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.     

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.     

Effect of potting mix microbial carrying capacity on biological control of rhizoctonia damping-off of radish and rhizoctonia crown and root rot of poinsettia

ABSTRACT Potting mixes prepared with dark, highly decomposed Sphagnum peat, with light, less decomposed Sphagnum peat, or with composted pine bark, all three of which were colonized by indigenous microorganisms, failed to consistently suppress Rhizoctonia damping-off of radish or Rhizoctonia crown and root rot of poinsettia. Inoculation of these mixes with Chryseobacterium gleum (C(299)R(2)) and Trichoderma hamatum 382 (T(382)) significantly reduced the severity of both diseases in the composted pine bark mix in which both biocontrol agents maintained high populations over 90 days. These microorganisms were less effective against damping-off in the light and dark peat mixes, respectively, in which populations of C(299)R(2) declined. In contrast, crown and root rot, a disease that is severe late in the crop, was suppressed in all three types of mixes. High populations of T(382) in all three mixes late during the cropping cycle may have contributed to control of this disease.     

Compost and compost water extract-induced systemic acquired resistance in cucumber and Arabidopsis

ABSTRACT A biocontrol agent-fortified compost mix, suppressive to several diseases caused by soilborne plant pathogens, induced systemic acquired resistance (SAR) in cucumber against anthracnose caused by Colletotrichum orbiculare and in Arabidopsis against bacterial speck caused by Pseudomonas syringae pv. maculicola KD4326. A peat mix conducive to soilborne diseases did not induce SAR. The population size of P. syringae pv. maculicola KD4326 was significantly lower in leaves of Arabidopsis plants grown in the compost mix compared to those grown in the peat mix. Autoclaving destroyed the SAR-inducing effect of the compost mix, and inoculation of the autoclaved mix with nonautoclaved compost mix or Pantoea agglomerans 278A restored the effect, suggesting the SAR-inducing activity of the compost mix was biological in nature. Topical sprays with water extract prepared from the compost mix reduced symptoms of bacterial speck and the population size of pathogenic KD4326 in Arabidopsis grown in the peat mix but not in the compost mix. The peat mix water extract applied as a spray did not control bacterial speck on plants grown in either mix. Topical sprays with salicylic acid (SA) reduced the severity of bacterial speck on plants in the peat mix but did not further reduce the severity of symptoms on plants in the compost mix. The activity of the compost water extract was heat-stable and passed through a 0.2-mum membrane filter. beta-1,3-Glucanase activity was low in cucumber plants grown in either mix, but when infected with C. orbiculare, this activity was induced to significantly higher levels in plants grown in the compost mix than in plants grown in the peat mix. Similar results were obtained for beta-D-glucuronidase (GUS) activity driven by a PR2 (beta-1,3-glucanase) gene promoter in transgenic Arabidopsis plants grown in the compost or peat mix. GUS activity was induced with topical sprays of the compost water extract or SA in plants not inoculated with the pathogen, suggesting that compost-induced disease suppression more than likely involved the potentiation of resistance responses rather than their activation and that compost-induced SAR differed from SAR induced by pathogens, SA, or compost water extract.     

Suppression of Phytophthora cinnamomi in Potting Mixes Amended with Uncomposted and Composted Animal Manures

ABSTRACT We examined the effects of fresh and composted animal manures on the development of root rot, dieback, and plant death caused by Phytophthora cinnamomi. Fresh chicken manure, or chicken manure composted for 5 weeks before incorporation into the potting mix (25%, vol/vol), significantly reduced pathogen survival and the development of symptoms on Lupinus albus seedlings. Chicken manure composted for 2 weeks was less suppressive. Cow, sheep, and horse manure, whether fresh or composted, did not consistently suppress populations of P. cinnamomi or disease symptoms at the rates used (25%, vol/vol). All composts increased organic matter content, total biological activity, and populations of actinomycetes, fluorescent pseudomonads, and fungi. Only chicken manure stimulated endospore-forming bacteria, a factor that was strongly associated with seedling survival. Fallowing the potting mix for an additional 8 weeks after the first harvest increased the survival of lupin seedlings in a second bioassay, with survival rates in chicken manure compost-amended potting mix exceeding 90%. These data suggest that the ability of composted manure to stimulate sustained biological activity, in particular the activity of endospore-forming bacteria, is the key factor in reducing disease symptoms caused by P. cinnamomi. Supporting these results, the survival of rooted cuttings of Thryptomene calycina was significantly higher in sand-peat potting mix following amendment with commercially available chicken manure (15% vol/vol). However, this protection was reduced if the potting mix was steam pasteurized before amendment, indicating that suppression was due to endogenous as well as introduced microbes. Chicken manure compost incorporated at 5% (vol/vol) or more was strongly phytotoxic to young Banksia spinulosa plants and is not suitable as an amendment for phosphorus-sensitive plants.     

A rapid bioassay for screening rhizosphere microorganisms for their ability to induce systemic resistance

ABSTRACT We developed a rapid and miniaturized bioassay for screening large numbers of rhizosphere microorganisms for their ability to induce systemic resistance to bacterial leaf spot of radish caused by Xanthomonas campestris pv. armoraciae. In this bioassay, Pantoea agglomerans strain E278Ar controlled symptoms of disease as effectively as 2,6-dichloroisonicotinic acid when applied to the roots of seedlings produced in growth pouches in a soilless system. E278Ar essentially did not migrate from seedling roots to the foliage. This suggests that induction of systemic resistance could best explain the observed reduction in disease severity. Three mini-Tn5Km-induced mutants of strain E278Ar were isolated that had lost the ability to induce resistance. The bioassay also was used to demonstrate that the fungal biocontrol agent Trichoderma hamatum strain 382 induces systemic resistance in radish. The bioassay required only 14 to 18 days from seeding until rating for disease severity, which is 10 to 14 days less than earlier bioassays.     

Disease Progression by Active Mycelial Growth and Biocontrol of Pythium ultimum var. ultimum Studied Using a Rhizobox System

ABSTRACT This study demonstrates that outward growth of mycelium from primary foci through bulk potting mix to roots of adjoining plants can be an important means of spread of damping-off and root rot caused by Pythium ultimum. The use of a rhizobox system, which confines plant roots, enabled us to study the spread of actively growing mycelium between root systems placed at precise distances from each other. In steamed potting mix, hyphae of P. ultimum on average grew 9.6 cm from diseased root tissue compared to 5.3 cm in raw potting mix. The density of mycelium was highest within the first 2 cm from the infected root tissue, decreasing with increasing distances from the roots. Accordingly, the disease on adjacent plants decreased as the distance from infected roots increased. The time required for damping-off of adjacent plants was 3 days slower in raw as compared to steamed potting mix and increased by 2 days for each additional centimeter between the rhizoboxes. The presence of Trichoderma harzianum diminished the production of secondary inoculum and reduced the ability of P. ultimum hyphae to extend through bulk potting mix. In conclusion, the concentration of the primary inoculum, the plant density, the distance separating diseased from healthy roots, the resident microflora, and the presence of an antagonist were shown to be important factors affecting disease spread by mycelial growth.     

Systemic Modulation of Gene Expression in Tomato by Trichoderma hamatum 382

ABSTRACT A light sphagnum peat mix inoculated with Trichoderma hamatum 382 consistently provided a significant (P = 0.05) degree of protection against bacterial spot of tomato and its pathogen Xanthomonas euvesicatoria 110c compared with the control peat mix, even though this biocontrol agent did not colonize aboveground plant parts. To gain insight into the mechanism by which T. hamatum 382 induced resistance in tomato, high-density oligonucleotide microarrays were used to determine its effect on the expression pattern of 15,925 genes in leaves just before they were inoculated with the pathogen. T. hamatum 382 consistently modulated the expression of genes in tomato leaves. We identified 45 genes to be differentially expressed across the replicated treatments, and 41 of these genes could be assigned to at least one of seven functional categories. T. hamatum 382-induced genes have functions associated with biotic or abiotic stress, as well as RNA, DNA, and protein metabolism. Four extensin and extensin-like proteins were induced. However, besides pathogenesis-related protein 5, the main markers of systemic acquired resistance were not significantly induced. This work showed that T. hamatum 382 actively induces systemic changes in plant physiology and disease resistance through systemic modulation of the expression of stress and metabolism genes.     

Suppression of fusarium wilt of radish by co-inoculation of fluorescentPseudomonas spp. and root-colonizing fungi

In an earlier study, treatment of radish seed with the bacteriumPseudomonas fluorescens WCS374 suppressed fusarium wilt of radish (Fusarium oxysporum f. sp.raphani) in a commercial greenhouse [Leemanet al., 1991b, 1995a]. In this greenhouse, the areas with fusarium wilt were localized or expanded very slowly, possibly due to disease suppressiveness of the soil. To study this phenomenon, fungi were isolated from radish roots collected from the greenhouse soil. Roots grown from seed treated with WCS374 were more abundantly colonized by fungi than were roots from nonbacterized plants. Among these were several species known for their antagonistic potential. Three of these fungi,Acremonium rutilum, Fusarium oxysporum andVerticillium lecanii, were evaluated further and found to suppress fusarium wilt of radish in a pot bioassay. In an induced resistance bioassay on rockwool,F. oxysporum andV. lecanii suppressed the disease by the apparent induction of systemic disease resistance. In pot bioassays with thePseudomonas spp. strains, the pseudobactin-minus mutant 358PSB^– did not suppress fusarium wilt, whereas its wild type strain (WCS358) suppressed disease presumably by siderophore-mediated competition for iron. The wild type strains of WCS374 and WCS417, as well as their pseudobactin-minus mutants 374PSB^– and 417PSB^– suppressed fusarium wilt. The latter is best explained by the fact that these strains are able to induce systemic resistance in radish, which operates as an additional mode of action. Co-inoculation in pot bioassays, ofA. rutilum, F. oxysporum orV. lecanii with thePseudomonas spp. WCS358, WCS374 or WCS417, or their pseudobactin-minus mutants, significantly suppressed disease (except forA. rutilum/417PSB^– and all combinations with 358PSB^–), compared with the control treatment, if the microorganisms were applied in inoculum densities which were ineffective in suppressing disease as separate inocula. If one or both of the microorganism(s) of each combination were applied as separate inocula in a density which suppressed disease, no additional suppression of disease was observed by the combination. The advantage of the co-inoculation is that combined populations significantly suppressed disease even when their individual population density was too low to do so. This may provide more consistent biological control. The co-inoculation effect obtained in the pot bioassays suggests that co-operation ofP. fluorescens WCS374 and indigenous antagonists could have been involved in the suppression of fusarium wilt of radish in the commercial greenhouse trials.Abbreviations CFU colony forming units - KB King's B - PGPR plant growth-promoting rhizobacteria - CQ colonization quotient     

Induction of systemic resistance byPseudomonas fluorescens in radish cultivars differing in susceptibility to fusarium wilt,using a novel bioassay

Pseudomonas fluorescens-mediated induction of systemic resistance in radish against fusarium wilt (Fusarium oxysporum f. sp.raphani) was studied in a newly developed bioassay using a rockwool system. In this bioassay the pathogen and bacterium were confirmed to be confined to spatially separate locations on the plant root, throughout the experiment. Pathogen inoculum obtained by mixing peat with microconidia and subsequent incubation for four days at 22 °C, yielded a better percentage of diseased plants than a microconidial suspension drench, an injection of a microconidial suspension into the hypocotyl, or a talcum inoculum.Pseudomonas fluorescens strain WCS374 applied in talcum or peat, but not as a suspension drench, induced systemic resistance. A minimal initial bacterial inoculum density of 10⁵ CFU WCS374 root^–1 was required to significantly reduce the percentage diseased plants. At least one day was necessary between bacterization of strain WCS374 in talcum on the root tips and inoculation of the pathogen in peat on the root base, for an optimal induction of systemic resistance. Strain WCS374 induced systemic resistance in six radish cultivars differing in their susceptibility toF. oxysporum f. sp.raphani. Significant suppression of disease by bacterial treatments was generally observed when disease incidence in the control treatment, depending on pathogen inoculum density, ranged between approximately 40 to 80%. Strains WCS374 and WCS417 ofPseudomonas fluorescens induced systemic resistance against fusarium wilt, whereasP. putida WCS358 did not. This suggests that the induction of systemic resistance byPseudomonas spp. is dependent on strain-specific traits.Abbreviations CFU colony forming units - IFC immunofluorescence colony-staining - ISR induced systemic resistance - PBS phosphate buffered saline - SAR systemic acquired resistance     

Reduced severity and impact of Fusarium wilt on strawberry by manipulation of soil pH, soil organic amendments and crop rotation

Strawberry (Fragaria?×?ananassa) is one of the most important berry crops worldwide. Fusarium wilt poses a serious threat to commercial strawberry production worldwide and causes severe economic losses. Our previous surveys suggested that soil pH, soil amendment with organic matter and/or crop rotation could offer opportunities for improved management of strawberry disease. Studies were conducted for the first time to determine the effects of soil pH, soil amendments with manure compost and crop residue, and crop rotation on the severity and impact of Fusarium wilt on strawberry. At soil pH 6.7, plants showed the least severe disease and the lowest reductions in shoot and root dry weight (DW) of plants from disease, significantly lower than those of plants in acidic soil at pH 5.2 or 5.8. In soil amendment with manure compost at 5.0?%, plants showed the least severe disease and the lowest reductions in shoot and root DW of plants from disease, significantly lower than those of plants in the other three levels of manure compost. In soil amendment with crop residue at 2.5?% or 5.0?%, shoot and root disease of plants and reductions in shoot and root DW of plants from disease were significantly lower than those of plants in soil without crop residue or excessive crop residue amendment at 10.0?%. Plants in soil rotated with tomato not only showed the least severe disease but also showed the lowest reductions in shoot and root DW of plants from disease, significantly lower than those of plants in soil continuously planted with strawberry without rotation or rotated with capsicum. Soil pH, soil amendment with manure compost or crop residue, and crop rotation, all significantly reduced the severity and impact of Fusarium wilt on strawberry. There is great potential for manipulating soil pH, adding soil organic amendments and utilizing crop rotation, not only to successfully manage Fusarium wilt on strawberry, but to do so in a sustainable way without current reliance upon chemical fumigants.     

Evaluation of organic amendments from agro‐industry waste for the control of verticillium wilt of olive

Biological control of plant diseases using soil amendments such as animal manure and composted materials can minimize organic waste and has been proposed as an effective strategy in crop protection. In this study, 35 organic amendments (OAs) and 16 compost mixtures were evaluated against Verticillium dahliae by assessing both the antagonistic effect on the mycelial growth of two representative isolates of V. dahliae and the effect on the reduction of microsclerotia viability of the pathogen in naturally infested soil. Eleven OAs and five compost mixtures showed a consistent inhibition effect in in vitro sensitivity tests, with solid olive‐oil waste compost one of the most effective. Therefore, a bioassay with olive plants was conducted to evaluate the suppressive effect against V. dahliae of these selected OAs and compost mixtures. Significant reduction in the severity of the symptoms of V. dahliae indicates the potential use of grape marc compost (100% disease severity reduction) and solid olive‐oil waste, combined with other OAs. Microorganism mixtures and dairy waste OAs had a potential suppressive effect when they were combined with compost, showing a 73% and 63% disease severity reduction, respectively. A mixture of agro‐industrial waste with other biological control agents is a promising strategy against verticillium wilt of olive. To the authors' knowledge, this is the first report on the effectiveness of compost extracts (compost teas) on the inhibition of natural microsclerotia of V. dahliae, and also on verticillium wilt suppression in olive with solid olive‐oil waste.     

Plant responses to plant growth-promoting rhizobacteria

Non-pathogenic soilborne microorganisms can promote plant growth, as well as suppress diseases. Plant growth promotion is taken to result from improved nutrient acquisition or hormonal stimulation. Disease suppression can occur through microbial antagonism or induction of resistance in the plant. Several rhizobacterial strains have been shown to act as plant growth-promoting bacteria through both stimulation of growth and induced systemic resistance (ISR), but it is not clear in how far both mechanisms are connected. Induced resistance is manifested as a reduction of the number of diseased plants or in disease severity upon subsequent infection by a pathogen. Such reduced disease susceptibility can be local or systemic, result from developmental or environmental factors and depend on multiple mechanisms. The spectrum of diseases to which PGPR-elicited ISR confers enhanced resistance overlaps partly with that of pathogen-induced systemic acquired resistance (SAR). Both ISR and SAR represent a state of enhanced basal resistance of the plant that depends on the signalling compounds jasmonic acid and salicylic acid, respectively, and pathogens are differentially sensitive to the resistances activated by each of these signalling pathways. Root-colonizing Pseudomonas bacteria have been shown to alter plant gene expression in roots and leaves to different extents, indicative of recognition of one or more bacterial determinants by specific plant receptors. Conversely, plants can alter root exudation and secrete compounds that interfere with quorum sensing (QS) regulation in the bacteria. Such two-way signalling resembles the interaction of root-nodulating Rhizobia with legumes and between mycorrhizal fungi and roots of the majority of plant species. Although ISR-eliciting rhizobacteria can induce typical early defence-related responses in cell suspensions, in plants they do not necessarily activate defence-related gene expression. Instead, they appear to act through priming of effective resistance mechanisms, as reflected by earlier and stronger defence reactions once infection occurs.     

Evaluation of compost amendments for suppressiveness against Verticillium wilt of eggplant and study of mode of action using a novel Arabidopsis pathosystem

The induced resistance potential of eleven compost samples that originated from four different countries (Greece, France, Netherlands and Israel) and were manufactured from various raw materials, was evaluated in an Arabidopsis thaliana–Verticillium dahliae pathosystem under greenhouse conditions using a novel Plexiglas chamber. Five out of eleven composts tested showed significant disease suppressiveness compared to the control treatment; three composts exhibited disease severity equal to the control, while in the other three composts, disease severity was higher than the control treatment. Two of the tested composts that showed strong or medium suppressiveness were further evaluated under field conditions against Verticillium wilt of eggplant. Neither of them significantly reduced disease severity or resulted in higher fruit yield in a semi-commercial field test although they could induce a systemic resistance response in the greenhouse. However, as a consequence of a growth-promoting effect, one of the compost samples tested in the field resulted in a significant yield increase compared with the other.     

Biological control of Fusarium wilt by <Emphasis Type="Italic">Bacillus amyloliquefaciens</Emphasis> IUMC7 isolated from mushroom compost

Bacillus amyloliquefaciens IUMC7 isolated from mushroom compost inhibited growth of Fusarium oxysporum f. sp. lycopersici (FOL) on culture plates, and a culture supernatant of IUMC7 inhibited in vitro germ tube elongation of FOL. When compared with control soils, mushroom compost inoculated with IUMC7 significantly reduced disease severity caused by FOL in tomato plants. PCR tests for expression of PR genes indicated that IUMC7 did not induce resistance in tomato plants. These results suggested that the suppression of disease was mainly caused by antimicrobial compounds produced by IUMC7.     

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.     

Effects of the mycoparasite Verticillium biguttatum on barley stunt disease caused by Rhizoctonia solani anastomosis group 8 in a model system

The height of barley stunted by Rhizoctonia solani anastomosis group 8 was significantly increased by up to 72·8% after incubation for 8 days at 20°C in seedling tray tests following application of the mycoparasite Verticillium biguttatum. The pathogen and mycoparasite were applied at the rate of 1% Perlite maizemeal inoculum (w/w potting mixture) resulting in propagule densities of approximately 24·0 and 6·6 × 10⁵ colony-forming units (cfu) per g potting mixture, respectively. Sieving (2 mm) R. solani inoculum prior to dilution in potting mixture increased the recovery of propagules from 1·2 × 2·1 × 10³ cfu per g inoculum compared with recovery when inoculum was sieved after dilution. Applications of a V. biguttatum isolate from the UK (vbl) and a Dutch isolate (M73) reduced stunting to a similar extent but did not stimulate the growth of healthy plants. The height of stunted plants was significantly increased after application of V. biguttatum inoculum after 6 days if inoculated trays were preincubated for 1 day prior to planting but a similar increase was only detected after 7 days if seeds were planted immediately. The number of stunted plants which emerged after 4 days was significantly increased by treatment with V. biguttatum but preincubation had no additional effect. These results suggested that control of R. solani was effected both before and after the initiation of disease.     

Relationships Between Verticillium dahliae Inoculum Density and Wilt Incidence, Severity, and Growth of Cauliflower

ABSTRACT Microplot and field experiments were conducted to evaluate the effects of inoculum density on Verticillium wilt and cauliflower growth. Soil containing Verticillium dahliae microsclerotia was mixed with various proportions of fumigated soil to establish different inoculum densities (fumigated soil was used as the noninfested control). Seven inoculum density treatments replicated four times were established, and the treatments were arranged in a randomized complete block design. Soil was collected from each microplot immediately after soil infestation for V. dahliae assay by plating onto sodium polypectate agar (NP-10) selective medium using the Anderson sampler technique. Five-week-old cauliflower was transplanted into two beds within each 1.2- by 1.2-m microplot. At the same time, several extra plants were also transplanted at the edge of each bed for destructive sampling to examine the disease onset (vascular discoloration) after planting. Cauliflower plants were monitored for Verticillium wilt development. Stomatal resistance in two visually healthy upper and two lower, diseased leaves in each microplot was measured three times at weekly intervals after initial wilt symptoms occurred. At maturity, all plants were uprooted, washed free of soil, and wilt incidence and severity, plant height, number of leaves, and dry weights of leaves and roots were determined. The higher the inoculum density, the earlier was disease onset. A density of 4 microsclerotia per g of dry soil caused 16% wilt incidence, but about 10 microsclerotia per g of soil caused 50% wilt incidence. Both wilt incidence and severity increased with increasing inoculum density up to about 20 microsclerotia per g of soil, and additional inoculum did not result in significantly higher disease incidence and severity. A negative exponential model described the disease relationships to inoculum levels under both microplot and field conditions. Stomatal resistance of diseased leaves was significantly higher at higher inoculum densities; in healthy leaves, however, no treatment differences occurred. The height, number of leaves, and dry weights of leaves and roots of plants in the fumigated control were significantly higher than in infested treatments, but the effects of inoculum density treatments were variable between years. Timing of cauliflower infection, crop physiological processes related to hydraulic conductance, and wilt intensity (incidence and severity) were thus affected by the inoculum density. Verticillium wilt management methods used in cauliflower should reduce inoculum density to less than four micro-sclerotia per g of soil to produce crops with the fewest number of infected plants.     

Suppression of Rhizoctonia solani in Potting Mixtures Amended with Compost Made from Organic Household Waste

ABSTRACT Compost made from organic household and garden waste was used to substitute part of the peat in potting mixtures used for growing woody ornamental nursery stock. The effects of amendment with compost on the colonization of potting mixture by Rhizoctonia solani (AG1) were studied in greenhouse experiments. A bioassay was developed using cucumber as a sensitive herbaceous test plant as a substitute for woody ornamental cuttings. Pathogen growth in the potting mixture was estimated by measuring the distance over which damping-off of seedlings occurred. Compost from two commercial composting facilities suppressed growth of R. solani in potting mixtures with 20% of the product when the compost was fresh (directly after delivery) or long matured (after 5 to 7 months of additional curing). In contrast, short-matured compost (1 month of additional curing) from the same batches stimulated pathogen growth. In vitro mycelial growth of R. solani on mixtures with mature compost was inhibited by microbial antagonism. Compost-amended potting mixtures responded differentially to the addition of cellulose powder; the effect on suppressiveness depended on curing time and origin of the compost. In long-matured compost, suppressiveness to R. solani was associated with high population densities of cellulolytic and oligotrophic actinomycetes. The ratio of the population density of actinomycetes to that of other bacteria was around 200-fold higher in mature suppressive compost than in conducive compost.     

Nematicidal potential of materials from <Emphasis Type="Italic">Medicago</Emphasis> spp.

The nematicidal effect of soil amendments with dry top and root material from Medicago sativa and/or Medicago arborea was evaluated on the root-knot nematode Meloidogyne incognita and on the cyst nematode Globodera rostochiensis in potting mixes. All amendments suppressed root and soil population densities of both nematode species compared to non-treated and chemical controls. The suppressiveness of M. sativa differed between top and root material and among the amendment rates. In field conditions soil amendments with 20 or 40 t ha⁻¹ of a pelleted M. sativa meal increased tomato crop yield and reduced soil population densities and root galling by M. incognita. It is suggested that saponins were at least partly responsible for the nematicidal activity.