Modeling and analysis of disease-induced host growth in the epidemiology of take-all

ABSTRACT Epidemiological modeling, together with parameter estimation to experimental data, was used to examine the contribution of disease-induced root growth to the spread of take-all in wheat. Production of roots from plants grown in the absence of disease was compared with production of those grown in the presence of disease and the precise form of diseaseinduced growth was examined by fitting a mechanistic model to data describing change in the number of infected and susceptible roots over time from a low and a high density of inoculum. During the early phase of the epidemic, diseased plants produced more roots than their noninfected counterparts. However, as the epidemic progressed, the rate of root production for infected plants slowed so that by the end of the epidemic, and depending on inoculum density, infected plants had fewer roots than uninfected plants. The dynamical change in the numbers of infected and susceptible roots over time could only be explained by the mechanistic model when allowance was made for disease-induced root growth. Analysis of the effect of disease-induced root production on the spread of disease using the model suggests that additional roots produced early in the epidemic serve only to reduce the proportion of diseased roots. However, as the epidemic switches from primary to secondary infection, these roots perform an active role in the transmission of disease. Some consequence of disease-induced root growth for field epidemics is discussed.     

Temporal Dynamics of Phytophthora Blight on Bell Pepper in Relation to the Mechanisms of Dispersal of Primary Inoculum of Phytophthora capsici in Soil

ABSTRACT The effect of components of primary inoculum dispersal in soil on the temporal dynamics of Phytophthora blight epidemics in bell pepper was evaluated in field and growth-chamber experiments. Phytophthora capsici may potentially be dispersed by one of several mechanisms in the soil, including inoculum movement to roots, root growth to inoculum, and root-to-root spread. Individual components of primary inoculum dispersal were manipulated in field plots by introducing (i) sporangia and mycelia directly in soil so that all three mechanisms of dispersal were possible, (ii) a plant with sporulating lesions on the soil surface in a plastic polyvinyl chloride (PVC) tube so inoculum movement to roots was possible, (iii) a wax-encased peat pot containing sporangia and mycelia in soil so root growth to inoculum was possible, (iv) a wax-encased peat pot containing infected roots in soil so root-to-root spread was possible, (v) noninfested V8 vermiculite media into soil directly as a control, or (vi) wax-encased noninfested soil as a control. In 1995 and 1996, final incidence of disease was highest in plots where sporangia and mycelia were buried directly in soil and all mechanisms of dispersal were operative (60 and 32%) and where infected plants were placed in PVC tubes on the soil surface and inoculum movement to roots occurred with rainfall (89 and 23%). Disease onset was delayed in 1995 and 1996, and final incidence was lower in plants in plots where wax-encased sporangia (6 and 22%) or wax-encased infected roots (22%) were buried in soil and root growth to inoculum or root-to-root spread occurred. Incidence of root infections was higher over time in plots where inoculum moved to roots or all mechanisms of dispersal were possible. In growth-chamber studies, ultimately all plants became diseased regardless of the dispersal mechanism of primary inoculum, but disease onset was delayed when plant roots had to grow through a wax layer to inoculum or infected roots in tension funnels that contained small volumes of soil. Our data from both field and growth-chamber studies demonstrate that the mechanism of dispersal of the primary inoculum in soil can have large effects on the temporal dynamics of disease.     

Population Dynamics of Fusarium Oxysporum f. Sp. Radicis-lycopersici in Relation to the Onset of Fusarium Crown and Root Rot of Tomato

Fusarium oxysporum f. sp. radicis-lycopersici the causal agent of crown and root rot in tomato comprises two overlapping separate phases: monocyclic and polycyclic. Oversummering inoculum is the source of primary infection (the monocyclic phase) and the spread from plant to plant via root-to-root contact is the source of the secondary infection (the polycyclic phase). In the present work, relationships between initial inoculum density, population dynamics of the pathogen in the root zone of diseased plants, and disease onset were studied. For the monocyclic phase, 55.1% of the variance of disease onset was attributed to the rate of pathogen proliferation in the root zone of plants, and only 12.8% of the variance was attributed to the amount of initial inoculum density. For the polycyclic phase, disease onset was not related to either initial inoculum density or the rate of pathogen proliferation in the root zone. At disease onset, the inoculum density of the pathogen in the root zone of plants infected from oversummering inoculum reached an average of 4.08 log cfu g soil^–1. The inoculum density of the pathogen in the root zone of plants infected by their diseased neighbors was 3.23 log cfu g soil^–1. A large variation in pathogen proliferation rate in the root zone was found among individual plants, suggesting that differences in the level of soil suppressiveness may occur not only between fields, but even in the same field over short distances.     

Demonstration of secondary infection by Pythium violae in epidemics of carrot cavity spot using root transplantation as a method of soil infestation

Cavity spot of carrot (CCS), one of the most important soilborne diseases of this crop worldwide, is characterized by small sunken elliptical lesions on the taproot caused by a complex of pathogens belonging to the genus Pythium , notably P. violae . In most soilborne diseases the soil is the source of inoculum for primary infections, with diseased plants then providing inoculum for secondary infections (both auto- and alloinfection). Using fragments of CCS lesions to infest soil, it was demonstrated that CCS lesions on carrot residues can cause primary infection of healthy roots. Using a novel soil infestation method, in which an artificially infected carrot root (the donor plant) was placed close to healthy roots (receptor plants) the formation of typical CCS lesions were induced more efficiently than the use of classical soil inoculum and showed that CCS can spread from root to root by alloinfection from transplanted diseased roots. The method also demonstrated the polycyclic nature of a CCS epidemic caused by P. violae in controlled conditions. Secondary infections caused symptoms and reduced root weight as early as two weeks after transplantation of the diseased carrot. This reproducible method may be used for delayed inoculation and for studying the effect of cropping factors and the efficacy of treatments against primary and secondary cavity spot infections.     

Suppression of Rhizoctonia solani on Impatiens by Enhanced Microbial Activity in Composted Swine Waste-Amended Potting Mixes

ABSTRACT Peat moss-based potting mix was amended with either of two composted swine wastes, CSW1 and CSW2, at rates from 4 to 20% (vol/vol) to evaluate suppression of pre-emergence damping-off of impatiens (Impatiens balsamina) caused by Rhizoctonia solani (anastomosis group-4). A cucumber bioassay was used prior to each impatiens experiment to monitor maturity of compost as the compost aged in a curing pile by evaluating disease suppression toward both Pythium ultimum and R. solani. At 16, 24, 32, and 37 weeks after composting, plug trays filled with compost-amended potting mix were seeded with impatiens and infested with R. solani to determine suppression of damping-off. Pre-emergence damping-off was lower for impatiens grown in potting mix amended with 20% CSW1 than in CSW2-amended and nonamended mixes. To identify relationships between disease suppression and microbial parameters, samples of mixes were collected to determine microbial activity, biomass carbon and nitrogen, functional diversity, and population density. Higher rates of microbial activity were observed with increasing rates of CSW1 amendment than with CSW2 amendments. Microbial biomass carbon and nitrogen also were higher in CSW1-amended mixes than in CSW2-amended potting mixes 1 day prior to seeding and 5 weeks after seeding. Principal component analysis of Biolog-GN2 profiles showed different functional diversities between CSW1- and CSW2-amended mixes. Furthermore, mixes amended with CSW1 had higher colony forming units of fungi, endospore-forming bacteria, and oligotrophic bacteria. Our results suggest that enhanced microbial activity, functional and population diversity of stable compost-amended mix were associated with suppressiveness to Rhizoctonia damping-off in impatiens.     

Rhizosphere Competence of Pythium oligandrum

ABSTRACT The associations of Pythium oligandrum with the root cortex, rhizoplane, and rhizosphere were measured with 11 crop species. This work was expedited by the use of a semiselective technique for isolation of P. oligandrum from soil and plant material. Cortical colonization of roots by P. oligandrum was not detected, and the rhizoplanes of the roots of most crops were free of the fungus. However, P. oligandrum was detected in large quantities with every crop tested when roots with adhering soil (rhizosphere soil) were assayed. Different crop species and cultivars of cantaloupe, cauliflower, and tomato varied in rhizosphere densities of P. oligandrum, but rhizosphere population densities of the fungus were consistently higher than in nonrhizosphere soils with plants grown in P. oligandrum-infested sterilized potting mix or an unsterilized mineral soil. After transplanting tomatoes into potting mix infested with P. oligandrum, increases in CFU occurred over time in the rhizosphere but not in the nonrhizosphere soil. In trials on delivery methods of inoculum of P. oligandrum, the rhizosphere populations of tomato plants grown in potting mix were about sixfold higher compared to seed-coat treatments when ground, alginate pelleted oospores were applied to seedlings growing in plug containers prior to transplanting or to pots containing potting mix before direct seeding.     

哈茨木霉菌(Trichoderma harzianum)和终极腐霉菌(Pythium ultimum)对玉米蛋白质组的影响(I)

 哈茨木霉(Trichoderma harzianum) T22菌株已普遍用于防治包括由终极腐霉(Pythium ultimum)引起的苗病或根腐病在内的各种病害。玉米自交系Mo17种子经T22处理后播种在接种腐霉或未接种的田间土壤内,5 d后取幼苗的根系或幼茎提取蛋白。结果表明:在接种腐霉菌的土壤内,未进行T22处理的5 d龄幼苗长势明显比对照差,而经T22处理的幼苗长势明显比对照好。T22和腐霉菌复合处理及T22单独处理对幼苗生长影响基本相同。本研究建立了蛋白质提取和双向电泳分离技术。通过双向电泳及相应的分析软件(PDQuest^TM 2-D softw are)可将不同处理的幼苗自交系蛋白进行分离。T22菌株处理的根系产生104种上游调控蛋白和164种下游调控蛋白,T22与腐霉菌复合处理可产生97种上游调控蛋白和150种下游调控蛋白,而用腐霉菌单一处理诱导的上游或下游蛋白的数量明显少于上述2个处理。T22或腐霉菌单一或复合处理的根系蛋白质组图谱与空白对照相比差异显著,它们与对照的蛋白质组图谱相似系数分别为0.72、0.51和0.49;T22与腐霉菌分别处理的蛋白质组图谱间也相差明显,两者的相似系数仅为0.65。进一步研究发现,T22菌体蛋白质组图谱与上述各种处理的蛋白质组图谱相似系数均很低,说明各种处理诱导后的幼苗根系蛋白质组组分主要来自植物本身,其变化主要因T22或腐霉菌的诱导所致。腐霉菌的侵染对寄主根系蛋白组图谱影响明显高于T22的作用。蛋白质组中各种蛋白质的质谱分析(M ALDI-TOF)与鉴定将另文发表。     

Control of damping-off in cress and sugar-beet by commercial seed-coating with Pythium oligandrum

Seeds of cress and sugar-beet were coated with oospores of Pythium oligandrum using commercial seed-pelleting or film-coating procedures. Following either procedure approximately 10⁴ oospores were recovered from both seed types, achieving 75.94% of the targeted dose. Oospore germination (9.19%) was unaffected by the coating treatments. Both types of treatment reduced damping-off of cress caused by P. ultimum in artificially infested sand and potting compost and by Rhizoctonia solani in artificially infested sand. In some cases, the level of control was equivalent to fungicide drenches. In general, pelleting of P. oligandrum on cress gave better control than film-coating treatments. P. oligandrum also reduced damping-off of sugar-beet in soil naturally infested with Aphanomyces cochlioides and Pythium spp. Control was equivalent to that achieved with hymexazol fungicide seed-coating treatments and was related to the inoculum potential of A. cochlioides in the soil; neither standard hymexazol coatings nor P. oligandrum treatments gave control at high inoculum potentials. P. oligandrum was not rhizosphere competent on cress or sugar-beet.     

An Epidemiological Analysis of the Role of Disease-Induced Root Growth in the Differential Response of Two Cultivars of Winter Wheat to Infection by Gaeumannomyces graminis var. tritici

ABSTRACT Epidemiological modeling combined with parameter estimation of experimental data was used to examine differences in the contribution of disease-induced root production to the spread of take-all on plants of two representative yet contrasting cultivars of winter wheat, Ghengis and Savannah. A mechanistic model, including terms for primary infection, secondary infection, inoculum decay, and intrinsic and disease-induced root growth, was fitted to data describing changes in the numbers of infected and susceptible roots over time at a low or high density of inoculum. Disease progress curves were characterized by consecutive phases of primary and secondary infection. No differences in root growth were detected between cultivars in the absence of disease and root production continued for the duration of the experiment. However, significant differences in disease-induced root production were detected between Savannah and Genghis. In the presence of disease, root production for both cultivars was characterized by stimulation when few roots were infected and inhibition when many roots were infected. At low inoculum density, the transition from stimulation to inhibition occurred when an average of 5.0 and 9.0 roots were infected for Genghis and Savannah, respectively. At high inoculum density, the transition from stimulation to inhibition occurred when an average of 4.5 and 6.7 roots were infected for Genghis and Savannah, respectively. Differences in the rates of primary and secondary infection between Savannah and Genghis also were detected. At a low inoculum density, Genghis was marginally more resistant to secondary infection whereas, at a high density of inoculum, Savannah was marginally more resistant to primary infection. The combined effects of differences in disease-induced root growth and differences in the rates of primary and secondary infection meant that the period of stimulated root production was extended by 7 and 15 days for Savannah at a low and high inoculum density, respectively. The contribution of this form of epidemiological modeling to the better management of take-all is discussed.     

Suppression of Seedling Damping-Off Caused by Pythium ultimum, P. irregulare, and Rhizoctonia solani in Container Media Amended with a Diverse Range of Pacific Northwest Compost Sources

ABSTRACT Suppression of seedling damping-off disease caused by Pythium spp. and Rhizoctonia solani is a potential benefit of formulating soilless container media with compost. Thirty-six compost samples from Pacific Northwest commercial composting facilities were analyzed for a number of physical, chemical, and biological properties, including suppression of damping-off caused by Pythium ultimum, P. irregulare, and R. solani. The samples were produced from diverse feedstocks and composting technol ogies; this was reflected in a large degree of variability in the measured properties. When mixed with sphagnum peat moss and inorganic aggregates, 67% of the compost samples significantly suppressed P. irregulare damping-off of cucumber, 64% suppressed P. ultimum damping-off of cucumber, and 17% suppressed damping-off of cabbage caused by R. solani. Suppression of Pythium damping-off was related to the potential of compost to support microbial activity and a qualitative index of ammonia volatilization. Suppression of Rhizoctonia damping-off was not related to any one compost factor. Currently available compost products potentially could provide commercially acceptable control of damping-off caused by Pythium spp., but it is necessary to fortify composts with microbial antagonists for the control of R. solani.     

Epidemiology and chemical control of take-all on seminal and adventitious roots of wheat

ABSTRACT Epidemiological modeling is used to examine the effect of silthiofam seed treatment on field epidemics of take-all in winter wheat. A simple compartmental model, including terms for primary infection, secondary infection, root production, and decay of inoculum, was fitted to data describing change in the number of diseased and susceptible roots per plant over thermal time obtained from replicated field trials. This produced a composite curve describing change in the proportion of diseased roots over time that increased monotonically to an initial plateau and then increased exponentially thereafter. The shape of this curve was consistent with consecutive phases of primary and secondary infection. The seed treatment reduced the proportion of diseased roots throughout both phases of the epidemic. However, analysis with the model detected a significant reduction in the rate of primary, but not secondary, infection. The potential for silthiofam to affect secondary infection from diseased seminal or adventitious roots was examined in further detail by extending the compartmental model and fitting to change in the number of diseased and susceptible seminal or adventitious roots. Rates of secondary infection from either source of infected roots were not affected. Seed treatment controlled primary infection of seminal roots from particulate inoculum but not secondary infection from either seminal or adventitious roots. The reduction in disease for silthiofam-treated plants observed following the secondary infection phase of the epidemic was not due to long-term activity of the chemical but to the manifestation of disease control early in the epidemic.     

Interactions Between Trichoderma harzianum Strain T22 and Maize Inbred Line Mo17 and Effects of These Interactions on Diseases Caused by Pythium ultimum and Colletotrichum graminicola

ABSTRACT Seed treatment with Trichoderma harzianum strain T22, which results in colonization of plant roots but little or no colonization of shoots or leaves, had substantial effects on growth of and disease expression in maize inbred line Mo17. Shoots and roots of 10-day-old seedlings grown in a sandy loam field soil were larger (roots were nearly twice as long) in the presence of T22 than in its absence. Both main and secondary roots were increased in size and area and the root hair area was greater with T22. However, root hair area per unit of root length was greater in control plants. Increased growth probably was due to direct stimulation of plant growth in addition to effects from biological control of deleterious microflora. Seedlings of Mo17 grown in autoclaved or mefenoxamtreated sandy loam field soil were larger than those produced in untreated soil. However, seedlings grown in the presence of T22, either in treated or untreated soil, were larger than those produced in its absence. Infestation of soil with Pythium ultimum had little effect upon growth of Mo17. The presence of T22 increased protein levels and activities of beta-1,3 glucanase, exochitinase, and endochitinase in both roots and shoots, even though T22 colonized roots well but colonized shoots hardly at all. With some enzymes, the combination of T22 plus P. ultimum gave the greatest activity. Plants grown from T22-treated seed had reduced symptoms of anthracnose following inoculation of leaves with Colletotrichum graminicola, which indicates that root colonization by T22 induces systemic resistance in maize.     

Compost Tea as a Container Medium Drench for Suppressing Seedling Damping-Off Caused by Pythium ultimum

ABSTRACT Compost tea is being used increasingly in agricultural production to control plant diseases. However, there has been limited investigation relating disease control efficacy to various compost tea production methods, particularly compost tea produced with active aeration and additives to increase microbial population densities in compost tea. Aerated compost tea (ACT) and nonaerated compost tea (NCT), produced with or without additives, was investigated for the suppression of damping-off of cucumber caused by Pythium ultimum. Compost tea was used to drench soilless container medium inoculated with P. ultimum; effect on damping-off ranged from not suppressive to consistently suppressive depending on the method used to produce the tea. The most consistent formulation for damping-off suppression was ACT produced with kelp and humic acid additives. Producing ACT with a molasses-based additive inconsistently suppressed damping-off; evidence suggests that residual nutrients can interfere with disease suppression. Heating or diluting compost tea negated suppression. Across all compost tea samples, there was no significant relationship of bacterial populations, measured as active cells, total cells, or CFU, to disease suppression. However, for all ACT produced without the molasses-based additive, there was a threshold of bacterial population density (6 log(10) active cells per ml, 7.48 log(10) total cells per ml, or 7 log(10) CFU per ml) above which compost teas were suppressive.     

Isolation and characterization of rhizobacteria from composts that suppress the severity of bacterial leaf spot of radish

ABSTRACT Composts can induce systemic resistance in plants to disease. Unfortunately, the degree of resistance induced seems highly variable and the basis for this effect is not understood. In this work, only 1 of 79 potting mixes prepared with different batches of mature, stabilized composts produced from several different types of solid wastes suppressed the severity of bacterial leaf spot of radish caused by Xanthomonas campestris pv. armoraciae compared with disease on plants produced in a nonamended sphagnum peat mix. An additional batch of compost-amended mix that had been inoculated with Trichoderma hamatum 382 (T(382)), which is known to induce systemic resistance in plants, also suppressed the disease. A total of 11 out of 538 rhizobacterial strains isolated from roots of radish seedlings grown in these two compostamended mixes that suppressed bacterial leaf spot were able to significantly suppress the severity of this disease when used as inoculum in the compost-amended mixes. The most effective strains were identified as Bacillus sp. based on partial sequencing of 16S rDNA. These strains were significantly less effective in reducing the severity of this disease than T(382). A combined inoculum consisting of T(382) and the most effective rhizobacterial Bacillus strain was less effective than T(382) alone. A drench applied to the potting mix with the systemic acquired resistance-inducing chemical acibenzolar-S-methyl was significantly more effective than T(382) in several, but not all tests. We conclude that systemic suppression of foliar diseases induced by compost amendments is a rare phenomenon. Furthermore, inoculation of compost-amended potting mixes with biocontrol agents such as T(382) that induce systemic resistance in plants can significantly increase the frequency of systemic disease control obtained with natural compost amendments.     

Survival of Phytophthora ramorum in Rhododendron root balls and in rootless substrates

This study assesses the survival of Phytophthora ramorum in the root ball of Rhododendron container plants as well as in different rootless forest substrates and a horticultural potting medium. Following inoculation of the root balls, the aboveground plant parts stayed symptomless, whilst the pathogen could be recovered with a novel non‐destructive baiting assay from the root balls until at least 8 months post‐inoculation. Plating of surface‐sterilized roots and direct microscopic analysis confirmed the presence of P. ramorum in the roots. Phytophthora ramorum could also be baited from the root balls of symptomless Rhododendron plants from commercial nurseries, even 2 years after acquisition. Survival of P. ramorum in rootless media was assessed after burying disks of infected leaf material below the soil surface in columns filled with four different undisturbed forest substrates or a potting medium, and incubated at an outdoor quarantine facility. Phytophthora ramorum could be recovered at least 33 months after burial from all substrates, with a significant increase in recovery after the winter period. These data suggest the possibility for long‐term symptomless presence of P. ramorum in root balls of commercial Rhododendron plants as well as survival in potting medium and different forest substrates under western European climate conditions. Symptomless presence in root balls can contribute to latent spread of this pathogen between nurseries. The novel baiting test, being non‐destructive, simple and applicable to a relatively large number of plants, can offer a valuable tool to test plants for the presence of Phytophthora species in root balls.     

Systemic Resistance Induced by Trichoderma spp.: Interactions Between the Host, the Pathogen, the Biocontrol Agent, and Soil Organic Matter Quality

ABSTRACT Several factors affect the ability of Trichoderma spp. to provide systemic disease control. This paper focuses on the role of the substrate in which plants are grown, resistance of the host to disease, and the ability of introduced Trichoderma inoculum to spread under commercial conditions. Several reports reveal that foliar disease control provided by Trichoderma spp. is more effective on plants grown in compost-amended media compared with in lower-in-microbial-carrying-capacity sphagnum peat media. In Rhododendron spp., host resistance affects control of Phytophthora dieback provided by Trichoderma spp. For example, T. hamatum 382 (T382) significantly (P = 0.05) suppressed the disease on susceptible cv. Roseum Elegans while plant vigor was increased. The disease was not suppressed, however, on highly susceptible cvs. Aglo and PJM Elite even though the vigor of these plants was increased. Using a strain-specific polymerase chain reaction assay under commercial conditions, it was demonstrated that introduced inoculum of T382 did not spread frequently from inoculated to control compost-amended media. Other Trichoderma isolates typically are abundant in control media within days after potting unless inoculated with a specific Trichoderma isolate. Thus, the low population of isolates that can induce systemic resistance in composting and potting mix environments may explain why most compost-amended substrates do not naturally suppress foliar diseases.     

Size, shape and intensity of aggregation of take-all disease during natural epidemics in second wheat crops

Point pattern analysis (fitting of the beta-binomial distribution and binary form of power law) was used to describe the spatial pattern of natural take-all epidemics (caused by Gaeumannomyces graminis var. tritici ) on a second consecutive crop of winter wheat in plots under different cropping practices that could have an impact on the quantity and spatial distribution of primary inoculum, and on the spread of the disease. The spatial pattern of take-all was aggregated in 48% of the datasets when disease incidence was assessed at the plant level and in 83% when it was assessed at the root level. Clusters of diseased roots were in general less than 1 m in diameter for crown roots and 1–1·5 m for seminal roots; when present, clusters of diseased plants were 2–2·5 m in diameter. Anisotropy of the spatial pattern was detected and could be linked to soil cultivation. Clusters did not increase in size over the cropping season, but increased spatial heterogeneity of the disease level was observed, corresponding to local disease amplification within clusters. The relative influences of autonomous spread and inoculum dispersal on the size and shape of clusters are discussed.     

Mutation of a degS homologue in Enterobacter cloacae decreases colonization and biological control of damping-off on cucumber

We have been using mutagenesis to determine how biocontrol bacteria such as Enterobacter cloacae 501R3 deal with complex nutritional environments found in association with plants. E. cloacae C10, a mutant of 501R3 with a transposon insertion in degS, was diminished in growth on synthetic cucumber root exudate (SRE), colonization of cucumber seed and roots, and control of damping-off of cucumber caused by Pythium ultimum. DegS, a periplasmic serine protease in the closely related bacterium Escherichia coli K12, is required for the RpoE-mediated stress response. C10 containing wild-type degS from 501R3 or from E. coli K12 on pBeloBAC11 was significantly increased in growth on SRE, colonization of cucumber roots, and control of P. ultimum relative to C10 containing pBeloBAC11 alone. C10 and 501R3 were similar in sensitivity to acidic conditions, plant-derived phenolic compounds, oxidative stress caused by hydrogen peroxide, dessication, and high osmoticum; stress conditions potentially associated with plants. This study demonstrates a role for degS in the spermosphere and rhizosphere during colonization and disease control by Enterobacter cloacae. This study implicates, for the first time, the involvement of DegS and, by extension, the RpoE-mediated stress response, in reducing stress on E. cloacae resulting from the complex nutritional environments in the spermosphere and rhizosphere.     

Differential interactions of <Emphasis Type="Italic">Verticillium longisporum</Emphasis> and <Emphasis Type="Italic">V. dahliae</Emphasis> with <Emphasis Type="Italic">Brassica napus</Emphasis> detected with molecular and histological techniques

The differential interactions of V. longisporum (VL) and V. dahliae (VD) on the root surface and in the root and shoot vascular system of Brassica napus were studied by confocal laser scanning microscopy (CLSM), using GFP tagging and conventional fluorescence dyes, acid fuchsin and acridin orange. VL and VD transformants expressing sGFP were generated by Agrobacterium-mediated transformation. GFP signals were less homogenous and GFP tagging performed less satisfactory than the conventional fluorescence staining when both were studied with CLSM. Interactions of both pathogens were largely restricted to the root hair zone. At 24 h post-inoculation (hpi), hyphae of VL and VD were found intensely interwoven with the root hairs. Hyphae of VL followed the root hairs towards the root surface. At 36 hpi, VL hyphae started to cover the roots with a hyphal net strictly following the grooves of the junctions of the epidermal cells. VL started to penetrate the root epidermal cells without any conspicuous infection structures. Subsequently, hyphae grew intracellularly and intercellularly through the root cortex towards the central cylinder, without inducing any visible plant responses. Colonisation of the xylem vessels in the shoot with VL was restricted to individual vessels entirely filled with mycelium and conidia, while adjacent vessels remained completely unaffected. This may explain why no wilt symptoms occur in B. napus infected with VL. Elevated amounts of fungal DNA were detectable in the hypocotyls 14 days post-inoculation (dpi) and in the leaves 35 dpi. Root penetration was also observed for VD, however, with no directed root surface growth and mainly an intercellular invasion of the root tissue. In contrast to VL, VD started ample formation of conidia on the roots, and was unable to spread systemically into the shoots. VD did not form microsclerotia in the root tissue as widely observed for VL. This study confirms that VD is non-pathogenic on B. napus and demonstrates that non-host resistance against this fungus materializes in restriction of systemic spread rather than inhibition of penetration.     

Biological control of damping-off of sugar beet by Pseudomonas putida applied to seed pellets

Pseudomonas putida 40RNF applied to seed pellets reduced the occurrence of Pythium damping-off of sugar beet. A density of 6 × 10⁷ 40RNF per pellet reduced Pythium damping-off from 70 to 26% when seeds were sown in artificially infested soil (250 propagules Pythium ultimum per g dry soil). The efficacy of 40RNF was dependent on its density in the seed pellet (in the range 2 × 10⁴–6 × 10⁸ per pellet) and on the number of propagules of Pythium in soil. 40RNF declined to or stabilized at approximately 1 × 10⁶ per pellet 3 days after planting, and this was independent of the inoculum density. This indicated that the crucial steps resulting in damping-off of sugar beet caused by Pythium ultimum must occur within 3–4 days of sowing. 40RNF reduced pericarp colonization by P. ultimum by 43% 48 h after planting and caused a 68% decrease in the number of sporangia of P. ultimum in the surrounding soil (0.0–5.0 mm). P. putida 40RNF also reduced pre and post-emergence damping-off (from 69.5 to 37.5%) caused by indigenous populations of Pythium species in an infested soil and this was as effective as the fungicide hymexazol (69.5 to 40%).