Tomato Fusarium wilt suppressiveness. The relationship between the organic plant growth media and their microbial communities as characterised by Biolog

Fusarium wilts are economically important diseases for which there are no effective chemical control measures. Biological control strategies are becoming efficient alternatives for controlling this disease. The suppressiveness to Fusarium oxysporum f. sp. lycopersici race 1 of grape marc compost and cork compost was evaluated in comparison to peat by using a susceptible cultivar of tomato (Lycopersicon esculentum cv. Marmande). Based on community level physiological profiles, different community structures were evident among the plant growth media evaluated. The peat microbial community, growth medium conducive to wilt, used mostly sugars, while those associated with both composts, the very suppressive grape marc and the moderately suppressive cork, used mostly carboxylic acids, amino acids, amines, phenolic compounds and polymers.     

Suppressiveness of 18 composts against 7 pathosystems: Variability in pathogen response

Compost is often reported as a substrate that is able to suppress soilborne plant pathogens, but suppression varies according to the type of compost and pathosystem. Reports often deal with a single pathogen while in reality crops are attacked by multiple plant pathogens. The goal of the present study was to evaluate the disease suppression ability of a wide range of composts for a range of plant pathogens. This study was conducted by a consortium of researchers from several European countries. Composts originated from different countries and source materials including green and yard waste, straw, bark, biowaste and municipal sewage. Suppressiveness of compost-amended (20% vol./vol.) peat-based potting soil was determined against Verticillium dahliae on eggplant, Rhizoctonia solani on cauliflower, Phytophthora nicotianae on tomato, Phytophthora cinnamomi on lupin and Cylindrocladium spathiphylli on Spathiphyllum sp., and of compost-amended loamy soil (20% vol./vol.) against R. solani on Pinus sylvestris and Fusarium oxysporum f. sp. lini on flax. From the 120 bioassays involving 18 composts and 7 pathosystems, significant disease suppression was found in 54% of the cases while only 3% of the cases showed significant disease enhancement. Pathogens were affected differently by the composts. In general, prediction of disease suppression was better when parameters derived from the compost mixes were used rather than those derived from the pure composts. Regression analyses of disease suppression of the individual pathogens with parameters of compost-amended peat-based mixes revealed the following groupings: (1) competition-sensitive: F. oxysporum and R. solani/cauliflower; (2) rhizosphere-affected: V. dahliae; (3) pH-related: P. nicotianae; and (4) specific/unknown: R. solani/pine, P. cinnamomi and C. spathiphylli. It was concluded that application of compost has in general a positive or no effect on disease suppression, and only rarely a disease stimulating effect.     

Identifying the characteristics of organic soil amendments that suppress soilborne plant diseases

Application of organic amendments has been proposed as a strategy for the management of diseases caused by soilborne pathogens. However, inconsistent results seriously hinder their practical use. In this work we use an extensive data set of 2423 studies derived from 252 papers to explore this strategy. First, we assess the capability of a specific organic amendment to control different diseases; second, we investigate the influence of organic matter (OM) decomposition on disease suppressiveness; and third, we search for physical, chemical and biological parameters able to identify suppressive OM. OM was found to be consistently suppressive to different pathogens in only a few studies where a limited number of pathogens were tested. In the majority of studies a material suppressive to a pathogen was ineffective or even conducive to other pathogens, suggesting that OM suppressiveness is often pathogen-specific. OM decomposition in many studies (73%, n = 426) emerged as a crucial process affecting suppressiveness. During decomposition, disease suppression either increased, decreased, was unchanged or showed more complex responses, such as ‘hump-shaped’ dynamics. Peat suppressiveness generally decreased during decomposition, while responses of composts and crop residues were more complex. However, due to the many interactions of contributing factors (OM quality, microbial community composition, pathosystem tested and decomposition time), it was difficult to identify specific predictors of disease suppression. Among the 81 parameters analysed, only some of the 643 correlations showed a consistent relationship with disease suppression. The response of pathogen populations to OM amendments was a reliable feature only for some organic matter types (e.g. crop residues and organic wastes with C-to-N ratio lower than ∼15) and for pathogens with a limited saprophytic ability (e.g., Thielaviopsis basicola and Verticillium dahliae). Instead, population responses of the pathogenic fungi Phytophthora spp., Rhizoctonia solani and Pythium spp. appeared unrelated to disease suppression. Overall, enzymatic and microbiological parameters, rather than chemical ones, were much more informative for predicting suppressiveness. The most useful features were FDA activity, substrate respiration, microbial biomass, total culturable bacteria, fluorescent pseudomonads and Trichoderma populations. We conclude that the integration of different parameters (e.g. FDA hydrolysis and chemical composition by ¹³C NMR) may be a promising approach for identification of suppressive amendments.     

Influence of soil type and indigenous pathogenic fungi on bean hypocotyl rot caused by Rhizoctonia solani AG4 HGI in Cuba

Calcisol, ferralsol and vertisol soils, representative of different bean production areas of Villa Clara province in Cuba, were selected to determine the impact of soil type on bean hypocotyl rot severity caused by Rhizoctonia solani AG4 HGI (isolate CuVC-Rs7). In inoculated autoclaved soil, hypocotyl rot was most severe in calcisol soil, followed by ferralsol soils and then vertisol soils. In inoculated natural soils, disease severity was lower in vertisol and calcisol soils and higher in ferralsol soil, indicating that biological factors are suppressing or stimulating the pathogenic efficiency of R. solani. Native binucleate Rhizoctonia AGF, Sclerotium rolfsii and R. solani AG 4 HGI were isolated from bean plants grown in natural calcisol, vertisol and ferralsol soils, respectively. Subsequent studies about the interaction between these fungi and R. solani indicated that they were involved in the variability of disease severity caused by R. solani. The addition of R. solani AG4 HGI (isolate CuVC-Rs7) into each autoclaved soil inoculated with binucleate Rhizoctonia or S. rolfsii resulted in a reduction of disease severity caused by this pathogen while in soils inoculated with native R. solani AG4 HGI, disease severity increased. Irrespective of fungal interactions, calcisol was always the most disease conducive soil and vertisol the most disease repressive soil. The mechanisms by which native pathogenic fungi could influence disease severity caused by R. solani are discussed.     

Effect of successive cauliflower plantings and Rhizoctonia solani AG 2-1 inoculations on disease suppressiveness of a suppressive and a conducive soil

Disease suppressiveness against Rhizoctonia solani AG 2-1 in cauliflower was studied in two marine clay soils with a sandy loam texture. The soils had a different cropping history. One soil had a long-term (40 years) cauliflower history and was suppressive, the other soil was conducive and came from a pear orchard not having a cauliflower crop for at least 40 years. These two soils were subjected to five successive cropping cycles with cauliflower or remaining fallow in a greenhouse experiment. Soils were inoculated with R. solani AG 2-1 only once or before every crop. Disease decline occurred in all treatments cropped with cauliflower, either because of a decreased pathogen population or increased suppressiveness of the soil. Disease suppressiveness tests indicated that the conducive soil became suppressive after five subsequent cauliflower crops inoculated each cycle with R. solani AG 2-1. Suppressiveness of all treatments was measured in a seed germination test (pre-emergence damping-off) as well as by measuring the spread of R. solani symptoms in young plants (post-emergence damping-off). Results showed that suppressiveness was significantly stimulated by the successive R. solani inoculations; presence of the cauliflower crop had less effect. Suppressiveness was of biological origin, since it disappeared after sterilization of the soil. Moreover, suppressiveness could be translocated by adding 10% suppressive soil into sterilized soil. The suppressive soil contained higher numbers of culturable filamentous actinomycetes than the conducive soil, but treatments enhancing suppressiveness did not show an increased actinomycetes population. The suppressiveness of the soil samples did also not correlate with the number of pseudomonads. Moreover, no correlation was found with the presence of different mycoparasitic fungi, i.e. Volutella spp., Gliocladium roseum, Verticillium biguttatum and Trichoderma spp. The suppressive soil contained a high percentage of bacteria with a strong in vitro inhibition of R. solani. These bacteria were identified as Lysobacter (56%), Streptomyces (23%) and Pseudomonas (21%) spp. A potential role of Lysobacter in soil suppressiveness was confirmed by quantitative PCR detection (TaqMan), since a larger Lysobacter population was present in suppressive cauliflower soil than in conducive pear orchard soil. Our experiments showed that successive cauliflower plantings can cause a decline of the damage caused by R. solani AG 2-1, and that natural disease suppressiveness was most pronounced after subsequent inoculations with the pathogen. The mode of action of the decline is not yet understood, but antagonistic Lysobacter spp. are potential key organisms.     

Screening of bacterial isolates from various European soils for in vitro antagonistic activity towards Rhizoctonia solani and Fusarium oxysporum: Site-dependent composition and diversity revealed

A cultivation-based approach was used to determine the in vitro antagonistic potential of soil bacteria towards Rhizoctonia solani AG3 and Fusarium oxysporum f. sp. lini (Foln3). Four composite soil samples were collected from four agricultural sites with previous documentation of disease suppression, located in France (FR), the Netherlands (NL), Sweden (SE) and the United Kingdom (UK). Similarly, two sites from Germany (Berlin, G-BR; and Braunschweig, G-BS) without documentation of disease suppression were sampled. Total bacterial counts were determined by plating serial dilutions from the composite soil samples onto R2A, AGS and King's B media. A total of 1,788 isolates (approximately 100 isolates per medium and site) was screened for antifungal activity, and in vitro antagonists (327 isolates) were found amongst the dominant culturable bacteria isolated from all six soils. The overall proportion of antagonists and the number of isolates with inhibitory activity against F. oxysporum were highest in three of the suppressive soils (FR, NL and SE). Characterization of antagonistic bacteria revealed a high phenotypic and genotypic diversity. Siderophore and protease activity were the most prominent phenotypic traits amongst the antagonists. The composition and diversity of antagonists in each soil was site-specific. Nevertheless, none of the antimicrobial traits of bacteria potentially contributing to soil suppressiveness analyzed in this study could be regarded as specific to a given site.     

Cause and duration of mustard incorporation effects on soil-borne plant pathogenic fungi

Two fungal plant pathogens, Rhizoctonia solani AG 2-2 and Fusarium oxysporum f.sp. lini, were studied in relation to general responses of soil fungi and bacteria following incorporation of Brassica juncea. Our aim was to understand to what extent the changes in the biological and physicochemical characteristics of the soil could explain the effects on the studied pathogens and diseases, and to determine the temporal nature of the responses. Short-term effects of mustard incorporation (up to 4 months) were investigated in a microcosm experiment, and compared with a treatment where composted plant material was incorporated. In a field experiment, the responses were followed up to 11 months after removal or incorporation of a mustard crop. In general, responses in the variables measured changed more after incorporation of fresh mustard material than after addition of similar amounts of composted plant material (microcosms) or after removal of the mustard crop (field). The soil inoculum potential of R. solani AG 2-2 decreased directly after incorporation of mustard, but increased later to disease levels above those in the untreated soil. Neither of these effects could be explained by changes in the population density of R. solani AG 2-2. Fusarium spp. were less influenced, although an increase in the suppressiveness to Fusarium wilt was observed after mustard incorporation as compared with the treatment where mustard was removed. The microbial responses to mustard incorporation were more pronounced for bacteria than for fungi. After an initial substantial increase, the bacterial density decreased but remained above the levels in the control treatment throughout the experimental periods. The bacterial community structure was modified up to 8 months after mustard incorporation. We conclude that incorporation of fresh mustard influences soil microbial communities, especially the bacteria, and has a potential to control the pathogenic activity of R. solani 2-2 on a short-term perspective. The time dependency in microbial responses is important and should be taken into consideration for the evaluation of the potential of Brassicas to control plant disease on a field scale.     

Brassica napus seed meal soil amendment modifies microbial community structure, nitric oxide production and incidence of Rhizoctonia root rot

A low glucosinolate content (21.8 μmol g⁻¹) Brassica napus seed meal (RSM) applied to orchard soils altered communities of both pathogenic and saprophytic soil micro-organisms. RSM amendment reduced infection by native and introduced isolates of Rhizoctonia spp. and recovery of Pratylenchus spp. from apple roots. Root infection by Rhizoctonia solani AG-5 was also suppressed in split-root assays where a portion of the root system was cultivated in RSM-amended soils and the remainder grown in the presence of the pathogen but lacking RSM. R. solani hyphal growth was not inhibited by RSM amendment. Suppression of Pratylenchus was attained to an equivalent extent by amending soils with either RSM or soybean meal (SM) when applied to provide a similar N content. Thus, glucosinolate hydrolysis products did not appear to have a significant role in the suppression of Rhizoctonia spp. or Pratylenchus spp. obtained via RSM amendment. RSM amendment elevated populations of Pythium spp. and of ammonia-oxidizing bacteria that release nitric oxide but suppressed fluorescent pseudomonad numbers. Streptomyces spp. soil populations increased significantly in response to RSM but not SM amendment. The vast majority of Streptomyces spp. recovered from the apple rhizosphere produced nitric oxide and possessed a nitric oxide synthase homolog. We propose that transformations in the bacterial community structure are associated with the observed control of Rhizoctonia root rot, with NO production by soil bacteria potentially having a role in the induction of plant systemic resistance.     

Variation of Pythium-induced cocoyam root rot severity in response to soil type

In Cameroon, andosols are suspected to be suppressive to cocoyam (Xanthosoma sagittifolium) root rot disease (CRRD) caused by the Oomycete pathogen Pythium myriotylum. To determine factors involved in disease suppressiveness, andosols were studied in comparison to ferralsols known to be disease-conducive. Soil samples were collected from six sites of which three were in andosols around Mount Cameroon (Boteva, Njonji, and Ekona) and the three others in ferralsols (Bakoa, Lapkwang, and Nko’o canane). Greenhouse plant experiments were used to assess soil suppressiveness. Soils were artificially infested with two levels of P. myriotylum inoculum (100 and 300 mycelia strands g⁻¹ soil) prior to planting cocoyam. Disease severity was significantly higher in ferralsols than in andosols. Andosols partly lost their suppressiveness as a result of autoclaving and could recover suppressiveness following recolonisation by their original microflora. Soil microbial groups implicated in the disease suppression were investigated by assessing the effect of fungicide, bactericide, and pasteurisation on andosol suppressiveness. Andosols suppressiveness was significantly reduced following pasteurisation and treatment with fungicide and bactericide. The possible influence of microbial biomass on andosol suppressiveness was investigated by comparing microbial populations of suppressive andosols to those in andosols that had lost suppressiveness. A comparative analysis of suppressive and conducive soil properties was performed to identify soil variables, which may contribute to soil suppressiveness. Soil chemical analysis results showed that organic matter content was higher in andosols than in ferralsols. In addition, the content of mineral nutrients such as Ca, K, Mg and N, was higher in andosols than in ferralsols. These soil variables negatively correlated with disease severity. By contrast, sand and clay, which were higher in ferralsols than in andosols, were positively related to disease severity. This study has confirmed the suppressive nature of andosols from Mount Cameroon to CRRD. The results suggest that high organic matter content is likely mediating P. myriotylum suppression in andosols by improving soil structure, increasing soil nutrient content and microbial biomass, and sustaining microbial activity.     

Re-establishment of suppressiveness to soil- and air-borne diseases by re-inoculation of soil microbial communities

The aim of this study was to investigate the potentials and limitations in restoring soil suppressiveness in disturbed soils. Soils from three sites in UK and Switzerland (STC, REC, THE) differing in their level of suppressiveness to soil-borne and air-borne diseases were γ-irradiated and this soil matrix was re-inoculated with 1% (w/w) of either parent native soil or native soil from the other sites (‘soil inoculum’). Suppressiveness to air-borne and soil-borne diseases was quantified by means of the host-pathogen systems Lepidium sativum (cress)-Pythium ultimum, an oomycete causing root rot and seedling damping-off, and Arabidopsis thaliana-Hyaloperonospora parasitica, an oomycete causing downy mildew. Soil microbial biomass, activity and community structure, as determined by phospholipid fatty acid (PLFA) profiles, were measured in native, γ-irradiated, and re-inoculated soils. Both, L. sativum and A. thaliana were highly susceptible to the pathogens if grown on γ-irradiated soils. Re-inoculation completely restored suppressiveness of soils to the foliar pathogen H. parasitica, independently of soil matrix or soil inoculum, whereas suppressiveness to P. ultimum depended on the soil matrix and, to a lesser extent, on the soil inoculum. However, the soil with the highest inherent suppressiveness did not reach the initial level of suppressiveness after re-inoculation. In addition, native microbial populations as defined by microbial biomass, activity and community structure, could not be fully restored in re-inoculated soils. As for suppressiveness to P. ultimum, the soil matrix, rather than the source of soil inoculum was identified as the key factor for re-establishing the microbial community structure. Our data show that soils do not or only slowly fully recover from sterilisation by γ-irradiation, indicating that agricultural soil management practices such as soil fumigation or heat treatments frequently used in vegetable cropping should be avoided.     

Compost suppressiveness against Fusarium oxysporum was not reduced after one-year storage under various moisture and temperature conditions

The effect of storage conditions on compost suppressiveness against fusarium wilt of melon, caused by Fusarium oxysporum f. sp. melonis (FOM) was studied in relation to the dynamics of compost microbial activity and biodegradability. For this purpose, mature suppressive compost, prepared from tomato plants and separated cow manure, was divided into four portions and stored for one year under cool/warm (12 or 28 °C) or dry/wet (15-35 or 55-65% moisture content) conditions, in four different combinations: cool-dry, warm-dry, cool-wet and warm-wet. All composts retained and even enhanced their suppressive capacity during storage, with no significant differences among them by the end of the storage period. However, significant differences were found in the dynamics of some of the measured chemical and microbial properties. The microbial activity of composts stored under wet conditions was higher than that of those stored under dry condition, which resulted in a substantial decrease in dissolved organic matter content (expressed as dissolved organic carbon; DOC) and increase in its recalcitrance to biological degradation, decrease in basal heat emission, slower response to added glucose or citric acid, and higher NO₃ concentration, indicating increased nitrification under wet conditions. The DOC significantly correlated with several microbial properties as well as with compost suppressiveness of fusarium wilt of melon seedlings, and may be regarded as a most suitable general index for compost maturity. A best-subset multiple linear regression analysis revealed that the three best predictors, namely dissolved organic carbon (DOC), basal heat, and mesophilic bacterial counts, could explain as much as 83% of the total variance in compost suppressiveness. The generally agreed association between compost maturity and suppressiveness was verified in this case. It appears that compost microbial populations might compete and interfere with the saprophytic stage of FOM conidia, between germination and host invasion. In conclusion, it was demonstrated that compost suppressiveness against fusarium wilt of melon can be maintained for at least one year under a wide range of storage conditions, without any loss of suppressive capacity. This fact has positive logistical implications for the use of suppressive composts against FOM.     

Evaluation of different Coniothyrium minitans inoculum sources and application rates on apothecial production and infection of Sclerotinia sclerotiorum sclerotia

The effects of three Coniothyrium minitans isolates (Conio, IVT1 and Contans^®), applied to soil as conidial suspensions or as maizemeal-perlite (MP) inocula (Conio), on apothecial production and infection of Sclerotinia sclerotiorum sclerotia were assessed in two soil pot bioassays and two novel box bioassays in the glasshouse at different times of the year. C. minitans isolate Conio applied as either MP or ground MP at full rate (10⁶-10⁷ cfu cm⁻³ soil) consistently decreased the carpogenic germination, recovery and viability of sclerotia and increased C. minitans infection of the sclerotia of S. sclerotiorum by in comparison with either MP or conidial suspension treatments applied at lower rates (10³-10⁴ cfu cm⁻³ soil). Additionally, when applied at the same rate, MP inoculum of C. minitans was consistently more effective at reducing carpogenic germination than a conidial suspension. The effect of MP and ground MP at full rate on carpogenic germination was expressed relatively early as those sclerotia recovered before apothecia appeared on the soil surface already had reduced numbers of apothecial initials. In general, there were few differences between the isolates of C. minitans applied as conidial suspensions. Box bioassays carried out at different times of the year indicated that temperature and soil moisture influenced both apothecial production and mycoparasitism. Inoculum concentration of C. minitans and time of application appear to be important factors in reducting apothecial production by S. sclerotiorum.     

Host status of 10 fungal isolates for two nematode species, Filenchus misellus and Aphelenchus avenae

The classification of nematodes in the family Tylenchidae into plant parasites, plant associates or fungal-feeders for community analyses, have been much discussed by nematode ecologists. For an appropriate classification, fungal-feeding habits in the family need to be studied. To evaluate the host status of 10 fungal isolates for Filenchus misellus (Tylenchidae) and Aphelenchus avenae (Aphelenchida, Aphelenchidae), population growth rates, body length and width and sex ratios of the nematodes were measured after 40-day culture on fungal colonies at 25 °C. For F. misellus, the fungi determined as good hosts were two Basidiomycota fungi (Agaricus bisporus, Coprinus cinereus), three Ascomycota fungi (Chaetomium cochlioides, Chaetomium funicola, Chaetomium globosum) and a plant-pathogenic fungus (Rhizoctonia solani) on the basis of nematode population growth rate and female body length. Interestingly Pleurotus ostreatus, known as a predaceous fungus for the other nematodes, was also a good host for F. misellus. While, for A. avenae, good hosts were four plant-pathogenic fungi (Fusarium oxysporum f. sp. conglutinans, F. oxysporum f. sp. cucumerinum, Pythium ultimum, R. solani) and A. bisporus. A. avenae was trapped and preyed upon by Pleurotus hyphae. In F. misellus, males were 7-21% of adults, but the ratio did not correlate significantly with the population growth rate. In A. avenae, no male occurred. Differences in habitat preference between Filenchus and Aphelenchus were explained on the basis of the host status and habitat preferences of the tested fungi.     

Increased pepper yields following incorporation of biofumigation cover crops and the effects on soilborne pathogen populations and pepper diseases

The use of brassica cover crops and their associated degradation compounds as biofumigants to manage soilborne pathogens could offer vegetable growers an alternative to the restricted broad-spectrum fumigant methyl bromide. Biofumigation was tested in two experiments to manage Rhizoctonia solani, Sclerotium rolfsii, and Pythium spp., and the diseases they cause on pepper. Field plots were seeded in fall 2010 and spring 2011 to oilseed radish (Raphanus sativus L.), ‘Pacific Gold’ mustard (Brassica juncea (L.) Czern), or ‘Dwarf Essex’ winter rapeseed (Brassica napus L.). Cover crops were disked into soil in spring 2011 and immediately covered with virtually impermeable film (VIF) to reduce the escape of volatile pesticidal compounds. Controls included fallow plots with (CVIF) and without (fallow) VIF. Green bell pepper was transplanted into all plots. Concentrations of isothiocyanates (ITCs), the brassica degradation compounds primarily responsible for pesticidal activity, were highest following incorporation of mustard. Rapeseed yielded the second highest ITC concentrations. Radish yielded very low ITC concentrations in experiment 1, and none during experiment 2. ITCs also were detected in low concentrations in CVIF treatments. All treatments that received VIF reduced populations of R. solani compared to fallow, with no differences between biofumigation treatments and CVIF. Biofumigation treatments did not reduce populations of Pythium spp. or S. rolfsii compared to CVIF. Pepper stunting was significantly lower in treatments that received VIF compared to fallow, with no consistently significant differences between biofumigation treatments and CVIF. Pythium isolated from roots of stunted peppers was identified as Pythium aphanidermatum. Biofumigation treatments did not reduce plant mortality. Pepper yields were highest in biofumigation treatments compared to CVIF, and CVIF yields were higher than fallow yields.     

Bio-indicators to assess impacts of heavy metals in land-applied sewage sludge

Soils from a pastoral farm that had received large amounts of heavy metal contaminated sewage sludge 6-10 years previously were investigated to determine the impact of heavy metals on Rhizobium. The 8 ha application area was originally divided into five different-sized blocks (blocks 1-5), which received sludge at different times between 1991 and 1994. The response of a lux biosensor based on R. leguminosarum bv. trifolii (Rhizotox-C), was compared with more traditional techniques for measuring the presence of effective strains of Rhizobium (MPN) and nitrogen fixation (δ¹⁵N natural abundance). Although population size (MPNs), nitrogen fixation and biosensor response varied between treatment blocks, linear regression analysis determined that this block effect could not be directly linked to soil heavy metal concentrations, but was probably due to biological, physical, chemical and environmental compounding factors at the site. In this type of uncontrolled field application, the lux bioassay may provide the most useful information as it measures toxicity to any microorganism exposed to the soil solution, for example, the free living rhizobia.     

Suppression of soil-borne pathogens of tomato by composts derived from agro-industrial wastes abundant in Mediterranean regions

We studied nine composts derived from wastes and by-products of the olive oil, wine, and Agaricus mushroom agro-industries. They were mixed with peat at 1:3 w w ⁻¹ ratios and comparatively evaluated in pot experiments to assess suppressiveness against soil-borne and foliar pathogens of tomato. All compost amendments demonstrated high levels of suppressiveness against Phytophthora nicotianae Breda de Haan in tomato, when they were applied directly after curing (T0) indicating the occurrence of a “general suppression phenomenon” (81–100% decrease in plant disease incidence). They were, however, relatively less effective when applied 9 months after curing (T1, 55–100% disease decrease). Suppressiveness against Fusarium oxysporum f.sp. radicis-lycopersici Jarvis & Shoemaker was relatively lower and varied widely among composts (8–95% and 22–87% decrease in plant disease incidence for T0 and T1, respectively). Three of the composts conferred induced systemic resistance against the foliar pathogen Septoria lycopersici Speg. Biotic properties were determined, including respiration, fluorescein diacetate hydrolysis, and β-glucosidase activity of composts. The comparative evaluation of the nine composts revealed no shared critical biotic or abiotic characteristics indicative of their suppressive effects on the soil-borne and foliar pathogens. The complex origin of compost suppressiveness is discussed and the implementation of individual evaluation of each compost product for a specific use is advocated.     

Calcium concentrations of soil affect suppressiveness against Aphanomyces root rot of pea

The potential for field soils to cause Aphanomyces root rot of pea (Pisum sativum) was estimated for a large number of samples from commercial pea fields over a period of 5 years, using a greenhouse bioassay. The aim of the research project was to gain a mechanistic understanding of soil suppressiveness to the disease. Regression analysis showed that of the measured soil variables (Ca, Mg, K, P, pH), soil Ca concentrations had the strongest (negative) correlation with disease prevalence, and also a significant negative correlation with disease severity in samples with confirmed presence of the disease. Greenhouse bioassays using a set of non-infested soils inoculated with artificially produced oospore inoculum of the casual organism Aphanomyces euteiches, showed a similar negative correlation between soil Ca content and disease severity. Disease severity was not consistently affected by soil sterilisation, but was lowered by the addition of two different Ca salts. In contrast, addition of sodium bicarbonate to two soils lowered the content of water-soluble Ca in the soils and increased disease severity. Studies of cultures of A. euteiches exposed to varying Ca concentrations in vitro showed that zoospore production was inhibited at submillimolar concentrations, while mycelial growth was stimulated or unaffected. We conclude that free Ca is a major variable controlling the degree of soil suppressiveness against A. euteiches, and that inhibition of zoospore production from oospores is a possible mechanism.     

Potential of Azotobacter spp. as biocontrol agents against Rhizoctonia solani and Fusarium oxysporum in cotton (Gossypium hirsutum), guar (Cyamopsis tetragonoloba) and tomato (Lycopersicum esculentum)

During the current investigation, 51 bacterial isolates/mutants of Azotobacter chroococcum, Azospirillum spp. and Gluconacetobacter diazotrophicus were tested for antifungal activity against three fungal pathogens, namely Rhizoctonia solani cotton, Rhizoctonia solani rice and Fusarium oxysporum tomato using a dual-culture technique under laboratory conditions. Ten isolates/mutants were found to be inhibitory against R. solani cotton, six against R. solani rice and twelve against F. oxysporum tomato. Nearly 50% of the isolates/mutants were siderophore positive. The antimicrobial substance was found to be extracellular and proteinaceous in nature, but in some strains it was found to be associated with other complex material also. Almost all the positive isolates/mutants showed production of antibiotic and antifungal compounds. On the basis of this study, in vitro results under laboratory conditions were reproduced in pot experiments in the greenhouse and chosen isolates/mutants of A. chroococcum were found to be effective biocontrol agents against R. solani cotton (disease index 16.7%) and R. solani rice (2.5%) in cotton crop, whereas in guar, the crop disease index was 2.5% against R. solani cotton, 13.3% against R. solani rice and 0% against F. oxysporum tomato in tomato crop compared with their respective checks.     

Microbial enrichment to enhance the disease suppressive activity of compost

Compost amended soil has been found to be suppressive against plant diseases in various cropping systems. The level and reproducibility of disease suppressive properties of compost might be increased by the addition of antagonists. In the present study, the establishment and suppressive activity of two fungal antagonists of soil-borne diseases was evaluated after their inoculation in potting soil and in compost produced from different types of organic waste and at different maturation stages. The fungal antagonists Verticillium biguttatum, a mycoparasite of Rhizoctonia solani, and a non-pathogenic isolate of Fusarium oxysporum antagonistic to fusarium wilt, survived at high levels (10³–10⁵ CFU g^–1) after 3 months incubation at room temperature in green waste compost and in potting soil. Their populations faded out in the organic household waste compost, especially in the matured product. In bioassays with R. solani on sugar beet and potato, the disease suppressiveness of compost increased or was similar after enrichment with V. biguttatum. The largest effects, however, were present in potting soil, which was very conducive for the disease as well as the antagonist. Similar results were found in the bioassay with F. oxysporum in carnation where enrichment with the antagonistic F. oxysporum had a positive or neutral effect. We foresee great potential for the application of antagonists in agriculture and horticulture through enrichment of compost or potting soil with antagonists or other beneficial micro-organisms.     

Biological control of damping-off caused by Rhizoctonia solani using chitinase-producing Paenibacillus illinoisensis KJA-424

A bacterium having strong chitinolytic activity was isolated from a coastal soil in Korea and identified as Paenibacillus illinoisensis KJA-424 on the basis of the nucleotide sequence of a 16S rRNA gene. By activity staining after SDS-PAGE, three major chitinase bands with chitinolytic activity, approximate molecular weight of 63, 54 and 38 kDa were detected. On co-culture Rhizoctonia solani with KJA-424, abnormal swelling and deformation of R. solani hyphae were observed, where the release of N-acetyl-d-glucosamine was detected. The bacterium suppressed the symptom of damping-off cucumber seedlings caused by R. solani, in greenhouse trial.