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.     

Temperature dependence of the pathogenicity of several isolates of Rhizoctonia solani in some bulb crops as an intrinsic property of the isolate

Several isolates ofRhizoctonia solani obtained from tulip, iris or lettuce, infected various bulb crops either only at soil temperatures of 13 °C or higher (‘warmth preferring isolates’), or mainly at soil temperatures below 13 °C (‘cold preferring isolates’). Infection of the host by cold preferring isolates at temperatures above 13 °C was possible when the inoculum was in close contact with the host. Pathogenic activity under unfavourable temperature conditions was relatively brief but could recur if the temperature became favourable. Differences in the rate of sprout growth due to preplanting temperature treatment of bulbs influenced infection, but the main characteristic of the isolates with respect to the temperature-pathogenicity relationship prevailed.     

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.     

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.     

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.     

Inactivation of sclerotia of Rhizoctonia solani on potato tubers by Verticillium biguttatum,a soil-borne mycoparasite

Experiments in the laboratory and on farms with potato tubers in storage are described in which sclerotia ofRhizoctonia solani were inactivated after inoculation of infected tubers with a suspension of conidia and hyphal fragments ofVerticillium biguttatum. Sclerotia on freshly harvested tubers can be killed in a period of six to eight weeks, provided that (1) a direct contact between sclerotia and conidia ofV. biguttatum is obtained, (2) the temperature during the storage period is at least 15 °C, but preferably closer to 20 °C during the first weeks, and (3) the relative humidity of the air between the tubers is at least 99%. Seed tubers are only certified as export quality if the infection withR. solani, visible as sclerotia on the tubers, is assessed as below a specified incidence. To restore the economical value of tubers with many sclerotia, living sclerotia can be inactivated byV. biguttatum. However, also dead sclerotia have to be removed, as dead and living sclerotia cannot be distinguished visually by inspectors. A satisfactory way to remove dead sclerotia from tubers has not yet been found.     

Effect of label and sublabel rates of metam sodium in combination with<Emphasis Type="Italic">Trichoderma hamatum,T. harzianum,T. virens,T. viride</Emphasis> on survival and growth of<Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

This work was undertaken to determine the effects ofTrichoderma spp. combined with label and sublabel rates of metam sodium on survival ofRhizoctonia solani in soil. Soils were infested with wheat bran preparations ofTrichoderma hamatum Tri-4,T. harzianum Th-58,T. virens Gl-3, andT. viride Ts-1-R3. Soil was also infested with sterile beet seeds that were colonized withR. solani. Beet seeds were later recovered, plated onto water agar plus antibiotics, and the growth ofR. solani was recorded. Preliminary experiments showed thatT. hamatum andT. virens reduced survival and saprophytic activity ofR. solani when the biocontrol fungi were incorporated into soil at 1.5% (w:w) or greater. Based on these data, biocontrol fungi in subsequent experiments were incorporated into soil at 2%. Metam sodium at label rate killed all biocontrol fungi andR. solani. At 1:2 and 1:5 dilutions, metam sodium reduced survival ofR. solani and allTrichoderma spp. When biocontrol fungi plus the label rate of metam sodium and 1:5, 1:10, 1:50 or 1:100 dilutions of the label rate were tested together, there were no interactions between any biocontrol agent and the fumigant with respect to colony diameter, reflecting that allTrichoderma isolates tested reacted similarly to increasing concentrations of metam sodium. At the label rate of metam sodium, allTrichoderma spp. significantly reduced colony diameter, but not growth rate, ofR. solani from beet seed. For the levels of metam sodium tested in combination withTrichoderma, it does not appear feasible to use a reduced rate of metam sodium to controlR. solani. However, the combination ofTrichoderma with metam sodium does reduce growth ofR. solani in comparison with that provided by metam sodium at the label rate. http://www.phytoparasitica.org posting Feb. 11, 2004.     

Pathogenicity,host specificity and anastomosis groups ofRhizoctonia spp. isolated from soils in Israel

One hundred and twenty-nine isolates ofRhizoctonia spp. were obtained from soil samples in Israel and from culture collections in the U.S.A. and Japan. The isolates varied in host range and disease severity when tested on six to eleven different host plants. Approximately 30% of the isolates were nonpathogenic to all the host plants tested. Mycelial growth rate of the nonpathogenic strains did not differ significantly from that of the virulent isolates. The 107 Israeli isolates represented anastomosis groups (AG) 1, 2, 3, 4, 5 and 6 ofR, solani, two groups ofR. zeae, and three groups of binucleateRhizoctonia AG: A, F and K.     

Infection process of Rhizoctonia solani on Solanum tuberosum and effects of granular nematicides

The infection process ofRhizoctonia solani AG-3 was studied on potato sprouts, cv. Bintje, in growth chamber trials at 15 °C. Initially hyphae ofR. solani grew predominantly in the longitudinal direction of the sprouts (runner hyphae). They tended to follow the junctions between epidermis cells as was observed by SEM. The hyphae formed side-branches mainly half-way of the subterranean parts of the sprouts. They branched several times with short swollen cells to form infection cushions. Lesions developed only underneath the infection cushions and were first observed five days after inoculation. The necrotic area was proportional to the area covered with infection cushions on the sprouts. Depth of the lesions could extend up to the vascular bundle. Sprouts were colonized only in healthy tissue in the epidermal layer underneath the infection cushion and in necrotic tissue. A few days after appearance of the lesions,R. solani formed brown, uninfective mycelium on and in the circumference of these lesions.Aldicarb did not influence any part of the infection process. Ethoprophos delayed the emergence of sprouts, but increased the number of sprouts per tuber. As soon as sprouts had emerged, growth was considerably promoted by ethoprophos. Ethoprophos delayed the appearance of lesions and reduced their size. Oxamyl showed the same effects to a smaller extent.As the size of lesions appears to be proportional to the size of the infection cushions, any agents that change the size of the infection cushions, such as pesticides or antagonists, may alter the severity of the disease.Samenvatting Het infectieproces vanRhizoctonia solani AG-3 werd bestudeerd op aardappelspruiten, cv. Bintje, in een klimaatcel bij 15C. Aanvankelijk groeide de schimmel met runnerhyfen voornamelijk in de lengterichting van de spruit. Via SEM kon waargenomen worden, dat de hyfen hierbij vooral over de begrenzingen van de epidermiscellen groeiden. Het mycelium vormde veel zijvertakkingen, bestaande uit iets gezwollen korte cellen, welke voornamelijk halverwege op het ondergrondse deel van de spruit gevormd werden. Een dichte massa van deze cellen vormde een infectiekussentje. Lesies, welke vanaf vijf dagen na inoculatie werden waargenomen, bevonden zich slechts onder spruitoppervlak bezet met infectiekussentjes. De lesiegrootte was recht evenredig met het spruitoppervlak dat bezet was met infectiekussentjes. De diepte van de lesies reikte tot aan de vaatbundels. De spruit werd alleen door de schimmel gekoloniseerd in gezond epidermisweefsel onder het infectiekussentje en in necrotisch weefsel. Enkele dagen na verschijning van lesies vormde R.solani bruin, niet infectieus, mycelium op en rondom de lesies.Aldicarb had geen effect op het infectieproces. Ethoprophos vertraagde de opkomst en verhoogde het aantal tot ontwikkeling gekomen spruiten per knol in gestoomd zand. Direct na opkomst had ethoprophos echter een sterk groeistimulerend effect. Ethoprophos vertraagde de lesievorming en reduceerde de lesiegrootte, vergeleken met onbehandelde planten. Oxamyl vertoonde deze effecten in geringere mate.Daar de lesiegrootte direct gecorreleerd blijkt met de grootte van het infectiekussentje, mag verwacht worden dat elke beïnvloeding van de ontwikkeling van het mycelium van R.solani, bijvoorbeeld door pesticiden of antagonisten, een verandering van de lesiegrootte ten gevolge heeft.     

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.     

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.     

Identification of Rhizoctonia solani AG1-IB as a causal agent of leaf blight in Cynanchum paniculatum in China

A severe disease causing leaf blight of Cynanchum paniculatum was observed in China. From the diseased plants, 24 fungal isolates were obtained. Morphological observations and nuclear conditions of all the isolates indicated characteristics of Rhizoctonia solani. The ribosomal DNA internal transcribed spacer sequences of the isolates showed more than 99 % sequence homology with published sequences of R. solani anastomosis group 1 subgroup IB (AG1-IB). All tested isolates were pathogenic and showed significant symptoms as observed in the fields. This is the first report of leaf blight of C. paniculatum caused by R. solani AG1-IB worldwide.     

Identification and pathogenicity of<Emphasis Type="Italic">Rhizoctonia solani</Emphasis> and binucleate<Emphasis Type="Italic">Rhizoctonia</Emphasis> anastomosis groups isolated from forage legumes in Erzurum,Turkey

Abstrast  Three-hundred-twenty-five isolates ofRhizoctonia (215R. solani and 110 binucleateRhizoctonia) were obtained from roots and crowns of alfalfa, sainfoin and common vetch grown in Erzurum, Turkey. The isolates were assigned to five anastomosis groups (AG) ofR. solani (AG-2-1, AG-3, AG-4, AG-5, and AG-10) and two anastomosis groups of binucleateRhizoctonia (AG-I and AG-K). In pathogenicity tests on alfalfa, sainfoin and common vetch, the highest disease severities were caused by isolates of AG-4 and AG-5. Isolates of AG-10 and AG-I were not pathogenic on the three tested forage legumes, whereas isolates of AG-K on alfalfa and sainfoin, and of AG-2-1 on sainfoin, were moderately virulent. Alfalfa isolate AG-3 was moderately virulent on sainfoin. This is the first report ofR. solani AG-3, AG-5, AG-10 and binucleateRhizoctonia AG-I on alfalfa. In addition, all theR. solani and binucleateRhizoctonia groups isolated from sainfoin and common vetch were recovered from these crops for the first time in Turkey. http://www.phytoparasitica.org posting Dec. 16, 2002.     

Biocontrol ofRhizoctonia damping-off of cucumber by non-pathogenic binucleateRhizoctonia

Three isolates of binucleateRhizoctonia (BNR) were tested for biological control of damping-off of cucumber seedlings caused byRhizoctonia solani AG 2-2 and AG 4. BNR isolates L2 (AG Ba) and W1 and W7 (AG A) provided protection of 58 to 71% against virulent isolate C4 of AG 4 and 64 to 75% protection against virulent isolate RH 65 of AG 2-2. Varying protection was provided to the seedlings by the BNR isolates against the virulentR. solani from the two AGs depending on their combination. The BNR isolates did not vary in providing protection to the seedling when tested against virulent C4 when both isolates were inoculated using three different methods,viz. in water agar, combination of water agar and soil and using soil alone. Protection of 58 to 71 % was provided by the isolates when inoculation was done on the hypocotyl using water agar, 62.8 to 75% using the combination of water agar and soil, and 75 to 85% when inoculation of both isolates was done in soil. Pre-incubation of BNR W7 or delayed inoculation of C4 (from 0.5 day to longer duration) using the different methods provided an increased protection to the seedlings to give complete inhibition of damping-off disease. Simultaneous inoculation of both BNR W7 and C4 using the three methods failed to provide protection to the seedlings. Among the BNR isolates, BNR W7 showed plant growth promotion in terms of significant increase in plant height (P=0.01) and fresh weight (P=0.05).     

Einsatz bakterieller Antagonisten zur Bekämpfung von Krankheiten verursacht durch Rhizoctonia solani

Rhizoctonia solani is an important soil-borne pathogen causing diseases in numerous economically important crops. The pathogen R. solani can be responsible for relevant yield losses as well as on lettuce and potato. To develop a biocontrol strategy, two bacterial strains, Pseudomonas fluorescens L13–6-12 and Serratia plymuthica 3Re4–18 were evaluated against R. solani causing black scurf in potato and bottom rot in lettuce. The disease suppression effect of the two antagonists was tested as well as in a climate chamber and in the field during two vegetation periods. The results of the climate chamber experiments showed, that R. solani can significantly reduced the lettuce growth. The dry mass losses on lettuce and the disease severity on potato sprouts were significantly limited through bacterization. The antagonist L13–6-12 showed best disease suppression effect in climate chamber experiments on both crops. A significant lower disease severity was to observed in treatments with bacterial antagonists as well as on lettuce plants and harvested potato tubers during both vegetation periods. Also the dry mass losses on lettuce were clear reduced in treatments with bacterial antagonists, whereas only a partly limitation of yield losses on potato was to achieved. In summary, the results supported that the use of bacterial antagonists can be part of a control strategy against R. solani.     

Possible Mechanisms Influencing the Dynamics of Rhizoctonia Disease of Tulips

looseness-1In two observation fields, where six sites were artificially infested with Rhizoctonia solani AG 2-t, bare patches developed. These patches did not re-occur at the site of infestation in three successive years. In fields with and without artificial infestation, natural infection of tulip bulbs by Rhizoctonia spp. occurred. The spatial distribution of infected tulip bulbs was visualised in maps after kriging. The influence of sampling intensity was evaluated by stepwise reduction obtained in the observed data set of the first year. Omnidirectional semivariogram characteristics did not change when sampling intensity was reduced down to 10%. The average maximum prediction error was minimised at sampling intensities varying from 7% to 25%. Naturally occurring bare patches slowly vanished during successive cropping of flower bulbs and did not re-appear in the fourth growing season. A high frequency of isolation of R. solani AG 2-t in one field (Lisse-2) in the fourth consecutive crop did not result in bare patches in that year. It is hypothesised that a reduction in aggressiveness may account for this observation. In contrast, bulb rot due to Rhizoctonia spp. increased during the observation period. R. solani AG 5 isolates were seldom isolated before the bulbs flowered, but were the dominant isolate from bulbs at harvest. In a growth chamber experiment, it was demonstrated that AG 5 did not account for replacement of AG 2-t. However, it was demonstrated that competition may partially explain replacement of AG 2-t isolates during the growing season. At 18 °C, but not at 9 °C, an AG 4 isolate prevented AG 2-t colonising and infecting iris bulbs when both isolates were introduced together to soil. Rhizoctonia populations develop in relation to soil temperature and plant development. It is hypothesised that a temporal niche differentiation may be one of the mechanisms affecting the dynamics of rhizoctonia bare patch of tulips.     

Zwartrot in sla,veroorzaakt doorRhizoctonia solani

Summary In the autumn of 1962, a so-called blackrot of lettuce occurred on a relatively large scale. The leaves of older and younger plants which lie on the soil, become black and rotten from the point of contact with the soil. The disease spreads over the whole leaf, leaving the remains as a black print on the soil surface. From diseased leavesRhizoctonia solani Kühn (stat. mycel. ofThanatephorus cucumeris (Frank) Donk) andBotrytis cinerea Pers. ex Fr. were isolated. Since the latter was expected to be a secondary invader, it was not used in the subsequent inoculation experiments. After inoculation with theR. solani isolate, the typical blackrot symptoms appeared within eight days. This isolate was not pathogenic to tomatoplants. Possibly the disease may be controlled with PCNB, an effective fungicide againstRhizoctonia. As far as the authors are aware, this is the first record of blackrot of lettuce, caused byR. solani.     

Induction of systemic resistance by a hypovirulent Rhizoctonia solani isolate in tomato

A polynucleate Rhizoctonia isolate (R3) was analysed for virulence, growth characteristics, enzyme production and presence of dsRNAs. Taxonomic position was assessed morphologically and by anastomosis group (AG) testing and ITS sequence analysis. Results indicated that R3 is a hypovirulent R. solani AG 4. Mechanisms underlying biocontrol towards virulent R. solani and Botrytis cinerea were investigated and plant-mediated resistance was followed using biochemical markers of defence (PR1, laminarinase, chitinase). Control apparently relies on spatial and nutrient competition in soil, and on systemic induced resistance. This is the first report on induction of systemic resistance and of defence markers by a hypovirulent strain of R. solani.     

Pathogenicity and Fungicide Sensitivity of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> and <Emphasis Type="Italic">R. cerealis</Emphasis> Isolates

The Rhizoctonia solani species consists of multinucleate isolates that belong to anastomosis groups AG1–AG3 and differ in virulence and host affinity. R. cerealis is a binucleate species of anastomosis group AG-D which causes sharp eyespot, a common plant disease in Poland. Rhizoctonia spp. is a ubiquitous soil pathogen that poses a significant threat for global crop production due to the absence of effective crop protection products. The aim of this study was to determine the virulence of R. solani and R. cerealis isolates towards Beta vulgaris, Zea mays, Triticum spelta and T. aestivum seedlings, to confirm the presence of endopolygalacturonase genes pg1 and pg5 in the genomes of the tested isolates and to evaluate the tested isolates’ sensitivity to triazole, strobilurin, imidazole and carboxamide fungicides. All tested isolates infected B. vulgaris seedlings. but none of them were virulent against Z. mays plants. R. solani isolates AG4 PL and AG2-2IIIB PL were characterized by the highest virulence (average infestation score of 2.37 and 2.53 points on a scale of 0–3 points) against sugar beet seedlings. The prevalence of infections caused by most of the analysed isolates (in particular R. solani AG4 J—11.8, and R. cerealis RC2—0.78) was higher in spelt than in bread wheat. The virulence of the analysed isolates was not correlated with the presence of pg1 and pg5 genes. The efficacy of the tested fungicides in controlling Rhizoctonia spp. infections was estimated at 100% (propiconazole + cyproconazole), 98.8% (penthiopyrad), 95.4% (tebuconazole) and 78.3% (azoxystrobin).     

Characterization of Japanese <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-2-1 isolates using rDNA-ITS sequences,culture morphology,and growth temperature

Rhizoctonia solani AG-2-1 is classified into three subsets (Subsets 1–3) based on the rDNA-ITS sequence. Few Japanese isolates, however, have been phylogenetically analyzed. To understand the distribution and diversification of AG-2-1 isolates in Japan, we examined 23 Japanese AG-2-1 isolates (15 from Hokkaido, the northernmost island, and eight from NARO Genebank) in terms of rDNA-ITS sequences, culture morphology, and temperature-dependent growth characteristics. Of these, 15 isolates were found to belong to Subset 1. One isolate, which formed a light brown colony with concentric rings and grew slowly at 25 °C, was classified into Subset 2. Six isolates had varied culture morphology and relatively faster growth than Subset 1 isolates at 30 °C. They formed a clade on the phylogenetic tree, designated clade HK, with cauliflower isolates from Belgium and the Netherlands, with a bootstrap value of 47%, and were separate from the three known subsets. Sequence similarity in the rDNA-ITS region for this clade ranged from 98.2 to 100%, whereas clade HK isolates had 96.7–98.6% similarity with the isolates in each subset. This result suggests that clade HK is likely an independent intragroup within AG-2-1, although the rDNA-ITS sequences in this clade were variable. One isolate was not assignable to any clade because it was intermediate between isolates in clade HK and Subset 2. This is the first report describing variation among rDNA-ITS sequences of Japanese AG-2-1 isolates.