sativus), caused by Podosphaera xanthii, is a major foliar disease worldwide and resistance is one of the main objectives in cucumber breeding programs. The resistance to PM in cucumber stem is important to the resistance for the whole plant. In this study, genetic analysis and gene mapping were implemented with cucumber inbred lines NCG-122 (with resistance to PM in the stem) and NCG-121 (with susceptibility in the stem). Genetic analysis showed that resistance to PM in the stem of NCG-122 was qualitative and controlled by a single-recessive nuclear gene (pm-s). Susceptibility was dominant to resistance. In the initial genetic mapping of the pm-s gene, 10 SSR markers were discovered to be linked to pm-s, which was mapped to chromosome 5 (Chr.5) of cucumber. The pm-s gene's closest flanking markers were SSR20486 and SSR06184/SSR13237 with genetic distances of 0.9 and 1.8 cM, respectively. One hundred and fifty-seven pairs of new SSR primers were exploited by the sequence information in the initial mapping region of pm-s. The analysis on the F2 mapping population using the new molecular markers showed that 17 SSR markers were confirmed to be linked to the pm-s gene. The two closest flanking markers, pmSSR27and pmSSR17, were 0.1 and 0.7 cM from pm-s, respectively, confirming the location of this gene on Chr.5. The physical length of the genomic region containing pm-s was 135.7 kb harboring 21 predicted genes. Among these genes, the gene Csa5G623470 annotated as encoding Mlo-related protein was defined as the most probable candidate gene for the pm-s. The results of this study will provide a basis for marker-assisted selection, and make the benefit for the cloning of the resistance gene.

Synthesis, Fungicidal Activity, and Structure Activity Relationship of ß-Acylaminocycloalkylsulfonamides against Botrytis cinerea.

In order to discover new antifungal agrochemicals that could have highly active and novel motifs, thirty-six new 2-acylaminocycloalkylsulfonamides (IV) were synthesized. Their structures were characterized and confirmed by 1H NMR, 13C NMR, IR, MS, elemental analysis and X-ray single crystal diffraction. In vitro and in vivo activities against various Botrytis cinerea strains were evaluated. Bioassay results revealed that most of the title compounds exhibited excellent in vitro fungicidal activity, in which compound IV-26 showed the highest activity against sensitive, low-resistant, moderate-resistant and high-resistant strains of B. cinerea compared with the positive fungicide procymidone. Meanwhile in vivo fungicidal activity of compound IV-31 was better than the commercial fungicides procymidone and chesulfamide in greenhouse trial. The structure activity relationship (SAR) was also discussed and the results were of importance to the structural optimization and development of more potent sulfonamides antifungal agents.

Differential Colonization Dynamics of Cucurbit Hosts by Erwinia tracheiphila.

Bacterial wilt is one of the most destructive diseases of cucurbits in the Midwestern and Northeastern United States. Although the disease has been studied since 1900, host colonization dynamics remain unclear. Cucumis- and Cucurbita-derived strains exhibit host preference for the cucurbit genus from which they were isolated. We constructed a bioluminescent strain of Erwinia tracheiphila (TedCu10-BL#9) and colonization of different cucurbit hosts was monitored. At the second-true-leaf stage, Cucumis melo plants were inoculated with TedCu10-BL#9 via wounded leaves, stems, and roots. Daily monitoring of colonization showed bioluminescent bacteria in the inoculated leaf and petiole beginning 1 day postinoculation (DPI). The bacteria spread to roots via the stem by 2 DPI, reached the plant extremities 4 DPI, and the plant wilted 6 DPI. However, Cucurbita plants inoculated with TedCu10-BL#9 did not wilt, even at 35 DPI. Bioluminescent bacteria were detected 6 DPI in the main stem of squash and pumpkin plants, which harbored approximately 10(4) and 10(1) CFU/g, respectively, of TedCu10-BL#9 without symptoms. Although significantly less systemic plant colonization was observed in nonpreferred host Cucurbita plants compared with preferred hosts, the mechanism of tolerance of Cucurbita plants to E. tracheiphila strains from Cucumis remains unknown.

Niche and host-associated functional signatures of the root surface microbiome.

Plant microbiomes are critical to host adaptation and impact plant productivity and health. Root-associated microbiomes vary by soil and host genotype, but the contribution of these factors to community structure and metabolic potential has not been fully addressed. Here we characterize root microbial communities of two disparate agricultural crops grown in the same natural soil in a controlled and replicated experimental system. Metagenomic (genetic potential) analysis identifies a core set of functional genes associated with root colonization in both plant hosts, and metatranscriptomic (functional expression) analysis revealed that most genes enriched in the root zones are expressed. Root colonization requires multiple functional capabilities, and these capabilities are enriched at the community level. Differences between the root-associated microbial communities from different plants are observed at the genus or species level, and are related to root-zone environmental factors.

Genetic and virulence variability among Erwinia tracheiphila strains recovered from different cucurbit hosts.

The causal agent of cucurbit bacterial wilt, Erwinia tracheiphila, has a wide host range in the family Cucurbitaceae, including economically important crops such as muskmelon (Cucumis melo), cucumber (C. sativus), and squash (Cucurbita spp.). Genetic variability of 69 E. tracheiphila strains was investigated by repetitive-element polymerase chain reaction (rep-PCR) using BOXA1R and ERIC1-2 primers. Fingerprint profiles revealed significant variability associated with crop host; strains isolated from Cucumis spp. were clearly distinguishable from Cucurbita spp.-isolated strains regardless of geographic origin. Twelve E. tracheiphila strains isolated from muskmelon, cucumber, or summer squash were inoculated onto muskmelon and summer squash seedlings, followed by incubation in a growth chamber. Wilt symptoms were assessed over 3 weeks, strains were reisolated, and rep-PCR profiles were compared with the inoculated strains. Wilting occurred significantly faster when seedlings were inoculated with strains that originated from the same crop host genus (P<0.001). In the first run of the experiment, cucumber and muskmelon strains caused wilting on muskmelon seedlings at a median of 7.8 and 5.6 days after inoculation (dai), respectively. Summer squash seedlings wilted 18.0, 15.7, and 5.7 dai when inoculated with muskmelon-, cucumber-, and squash-origin strains, respectively. In a second run of the experiment, cucumber and muskmelon strains caused wilting on muskmelon at 7.0 and 6.9 dai, respectively, whereas summer squash seedlings wilted at 23.6, 29.0 and 9.0 dai when inoculated with muskmelon-, cucumber-, and squash-origin strains, respectively. Our results provide the first evidence of genetic diversity within E. tracheiphila and suggest that strain specificity is associated with plant host. This advance is a first step toward understanding the genetic and population structure of E. tracheiphila.

The corky root rot pathogen Pyrenochaeta lycopersici secretes a proteinaceous inducer of cell death affecting host plants differentially.

Pathogenic isolates of Pyrenochaeta lycopersici, the causal agent of corky root rot of tomato, secrete cell death in tomato 1 (CDiT1), a homodimeric protein of 35 kDa inducing cell death after infiltration into the leaf apoplast of tomato. CDiT1 was purified by fast protein liquid chromatography, characterized by mass spectrometry and cDNA cloning. Its activity was confirmed after infiltration of an affinity-purified recombinant fusion of the protein with a C-terminal polyhistidine tag. CDiT1 is highly expressed during tomato root infection compared with axenic culture, and has a putative ortholog in other pathogenic Pleosporales species producing proteinaceous toxins that contribute to virulence. Infiltration of CDiT1 into leaves of other plants susceptible to P. lycopersici revealed that the protein affects them differentially. All varieties of cultivated tomato (Solanum lycopersicum) tested were more sensitive to CDiT1 than those of currant tomato (S. pimpinellifolium). Root infection assays showed that varieties of currant tomato are also significantly less prone to intracellular colonization of their root cells by hyphae of P. lycopersici than varieties of cultivated tomato. Therefore, secretion of this novel type of inducer of cell death during penetration of the fungus inside root cells might favor infection of host species that are highly sensitive to this molecule.

Cell death of Nicotiana benthamiana is induced by secreted protein NIS1 of Colletotrichum orbiculare and is suppressed by a homologue of CgDN3.

Colletotrichum orbiculare, the causal agent of cucumber anthracnose, infects Nicotiana benthamiana. Functional screening of C. orbiculare cDNAs in a virus vector-based plant expression system identified a novel secreted protein gene, NIS1, whose product induces cell death in N. benthamiana. Putative homologues of NIS1 are present in selected members of fungi belonging to class Sordariomycetes, Dothideomycetes, or Orbiliomycetes. Green fluorescent protein-based expression studies suggested that NIS1 is preferentially expressed in biotrophic invasive hyphae. NIS1 lacking signal peptide did not induce NIS1-triggered cell death (NCD), suggesting apoplastic recognition of NIS1. NCD was prevented by virus-induced gene silencing of SGT1 and HSP90, indicating the dependency of NCD on SGT1 and HSP90. Deletion of NIS1 had little effect on the virulence of C. orbiculare against N. benthamiana, suggesting possible suppression of NCD by C. orbiculare at the postinvasive stage. The CgDN3 gene of C. gloeosporioides was previously identified as a secreted protein gene involved in suppression of hypersensitive-like response in Stylosanthes guianensis. Notably, we found that NCD was suppressed by the expression of a CgDN3 homologue of C. orbiculare. Our findings indicate that C. orbiculare expresses NIS1 at the postinvasive stage and suggest that NCD could be repressed via other effectors, including the CgDN3 homologue.

Evaluation of the combination of 1,3-dichloropropene and dazomet as an efficient alternative to methyl bromide for cucumber production in China.

BACKGROUND: The combination of 1,3-dichloropropene (1,3-D) and dazomet (DZ) offers a potential alternative to methyl bromide (MB) for soil disinfection. MB is scheduled to be withdrawn from routine use by 2015 in developing countries. Combination treatments of 1,3-D + DZ were evaluated in a laboratory study and in two commercial cucumber fields. RESULTS: Laboratory studies found that nearly all of the tested combinations of 1,3-D and DZ displayed positive synergistic activity on root-knot nematodes (Meloidogyne spp.), two major soilborne fungi (Fusarium spp. and Phytophthora spp.) and the seeds of two major weed species (Digitaria sanguinalis and Abutilon theophrasti). Field trials revealed that the combination of 1,3-D and DZ (at 10 + 25 g m(-2) ) successfully suppressed Meloidogyne spp. root galling, sharply reduced Fusarium spp. and Phytophthora spp. and maintained high cucumber yields. The combination treatment of 1,3-D + DZ was more effective than 1,3-D or DZ alone and provided results similar to methyl bromide with respect to pest control, plant mortality, plant height, yield and income. All of the treatments were significantly better than the non-treated control. CONCLUSION: The results indicate that the tested combination of 1,3-D and DZ offers an efficient alternative to methyl bromide for cucumber production.

[Field resistance of Phytophthora melonis to metalaxyl in South China].

OBJECTIVE: Phytophthora melonis is the casual agent of wax gourd and cucumber Phytophthora blight which becomes a constraint for sustainable production of the related crops. Metalaxyl is one of the principal fungicides for controlling the disease now. The objectives of the present study were: (1) to investigate the baseline sensitivity and field resistance of P. melonis to metalaxyl in South China; (2) to test the occurrence of metalaxyl-resistant mutants from metalaxyl-sensitive wild type strains exposed to the fungicide; and (3) to monitor the development of metalaxyl resistance in P. melonis population. METHODS: Over 400 samples of wax gourd and cucumber Phytophthora blight were collected from Guangxi Zhuang Autonomous Region and Guangdong province during 2007-2010, and 193 strains of P. melonis were isolated and purified. The sensitivity of the isolated strains to metalaxyl was tested using mycelial growth rate method in vitro and floating-leaf-disk method in vivo, respectively. The metalaxyl-sensitive strains were induced on PDA plates containing 10 microg/mL metalaxyl. RESULTS: The sensitive, moderately resistant and resistant strains were recorded as 29.0% , 18.1% and 52.8%, respectively, among 193 tested strains. The frequency and level of resistance of P. melonis from Guangdong were higher than that from Guangxi. The strains from cucumber was generally more resistant to metalaxyl than those from wax gourd. The metalaxyl-resistant strains were frequently detected as predominant populations in most of the sampling sites and the highest resistance index (4226.9) was confirmed. Metalaxyl-resistant (M1r) mutants could be isolated from approximately 60% of the sensitive wild-type strains. The resistance level of the M mutants was 189-407 times higher than that of their sensitive parental strains. The EC50 values of 9 sensitive strains from a sampling site without a record of phenylamide fungicide application ranged from 0.0429 to 0.5461 microg/mL. Their mean EC50 value (0.3200 +/- 0.1617 microg/mL) was considered as the baseline sensitivity of P. melonis to metalaxyl in South China. CONCLUSION: Metalaxyl-resistant strains universally occur in South China, especially in the vegetable-growing areas with a longer history of metalaxyl application. The establishment of the baseline sensitivity of P. melonis to metalaxyl will provide a science-based guide for evaluating and further monitoring resistance of the pathogen to the fungicide.

Molecular characterization of the extracellular matrix in a Pseudomonas putida dsbA mutant: implications for acidic stress defense and plant growth promotion.

The Pseudomonas putida dsbA mutant displays enhanced extracellular matrix production, which promotes biofilm formation. Here we confirmed that the extracellular matrix consists of both capsular polysaccharides and exopolysaccharides. However, the carbohydrate composition of the P. putida dsbA mutant matrix was shown to be similar to that of the wild-type strain. Our data indicate that the overproduced matrix itself, rather than alterations in the matrix composition, promotes biofilm formation in the P. putida dsbA mutant. Moreover, the mutant was more sensitive than the wild-type to alkali stress (pH 9.0 to 10.0), but not to acidic stress (pH 5.0). Interestingly, acidic stress stimulated polysaccharide production and pellicle formation, while these changes were recovered to the level of the wild-type under alkali conditions in the P. putida dsbA mutant. Enhanced biofilm formation of the dsbA mutant increased the efficiency with which P. putida attached to tomato and pepper seeds, which have longer germinated roots than the wild-type strain. This phenomenon could not be observed in the cucumber plant, which suggests that each plant seed has a different effect on the attachment of P. putida. Interestingly, this increased attachment to plant seeds resulted in more root colonization and plant growth promotion. The findings of this study suggested that the overproduced extracellular matrix caused by deletion of the dsbA gene could have pleiotropic effect on P. putida phenotypes, including acidic stress defense and plant growth promotion.

A yeast STE11 homologue CoMEKK1 is essential for pathogenesis-related morphogenesis in Colletotrichum orbiculare.

Several signal transduction pathways, including mitogen-activated protein kinase (MAPK) pathways, are involved in appressorium development in Colletotrichum orbiculare, the causal agent of cucumber anthracnose disease. In this study, CoMEKK1, a yeast MAPK kinases (MAPKK) kinase STE11 homolog, was identified as a disrupted gene in an Agrobacterium tumefaciens-mediated transformation mutant. The phenotype of comekk1 disruptant was similar to that of cmk1, a Saccharomyces cerevisiae Fus3/Kss1 MAPK homolog mutant. Moreover, comekk1 and cmk1 mutants were sensitive to high osmotic and salinity stresses, indicating that Comekk1p/Cmk1p signal transduction is involved in stress tolerance. The transformants of the wild type and the comekk1 mutant expressing a constitutively active form of the CoMEKK1 showed slower hyphal growth and abnormal appressorium formation, whereas those of the cmk1 disruptant did not. A Cmk1p-green fluorescent protein (GFP) intracellular localization experiment indicated that nuclear localization of the Cmk1p-GFP fusion protein induced by salt stress was diminished in comekk1 mutants. These results indicate that Comekk1p functions upstream of Cmk1p.

Dispersal and movement mechanisms of Phytophthora capsici sporangia.

Understanding the mechanisms of Phytophthora capsici sporangial dissemination is paramount to understanding epidemic initiation and development. Direct laboratory observations showed P. capsici sporangial dispersal occurred in water with capillary force, but did not occur in response to wind or a reduction in relative humidity. Atmospheric sporangial concentrations were monitored under field conditions using a volumetric spore sampler in a commercial cucurbit field and in an experimental setting where copious sporangia were continuously available in close proximity to the spore trap. Dispersal was infrequent (0.7% of total hours monitored) during sampling in a commercial field; 14 sporangia were detected during a 7.5-week sampling period. In the experimental field situation, dispersal occurred in 4.6% of the hours sampled and 438 sporangia were impacted onto tapes during a 7-week sampling period. Airborne sporangial concentrations were positively associated with rainfall at both sites, but not vapor pressure deficit. Furthermore, in the experimental field situation, wind speed was not significant in regression analysis. Wind speed was not measured in the commercial field. Hence, both direct laboratory observations and volumetric spore sampling indicate that dispersal of sporangia via wind currents is infrequent, and sporangia are unlikely to be naturally dispersed among fields by wind alone.

Alternative sanitizers to chlorine for use on fresh-cut "Galia" (Cucumis melo var. catalupensis) melon.

Chlorine is commonly used to reduce microbial load in fresh-cut vegetables. However, the production of chlorinated organic compounds, such as trihalomethanes, which are potential carcinogens, has created the need to investigate the efficiency of nontraditional sanitizers and alternative techniques. The effects of 4 novel sanitizers were tested in fresh-cut "Galia" melon: chlorine dioxide (ClO(2)) at 3 mg/L, peracetic acid (PAA) at 80 mg/L, hydrogen peroxide (H(2)O(2)) at 50 mg/L, and nisin at 250 mg/L plus EDTA 100 mg/L (nisin + EDTA). A chlorine treatment (NaOCl at 150 mg/L) was used as a control. Pieces of melon were packed in polypropylene trays under passive modified atmosphere (3 to 4 kPa of O(2) and 10 to 11 kPa of CO(2)) and stored up to 10 d at 5 degrees C. Microbial growth, firmness, respiration rate, gas composition, sensory evaluation, color, total soluble solids (TSS), and tritable acidity (TA) were evaluated at days 0, 7, and 10. The novel sanitizers PAA, H(2)O(2), and nisin + EDTA, in the studied concentrations, reduced the microbial growth to a more efficient range than chlorine and ClO(2). In addition, those sanitizers delayed softness, did not affect the respiration rate, SST, or AT. The sensorial parameters were kept above the upper limit of marketability and they did not impart an "off flavor." These sanitizers maintained quality and shelf life of fresh-cut Galia melon for 10 d of storage at 5 degrees C. Nevertheless, other concentrations, in particular for ClO(2,) could be tested to study an extended shelf life in melon pieces.

The life history of the plant pathogen Pseudomonas syringae is linked to the water cycle.

Pseudomonas syringae is a plant pathogen well known for its capacity to grow epiphytically on diverse plants and for its ice-nucleation activity. The ensemble of its known biology and ecology led us to postulate that this bacterium is also present in non-agricultural habitats, particularly those associated with water. Here, we report the abundance of P. syringae in rain, snow, alpine streams and lakes and in wild plants, in addition to the previously reported abundance in epilithic biofilms. Each of these substrates harbored strains that corresponded to P. syringae in terms of biochemical traits, pathogenicity and pathogenicity-related factors and that were ice-nucleation active. Phylogenetic comparisons of sequences of four housekeeping genes of the non-agricultural strains with strains of P. syringae from disease epidemics confirmed their identity as P. syringae. Moreover, strains belonging to the same clonal lineage were isolated from snow, irrigation water and a diseased crop plant. Our data suggest that the different substrates harboring P. syringae modify the structure of the associated populations. Here, we propose a comprehensive life cycle for P. syringae--in agricultural and non-agricultural habitats--driven by the environmental cycle of water. This cycle opens the opportunity to evaluate the importance of non-agricultural habitats in the evolution of a plant pathogen and the emergence of virulence. The ice-nucleation activity of all strains from snow, unlike from other substrates, strongly suggests that P. syringae plays an active role in the water cycle as an ice nucleus in clouds.

Structural determination of hypnosin, a spore germination inhibitor of phytopathogenic Streptomyces sp. causing root tumor in melon (Cucumis sp.).

The structure of a germination inhibitor, hypnosin, isolated from phytopathogenic Streptomyces sp. causing root tumor of melon was determined to be 3-acetylaminopyrazine-2-carboxylic acid (1) by mass spectrometry, computational chemical prediction of UV spectrum, and synthesis of candidates. The structure-activity relationship of hypnosin and anthranilic acid was examined, and it was concluded that pyrazinecarboxylic acid or pyridine-2-carboxylic acid was the fundamental structure with activity, that methylation of the carboxyl group or decarboxylation destroyed activity, and that the presence of an amino group was inhibitory to the activity, whereas acetylation or deletion of an amino group enhanced activity. Hypnosin inhibited spore germination of some Streptomyces spp. in addition to the species with which it was isolated.

Co-occurrence of Arum- and Paris-type morphologies of arbuscular mycorrhizae in cucumber and tomato.

Colonization by arbuscular mycorrhizal (AM) fungi was investigated in cucumber (Cucumis sativus), tomato (Lycopersicon esculentum) and Clethra barbinervis (Ericales) grown in field-collected soil known from previous studies to generate Paris-type arbuscular mycorrhizae in C. barbinervis. Spores of Paraglomus, Acaulospora, Glomus, and Gigaspora were found in the soil. Formation of hyphal coils and arbusculate coils of Paris-type mycorrhizae and of arbuscules of Arum-type mycorrhizae in roots raised in this soil in the growth chamber were compared with the detection of DNA of AM fungi from the same root systems using Glomales-specific primers. Only Paris-type mycorrhizae with extensive arbusculate coils developed in C. barbinervis, but cucumber and tomato developed both Paris- and Arum-types in the same root systems. Glomaceae and Archaeosporaceae and/or Paraglomaceae were detected strongly in the DNA from both cucumber and tomato roots, in which Arum-type mycorrhizae were observed. In contrast, DNA of Glomaceae was detected more sparingly in C. barbinervis, in which Paris-type mycorrhizae dominated. Acaulosporaceae and Gigasporaceae were strongly detected in the DNA from both C. barbinervis and tomato, whereas they were more weakly detected in cucumber. These results indicate that the morphology of colonization is strongly influenced by the selection of fungi to colonize the host plant from among those in the soil environment.