Rhizoctonia spp. Causing Root and Hypocotyl Rot in Phaseolus vulgaris in Cuba

Sixty isolates of Rhizoctonia spp. were obtained from Cuban bean fields during the period 2004–2007. Isolates were characterized with different techniques, including nuclei staining, pectic zymogram, PCR–RFLP analysis of the rDNA–ITS region and sequencing of the rDNA–ITS region. The majority of the isolates were identified as multinucleate Rhizoctonia solani isolates, representing two different anastomosis groups (AGs), AG 2‐2 WB and AG 4 HGI; the remaining isolates were binucleate Rhizoctonia isolates and belonged to AG F and AG A. AG 4 HGI isolates were equally distributed in all soil types; AG 2‐2 isolates were more frequently isolated from cambisols, whereas AG F isolates were related to calcisols. Pathogenicity experiments in vitro and in the greenhouse, revealed that binucleate isolates only caused root rot, whereas R. solani isolates were able to cause root rot and hypocotyl rot. Furthermore, differences in virulence level were observed between R. solani and binucleate isolates and among different AGs. Isolates of R. solani AG 4 HGI and R. solani AG 2‐2 WB were the most aggressive, binucleate isolates of AG F were intermediate aggressive, whereas a binucleate isolate of AG A was weakly aggressive. In contrast with other reports about R. solani in bean, web blight symptoms were never observed during this study.     

Molecular characterization of Rhizoctonia solani isolates the incitant of soybean root rot

Rhizoctonia solani isolates used in this investigation were identified as anastomosis-4 (AG-40), collected from different localities from Assiut governorate in Egypt. Pathogenicity test of seven isolates of R. solani was evaluated on soybean Giza 111 cultivar under greenhouse conditions. All tested isolates were able to infect soybean plants causing root rot with different degrees of severities, isolate No. 1, 2 and 3 showed significantly highest root rot severity, while isolate No. 5 gave the lowest percentage of root rot rating. The sodium dodecyl sulphate polyacrylamide gel electrophoresis patterns were used to compare three isolates of R. solani. There are no variations among R. solani isolates except a few exceptions according to their protein patterns. DNA markers obtained from all isolates showed genetic similarity among different isolates obtained from different geographical regions barring few exceptions. Correlation between DNA patterns of R. solani isolates and their virulence was detected, but no correlation with anastomosis groups (AG).     

Genetic Structure and Aggressiveness of Rhizoctonia solani AG1‐IA,the Cause of Sheath Blight of Rice in Southern China

One hundred and eighty isolates of Rhizoctonia solani AG1‐IA, the causal agent of rice sheath blight, were obtained from six locations in southern China. The genetic structure of R. solani isolates was investigated using random amplified polymorphic DNA (RAPD) markers, and a considerable genetic variation among R. solani isolates was observed. Most of the genetic diversity was distributed within populations, rather than among them. The distribution pattern of the genetic variation of R. solani appears to be the result of high gene flow (Nm) and low‐genetic differentiation among populations. The aggressiveness of R. solani was visually assessed by rice seedlings of five different cultivars in the glasshouse. All isolates tested were found to induce significantly different levels of disease severity, reflecting considerable variation in aggressiveness. The isolates were divided into highly virulent, moderately virulent and weakly virulent groups, and the moderately virulent isolates were dominant in R. solani population. No significant correlation was observed among the genetic similarity, pathogenic aggressiveness and geographical origins of the isolates. Information obtained from this study may be useful for breeding for improved resistance to sheath blight.     

First Report of Rhizoctonia solani AG‐7 on Cotton in Egypt

Eighty‐two isolates of Rhizoctonia solani were recorded from roots of naturally‐infected seedlings of the Egyptian cotton (Gossypium barbadense L.). Anastomosis groups (AGs) of the isolates were determined by using 13 different AGs testers. Three (3.7%) of the isolates were identified as R. solani AG7, while the remaining isolates were belonging to the AG 2‐1, AG4 and AG5. The identification of the three isolates was based on the frequency of the C2 reaction with the AG7 tester isolate. No fusion was observed between AG7 and isolates representing the other 13 AGs. Colonies of AG7 isolates grown on potato dextrose agar (PDA), malt yeast agar (MYA) and melt peptone agar (MPA) were brown to dark brown with aerial mycelium and sclerotia. The isolates had pitted sclerotial clusters and brownish exudates after 21 days of culturing on PDA, but without clear zonation. Pathogenicity test under greenhouse conditions revealed that AG7 caused the common symptoms of damping–off, which included seed rot, lesions on the hypocotyls and root rot.     

Characterization of Rhizoctonia solani AG 4HG-III Causing Stem Canker and Wirestem on Green Amaranth and Chinese Amaranth

During December 2003, stem canker and wirestem were observed on the stems of green amaranth (Amaranthus viridis) and Chinese amaranth (Amaranthus tricolor) in greenhouses at Ximao district in Yunnnan Province, China. Isolates of Rhizoctonia solani obtained from the two amaranths with stem canker and wirestem, were identical to anastomosis group (AG)‐4. The isolates from diseased plant showed high virulence on young seedlings of two amaranths. Results of sequence analysis of 5.8s rDNA‐ITS of Chinese isolates showed 99–100% sequence similarity with AG‐4HG‐III tester isolates. When compared with other subgroups of AG‐4, Chinese isolates showed similarity levels of 94%. This is the first report of stem canker and wirestem of Green amaranth and Chinese amaranth caused by AG‐4HG‐III and AG‐4HG‐III in China.     

Interactions Between Rhizoctonia Root Rot and the Foliar Common Bean Diseases Anthracnose and Rust

The effects of co‐inoculation of Rhizoctonia solani and Colletotrichum lindemuthianum or Uromyces appendiculatus at different inoculum levels were studied on the disease dynamics and on the growth of bean plants under greenhouse conditions. Bean seeds were sown in R. solani‐infested soil. Additional experiments in which seedlings were transplanted to infested soil were also carried out. Conidial suspensions of C. lindemuthianum or uredospores of U. appendiculatus were inoculated onto leaves at plant developmental stages V2 and V3, respectively. Interactions between root rot and the aerial diseases were observed depending on the inoculum levels and on the timing of R. solani inoculation. Anthracnose severity tended to be higher on R. solani‐infected plants. Conversely, R. solani infection significantly reduced diameter of pustules and rust severity. When seedlings were transplanted to soil infested with low levels of R. solani, root rot severity and density of R. solani in the soil were magnified at high levels of C. lindemuthianum or U. appendiculatus. In these experiments, a synergistic interaction between root rot and anthracnose was observed to affect the plant dry weight. Antagonistic effects on the plant dry weight were found for the combination root rot/rust only when seeds were sown in infested soil.     

Genetic Characterization and Pathogenicity of Rhizoctonia solani Associated with Common Bean Web Blight in the Main Bean Growing area of Argentina

Common bean web blight (WB), caused by the fungus Rhizoctonia solani (teleomorph Thanatephorus cucumeris), is among the endemic fungal diseases of major impact in north‐western Argentina (NWA). This study aimed to analyse the genetic and pathogenic diversity of R. solani in Salta, NWA, where 97 isolates were recovered from commercial bean cultivars and wild beans showing WB symptoms in a major bean production area. The isolates were characterized on the basis of specific primers, rDNA‐ITS sequences and morphological characteristics. All the isolates were identified as R. solani AG 2‐2WB, and they exhibited considerable intragroup variation. The phylogenetic tree generated with the ITS sequences confirmed the isolates identification. Aggressiveness of the isolates towards bean seedlings was assessed in the greenhouse. A great variability in virulence was observed among the isolates analysed. On the basis of the disease reaction on foliar tissues, the isolates were grouped into three virulence categories as follows: weakly virulent (30%), moderately virulent (38%) and highly virulent (32%). However, no correlation between virulence and geographical origin was detected. The information generated in this study provides initial data on the population variability of the WB pathogen in north‐western Argentina and represents a valuable contribution to regional breeding programmes aimed to obtain cultivars with durable resistance.     

Guava Decline: A Complex Disease Involving Meloidogyne mayaguensis and Fusarium solani

In Brazil, Meloidogyne mayaguensis has become a threat to guava production. Approximately a third of the cultivated area is infested, leading almost inevitably to the decimation of the orchards. Because parasitized trees develop rotten roots as the disease progresses, the possibility that a soil‐borne pathogen could be involved was investigated. From several nematode‐free or nematode‐infested orchards, nearly 2000 root fragments were tested for bacteria and fungi. Positive isolations were obtained from nematode‐infested areas only and were predominantly identified as Fusarium sp. In a 5‐month microplot experiment, guava seedlings were uninoculated (control) or were inoculated with M. mayaguensis only or with this nematode and 21 days later with one of 11 Fusarium sp. isolates. A Scott–Knot analysis of several vegetative variables and of the extent of root rot allowed the generation of a dissimilarity dendrogram that indicated that four Fusarium sp. isolates were particularly associated with damage to the seedlings. Upon identification of these isolates as Fusarium solani, a 6‐month microplot experiment was set up, in which guava seedlings were uninoculated or were inoculated with one of the following: (i) M. mayaguensis only, (ii) four F. solani isolates, separately, (iii) four F. solani isolates separately, combined with physical injury of the roots with a knife, (iv) M. mayaguensis, and 21 days later with four F. solani isolates, separately. No root rot and virtually no effect on all variables were observed in the seedlings inoculated with the fungus isolates, with or without physical injury. Major root rot and a negative effect on all variables were observed in the seedlings inoculated with M. mayaguensis and all four F. solani isolates. This characterizes guava decline as a complex disease caused by the synergistic effect of these organisms, in which parasitism by the nematode predisposes the plants to root decay caused by the fungus.     

Impact of Continuous Cropping of Lettuce on the Disease Dynamics of Bottom Rot and Genotypic Diversity of Rhizoctonia solani AG 1‐IB

The impact of continuous cropping of lettuce on the disease dynamics of bottom rot and genotypic diversity of the causal pathogen Rhizoctonia solani AG 1‐IB was studied over 3 years with two crops per year within a field naturally infested with R. solani the pathogen. This field had not had lettuce cultivated in it for 7 years. The disease incidence (DI) and disease severity (DS) were assessed at each harvest and mapped. Surprisingly, a high DI was already observed in the first crop of year one of this field study. In addition, the pathogen was also found to be evenly distributed. Severely infected plants occurred mainly in patches, and the position varied between crops. A significant increase in DS was already observed in the second year, and both temperature conditions and continuous cropping influenced the DS on average over time. Rhizoctonia isolates were randomly collected from the first crop in 1999 and the sixth crop in 2001. The genotypic diversity within the subgroup of R. solani AG 1‐IB was analysed by BOX‐PCR genomic fingerprinting and the aggressiveness of isolates by bioassay. The fingerprints revealed a high level of genotypic diversity within the AG 1‐IB field population. However, continuous cropping was found not to have an impact on genotypic diversity and aggressiveness.     

Anastomosis Group Determination of Rhizoctonia solani Kühn (Telemorph: Thanatephorus cucumeris) Isolates from Tomatoes Grown in Aydin,Turkey and their Disease Reaction on Various Tomato Cultivars

In the Province of Aydin‐Turkey most frequently Fusarium spp. and secondly Rhizoctonia solani Kühn were isolated from the roots and crown of tomato plants. Based on affinities for hyphal fusion with test isolates, all R. solani isolates were identified as AG‐4. The tomato cultivars which were grown in soil infested with R. solani AG‐4 exhibited different reactions. From the point of symptom expression and the rate of seedling emergence Sunny 6066 F1 was found to be the most resistant cultivar, whereas Rio Grande, Rio Fuego, NDM 725, Interpeel and Konia were the most susceptible cultivars.     

Identification and Pathogenicity of Rhizoctonia spp. Causing Wirestem of Betula nigra in China

During July 2004, wirestem was frequently observed on the seedlings of Betula nigra at Dehong district in Yunnan Province, China. Isolates of Rhizoctonia spp. consistently obtained from their diseased leaves, roots and stems were identified as belonging to binucleate Rhizoctonia anastomosis groups (AG) AG‐P and AG‐R, and R. solani AG‐I IB and AG‐4 HG‐I, based on cultural characteristics, nuclear staining, anastomisis reaction and analysis of their ITS rDNA region. The percentage of recovery of AG‐P, AG‐1, AG‐R and AG‐4 was 48%, 39%, 8% and 3%, respectively. This is the first report of wirestem of red birch cause by binucleate Rhizoctonia AG‐P and AG‐R, and R. solani AG‐1 IB and AG‐4 HG‐I in China.     

Interactions between a fluorescent pseudomonad,an arbuscular mycorrhizal fungus and a hypovirulent isolate of Rhizoctonia solani affect plant growth and root architecture of tomato plants

Abstract

Although Rhizoctonia solani is a cosmopolitan soilborne pathogen, the genus includes isolates with different pathogenicity ranging from high virulence to avirulence. The biocontrol strain Pseudomonas fluorescens P190r and the arbuscular mycorrhizal (AM) fungus Glomus mosseae BEG12 were inoculated alone or in combination in tomato plants infested by the mildly virulent pathogen R. solani #235. Plant growth as well as root morphometric and topological parameters were evaluated. The infection of R. solani was significantly reduced by all the combinations of the beneficial microorganisms. Root systems of R. solani‐infected plants were weakly developed but highly branched with a herring‐bone pattern, while those inoculated with the AM fungus, alone or in combination with the bacterial strain, were longer and more developed, and displayed a dichotomous pattern. The interactions among these three microorganisms affected plant growth and root architecture of tomato plants.     

Diversity of Rhizoctonia solani associated with pulse crops in different agro-ecological regions of India

Four hundred seventy Rhizoctonia solani isolates from different leguminous hosts originating from 16 agro-ecological regions of India covering 21 states and 72 districts were collected. The disease incidence caused by R. solani varied from 6.8 to 22.2 % in the areas surveyed. Deccan plateau and central highlands, hot sub-humid ecoregion followed by northern plain and central highlands and hot semi-arid ecoregion showed the highest disease incidence. R. solani isolates were highly variable in growth diameter, number, size and pattern of sclerotia formation as well as hyphal width. The isolates obtained from aerial part of the infected plants showing web blight symptoms produced sclerotia of 1–2 mm in size whereas, the isolates obtained from infected root of the plants showing wet root rot symptoms produced microsclerotia (<1 mm). Majority of R. solani isolates showed <8 μm hyphal diameter. Based on morphological characters the isolates were categorized into 49 groups. Seven anastomosis groups (AGs) were identified among the populations of R. solani associated with the pulse crops. The frequency (25.6 %) of AG3 was the highest followed by AG2–3 (20.9 %) and AG5 (17.4 %). The cropping sequence of rice/sorghum/wheat-chickpea/mungbean/urdbean/cowpea/ricebean influenced the dominance of AG1 (16.3 %). Phylogenetic analysis utilizing ITS-5.8S rDNA gene sequences indicated high level of genetic similarity among isolates representing different AGs, crops and regions. ITS groups did not correspond to the morphological characters. The sequence data from this article has been deposited with NCBI data libraries with JF701707 to JF701795 accession numbers.     

Bulb and Root Rot in Lily (Lilium longiflorum) and Onion (Allium cepa) in Israel

In the past 10 years, there has been a substantial increase in reports, from growers and extension personnel, on bulb and root rots in lily (Lilium longiflorum) in Israel. Rot in these plants, when grown as cut flowers, caused serious economic damage expressed in reduction in yield and quality. In lily, the fungal pathogens involved in the rot were characterized as binucleate Rhizoctonia AG‐A, Rhizoctonia solani, Pythium oligandrum, Fusarium proliferatum (white and purple isolates) and F. oxysporum, using morphological and molecular criteria. These fungi were the prevalent pathogens in diseased plants collected from commercial greenhouses. Pathogenicity trials were conducted on lily bulbs and onion seedlings under controlled conditions in a greenhouse to complete Koch's postulates. Disease symptoms on lily were most severe in treatments inoculated with binucleate Rhizoctonia AG‐A, P. oligandrum and F. proliferatum. Plant height was lower in the above treatments compared with the control plants. The least aggressive fungus was R. solani. In artificial inoculations of onion, seedling survival was significantly affected by all fungi. The most pathogenic fungus was F. proliferatum w and the least were isolates of F. oxysporum (II and III). All fungi were successfully re‐isolated from the inoculated plants.     

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

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.     

Characterization and Pathogenicity of Rhizoctonia spp. from Onion in Amasya, Turkey

Forty‐two isolates of Rhizoctonia spp. were obtained from onion in Amasya, Turkey. Of these, 29% were Rhizoctonia solani (AG‐4), 69% were Waitea circinata var. zeae (Rhizoctonia zeae) and 2% were binucleate Rhizoctonia (AG‐B). Most of the isolates were recovered from rhizosphere soil. In pathogenicity tests on onion, R. solani AG‐4 caused the greatest disease severity, those of W. circinata var. zeae were moderately virulent but binucleate Rhizoctonia isolates were of low virulence. This is the first report of binucleate Rhizoctonia AG‐B and W. circinata var. zeae occurring on onion in Turkey.     

Effects of the Plant Defence Inducer, Acibenzolar-S-Methyl, on Hypocotyl Rot of Soybean Caused by Rhizoctonia solani AG-4

The plant defence inducer, acibenzolar‐S‐methyl (ASM) was tested for its ability to protect soybean against hypocotyl rot caused by Rhizoctonia solani Kühn AG‐4. ASM in vitro exhibited an antifungal dose‐dependant activity in the form of reduced mycelial growth. This inhibition reached 40% in comparison with the control at a concentration of 0.5 g/l ASM in the growth media. Seed imbibition with ASM at a concentration of 0.08 g/l and 0.5 g/l significantly induced a reduction of the intensity of hypocotyl rot symptoms caused by R. solani AG‐4 that was correlated with a stimulation of chitinase activity. The protective effect of ASM against R. solani AG‐4 is probably due to the combination of induced resistance and its effect on pathogen growth. Seed treatment with ASM affected also the growth of 2‐day‐old seedlings. A dose‐dependant inhibition of the seminal root growth was observed which reached 53% at a concentration of 0.5 g/l ASM. This growth reduction of soybean was transitional and was rapidly recovered in optimal growth conditions except at 0.5 g/l of ASM.     

Study on Rosellinia necatrix Isolates Causing White Root Rot Disease in Taiwan

White root rot is a serious soil‐borne disease of several woods and crops. Recently, white root rot of tea shrubs and ornamental trees has increasingly been observed in Taiwan. Thirty‐six isolates of white root rot pathogen, showing pear‐shape swellings adjacent to the hyphal septa, had been isolated from samples of white root rot collected from Taiwan for about 4 years. The pathogen isolates produced Dematophora anamorph. Conidia of the pathogen were one‐celled, hyaline, subglobal, with truncate base, 2.9–5.8 × 1.9–3.5 μm . Ascospore dimensions were in the range of 37.0–55.0 × 5.4–7.9 μm with a short, longitudinal and straight germ slit, which complied with Rosellinia necatrix. Based on molecular studies, the pathogen isolates collected from Taiwan except R701 were identified as R. nectarix. Isolate R701, which was relatively polymorphic in internal transcribed spacer DNA sequence than other isolates, was temporarily considered as R. necatrix‐related pathogenic Rosellinia spp. All the tea cuttings (Camellia sinensis) inoculated with isolates developed typical white root rot symptoms. Pathogenicity tests demonstrated the presence of variation in virulence among the Rosellina isolates. Most of the R. necatrix isolates originating from Acer morrisonense were less virulent than those that originated from other hosts. The pathogenic Rosellinia spp., isolate R701, was also highly virulent to both cultivars of tea cuttings.     

Advances in molecular interactions on the Rhizoctonia solani-sugar beet pathosystem

Rhizoctonia solani is a soilborne pathogen with a broad host range. An anastomosis group (AG) system based on hyphal fusions has been established to distinguish between different R. solani subgroups in this species complex. Members of the AG2-2IIIB subgroup can cause serious problems in sugar beet production, resulting in Rhizoctonia root and crown rot. In this review, we summarize the current molecular advances in the R. solani sugar beet pathosystem. The draft genome of R. solani AG2-2IIIB has an estimated size of 56.02 Mb, larger than any of the R. solani AGs sequenced to date. The genome of AG2-2IIIB has been predicted to harbor 11,897 protein-encoding genes, including a high number of carbohydrate-active enzymes (CAZymes). The highest number of CAZymes was observed for polysaccharide lyase family 1 (PL-1), glycoside hydrolase family 43 (GH-43), and carbohydrate esterase family 12 (CE-12). Eleven single-effector candidates were predicted based on AG2-2IIIB genome data. The RsLysM, RsRlpA, and RsCRP1 genes were highly induced upon early-stage infection of sugar beet seedlings, and heterologous expression in Cercospora beticola and model plant species demonstrated their involvement in virulence. However, despite the progress achieved thus far on the molecular interactions in this pathosystem, many aspects remain to be elucidated, including the development of efficient transformation systems, important for functional studies, and the silencing of undesirable traits in the sugar beet crop.     

Comparative and population genomic landscape of Phellinus noxius: A hypervariable fungus causing root rot in trees

The order Hymenochaetales of white rot fungi contain some of the most aggressive wood decayers causing tree deaths around the world. Despite their ecological importance and the impact of diseases they cause, little is known about the evolution and transmission patterns of these pathogens. Here, we sequenced and undertook comparative genomic analyses of Hymenochaetales genomes using brown root rot fungus Phellinus noxius, wood‐decomposing fungus Phellinus lamaensis, laminated root rot fungus Phellinus sulphurascens and trunk pathogen Porodaedalea pini. Many gene families of lignin‐degrading enzymes were identified from these fungi, reflecting their ability as white rot fungi. Comparing against distant fungi highlighted the expansion of 1,3‐beta‐glucan synthases in P. noxius, which may account for its fast‐growing attribute. We identified 13 linkage groups conserved within Agaricomycetes, suggesting the evolution of stable karyotypes. We determined that P. noxius has a bipolar heterothallic mating system, with unusual highly expanded ~60 kb A locus as a result of accumulating gene transposition. We investigated the population genomics of 60 P. noxius isolates across multiple islands of the Asia Pacific region. Whole‐genome sequencing showed this multinucleate species contains abundant poly‐allelic single nucleotide polymorphisms with atypical allele frequencies. Different patterns of intra‐isolate polymorphism reflect mono‐/heterokaryotic states which are both prevalent in nature. We have shown two genetically separated lineages with one spanning across many islands despite the geographical barriers. Both populations possess extraordinary genetic diversity and show contrasting evolutionary scenarios. These results provide a framework to further investigate the genetic basis underlying the fitness and virulence of white rot fungi.