Biological control of Rhizoctonia solani on potatoes by antagonists. 1. Preliminary experiments with Verticillium biguttatum,a sclerotium-inhabiting fungus

A common mycoparasite,Verticillium biguttatum, was found to kill sclerotia ofRhizoctonia solani placed on an inert material (perlite) as well as in soil at 15°C and 20°C, but not at 10°C. Compared with the effectivity ofV. biguttatum, that ofGliocladium roseum, Gliocladium nigrovirens, Hormiactis fimicola andTrichoderma hamatum on sclerotia was only low. In laboratory experiments, treatment of sclerotia-bearing seed potatoes withV. biguttatum reduced disease symptoms in the first stage of growth of the potato plant.V. biguttatum was found to occur on the subterranean part of the potato plant. On untreated plants the surface of the sprouts was colonised byV. biguttatum originating from the soil, presumably partly in response to the presence ofR. solani mycelium. In a preliminary field experiment,Verticillium treatment did not reduce symptoms on the stem. However, there was a marked reduction in sclerotium formation on the newly formed potato tubers. This offers perspectives for a commercial use ofV. biguttatum in the control ofR. solani.     

Nutritional requirements of the mycoparasitic fungus Verticillium biguttatum

Verticillium biguttatum was able to grow axenically in a synthetic liquid medium with a compound containing ammonium or amino group as nitrogen source, glucose as carbon source and biotin as growth factor. Among various carbon and nitrogen compounds tested, highest mycelial production was achieved with mannitol and with two ammonium salts and glutamine; sporulation reached highest values with galactose and glutamine. Highest yields of mycelium and conidia were obtained at pH 4.3 and 5.1, respectively. Although neutral and alkaline conditions were growth-limiting in the synthetic medium some growth ofV.biguttatum occurred on solid media at pH 7.0 and on sclerotia ofRhizoctonia solani in natural soil at pH 7.2–7.3.Samenvatting Een synthetisch vloeibaar medium met glucose als koolstofbron, een ammonium- of aminogroep bevattende verbinding als stikstofbron en biotine als groeifactor voldeed aan de voedingseisen vanVerticillium biguttatum. Van de verschillende koolstof- en stikstofbronnen leverden mannitol en twee ammoniumzouten en glutamine de hoogste opbrengsten aan mycelium; de opbrengst aan conidiën was het hoogst met galactose en glutamine. De hoogste opbrengsten aan mycelium en conidiën werden bereikt bij respectievelijk pH 4,3 en 5.1. Ofschoon neutrale en alkalische omstandigheden de groei vanV. biguttatum in het synthetische medium beperkten, werd enige groei vanV. biguttatum waargenomen op vaste voedingsbodems bij pH 7,0 en op sclerotia vanR. solani in natuurlijke grond bij pH 7,2–7,3.     

Dynamics ofRhizoctonia solani (black scurf) in successive potato crops

The position of plants withRhizoctonia solani sclerotia (black scurf) on progeny tubers was mapped for an experimental field at Haren where potatoes were grown continuously and in rotation with other crops for five successive years, and for another field at Borgercompagnie with a 12 frequency of potatoes during three potato crops. Initially, the distribution of plants with black scurf on both fields was rather dense and homogeneous. In the following years the distribution became heterogeneous and patchy. The local decline ofR. solani AG 3 (the common potato pathogen) in Haren was apparently caused by an unknown factor selectively suppressingR. solani AG 3, while simultaneouslyR. solani AG 5 increased in mass. This AG 5 type proved to be an inferior competitor of AG 3 on the potato plant in a laboratory experiment. The specificR. solani antagonistVerticillium biguttatum did not play a role. A similar factor could have reduced the formation of black scurf in the experimental field at Borgercompagnie, whereV. biguttatum was also too infrequent to account for the decline.R. solani AG 5 was not present here and could not indicate the presence of a selective factor against AG 3.     

Biological control of Rhizoctonia solani on potatoes by antagonists. 3. Inoculation of seed potatoes with different fungi

Inoculation of seed potatoes withVerticillium biguttatum and three other hyperparasitic fungi, alone or in combination, resulted in statistically significant reduction of infestation of potato plants byRhizoctonia solani. Gliocladium roseum, Trichoderma hamatum andHormiactis fimicola did not show prolonged protection againstR. solani under farming conditions.H. fimicola, however, inhibited mycelial growth ofR. solani in vitro, particularly in the lower temperature range whereV. biguttatum did not show any growth. Combining these two antagonistic fungi may be advantageous as they cover the entire temperature range in whichR. solani is active. At the end of the vegetation period,V. biguttatum was superseded byG. roseum.Production of sclerotia on newly formed tubers from seed potatoes inoculated withV. biguttatum (alone or in combination with the other three antagonists) was significantly reduced.Samenvatting Inoculative van pootaardappelen metVerticillium biguttatum, apart of sament met drie andere hyperparasieten, had een gunstig effect op het onderdrukken vanRhizoctonia solani op de plant;Trichoderma hamatum, Gliocaldium roseum enHormiactis fimicola, ieder apart toegediend, boden de plant op lange termijn geen bescherming tegenR. solani onder praktijkomstandigheden.H. fimicola bleek bij lage temperatuur, waarbijV. biguttatum geen groei-activiteit meer vertoonde, op hyfen en sclerotiën vanR. solani te kunnen groeien. Toepassing van deze schimmel enV. biguttatum in een gemengde inoculatie zou over een breder temperatuurtraject effectief kunnen zijn.Tegen het einde van het groeiseizoen vond er op de ondergrondse plantedelen een verschuiving plaats, waarbijG. roseum meer op de voorgrond trad. Verondersteld wordt dat de afname vanV. biguttatum op stolonen hiervan een gevolg was.De produktie van sclerotia op nieuwe aardappelen afkomstig van metV. biguttatum behandeld pootgoed (alleen of met andere hyperparasitaire schimmels) bleek sterk verminderd te zijn. Vooral dit gegeven maakt de biologische bestrijding vanR. solani interessant:V. biguttatum blijkt ook op lange termijn effectief te zijn.     

Biological control of Rhizoctonia solani on potatoes by antagonists. 2. Sprout protection against soil-borne R. solani through seed inoculation with Verticilium biguttatum

Verticillium biguttatum isolate M73, when inoculated on the seed tuber, was found to protect young potato sprouts againstRhizoctonia infection from the soil and also, as was demonstrated before, against infection from sclerotia on seed tubers. The experiments were performed under laboratory conditions and lasted a few weeks. V. biguttatum colonizes the surface of the sprouts and may be regarded as a natural inhabitant of the surface of sprouts and presumably also of stems and stolons in a later stage. This fungus may be of value for biological control ofR. solani.Samenvatting Isolaat M73 van de schimmelVerticillium biguttatum, indien geënt op de pootknol, blijkt de aantasting van aardappelspruiten doorRhizoctonia solani te kunnen verminderen. Zowel de aantasting vanuit de grond en, zoals reeds eerder aangetoond, vanaf sclerotiën op de pootknol worden minder. Verticillium biguttatum koloniseert het oppervlak van de spruiten en kan beschouwd worden als een natuurlijke bewoner van het oppervlak van spruiten en wellicht ook van stengels en stolonen in een later stadium. In kort durende proeven, als hier beschreven, lijktV. biguttatum goed bruikbaar voor een biologische bestrijding vanR. solani.     

Biological destruction of conidia ofVerticillium biguttatum

Because biological control ofRhizoctonia solani in potato with conidial suspensions of the mycoparasiteVerticillium biguttatum was often less successful in sandy soils than in loamy soils, we examined soils of potato fields for the presence of organisms destructive to conidia ofV. biguttatum.Representatives of conidiophagous testate amoebae were frequently present on sclerotium disks ofR. solani infected withV. biguttatum in all soils studied and were most active under moist conditions. Conidiophagous naked amoebae were also numerous, except for two loam soils, and were not sensitive to moist conditions. Conidiophagous ciliates were found in rather low numbers and were most frequently isolated from coarsely structured soils under moist conditions. Conidiophagous flagellates were very infrequently observed.A bacterial type, parasitizing and killing conidia and hyphae ofV. biguttatum, was observed in all soils studied. It produced clusters of cocci fixed to the outside of conidia and hyphae and was most active under moist soil conditions.The possible role of protozoan predators and bacterial parasites in the biological control ofR. solani in potato withV. biguttatum applied at planting is discussed.     

Biotrophic mycoparasitism byVerticillium biguttatum onRhizoctonia solani

Verticillium biguttatum cannot utilise cellulose or nitrate-nitrogen and it requires biotin for growth, yet it grew and sporulated abundantly onRhizoctonia solani on cellulose, obtaining at least organic carbon, nitrogen and biotin fromR. solani. Videomicroscopy of inter-hyphal interactions on films of water agar showed thatV. biguttatum behaved as a biotrophic mycoparasite. From germinating spores, it penetrated the hyphae ofR. solani and formed haustorium-like branches without killing the host cells, and the haustoria supported an external mycelial network of the mycoparasite. Later the mycoparasite sporulated, and the infected host cells died. On cellulosic substrataV. biguttatum did not reduce the growth ofR. solani, and often enhanced the rate of cellulose degradation. However,V. biguttatum drastically reduced the production of sclerotia byR. solani, often completely suppressing sclerotium production when the mycoparasite infected only a localized region of the host colony. This is ascribed to the creation of a nutrient sink by the parasite, consistent with biotrophy. On plates of cellulose agar the suppression of sclerotia was not confined to parasitized colonies but extended to adjacent colonies ofR. solani that had successfully anastomosed with the parasitized colony. There was no effect on adjacent vegetatively incompatible colonies, where attempted anastomoses caused cytoplasmic death. In comparable experiments the necrotrophic mycoparasiteGliocladium roseum had no long-distance effect on sclerotium production byR. solani.Suppression of sclerotium production may explain the reported success ofV. biguttatum in biocontrol of black scurf of potato in experimental field conditions.     

Biological control of Rhizoctonia solani on potatoes by antagonists. 4. Inoculation of seed tubers with Verticillium biguttatum and other antagonists in field experiments

Inoculation of seed potatoes with the mycoparasiteVerticillium biguttatum, isolate M73 (combined withGliocladium roseum in 1981, either alone or mixed with isolate M180 plus antibiotics-producing isolates ofAzotobacter chroococcum in 1982) repeatedly proved successful in reducingRhizoctonia solani on stolons and stems. In field experiments, this ultimately led to a reduced formation of sclerotia on new tubers, particularly in neutral sandy loam and clay loam soils. In 1981 inoculation with antagonists led, when compared with no inoculation, to average reductions of 22 and 42% for the harvest from clean, and 15 and 26% for the harvest from infected seed tubers grown on slightly acid sandy soils and on neutral loam soils, respectively. The harvest from clean, inoculated seed tubers had the lowest sclerotium index. In 1982 inoculation of seed tubers planted in slightly acid sandy soils gave reductions of the sclerotium index of up to 22%. In the neutral marine loam soils considerable reductions were often achieved, viz., in slightly infected loams 51–68% and in rather heavily infected ones 4–43%. Chemical disinfection of seed tubers proved effective only in loam soils that were slightly infested withR. solani. In both years inoculation of seed tubers with antagonists led to significantly lower sclerotium indices of the harvest (p=0.1% in 1981; p=5% in 1982). V. biguttatum was present more frequently and in greater densities on stems and stolons of plants from inoculated than from non-inoculated seed tubers. The latter were colonized by wildV. biguttatum strains from the soil, apparently less effective antagonists.Early in the season, the soil temperature was too low for growth ofV. biguttatum. Nevertheless, inoculation of tubers that were planted early resulted in a considerable cotrol ofR. solani.Samenvatting Het beënten van poters met de opRhizoctonia solani parasiterende schimmelVerticillium biguttatum isolaat M73 in combinatie metGliocladium roseum (1981) of metV. biguttatum M73 alleen of in combinatie met isolaat M180 plus antibiotische isolaten van de bacterieAzotobacter chroococcum (1982), bleek effectief in het terugdringen of het onderdrukken vanR. solani op stengels en stolonen en het verminderen van de aantasting.Beënting van het pootgoed leidde tot een vermindering van de sclerotium (lakschurft)-vorming op de nieuwe knollen, vooral in klei-en zavelgronden. In 1981 leidde beënting van poters tot reductie in de sclerotiumvorming van gemiddeld 22 en 42% voor de oogst uit schoon en 15 en 26% voor de oogst uit besmet pootgoed geteeld op respectievelijk zandgrond en klei- en zavelgrond.In 1982 leidde beënten van de poters uitgeplant in licht zure zandgrond tot een gemiddelde reductie van de sclerotiumindex van de oogst van 22%. In zwaar besmette zandgrond trad evenwel geen reductie op; de infectiedruk was hier te groot. In de neutrale zavel- en kleigronden, vaak ook in de zwaarder besmette percelen werden aanzienlijke reducties bereikt, in de licht besmette gemiddeld 51–68% en in de zwaarder besmette 4–43%. Ontsmetten van pootgoed bleek alleen effectief in percelen die licht metR. solani waren besmet.In beide jaren bleek beënten van pootgoed met antagonisten te resulteren in een significant lagere sclerotiumindex van de oogst (p=0,1% in 1981; p=5% in 1982). V. biguttatum was veel vaker en meer aanwezig op de ondergrondse stengeldelen en stolonen van planten uit beënt pootgoed dan op die van niet beënte poters. De laatsten werden gekoloniseerd door wilde stammen vanV. biguttatum uit de grond, die vaak minder effectieve antagonisten waren. Beënting van vroeg gepote knollen — als de temperatuur nog te laag is voor de groei vanV. biguttatum — leverde toch gunstige resultaten op.     

Suppressive effect of Azotobacter chroococcum on Rhizoctonia solani infestation of potatoes

Isolates ofAzotobacter chroococcum were found to be promising for the control of infestation of potato plants withRhizoctonia solani. Inoculation with an isolate ofVerticillium biguttatum in combination with isolates ofA. chroococcum effectively protected sprouts, stems and stolons against infestation withR. solani. The effect of inoculation varied with soil temperature.No sclerotia were formed on potatoes harvested from soil in pots inoculated with isolates ofA. chroococcum plusV. biguttatum under glasshouse conditions; the yield increased significantly over the control.Samenvatting Enkele isolaten vanAzotobacter chroococcum bleken sterk antagonistisch ten opzichte vanRhizoctonia solani en leken goed bruikbaar voor een biologische bestrijding van deze ziekteverwekker van de aardappel.Beënting van de grond metVerticillium biguttatum enA. chroococcum gaf — in potproeven — een effective bescherming van spruiten, stengels en stolonen tegen aantasting doorR. solani.De vorming van sclerotiën bleef achterwege op knollen die gevormd waren in grond die was beënt metA. chroococcum plusV. biguttatum. De opbrengst uit behandelde grond was hoger dan die uit onbehandelde.De effectiviteit van de behandeling is afhankelijk van de temperatuur.     

Growth of the mycoparasitic fungus Verticillium biguttatum from different geographical origins at near-minimum temperatures

Sclerotia ofRhizoctonia solani collected from potato tubers from different countries were assayed for the presence of mycoparasites. Among the mycoparasites observedVerticillium biguttatum predominated. Its geographical distribution was not restricted to certain latitudes or soil types;V. biguttatum occurred worldwide in potato fields.The minimum growth temperature of 57V. biguttatum isolates was found to be in the narrow range from 10 to 13°C, irrespective of their geographical origin. A non-linear logistic growth model was used to describe the radial growth onRhizoctonia mycelium and nutrient agar plates. At near-minimum temperature the maximum colony radii varied considerably; they were up to 3.8 times that of the reference isolate M73. Based on parameter values for logistic growth, fast-and slow-growing isolates could be distinguished. Although the growth properties ofV. biguttatum isolates from different locations varied, the presence of fast- and slow-growing isolates was not restricted to particular areas and both types could be found in the same field. However, bioassays with selected fast- and slow-growing isolates do not support the assumption that growth at near-minimum temperatures is a relevant criterion for screening isolates ofV. biguttatum in terms of effectiveness for biological control ofR. solani.     

Biological control of Rhizoctonia solani on potatoes by antagonists. 5. The effectiveness of three isolates of verticillium biguttatum as inoculum for seed tubers and of a soil treatment with a low dosage of pencycuron

In field experiments in 1983, inoculation of seed tubers with three isolates ofVerticillium biguttatum, separately or mixed, was successful in reducing the amount of sclerotia ofRhizoctonia solani formed on new tubers. In holocene marine soils the sclerotium index (s.i.) was on average 50% of that of the harvest from untreated tubers.In two slightly acid pleistocene sands an additional soil treatment with pencycuron at 20% of the recommended dosage improved the results of seed tuber inoculation in one soil (further reduction of the s.i. from 60 to 32%), but not in the other (reduction to 55%). The three isolates ofV. biguttatum used in these experiments did not differ in their efficacy to suppressR. solani.Lowering the number of spores in the inoculum fluid to 20% of the initial density (6×10⁵ per ml instead of 3×10⁶ did not influence the results.Samenvatting Het beënten van voorgekiemde pootaardappelen met drie isolaten vanVerticillium biguttatum, elk apart of als mengsel, bleek in veldproeven in 1983 gunstige resultaten te geven: de hoeveelheid lakschurft gevormd op nieuwe knollen was duidelijk minder dan in de onbehandelde controle. In zavelgronden werd de sclerotiumindex (maat voor de hoeveelheid lakschurft) gemiddeld teruggebracht tot de helft van de hoeveelheid die voorkwam op de oogst uit onbehandelde, niet ontsmette, poters.In twee licht zure zandgronden werd een aanvullende grondbehandeling gegeven (volvelds) met pencycuron in een concentratie die 20% was van de aanbevolen dosering. In één grond werden de resultaten van de enting hierdoor verbeterd en trad een verdere reductie van de sclerotiumindex op van 60% (met alleenV. biguttatum) tot 30%. In de andere grond bleef de reductie beperkt tot het niveau van 55%. De dosering was hier waarschijnlijk te laag.Het effect van enting metV. biguttatum op het verlagen van de hoeveelheid sclerotiën op de jonge knollen bleek minstens even goed als een chemische ontsmetting van het pootgoed en in de meeste gevallen zelfs beter.De drie isolaten vanV. biguttatum die in deze proeven gebruikt werden waren statistisch niet verschillend in hun vermogenR. solani terug te dringen. Evenmin was een verlaging van de sporendichtheid in de entvloeistof tot 20% (6×10⁵ sporen ml^–1) van de oorspronkelijk gebruikte dichtheid van invloed op de resultaten.     

Suppression of Rhizoctonia solani in potato fields. II. Effect of origin and degree of infection with Rhizoctonia solani of seed potatoes on subsequent infestation and on formation of sclerotia

The seed potatoes used in these experiments had been grown in a slightly acid pleistocene sandy soil or in a marine, holocene sandy loam. They were free of sclerotia ofR. solani or lightly or moderately speckled with them. Seed potatoes from the sandy soil produced plants that suffered less fromRhizoctonia than plants from seed potatoes that had been grown on the marine sandy loam. Similarly harvested tubers had, in a non-conducive soil and in conducive soils with a (very) low inoculum density ofR. solani, fewer sclerotia when they came from seed potatoes grown in an acid sand. In each soil, the degree of infestation of the crop not only depended on the severity of infection of the seed potatoes, but also on their origin. With regard to sclerotia production on tubers, three types of soil were distinguished: suppressive, conducive with a high, and conducive with a very low inoculum density ofR. solani. The differences in infestation and in the amounts of sclerotia on tubers between the crop grown from seed potatoes from the sandy soil and that from seed potatoes from the marine sandy loam soil, is attributed to a richer load of antagonists on the former and possibly to a larger proportion of saprophyticRhizoctonia strains among their sclerotia. The antagonists seem to be inhabitants of the subterranean parts of the plant and to function independently of the soil. This implies possibilities for their use in biological control.     

Increased susceptibility of potato to <Emphasis Type="Italic">Rhizoctonia</Emphasis> diseases in <Emphasis Type="Italic">Potato leafroll virus</Emphasis>-infected plants

In Hokkaido potato fields, tubers produced from the plants with leaf curl symptoms caused by potato leaf roll virus (PLRV) were noted to be more densely covered with Rhizoctonia sclerotia. This observation led us to hypothesize that potato infected with PLRV would have an increased susceptibility to Rhizoctonia solani. To test this hypothesis, in a pot experiment, we inoculated PLRV-infected mother tubers with Rhizoctonia. As a result, PLRV-infected plants produced significantly fewer and smaller tubers than virus-free plants did, suggesting that PLRV-infected plants are more susceptible than virus-free plants to R. solani. Virus-free seed tubers should thus be used to reduce Rhizoctonia diseases.     

Host and biotic factors affecting sporulation ofstemphylium botryosum f. sp. lycopersici on tomatoes and ofalternaria porri f. sp. sol ani on potatoes

Factors known to inhibit sporulation of bio trophic fungal pathogens were found to enhance sporulation of two necrotrophic fungi. The sporulating potential ofStemphyliurn botryosum f. sp.lycopersici on tomatoes and ofAlternaria porri f. sp.solani on potatoes increased with necrotization, reaching a maximum on dead leaves. Wetting the dead leaves for the whole period of incubation with increasing concentrations of glucose resulted in progressively decreasing sporulation of both pathogens. However, application of glucose during the first half of the incubation period inhibited sporulation ofS. botryosum f. sp.lycopersici on tomatoes only a little, and increased that ofA. porri f. sp.solani on potatoes. The capacity ofS. botryosum f. sp.lycopersici to sporulate on leaves lasted for 12 weeks at 29°C, and ofA. porri f. sp.solani for 12 weeks at 29°C and for over 21 weeks at 20°C. The results emphasized basic differences in sporulation between biotrophic and necrotrophic parasites. Specific techniques useful for studying sporulationin vivo are discussed.     

Compatible Biological and Chemical Control Systems for Rhizoctonia solani in Potato

A series of chemical and biological control agents were tested for compatibility with the Rhizoctonia-specific biocontrol fungus Verticillium biguttatum aimed at designing novel control strategies for black scurf (Rhizoctonia solani) and other tuber diseases in potato. The efficacy of chemicals, alone and in combination with V. biguttatum was tested in in vitro assays on nutrient agar plates, in bio-assays with minitubers and in the field. Generally, there were both antagonistic, neutral and additive interactions with V. biguttatum among the combinations tested; there were no indications for synergistic interactions. Broad-spectrum fungicides (azoxystrobin, chlorothalonil, thiabendazole) were fungitoxic to V. biguttatum as shown in in vitro assays, and hampered black scurf control by V. biguttatum in bio-assays. Oomycete-specific chemicals (cymoxanil and propamocarb) and various biocontrol strains (Gliocladium spp., Pseudomonas spp. and Trichoderma spp.) did not interfere with the growth of V. biguttatum on agar nutrient plates and did not affect black scurf control by V. biguttatum in co-applied treatments in the minituber bio-assay. Rhizoctonia-specific (pencycuron, flutalonil) fungicides co-applied with V. biguttatum showed additive effects on black scurf control. When combinations of V. biguttatum and cymoxanil or propamocarb were applied to immature potato tubers at green crop lifting, a reduction of both black scurf and Pythium- or Phytophthora-incited tuber rot was observed at harvest. In conclusion, the biocontrol fungus V. biguttatum is compatible with selected chemical control systems and may improve control efficacy in combination with Rhizoctonia-specific fungicides or may extend control spectrum in combination with Oomycete-specific fungicides.     

Serological recognition of Streptomyces species causing scab on potato tubers

Antisera were prepared against extracts of two tyrosinase-positiveStreptomyces spp., one of which caused a “deep” and the other a “russet” scab. Tyrosinase-positiveStreptomyces isolates not reacting with either of these antisera proved to be nonpathogenic to potato tubers, with few exceptions only. Not all isolates reacting with one or both antisera, however, were pathogenic and so all the serological positive ones had to be tested for pathogenicity to potato tubers. To obtain this relative specificity the antisera had to be absorbed with an extract of a non-pathogenic tyrosinase-positive isolate.     

Use of sclerotia of<Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis> for efficient production of conidia of<Emphasis Type="Italic">Coniothyrium minitans</Emphasis> in liquid culture

A study was conducted to determine the feasibility of using sclerotia ofSclerotinia sclerotiorum for producing conidia ofConiothyrium minitans in liquid culture. The medium (SST) was made of water containing 2.0, 1.5, 1.0 or 0.5% (w/v) ground sclerotia ofS. sclerotiorum and 100 μgl ⁻¹ thiamine hydrochloride (HCl). One ml of conidial suspension (2 × 10⁷ conidia ml⁻¹) ofC. minitans LRC 2534 was inoculated into 100 ml of SST medium or control (thiamine HCl in water) and incubated at 20 ± 2°C on a shaker at 200 rpm. Subsamples were removed periodically and examined under a compound microscope. Conidia in the SST media germinated within 24 h, developed into branched hyphae within 48 h, produced pycnidia after 3–4 days, and the pycnidia released mature conidia after 7 days. Production of conidia varied with the concentration of sclerotia in the SST medium. It was lower (3.6 × 10⁶ conidia ml⁻¹) at 0.5% but higher (1.2 × 10⁸ conidia ml⁻¹) at 2%. The new conidia were viable and the colonies developing from them showed the original morphological characteristics. It was concluded that using SST liquid medium as a substrate for mass production of conidia ofC. minitans has potential for use in commercial development of this mycoparasite as a biocontrol product. http:www.phytoparasitica.org posting Jan. 23, 2007.     

Effects of granular nematicides on growth and microbial antagonism to Rhizoctonia solani

Effects of nematicides on growth and microbial antagonism toRhizoctonia solani were investigated as part of a study on the mechanisms involved in the increased incidence of this pathogen in nematicide-treated potato crops.Ethoprophos inhibited mycelial growth ofR. solani on potato dextrose agar (PDA), Czapek Dox agar (CDA) and on water agar (WA). Aldicarb stimulated its growth on PDA up to 14% but not on CDA and WA. Oxamyl inhibited mycelial growth on CDA and WA, but not on PDA.Ethoprophos and aldicarb stimulated development of the mycoparasiteVerticillium biguttatum on cultures ofR. solani. The effect was dependent on the medium on which the host fungus was grown. ForRhizoctonia cultures on PDA, growth of the mycoparasite was highly promoted by aldicarb and to a lesser extent by ethoprophos. WhenR. solani was grown on CDA, the development of the mycoparasite was not affected by aldicarb, slightly stimulated by ethoprophos and slightly inhibited by oxamyl. On water agar, its development on the host mycelium was not affected.In field trials on sandy soil, nematicides encouragedV. biguttatum probably by increased availability of substrate (i.e.Rhizoctonia mycelium) perhaps through reduced activity of the mycophagous fauna.Soil fungistasis was increased by ethoprophos and to a lesser extent by aldicarb at very high doses. At normal field rates, no effects can be expected on fungistasis. So the increased stem and stolon infection of potatoes in nematicide-treated fields was not caused by a direct effect of the nematicides on growth ofR. solani or by suppressing the microbial antagonism.Samenvatting De invloed van granulaire nematiciden op de groei vanRhizoctonia solani en op het microbiële antagonisme tegen deze schimmel werd onderzocht in het kader van een studie over de mechanismen die een rol spelen bij de toename van de aantasting in een met nematiciden behandeld aardappelgewas.Ethoprofos remde de myceliumgroei vanR. solani op aardappeldextrose agar (PDA), Czapek Dox agar (CDA) en op wateragar (WA). Aldicarb stimuleerde op PDA de groei met maximaal 14%. Op CDA en WA werd geen effect van aldicarb waargenomen. Oxamyl veroorzaakte groeiremming op CDA en WA, maar niet op PDA.Ethoprofos en aldicarb stimuleerden de ontwikkeling van de mycoparasietVerticillium biguttatum op cultures vanR. solani. De mate van groeistimulering was afhankelijk van de voedingsbodem waarop de waard,R. solani, werd gekweekt. De groei vanV. biguttatum werd sterk gestimuleerd door aldicarb en in geringere mate door ethoprofos, wanneer de waard gekweekt werd op PDA. Aldicarb had geen effect op de mycoparasiet wanneerR. solani op CDA gekweekt werd, terwijl ethoprofos de groei wel iets stimuleerde en oxamyl een gering remmend effect had. Op WA werd geen effect van de nematiciden op het mycoparasitisme vastgesteld.In veldproeven op zandgrond stimuleerden de nematiciden het voorkomen vanV. biguttatum op de stolonen. Het effect werd waarschijnlijk veroorzaakt door een verhoogde substraat beschikbaarheid (d.w.z. mycelium vanR. solani). De verhoogde beschikbaarheid van dit mycelium kan samenhangen met een door nematiciden gereduceerde activiteit van de fungivore bodemfauna.De bodemfungistase werd verhoogd door ethoprofos en, in geringere mate, door aldicarb bij hogere doseringen. Bij de in de praktijk aanbevolen doseringen kan echter geen effect op de fungistase verwacht worden. De toename in stengel- en stolonaantasting van aardappelen, geteeld in met granulaire nematiciden behandelde percelen, kon niet worden toegeschreven aan een direct effect van de nematiciden op de groei vanR. solani of aan een vermindering van het microbiële antagonisme.     

Accumulation of hyperparasites of Rhizoctonia solani by addition of live mycelium of R. solani to soil

Addition of live mycelium ofRhizoctonia solani to three slightly, acid sands collected in winter from a field after a previous potato crop resulted in accumulation of hyperparasites and less infestation of sprouts of infected seed potatoes. A similar effect was observed in alkaline marine sandy loam soils, but only after a second addition of live mycelium. The predominant hyperparasite in these experiments wasVerticillium biguttatum.     

Antagonism of Azotobacter chroococcum isolates to Rhizoctonia solani

Antagonism of isolates ofAzotobacter chroococcum toRhizoctonia solani on agar plates was studied, and isolates were tested for their ability to controlR. solani infection of potato sprouts in sterilized and unsterilized soil. The degree of antagonism exhibited varied strongly among the isolates and was also found to be temperature-dependent. At 25, 20 and 15°C, all but one strongly antagonisticAzobacter isolate effectively prevented infection of sprouts of potatoes planted in a soil heavily infected with a pathogenic isolate ofR. solani. At 10°C none was effective.