Solani Anastomosis Group Research Articles

Divergence Between Sympatric Rice- and Soybean-Infecting Populations of Rhizoctonia solani Anastomosis Group-1 IA

Rhizoctonia solani anastomosis group (AG)-1 IA causes soybean foliar blighting (aerial blight) and rice sheath blight diseases. Although taxonomically related within the AG-1 complex, sister populations of R. solani AG-1 IA infecting Poaceae (rice) and Fabaceae (soybean) are genetically distinct based on internal transcribed spacer rDNA. However, there is currently no information available regarding the extent of genetic differentiation and host specialization between rice- and soybean-infecting populations of R. solani AG-1 IA. We used 10 microsatellite loci to compare sympatric R. solani AG-1 IA populations infecting rice and soybeans in Louisiana and one allopatric rice-infecting population from Texas. None of the 154 multilocus genotypes found among the 223 isolates were shared among the three populations. Partitioning of genetic diversity showed significant differentiation among sympatric populations from different host species (Phi(ST) = 0.39 to 0.41). Historical migration patterns between sympatric rice- and soybean-infecting populations from Louisiana were asymmetrical. Rice- and soybean-derived isolates of R. solani AG-1 IA were able to infect both rice and soybean, but were significantly more aggressive on their host of origin, consistent with host specialization. The soybean-infecting population from Louisiana was more clonal than the sympatric rice-infecting population. Most of the loci in the soybean-infecting populations were out of Hardy-Weinberg equilibrium (HWE), but the sympatric rice-infecting population from Louisiana was mainly in HWE. All populations presented evidence for a mixed reproductive system.

Genetic Structure of Populations of Rhizoctonia solani Anastomosis Group-1 IA from Soybean in Brazil

The Basidiomycete fungus Rhizoctonia solani anastomosis group (AG)-1 IA is a major pathogen of soybean in Brazil, where the average yield losses have reached 30 to 60% in some states in Northern Brazil. No information is currently available concerning levels of genetic diversity and population structure for this pathogen in Brazil. A total of 232 isolates of R. solani AG1 IA were collected from five soybean fields in the most important soybean production areas in central-western, northern, and northeastern Brazil. These isolates were genotyped using 10 microsatellite loci. Most of the multilocus genotypes (MLGTs) were site-specific, with few MLGTs shared among populations. Significant population subdivision was evident. High levels of admixture were observed for populations from Mato Grosso and Tocantins. After removing admixed genotypes, three out of five field populations (Maranhao, Mato Grosso, and Tocantins), were in Hardy-Weinberg (HW) equilibrium, consistent with sexual recombination. HW and gametic disequilibrium were found for the remaining soybean-infecting populations. The findings of low genotypic diversity, departures from HW equilibrium, gametic disequilibrium, and high degree of population subdivision in these R. solani AG-1 IA populations from Brazil are consistent with predominantly asexual reproduction, short-distance dispersal of vegetative propagules (mycelium or sclerotia), and limited long-distance dispersal, possibly via contaminated seed. None of the soybean-infecting populations showed a reduction in population size (bottleneck effect). We detected asymmetric historical migration among the soybean-infecting populations, which could explain the observed levels of subdivision.

Characterization and pathogenicity of Rhizoctonia isolates associated with cauliflower in Belgium

Sixty‐two isolates of Rhizoctonia spp. were collected from Belgian cauliflower fields during 2005 and 2006. The majority of the isolates (60 out of 62) had multinucleate cells and were identified as Rhizoctonia solani. Characterization of anastomosis groups (AGs) was performed using pectic zymograms, PCR‐RFLP and sequencing of the rDNA‐ITS region. The most prevalent AG was AG 2‐1 (55% of isolates), followed by AG 2‐1 subset Nt (11%), AG 1‐1C (8%), AG 5 (8%), AG 4 HGII (6%), AG 3 (5%) and AG 1‐1B (3%). Pathogenic potential towards different vegetable crops and towards maize was determined. Damage to cauliflower and endive was caused by different AGs, with the isolates aggressive towards cauliflower belonging to AG 2‐1, AG 2‐1 subset Nt, AG 4 HGII, AG 1‐1C, AG 1‐1B and AG 2‐2, and those aggressive towards endive belonging to AG 1‐1B, AG 1‐1C, AG 2‐1 subset Nt, AG 2‐2, AG 4 HGII and AG 5. The most aggressive isolates towards bean belonged to AG 2‐1 subset Nt and AG 2‐2, for lettuce to AG 1‐1B and AG 2‐1, on carrot to AG 4 HGII and towards maize to AG 2‐2. Within the isolates of AG 2‐1, variability was observed in PCR‐RFLP pattern and in aggressiveness towards several crops, indicating this subgroup to be heterogeneous. This is the first study concerning the occurrence of R. solani AGs causing wirestem in Belgian cauliflower fields and the first report of aggressive isolates of AG 1‐1C, AG 2‐1 subset Nt and AG 4 HGII associated with cauliflower.

Highly polymorphic microsatellite loci in the rice‐ and maize‐infecting fungal pathogen Rhizoctonia solani anastomosis group 1 IA

Ten polymorphic microsatellite loci were isolated and characterized from the rice- and maize-infecting Basidiomycete fungus Rhizoctonia solani anastomosis group AG-1 IA. All loci were polymorphic in two populations from Louisiana in USA and Venezuela. The total number of alleles per locus ranged from four to eight. All 10 loci were also useful for genotyping soybean-infecting R. solani AG-1 isolates from Brazil and USA. One locus, TC06, amplified across two other AG groups representing different species, showing species-specific repeat length polymorphism. This marker suite will be used to determine the global population structure of this important pathogenic fungus.

O papel de <em>Rhizoctonia</em> spp. binucleadas na indução de resistência a mela da soja

The role of non-pathogenic binucleate Rhizoctonia spp. (BNR) on the biocontrol of diseases caused by R. solani on many crops has been reported in the literature. However, in Brazil, there is no information about the potential of BNR as biocontrol agents against Rhizoctonia diseases on soybean. On this research we tested the hypothesis that BNR can induce resistance on soybean against the foliar blight caused by R. solani anastomosis group (AG) 1 IA. Thus, the objective of this research was to evaluate BNR isolates isolated from peanuts, snapbeans and soybean according to their ability for inducing resistance on soybean against the foliar blight disease, under greenhouse conditions. This research evidenced the role of BNR inducing resistance on soybeans against the foliar blight. However, both the occurrence and effectiveness of the phenomenon of induced resistance are dependent on the soybean cultivation season

First Report of Rhizoctonia solani Subgroup AG 1-ID Causing Leaf Blight on Durian in Vietnam.

During the rainy season in Vietnam, leaf blight disease caused by Rhizoctonia solani often occurs on 3- to 5-year-old durian (Durio zibethinus). Symptoms appear as large, pale brown, blighted lesions with an irregular border. In excessive moisture conditions, yellowish white hyphae appear on the lesions, and the affected leaves turn dark brown and wilt. There are no reports describing the anastomosis groups (AG) and subgroups of Rhizoctonia solani occurring in durian. In June of 2004, two isolates of R. solani were obtained from leaf blight lesions on durian growing in Binh Duong and Dong Nai provinces. The durian isolates were identified as AG 1 based on hyphal anastomosis. In pathogenicity tests, the durian isolates infected cucumber, mung bean, and leaf mustard seedlings grown on water agar in petri dishes. The rDNA-ITS sequence of the durian isolates was determined (GenBank Accession Nos. EF197797 and EF197798) and aligned with those of AG 1-IA, AG 1-IB, AG 1-IC, and AG 1-ID available in the GenBank database. The sequence similarity of the total rDNA-ITS region (including 5.8S) within the durian isolates was 99.9%. The sequence similarity of the durian isolates and AG 1-ID isolates was 99.1 to 100%, but similarity with other AG 1 subgroups was 89.1 to 94.0%. The results suggest that the two Vietnam durian isolates of R. solani are members of AG 1-ID. AG 1-ID has only been reported causing necrotic leaf spots on coffees in the Philippines (1). To our knowledge, this is the first report of R. solani AG 1-ID on durian and the first report of the presence of R. solani AG 1-ID in Vietnam. Reference: (1) A. Priyatmojo et al. Phytopathology 91:1054, 2001.

Transmission of the M2 double-stranded RNA in Rhizoctonia solani anastomosis group 3 (AG-3)

Horizontal transmission of the 3.57 kb M2 double-stranded RNA (dsRNA) between mycelia of somatically incompatible isolates of Rhizoctonia solani anastomosis group 3 (AG-3), an economically important pathogen of cultivated plants in the family Solanaceae, was investigated. Nine donor isolates of R. solani AG-3 containing the M2 dsRNA were paired on potato-dextrose agar with each of three different recipient isolates where the M2 dsRNA was absent. Reverse-transcription PCR (RT-PCR) was used to detect horizontal transmission of the M2 dsRNA via hyphal anastomosis from donor to recipient isolates by examining hyphal explants taken 3 cm from the hyphal interaction zone. PCR-RFLP genetic-based markers of two nuclear loci and one mitochondrial locus were used to confirm identity and transmission between donor and recipient isolates of R. solani AG-3. The frequency of transmission observed between 72 pairings of the eight donor and three recipient isolates was approximately 4% of the total pairings, and differences in the phenotype of the recipient isolates after acquisition of the M2 dsRNA via horizontal transmission were observed. To our knowledge this represents the first demonstration of transmission of dsRNA between genetically different individuals of R. solani confirmed with nuclear and mitochondrial markers. These results suggest that transmission can occur between somatically incompatible isolates of R. solani AG-3 but that maintenance of the dsRNA in the recipient isolates was not stable after repeated subculturing on nutrient medium.

Transmission of the M2 double-stranded RNA in Rhizoctonia solani anastomosis group 3 (AG-3)

Horizontal transmission of the 3.57 kb M2 double-stranded RNA (dsRNA) between mycelia of somatically incompatible isolates of Rhizoctonia solani anastomosis group 3 (AG-3), an economically important pathogen of cultivated plants in the family Solanaceae, was investigated. Nine donor isolates of R. solani AG-3 containing the M2 dsRNA were paired on potato-dextrose agar with each of three different recipient isolates where the M2 dsRNA was absent. Reverse-transcription PCR (RT-PCR) was used to detect horizontal transmission of the M2 dsRNA via hyphal anastomosis from donor to recipient isolates by examining hyphal explants taken 3 cm from the hyphal interaction zone. PCR-RFLP genetic-based markers of two nuclear loci and one mitochondrial locus were used to confirm identity and transmission between donor and recipient isolates of R. solani AG-3. The frequency of transmission observed between 72 pairings of the eight donor and three recipient isolates was approximately 4% of the total pairings, and differences in the phenotype of the recipient isolates after acquisition of the M2 dsRNA via horizontal transmission were observed. To our knowledge this represents the first demonstration of transmission of dsRNA between genetically different individuals of R. solani confirmed with nuclear and mitochondrial markers. These results suggest that transmission can occur between somatically incompatible isolates of R. solani AG-3 but that maintenance of the dsRNA in the recipient isolates was not stable after repeated subculturing on nutrient medium.

Phylogeography of the Solanaceae-infecting Basidiomycota fungus Rhizoctonia solani AG-3 based on sequence analysis of two nuclear DNA loci

BackgroundThe soil fungus Rhizoctonia solani anastomosis group 3 (AG-3) is an important pathogen of cultivated plants in the family Solanaceae. Isolates of R. solani AG-3 are taxonomically related based on the composition of cellular fatty acids, phylogenetic analysis of nuclear ribosomal DNA (rDNA) and beta-tubulin gene sequences, and somatic hyphal interactions. Despite the close genetic relationship among isolates of R. solani AG-3, field populations from potato and tobacco exhibit comparative differences in their disease biology, dispersal ecology, host specialization, genetic diversity and population structure. However, little information is available on how field populations of R. solani AG-3 on potato and tobacco are shaped by population genetic processes. In this study, two field populations of R. solani AG-3 from potato in North Carolina (NC) and the Northern USA; and two field populations from tobacco in NC and Southern Brazil were examined using sequence analysis of two cloned regions of nuclear DNA (pP42F and pP89).ResultsPopulations of R. solani AG-3 from potato were genetically diverse with a high frequency of heterozygosity, while limited or no genetic diversity was observed within the highly homozygous tobacco populations from NC and Brazil. Except for one isolate (TBR24), all NC and Brazilian isolates from tobacco shared the same alleles. No alleles were shared between potato and tobacco populations of R. solani AG-3, indicating no gene flow between them. To infer historical events that influenced current geographical patterns observed for populations of R. solani AG-3 from potato, we performed an analysis of molecular variance (AMOVA) and a nested clade analysis (NCA). Population differentiation was detected for locus pP89 (ΦST = 0.257, significant at P < 0.05) but not for locus pP42F (ΦST = 0.034, not significant). Results based on NCA of the pP89 locus suggest that historical restricted gene flow is a plausible explanation for the geographical association of clades. Coalescent-based simulations of genealogical relationships between populations of R. solani AG-3 from potato and tobacco were used to estimate the amount and directionality of historical migration patterns in time, and the ages of mutations of populations. Low rates of historical movement of genes were observed between the potato and tobacco populations of R. solani AG-3.ConclusionThe two sisters populations of the basidiomycete fungus R. solani AG-3 from potato and tobacco represent two genetically distinct and historically divergent lineages that have probably evolved within the range of their particular related Solanaceae hosts as sympatric species.

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.

Incompatibilidade somática em Rhizoctonia solani AG-1 IA da soja

O fungo Rhizoctonia solani AG-1 IA é um dos patógenos mais importantes que afeta a cultura da soja no Brasil, causando a mela ou queima foliar. A doença está associada com a fase teleomórfica de R. solani, o basidiomiceto Thanatephorus cucumeris. Neste estudo, baseando em conhecimento prévio sobre a biologia de R. solani AG-1 IA, duas hipóteses foram testadas. Na primeira hipótese postulou-se a ocorrência de incompatibilidade somática em populações de R. solani AG-1 IA. A segunda hipótese testada foi de que esta população de R. solani AG-1 IA da soja apresenta indicações de estrutura sexual clonal. Duas amostras de isolados de R. solani AG-1 IA da soja obtidas no Maranhão e no Mato Grosso foram utilizadas. Na primeira amostra, foram selecionados isolados apresentando diferentes perfis de RAPD (Random Amplified Polymorphic DNA), procurando maximizar a diversidade dos isolados, e evitando a introdução de possíveis clones no teste. Os isolados foram pareados em todas as combinações possíveis em meio de BDA mais carvão ativado e examinados quanto às interações somáticas resultantes. Seis grupos de incompatibilidade somática (GCS) foram detectados entre 24 isolados do AG-1 IA. Entretanto, análises microscópicas dos pareamentos entre isolados indicaram maior freqüência de incompatibilidade somática, impossibilitando o grupamento em GCS. No geral, a metodologia de avaliação das interações somáticas macroscópicas em meio BDA + carvão ativado, não se mostrou totalmente apropriada para discriminação das categorias de reações de compatibilidade entre isolados de R. solani AG-1 IA. Com a segunda amostra procurou-se determinar a ocorrência de clones na população do patógeno, ou seja, isolados que compartilham o mesmo padrão fenotípico de RAPD e somaticamente compatíveis. No caso de R. solani AG 1 IA da soja, a gama de interações somáticas entre pareamentos de isolados e, principalmente, os desvios na associação estrita entre os GCS detectados neste trabalho, conjuntamente com os perfis de RAPD observados anteriormente por Fenille (11) e Meyer (20), são consistentes com recombinação. Entretanto, o patógeno ainda apresenta um componente clonal expressivo na população. De um total de 43 isolados, os exemplos de prováveis clones na população do patógeno totalizaram 16 isolados.

Effects of water potential on mycelial growth, sclerotial production, and germination of Rhizoctonia solani from potato

The effects of osmotic and matric potential on mycelial growth, sclerotial production and germination of isolates of Rhizoctonia solani [anastomosis groups (AGs) 2-1 and 3] from potato were studied on potato dextrose agar (PDA) adjusted osmotically with sodium chloride, potassium chloride, glycerol, and matrically with polyethylene glycol (PEG) 6000. All isolates from AGs 2-1 and AG-3 exhibited fastest mycelial growth on unamended PDA (−0.4 MPa), and growth generally declined with decreasing osmotic and matric potentials. Growth ceased between −3.5 and −4.0 MPa on osmotically adjusted media, and at −2.0 MPa on matrically adjusted media, with slight differences between isolates and osmotica. Sclerotium yield declined with decreasing osmotic potential, and formation by AG 2-1 and AG-3 isolates ceased between −1.5 and −3.0 MPa and −2.5 and −3.5 MPa, respectively. On matrically adjusted media, sclerotial formation by AG 2-1 isolates ceased at −0.8 MPa, whereas formation by AG-3 isolates ceased at the lower matric potential of −1.5 MPa. Sclerotial germination also declined with decreasing osmotic and matric potential, with total inhibition occurring over the range −3.0 to −4.0 MPa on osmotically adjusted media, and at −2.0 MPa on matrically adjusted media. In soil, mycelial growth and sclerotial germination of AG-3 isolates declined with decreasing total water potential, with a minimum potential of −6.3 MPa permitting both growth and germination. The relevance of these results to the behaviour of R. solani AGs in soil and their pathogenicity on potato is discussed.

The rmc locus does not affect plant interactions or defence-related gene expression when tomato (Solanum lycopersicum) is infected with the root fungal parasite, Rhizoctonia.

A tomato mutant with reduced mycorrhizal colonisation, rmc, confers resistance to almost all arbuscular mycorrhizal (AM) fungal species tested, although there is variation in colonisation of different root cell layers by different fungi and one species of AM fungus can colonise this mutant relatively normally. These variations indicate a high degree of specificity in relation to AM colonisation. We explored the possibility of specificity or otherwise in interactions between rmc and three non-AM root-infecting fungi, Rhizoctonia solani anastomosis groups (AG) 4 and AG8, and binucleate Rhizoctonia (BNR). There were no differences between the wild type tomato 76R and rmc in the speed or extent to which these fungi infected roots or caused disease. Infection by R. solani induced high levels of defence-related gene expression in both tomato genotypes relative to non-infected plants. In contrast, with BNR the expression of these genes was not induced or induced to a much lower extent than with R. solani. The expression of defence-related genes with these two non-AM fungi was very similar in the two plant genotypes. It was different from effects observed during colonisation by AM fungi, which enhanced expression of defence-related genes in rmc compared with the wild type tomato. The specificity and molecular mechanisms of rmc in control of AM colonisation are discussed.

A New Method for the Quantification of Rhizoctonia solani and R. oryzae from Soil.

Rhizoctonia solani anastomosis group (AG) 8 and R. oryzae are important root pathogens on wheat and barley in the dryland production areas of the inland Pacific Northwest. R. solani AG-8 is difficult to isolate from root systems and quantify in soil because of slow growth and low population densities. However, both pathogens form extensive hyphal networks in the soil and can grow a considerable distance from a food base. A quantitative assay of active hyphae was developed, using wooden toothpicks as baits inserted into sample soils. After 2 days in soil, toothpicks were placed on a selective medium, and the numbers of colonies that grew after 24 h were counted under a dissecting microscope. R. solani and R. oryzae could be distinguished from other fungi based on hyphal morphology. This method was tested in natural soils amended with known inoculum densities of R. solani AG-8 and R. oryzae. Regressions were used to compare the inoculum density or toothpick colonization curves to a predicted curve based on the volume of the toothpicks. The slopes and y intercept of log-log transformed regressions did not differ from the predicted curves in most cases. This technique was used to assess the hyphal activity of R. solani AG-8 and R. oryzae from soil cores taken from various positions in and around Rhizoctonia bare patches at two locations. Activity of R. solani was highest in the center and inside edge of the patch, but there was no effect of patch position on R. oryzae. This simple and inexpensive technique can be used for detection and diagnosis in grower fields and to study the ecology and epidemiology of Rhizoctonia spp.

A One-Step, Immunochromatographic Lateral Flow Device Specific toRhizoctonia solaniand Certain Related Species, and Its Use to Detect and QuantifyR. solaniin 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).

Root growth altered by compaction of a sandy loam soil affects severity of rhizoctonia root rot of wheat seedlings

Rhizoctonia solani Anastomosis Group 8 damages seedling roots of wheat, causing the 'bare-patch' disease. This makes the first 4 weeks after germination the most critical period for disease development. As the field inoculum of the pathogen is mainly concentrated in the surface 10 cm of soil, the rate of root growth becomes critical for the vulnerable tissues of the root to escape the attack from the inoculum zone. To evaluate the effect of alteration of root growth by soil compaction on disease severity, a study was undertaken in 40-cm-deep pots made from PVC pipes (8.7 cm diameter). Four depths of soil compaction (whole soil profile compacted, whole soil loose, upper 10 cm loose and compacted below, upper 20 cm loose and compacted below) were tested using sieved soil. Effective root length of infected seedlings was higher in the pots where the whole soil profile was compacted than others. Reduction in dry root weights, where soil was compacted to heights of 0, 17.5, 27.5 or 37.5 cm following inoculation, were 68, 30, 74 and 56%, respectively. Reduction in shoot weights was 52, 22, 66 and 44%, respectively. Eight days after incubation, microbial activity was greater where the soil was highly compacted than where there was a low level of soil compaction. Saprophytic growth in soil of Rhizoctonia solani Anastomosis Group 8 was higher in loosely packed soil than in compacted soil. This shows that higher impact of disease under compacted soil conditions is due to reduced root growth and that disturbing the soil below seeds can reduce the impact of disease.

Genetic Diversity Among Iranian Isolates of Rhizoctonia solani Kühn Anastomosis Group1 Subgroups Based on Isozyme Analysis and Total Soluble Protein Pattern

Abstract Rhizoctonia solani is a destructive fungal pathogen with a wide host range. The R. solani complex species includes several divergent groups delimited by affinities for hyphal anastomosis. In this study, genetic variation among 20 isolates of R. solani anastomosis group 1 (AG1) subgroups (AG1‐IA and AG1‐IB) collected from Mâzandaran province, Iran, and standard isolates of these subgroups, was determined by isozyme analysis and total soluble protein profile. Mycelial protein pattern and isozyme analysis were studied using denaturing and non‐denaturing polyacrylamide gel electrophoresis, respectively. A total of 15 enzyme systems were tested, among which six enzymes including esterase, alkaline phosphatase, superoxide dismutase, octanol dehydrogenase, lactate dehydrogenase and mannitol dehydrogenase generated distinct and reproducible results. The soluble protein patterns were similar among the R. solani isolates examined; however, minor differences in banding pattern were observed between the two subgroups. In isozyme analysis, a total of 64 electrophoretic phenotypes were detected for all six enzymes used. Based on cluster analysis and similarity matrix, the fungal isolates were divided into two genetically distinct groups of I and II consistent with the previously reported AG1‐IA and AG1‐IB subgroups in AG1. Group I represented all isolates belonging to AG1‐IA subgroup, whereas group II represented all isolates belonging to AG1‐IB subgroup. Results from isozyme analysis suggest that the subgrouping concept within AGs is genetically based.

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.

Prolific production of sclerotia in soil by Rhizoctonia solani anastomosis group (AG) 11 pathogenic on lupin

SummaryRhizoctonia solani anastomosis group (AG) 11 causes serious damping‐off and hypocotyl rot of narrow‐leafed lupins (Lupinus angustifolius) in the northern grain‐belt of Western Australia. R. solani AG‐11 produced abundant sclerotia in sand overlaid on potato dextrose agar. Sclerotia were produced in larger numbers in natural Lancelin sand than in Geraldton loamy sand collected from the northern grain‐belt of Western Australia. The majority of the sclerotia produced were in &gt;250 to &lt;500 μam size range. The germination levels of sclerotia in the first two cycles of drying and germination were not significantly different. Sclerotia still retained 50% germination after four such cycles, indicating that they may have the ability to withstand the climatic cycles of the Mediterranean environment of southwestern Western Australia. The radial growth of the mycelium from sclerotia, however, declined with each drying and germination cycle. Inoculum potential of the pathogen increased with the size of sclerotia resulting in more severe lupin hypocotyl rot with larger sclerotia. The number of sclerotia produced in soil increased with increasing density of lupin seedlings. The results also indicate that R. solani AG‐11 can produce sclerotia on infected plant tissues as well as in soil. This is the first report of the prolific production of sclerotia by AG‐11 and their significant role in infection of lupins in soil in Western Australia.

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.