Wheat Genotype-Specific Induction of Soil Microbial Communities Suppressive to Disease Incited by Rhizoctonia solani Anastomosis Group (AG)-5 and AG-8

ABSTRACT The induction of disease-suppressive soils in response to specific cropping sequences has been demonstrated for numerous plant-pathogen systems. The role of host genotype in elicitation of the essential transformations in soil microbial community structure that lead to disease suppression has not been fully recognized. Apple orchard soils were planted with three successive 28-day cycles of specific wheat cultivars in the greenhouse prior to infestation with Rhizoctonia solani anastomosis group (AG)-5 or AG-8. Suppressiveness to Rhizoctonia root rot of apple caused by the introduced isolate of R. solani AG-5 was induced in a wheat cultivar-specific manner. Pasteurization of soils after wheat cultivation and prior to pathogen introduction eliminated the disease suppressive potential of the soil. Wheat cultivars that induced disease suppression enhanced populations of specific fluorescent pseudomonad genotypes with antagonistic activity toward R. solani AG-5 and AG-8, but cultivars that did not elicit a disease suppressive soil did not modify the antagonistic capacity of this bacterial community. When soils were infested prior to the initial wheat planting, all cultivars were uniformly susceptible to R. solani AG-8. However, when pathogen inoculum was added after three growth-cycles, wheat root infection during the fourth growth-cycle varied in a cultivar specific manner. The same wheat cultivar-specific response in terms of transformation of the fluorescent pseudomonad community and subsequent suppression of Rhizoctonia root rot of apple was observed in three different orchard soils. These results demonstrate the importance of host genotype in modification of indigenous saprophytic microbial communities and suggest an important role for host genotype in the success of biological control.     

Anastomosis group and pathogenicity of isolates of Rhizoctonia solani from potato crops in South Australia

Isolates of Rhizoctonia collected from the stems, roots, tuber sclerotia and soil of potato crops in Virginia and Lenswood, South Australia, were identified to anastomosis groups (AG). Of the 301 multinucleate isolates of Rhizoctonia solani tested, 90% were AG-3, 7% were AG-4 and 2% were AG-5; 12 isolates were binucleate Rhizoctonia spp. This is the first report of isolates of AG-4 and AG-5 causing disease in potato crops in South Australia. All AG-3, AG-4 and AG-5 isolates tested caused rhizoctonia disease symptoms on the potato cultivar Coliban in pathogenicity trials conducted under glasshotise conditions. Both AG-3 and AG-5 isolates caused black scurf and stem cankers, although symptoms of black scurf were less severe with AG-5. AG-4 isolates produced the most severe stem and stolon cankers of all isolates tested. The pathogenicity of tuber-borne inoculum was confirmed by growing plants from sclerotia-infested tubers. AG-8 isolates from diseased barley and wheat produced severe root cankers and caused loss of feeder roots on inoculated potato plants. Results suggest that rhizoctonia disease in potato fields in South Australia is caused by a combination of different anastomosis groups and this has important implications for crop rotations.     

Identification and quantification of Rhizoctonia solani and R. oryzae using real-time polymerase chain reaction

Rhizoctonia solani and R. oryzae are the principal causal agents of Rhizoctonia root rot in dryland cereal production systems of the Pacific Northwest. To facilitate the identification and quantification of these pathogens in agricultural samples, we developed SYBR Green I-based real-time quantitative-polymerase chain reaction (Q-PCR) assays specific to internal transcribed spacers ITS1 and ITS2 of the nuclear ribosomal DNA of R. solani and R. oryzae. The assays were diagnostic for R. solani AG-2-1, AG-8, and AG-10, three genotypes of R. oryzae, and an AG-I-like binucleate Rhizoctonia species. Quantification was reproducible at or below a cycle threshold (Ct) of 33, or 2 to 10 fg of mycelial DNA from cultured fungi, 200 to 500 fg of pathogen DNA from root extracts, and 20 to 50 fg of pathogen DNA from soil extracts. However, pathogen DNA could be specifically detected in all types of extracts at about 100-fold below the quantification levels. Soils from Ritzville, WA, showing acute Rhizoctonia bare patch harbored 9.4 to 780 pg of R. solani AG-8 DNA per gram of soil.. Blastn, primer-template duplex stability, and phylogenetic analyses predicted that the Q-PCR assays will be diagnostic for isolates from Australia, Israel, Japan, and other countries.     

Rhizoctonia species and anastomosis groups isolated from barley and wheat in Erzurum, Turkey

Ninety-eight isolates of Rhizoctonia spp. were obtained from barley and wheat grown in Erzurum, Turkey. Of these, 78% were Rhizoctonia solani (AG-2 type 1, AG-3, AG-4, AG-5 and AG-11), 10% were binucleate Rhizoctonia (AG-I and AG-K) and the remainder were Waitea circinata var circinata ( Rhizoctonia sp.). Among the binucleate Rhizoctonia , AG-I was not recovered from barley. In pathogenicity tests on barley and wheat, the highest disease severity was caused by isolates of AG-4 and AG-11, whereas isolates of AG-2 type 1, AG-3, AG-5 and W. c . var  circinata were moderately virulent. Isolates of binucleate Rhizoctonia were all nonpathogenic. This is the first report of R. solani AG-11 and W. c . var  circinata from Turkey.     

Molecular tools to investigate Rhizoctonia solani distribution in soil

Real-time PCR protocols were developed to detect and discriminate 11 anastomosis groups (AGs) of Rhizoctonia solani using ribosomal internal transcribed spacer (ITS) regions (AG-1-IA, AG-1-IC, AG-2-1, AG-2-2, AG-4HGI+II, AG-4HGIII, AG-8) or β-tubulin (AG-3, AG-4HGII, AG-5 and AG-9) sequences. All real-time assays were target group specific, except AG-2-2, which showed a weak cross-reaction with AG-2^tabac. In addition, methods were developed for the high throughput extraction of DNA from soil and compost samples. The DNA extraction method was used with the AG-2-1 assay and shown to be quantitative with a detection threshold of 10⁻⁷ g of R. solani per g of soil. A similar DNA extraction efficiency was observed for samples from three contrasting soil types. The developed methods were then used to investigate the spatial distribution of R. solani AG-2-1 in field soils. Soil from shallow depths of a field planted with Brassica oleracea tested positive for R. solani AG-2-1 more frequently than soil collected from greater depths. Quantification of R. solani inoculum in field samples proved challenging due to low levels of inoculum in naturally occurring soils. The potential uses of real-time PCR and DNA extraction protocols to investigate the epidemiology of R. solani are discussed.     

Effect of soil moisture content on the suppression of Rhizoctonia stem canker on potato by the nematode Aphelenchus avenae and the springtail Folsomia fimetaria

The effect of soil moisture content on the suppression of Rhizoctonia stem canker on potato by mycophagous soil animals was studied in growth chambers. Three soil moisture levels were established in two bioassays, in which potato sprouts grew through a 15-cm soil layer inoculated with sclerotia of Rhizoctonia solani (AG-3). In one experiment two levels of R. solani inoculum were applied. The effect on plant disease of mycophagous soil fauna was assessed by adding the springtail Folsomia fimetaria and/or the nematode Aphelenchus avenae to the soil. In the absence of mycophagous organisms, Rhizoctonia disease severity on potato stems was highest in dry soil. A. avenae and F. fimetaria reduced Rhizoctonia stem canker when applied at populations found in the field. They were effective over a broad range of soil moistures. The stimulatory effect of dry soil conditions on Rhizoctonia stem canker was counteracted by a greater efficacy of the mycophagous soil fauna under these conditions. Mild drought stress did not seem to be a limiting factor in the biological control of stem canker by these two organisms.     

Mechanism of action and efficacy of seed meal-induced pathogen suppression differ in a brassicaceae species and time-dependent manner

ABSTRACT The effect of seed meals derived from Brassica juncea, B. napus, or Sinapis alba on suppression of soilborne pathogens inciting replant disease of apple was evaluated in greenhouse trials. Regardless of plant source, seed meal amendment significantly improved apple growth in all orchard soils; however, relative differences in pathogen suppression were observed. All seed meals suppressed root infection by native Rhizoctonia spp. and an introduced isolate of Rhizoctonia solani AG-5, though B. juncea seed meal often generated a lower level of disease control relative to other seed meal types. When introduction of the pathogen was delayed until 4 to 8 weeks post seed meal amendment, disease suppression was associated with proliferation of resident Streptomyces spp. and not qualitative or quantitative attributes of seed meal glucosinolate content. Using the same experimental system, when soils were pasteurized prior to pathogen infestation, control of R. solani was eliminated regardless of seed meal type. In the case of B. juncea seed meal amendment, the mechanism of R. solani suppression varied in a temporal manner, which initially was associated with the generation of allylisothiocyanate and was not affected by soil pasteurization. Among those tested, only B. juncea seed meal did not stimulate orchard soil populations of Pythium spp. and infection of apple roots by these oomycetes. Although application of B. napus seed meal alone consistently induced an increase in Pythium spp. populations, no significant increase in Pythium spp. populations was observed in response to a composite B. juncea and B. napus seed meal amendment. Suppression of soil populations and root infestation by Pratylenchus spp. was dependent upon seed meal type, with only B. juncea providing sustained nematode control. Collectively, these studies suggest that use of a composite B. juncea and B. napus seed meal mixture can provide superior control of the pathogen complex inciting apple replant disease relative to either seed meal used alone.     

A One-Step, Immunochromatographic Lateral Flow Device Specific to Rhizoctonia solani and Certain Related Species, and Its Use to Detect and Quantify R. solani in Soil

ABSTRACT A murine hybridoma cell line GD2 secreting an immunoglobulin (Ig)M monoclonal antibody (MAb) was produced against surface antigens from an anastomosis group (AG) 4 isolate of Rhizoctonia solani (teleomorph: Thanatephorus cucumeris). Ascites were produced in mice using GD2 hybridoma cells and used to develop a rapid immunochromatographic lateral flow device (LFD) for the detection of antigens from R. solani and certain related Rhizoctonia spp. The LFD was tested for specificity against surface antigens from related and unrelated soil fungi. Antigens from representative isolates of R. solani AGs 1, 2-1, 2-3, 2-t, 3, 4, 5, 6, 7, 8, 9, 10, 11, and BI gave a positive response in LFD tests, as did antigens from Thanatephorus orchidicola, T. praticola, R. fragariae (teleomorph: Ceratorhiza fragariae), Ceratorhiza goodyerae-repentis, Ceratobasidium cornigerum, and binucleate AGE. Antigens from R. solani AGs 2-2, 2-2IIIB, and 2-2IV and from the related fungi R. carotae, R. cerealis (teleomorph: Ceratobasium cereale), R. crocorum (teleomorph: Helicobasidium brebissonii), R. oryzae (teleomorph Waitea circinata), and R. zeae gave negative responses, as did antigens from a range of unrelated fungi and oomycetes including Fusarium, Gliocladium, Trichoderma, Pythium, and Phytophthora spp. The usefulness of the LFD to detect R. solani was demonstrated in soils naturally infested with R. solani AG3. There was close agreement between results of LFD tests and conventional plate enrichment tests employing selective medium. The specificity of the technique was confirmed by polymerase chain reaction PCR using R. solani AG3-specific primers and by analyses based on sequences of the internal transcribed spacer (ITS)1-5.8S-ITS2 rRNA-encoding regions of unrelated fungi recovered from soil samples. The LFD was used to quantify R. solani AG4 in artificially infested soil samples (chopped potato soil inoculum). Estimates of CFU per gram of soil were derived using a most-probable number technique, which was based on the presence or absence of a detectable signal in the LFD. Estimates of CFU obtained in LFD tests and those obtained in a plate-trapped antigen enzyme-linked immunosorbent assay incorporating MAb GD2 were identical (449 CFU g(-1) of soil).     

Detecting Migrants in Populations of Rhizoctonia solani Anastomosis Group 3 from Potato in North Carolina Using Multilocus Genotype Probabilities

ABSTRACT The relative contribution of migration of Rhizoctonia solani anastomosis group 3 (AG-3) on infested potato seed tubers originating from production areas in Canada, Maine, and Wisconsin (source population) to the genetic diversity and structure of populations of R. solani AG-3 in North Carolina (NC) soil (recipient population) was examined. The frequency of alleles detected by multilocus polymerase chain reaction-restriction fragment length polymorphisms, heterozygosity at individual loci, and gametic phase disequilibrium between all pairs of loci were determined for subpopulations of R. solani AG-3 from eight sources of potato seed tubers and from five soils in NC. Analysis of molecular variation revealed little variation between seed source and NC recipient soil populations or between subpopulations within each region. Analysis of population data with a Bayesian-based statistical method previously developed for detecting migration in human populations suggested that six multilocus genotypes from the NC soil population had a statistically significant probability of being migrants from the northern source population. The one-way (unidirectional) migration of genotypes of R. solani AG-3 into NC on infested potato seed tubers from Canada, Maine, and Wisconsin provides a plausible explanation for the lack of genetic subdivision (differentiation) between populations of the pathogen in NC soils or between the northern source and the NC recipient soil populations.     

Characterization and origin of infection of Rhizoctonia solani associated with Brassica oleracea crops in the UK

An extensive study was conducted to determine where in the production chain Rhizoctonia solani became associated with UK module-raised Brassica oleracea plants. In total, 2600 plants from 52 crops were sampled directly from propagators and repeat sampled from the field. Additional soil, compost and water samples were collected from propagation nurseries and screened using conventional agar isolation methods. No isolates of R. solani were recovered from any samples collected from propagation nurseries. Furthermore, nucleic acid preparations from samples of soil and compost from propagation nurseries gave negative results when tested for R. solani using real-time PCR. Conversely, R. solani was recovered from 116 of 1300 stem bases collected from field crops. All the data collected suggested R. solani became associated with B. oleracea in the field rather than during propagation. Parsimony and Bayesian phylogenetic studies of ribosomal DNA suggested the majority of further classified isolates belonged to anastomosis groups 2-1 (48/57) and AG-4HGII (8/57), groups known to be pathogenic on Brassica spp. in other countries. Many R. solani isolates were recovered from symptomless plant material and the possibilities for such an association are discussed.     

新疆北疆棉田立枯丝核菌不同菌丝融合群致病力的研究

从新疆北疆棉区采集了典型的棉花立枯病病苗及棉田土标样686份,按常规分离方法分离得到399个分离物,从中鉴定出272个纯化的立枯丝核菌(Rhizoctonia solani Kühn)菌株。用标准菌株,通过载玻片菌丝融合试验测定,将纯化的272个菌株划归为3个菌丝融合群:即AG-2、AG-4和AG-5,分别占总菌株的6.24%、84.2%和1.1%。另有23个菌株不与任何标准菌株融合,占8.46%,说明新疆北疆棉田立枯丝核菌的优势菌系是多核丝核菌的AG-4融合群。通过从10种不同配方培养基中筛选效果好的麦芽蛋白胨(MPDA)配方培养基(Ⅱ)进行对峙培养,将纯化获得的272个丝核菌菌株,划分为6个不同的营养亲和群。将多核的不同菌丝融合群及其各营养亲和群代表菌株在3种不同主栽棉花品种上(每品种都加不接菌的对照)进行温室盆栽致病力测定,结果以AG-4对棉花的致病力最强,其次是非融合类,AG-2和AG-5虽致病,但并不致死苗,其平均病指数分别为94.9、81.4、53.1、43.5。     

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.     

Soybean Brown Stem Rot, Phytophthora sojae, and Heterodera glycines Affected by Soil Texture and Tillage Relations

ABSTRACT Investigations were conducted to determine whether the effects of tillage practices on the prevalence of brown stem rot of soybean (caused by Phialophora gregata), Heterodera glycines, and Phytophthora sojae were confounded by soil texture in samples collected in the fall of 1995 and 1996. Soil and soybean stem samples, along with tillage information, were collected from 1,462 randomly selected fields in Illinois, Iowa, Minnesota, Missouri, and Ohio in collaboration with the National Agricultural Statistics Service. The incidence of brown stem rot was determined from 20 soybean stem pieces collected from each field in a zigzag pattern. The detection frequency of P. sojae (expressed as percent leaf disks colonized) and population densities of H. glycines were determined from soil cores also collected in a zigzag pattern. The soil samples were grouped into various textural classes, and the effect of soil texture and tillage relations on the activities of each pathogen were determined. Both tillage and soil texture affected the incidence of brown stem rot; however, there was no interaction between tillage and soil texture. Conservation tillage had a greater (P < 0.05) incidence of brown stem rot in clay loam and silty clay loam than did conventional tillage. The detection frequency of P. sojae was not affected by tillage, but a tillage x texture interaction (P = 0.013) indicated that the effect of tillage depended on soil texture. There was a greater (P < 0.05) detection frequency of P. sojae in conservation tillage than in conventional tillage in silt loam and loam soils. However, in sandy loam, the detection frequency of P. sojae was greater (P = 0.0099) in conventional tillage than in conservation tillage. Population densities of H. glycines were significantly affected by both tillage and soil texture, but overall, there was no tillage x texture interaction. There was an inverse relationship between population densities of H. glycines and percent clay (r = -0.81, P = 0.01) in no-till fields, but little or no change in nematode densities was observed with increasing clay content in tilled fields. Population densities of H. glycines were less (P < 0.05) in no-till fields than in tilled fields in silty clay loam and clay soils. There was no difference in H. glycines densities between the tillage categories in soils sandier than silty clay loam or clay. The findings emphasize the need for cautious interpretation of the effects of tillage practices on diseases and pathogens in the absence of information on soil texture.     

新疆11种豆科作物立枯丝核菌菌丝融合群及营养亲和群研究

从新疆11种豆科作物病株上或病株根围土样中分离纯化出250个立枯丝核菌（Rhizoctonia DC）,番红O KOH染色后观察细胞核数目,经测试全部菌株均为多核,用标准菌株测定融合群, 250个菌株分属为AG 1、AG 2、AG 3、AG 4和AG 5共5个融合群,出现频率分别为16.4%、33.2%、0.4%、32.4%和17.6%,营养亲合群判别结果表明,AG 1、AG 2、AG 4和AG 5下各有2个VCG,说明新疆豆科作物立枯丝核菌各主要菌丝融合群内均有不同程度的分化。     

烟草靶斑病菌菌丝融合群及ITS序列分析

 烟草靶斑病是2006年我国新报道发生的一种叶部病害^［1］,其病原的无性世代为立枯丝核菌(Rhizoctonia solani Kühn),有性世代为瓜亡革菌(Thanatephorus cucumeris (Frank)Donk)。该病菌主要危害叶部形成病斑,对烟草的产量和品质影响显著,目前该病害主要分布在辽宁省丹东和铁岭地区,并呈现出迅速蔓延趋势。烟草靶斑病最早由巴西报道,此后,哥斯达黎加、美国、南非和津巴布韦也相继发生^［2,3］。     

Characterization of AG-13, a Newly Reported Anastomosis Group of Rhizoctonia solani

ABSTRACT Rhizoctonia solani anastomosis group (AG)-13 was collected from diseased roots of field grown cotton plants in Georgia in the United States. Isolates of AG-13 did not anastomose with tester isolates of AG-1 through AG-12. Mycelium of all isolates of AG-13 were light brown but darkened as cultures aged. All isolates produced aerial mycelium. Concentric rings were visible after 3 to 4 days of growth but disappeared as cultures aged and darkened. Individual sclerotia were up to 1.5 mm in diameter, similar in color to the mycelium, and generally embedded in the agar. Clumps of sclerotia up to 5 mm in diameter were produced on the agar surface. All attempts to induce basidiospore production were unsuccessful. The 5.8S region of the rDNA from isolates of AG-13 was identical in length and sequence to isolates of all other AGs of R. solani. Length and sequence of the internal transcribed spacer (ITS) regions of rDNA from isolates of AG-13 were unique among AGs of R. solani. Similarity between AG-13 and other AGs of R. solani ranged from 68 to 85% for ITS region 1 and 85 to 95% for ITS region 2. Selected isolates of AG-13 caused minor or no damage to barley, cauliflower, cotton, lettuce, potato, and radish in laboratory or greenhouse studies.     

Effect of Seed Quality and Combination Fungicide-Trichoderma spp. Seed Treatments on Pre- and Postemergence Damping-Off in Cotton

ABSTRACT Good quality seeds of cotton cultivars often escaped pre-emergence damping-off incited by Pythium spp. and Rhizopus oryzae, and they were resistant to postemergence damping-off incited by Rhizoctonia solani. Poor quality seeds, however, were highly susceptible to both phases of seedling disease and required seed treatment in order to survive. Pre-emergence damping-off incited by Pythium spp. and Rhizopus oryzae could be controlled by seed treatment with biocontrol preparations of a number of Trichoderma spp., but these treatments were much less effective in controlling postemergence disease incited by Rhizoctonia solani. Postemergence seedling disease can be controlled by fungicides, but they were much less effective in controlling the pre-emergence phase of the disease. Combination seed treatments of poor quality cotton seeds with fungicides and Trichoderma spp. preparations, followed by planting in pathogen-infested soil, indicated that this technique will control both phases of seedling disease. Seed treatment with either the fungicides or the biocontrol agents alone did not achieve this goal. The optimum combination treatment for disease control was that of chloroneb plus Trichoderma spp., followed by chloroneb plus metalaxyl (Deltacoat AD) plus T. virens strain G-6.     

Protection of potato from Rhizoctonia canker with binucleate Rhizoctonia fungi

Fifteen isolates of binucleate Rhizoctonia fungi (BNR) were studied as potential biocontrol agents for protection of potato from Rhizoctonia canker in artificially infested greenhouse soil and potato fields naturally infested with Rhizoctonia solani (AG-3). Eight of the BNR reduced incidence and severity of Rhizoctonia stem canker in greenhouse experiments by an average of 78 and 85%, respectively. In a field naturally infested with R. solani, selected isolates of BNR and the fungicide Tops 2.5D (thiophanate-methyl) were equally protective of potato from Rhizoctonia stem canker. BNR isolates gave protection of potato from Rhizoctonia stolon canker similar to PCNB and superior to Tops 2.5D. Cultivars Atlantic, Irish Cobbler, Kennebec, Norchip, Russet Burbank, and Superior were protected equally from Rhizoctonia stem canker by selected isolates of BNR under field conditions. Isolates of BNR show potential as biocontrol agents for protection of potato from Rhizoctonia canker.     

Mapping rhizoctonia patch in consecutive cereal crops in Western Australia

Rhizoctonia patches in a field were mapped during consecutive seasons of cropping with wheat, oats or barley sown directly without tillage into the previous season's stubble. Distribution, size, shape and number of patches varied considerably between seasons. Less than a quarter of patches were circular and the others tended to be elongated parallel to seed-rows. Barley was most severely affected, followed by wheat and then oats.
It is proposed that changes in patch distribution, configuration and number between seasons is due to shifts in a balance of suppressive or conducive soil factors interacting with Rhizoctonia solani inoculum.     

Isolation and identification of hypovirulent Rhizoctonia spp. from soil

Two-hundred and forty-eight isolates of Rhizoctonia spp, were obtained from 13 locations in Gifu Prefecture in Japan using the plant debris particles isolation, colonization of bait tissue, and soil-clump plating methods. Of the isolates, 143 were binucleate Rhizoctonia spp., 60 were R. solani and 45 were R. zeae. Three isolates of R. solani and 54 of binucleate Rhizoctonia spp, were hypovirulent on radish, whilst all isolates of R. zeae were highly virulent, Hypovirulent strains were isolated most frequently by the plant debris particles isolation method, Hypovirulent isolates of R. solani belonged to anastomosis group 4, whilst the hypovirulent binucleate Rhizoctonia isolates belonged to AG A, AG Ba, AG G, and AG O.
Thirty-two isolates of Rhizoctotria spp, selected for hypovirulence on radish were tested on cucumber in vitro. Only five binucleate Rhizoctonia isolates and one R. solani isolate were hypovirulent on both species, and these isolates were also hypovirulent on seven other crop species. Cucumber showed wide variation in disease susceptibility to different isolates but hypovirulent isolates exhibited a consistent reaction on five different host cultivars, Pathogenicity tests using cucumber grown in soil also showed consistent reactions with isolates selected either for hypovirulence or virulence. The results support the use of cucumber in bioassays for identifying hypovirulent isolates of binucleate Rhizoctonia spp.