Effect of temperature on virulence of Rhizoctonia solani on soybean leaves and seedlings

Isolates representing 11 anastomosis groups (AGs) of Rhizoctonia solani from various geographic locations and host plants were tested for virulence on soybean leaves at 15, 20, 25, 30, and 35°C, and on soybean seedlings at 20, 25, and 30°C. Numbers of infection cushions formed on soybean leaves were determined using light microscopy. Isolates of AG-1 IA, AG-1 IB and AG-5 were more virulent on soybean leaves at 20, 25, and 30°C than isolates of AG-1 IC and AG-4. Maximum numbers of infection cushions were formed on soybean leaves by AG-1 (IA, IB, and IC), AG-4, and AG-5 at 25 and 30°C. The other AGs tested did not form infection cushions on soybean leaves although some caused minimal disease severity. Isolates of AG-1 IA formed significantly more infection cushions and caused greater disease severity than AG-1 IB and other isolates at 35°C. Maximum seedling infection, based on per cent area of hypocotyl region covered by lesions occurred at 25 C for AG-1 (IA, IB, and IC) and AG-4. Isolates of AG-5 caused greater seedling infection at 20°C than at 25 and 30°C. The other AGs tested caused only minimal damage to the seedlings. Isolates of AG-4 and AG-5 are not known to cause Rhizoctonia foliar blights of soybean in Louisiana, but their potential to be destructive foliar pathogens is confirmed.     

Association of Binucleate Rhizoctonia with Soybean and Mechanism of Biocontrol of Rhizoctonia solani

ABSTRACT The association of binucleate Rhizoctonia (BNR) AG-K with soybean and the interaction of BNR, R. solani AG-4, and soybean seedlings were investigated to elucidate the mechanism of biocontrol of R. solani by BNR. Sixty-hour-old seedlings were inoculated and incubated in a growth chamber at 24 degrees C; plants were examined with light microscopy and with scanning and transmission electron microscopy at various times following inoculation. BNR grew over hypocotyls, roots, and root hairs, but only colonized epidermal cells. Hyphae of BNR appeared to attach to the epidermis and, 5.5 h following inoculation, began penetrating cells by means of penetration pegs without forming distinct appressoria or infection cushions. There was evidence of cuticle degradation at the point of penetration. Infection hyphae moved to adjacent epidermal cells by direct penetration of epidermal radial walls. There were epidermal and cortical cell necrosis, beginning with the fragmentation of the tonoplast and followed by the disintegration of cytoplasm, organelles, and plasma membranes. Cell necrosis was also observed in adjacent cells where there was no evidence of BNR hyphae. Cell walls were not destroyed. After 144 h, there was noevidence of BNR hyphae in cortical cells. Attempted penetrations were observed, but papillae formed on the inside of cortical cell walls. Pre-inoculation of soybean seedlings with BNR 24 or 48 h before inoculation with R. solani (1 cm between inocula) affected the growth of R. solani on soybean tissue. There were fewer hyphae of R. solani, the hyphae branched sparingly, and infection cushions were rare when compared with hyphal growth on soybean inoculated only with R. solani. These effects were observed before the BNR hyphae began to intermingle with the hyphae of R. solani on the surface of the inoculated host. Preinoculation of soybean seedlings 24 h before inoculation with R. solani significantly (P = 0.05) reduced disease incidence and severity caused by R. solani AG-4. The lesions caused by R. solani always appeared distally, not proximally, to the BNR inoculum. The interactions of intermingling hyphae of BNR and R. solani were examined in vitro and on the surface of the host. There was no evidence of lysis, mycoparasitism, inhibition of growth, or any other form of antagonism between hyphae. The results of these studies strongly suggest that induced resistance is the mechanism of biocontrol of R. solani on soybean by BNR. The inhibition of hyphal growth of R. solani on the surface of soybean tissue preinoculated with BNR appears to be a novel characteristic of induced resistance.     

Characterization of Rhizoctonia solani Associated with Soybean in Brazil

Rhizoctonia solani causes pre- and post-emergence damping-off, root and hypocotyl rot and foliar blight in soybean. Foliar blight has resulted in yield losses of 31–60% in north and northeast Brazil. The aim of this study was to characterize isolates of R. solani associated with soybean in Brazil. Among 73 Rhizoctonia isolates examined, six were binucleate and 67 were multinucleate. The multinucleate iso1ates were characterized according to hyphal anastomosis reaction, mycelial growth rate, thiamine requirement, sclerotia production, and RAPD molecular markers. Four isolates that caused hypocotyl rot belonged to AG-4 and using RAPD analysis they grouped together with the HGI subgroup. Another isolate that caused root and hypocotyl rots was thiamine auxotrophic, grew at 35°C, and belonged to AG-2-2 IIIB. All 62 isolates that caused foliar blight belonged to AG-1 IA. RAPD analysis of R. solani AG-1 IA soybean isolates showed high genetic similarity to a tester strain of AG-1 IA, confirming their classification. The teleomorph of R. solani, Thanatephorus cucumeris was produced in vitro by one AG-1 IA isolate from soybean. The AG-4 and AG-2-2 IIIB isolates caused damping-off and root and hypocotyl rots of soybean seedlings cv. FT-Cristalina, under greenhouse conditions. The AG-2-2 IIIB isolate caused large lesions on the cortex tissue, that was distinct from the symptoms caused by AG-4 isolates. The AG-1 IA isolates caused foliar blight in adult soybean plants cv. Xingu under the greenhouse and also in a detached-leaf assay.     

Integrated control of Rhizoctonia solani and fusarium solani in tobacco transplants with Trichoderma hurziunum and triadimenol

Trichoderma cultures were isolated locally from soils and screened on potato-dextrose agar against Rhizoctonia solani. Several isolates of T. harzianum reduced the growth and build up of populations of R. solani and to a lesser extent of Fusarium solani in sterilized soil. In field experiments, biological control of R. solani and F. solani infections in tobacco transplants was achieved by adding T. harzianum (T77) to methyl-bromide-fumigated seedbeds before seed was sown. There was also increased growth and leaf yield that was not directly correlated with disease control. Triadimenol fungicide, integrated with the Trichoderma treatment, enhanced disease control.     

稻曲病菌和水稻纹枯病菌真菌病毒的研究进展

由稻绿核菌(Ustilaginoidea virens)和立枯丝核菌(Rhizoctonia solani)引起的稻曲病和水稻纹枯病是水稻上的2种重要真菌病害.真菌病毒是一类以真菌和卵菌为寄主的病毒,一些引起寄主真菌低毒力的真菌病毒具有生防潜力,可用来防治植物真菌病害.评述了从稻曲病菌和水稻纹枯病菌中分别发现并完成测序...     

Uptake of benodanil by tobacco seedlings and its relation to the rate of application and control of Rhizoctonia solani

Damage to the underground part of the stems of tobacco seedlings, drenched with benodanil in the seedbeds and transplanted into field soil containing Rhizoctonia solani Kühn, was negatively correlated with log(amount of benodanil applied). During the first 3 weeks of growth, the dry mass per seedling was positively correlated with the rate of benodanil applied and negatively correlated with stem damage. Concentrations of the fungicide extractable from roots, stems and leaves at transplanting and after 21 days were positively correlated with the rates of application; in 21 days they decreased by 94% in stems and roots. Stem damage at 21 days after planting was negatively correlated with log(benodanil concentration) at planting and 21 days later.     

广西稻瘟病和纹枯病的发生与综合防治

广西水稻年种植面积约3300万亩,近年水稻病虫发生为害呈逐年加重态势,年发生面积达9000万亩次,总体发生程度达4(5)级,尤其是稻瘟病和纹枯病的发生,给水稻造成了巨大的损失。稻瘟病是广西水稻上发生为害仅次于水稻纹枯病的病害,全区总体发生程度一般为中等,局部中等偏重发生,近几年年发生面积在800多万亩次左右,约占水稻种植面积的1/4多。其发生特点是:(1)以历史病区和种植感病品种的稻区,特别是桂东北、桂东南及沿海地区局部稻田发生较重;(2)个别感病的优质稻、常规稻发病后,往往易造成落窝,产量损失较大;     

A new quantitative real-time PCR assay for Rhizoctonia solani AG3-PT and the detection of AGs of Rhizoctonia solani associated with potato in soil and tuber samples in Great Britain

Rhizoctonia solani is an important pathogen of potatoes causing stem canker and black scurf. The fungus is a species complex comprised of 13 known anastomosis groups (AGs). AG3-PT is the anastomosis group frequently associated with disease in potatoes. A real-time PCR assay was designed to the rDNA ITS region of AG3-PT isolates to enable the pathogen to be detected directly in tuber and soil samples. The resulting assay was highly specific for AG3-PT, and did not amplify DNA from isolates from other AGs or subgroups of AG3. Using a bulk DNA extraction method capable of extracting from up to 250 g of soil, the assay could detect one individual sclerotium of AG3-PT (weighing 200 μg) in 250 g of soil. The AG3-PT assay was used, with assays for AG2-1, AG5 and AG8 to determine the prevalence of those AGs in UK potato soils and tubers. AG2-1 and AG3-PT were the predominant groups in tubers and soils, although AG3-PT was more frequently isolated from tubers, highlighting its importance as a potato pathogen. AG3-PT was also detected in more than half of the tuber samples tested suggesting the importance of seed borne inoculum.     

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.     

云南香石竹立枯病病原的鉴定及致病性研究

 27 Rhizoctonia solani isolates from carnation with damping-off symptoms were identified as anastomosis group AG-4 based on cultural characteristics and anastomosis in Yunnan.Sequence analysis of 5.8S rDNA-ITS of yx and tx presented 99% sequence similarity with AG-4 HGⅠtester isolate.Pathogenicity tests showed that isolates of AG-4 HG Ⅰ were pathogenic to carnation.This is the first record of damping-off of carnation caused by AG-4 HG Ⅰ in China.     

Genetic variability and pathogenicity of Rhizoctonia solani associated with black scurf of potato in New Zealand

Isolates (a total of 129) of Rhizoctonia solani were collected from black scurf on potato tubers from different potato‐growing regions in New Zealand. Sequence analysis of the nuclear ribosomal DNA internal transcribed spacer (rDNA–ITS) regions from these isolates identified three anastomosis groups (AGs), AG‐3PT, AG‐2‐1 and AG‐5. Isolates classified as AG‐3PT were widely distributed, whereas AG‐2‐1 and AG‐5 were confined to distinct locations. Sequence heterogeneity was identified in the ITS regions of 100 AG‐3PT and AG‐2‐1 isolates. Variation in the sequence and length of the rDNA–IGS1 region was also observed for selected isolates of AG‐3PT and AG‐2‐1. Phylogenetic studies found all AG‐2‐1 isolates belong to AG‐2Nt, a subset of AG‐2‐1 previously associated with solanaceous crops in other countries. AG‐2‐1 isolates were consistently more aggressive than those of AG‐3PT. Delayed emergence, severe infection on stolons, formation of aerial tubers and considerable yield losses were associated with AG‐2‐1, but they caused negligible black scurf. In contrast, AG‐3PT caused black scurf on progeny tubers but variable effects on stem emergence and stolons. Furthermore, AG‐2‐1 isolates caused severe tuber malformation, but isolates of other AGs did not. This is the first report on the AG composition, genetic variability and pathogenicity of R. solani isolates associated with black scurf of New Zealand potatoes.     

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.     

四川省不同寄主立枯丝核菌的遗传分化和致病力研究

 在四川省生态条件下,从不同水稻和玉米植株上分别分离到来源不同的立枯丝核致病菌15株和7株。致病力、菌丝融合实验结果表明,菌株均属于AG-11A群,各菌株间致病力差异显著。对分离菌株进行RAPD分析,结果显示,相似系数为0.92处菌株可聚合为5类,聚类分组和寄主来源有一定的相关性,来自相同寄主菌株的亲缘关系较近,不同寄主对立枯丝核菌的遗传分化有一定的影响,与病原菌的致病力差异没有直接的相关性。     

Identification of Rhizoctonia solani associated with field-grown tulips using ITS rDNA polymorphism and pectic zymograms

Methods based on internal transcribed spacers (ITS) ribosomal DNA (rDNA) polymorphism and pectic zymograms (ZG) were compared for their use in routine identification of Rhizoctonia solani isolates occurring in flower bulb fields. Thirty three AG 2-t isolates, pathogenic to tulips, could be distinguished from AG 1-IC, AG 2-2IIIB and AG 2-2IV, AG 3 and AG 5 by means of ITS rDNA fragment length and after digestion with EcoR I from AG 4 and AG 5. AG 2-t isolates and two Japanese isolates, pathogenic to crucifers and tulips, had an estimated fragment size of 710 bp, whereas Dutch AG 2-1 isolates, non-pathogenic to tulips, showed an estimated fragment size of 705 bp on agarose gel. Digestion of AG 2-t and AG 2-1 isolates with EcoR I, Sau3A I, Hae III and Hinc II revealed four and five distinct ITS rDNA digestion patterns, respectively. In AG 2 isolates 2tR114, 21R14 and 21R61 a double digestion pattern, indicating different ITS sequences within an isolate, was found. The observed ITS fragment length polymorphism between isolates pathogenic and non-pathogenic to tulips were considered too small to be used in routine screening of field isolates. Sequencing of AG 2 isolates 21R01, 21R06, 2tR002 and 2tR144 showed a total ITS rDNA fragment length of 715, 713, 714, and 728 bp. As an alternative to ITS rDNA fragment length polymorphism, pectic enzyme patterns were studied using a commercially available vertical gel-electrophoresis system and non-denaturing polyacrylamide gels amended with pectin. Anastomosis tester isolates AG 1 to AG 11 revealed different ZG. Fifty AG 2-t isolates and five AG 2-1 isolates belonged to a homogeneous pectic zymogram group. We propose to assign AG 2 isolates pathogenic to crucifers and tulip to ZG5-1. AG 2-1 isolates, non-pathogenic to tulip, formed a heterogeneous group with 4 distinct ZG. Pectic zymography provides an easy, quick and unambiguous method for routine identification of large numbers of field isolates. Such a technique is needed for research on the dynamics of Rhizoctonia populations to develop environmentally friendly control measures of rhizoctonia disease in field-grown flower bulbs.     

Suppression of Rhizoctonia solani in Potting Mixtures Amended with Compost Made from Organic Household Waste

ABSTRACT Compost made from organic household and garden waste was used to substitute part of the peat in potting mixtures used for growing woody ornamental nursery stock. The effects of amendment with compost on the colonization of potting mixture by Rhizoctonia solani (AG1) were studied in greenhouse experiments. A bioassay was developed using cucumber as a sensitive herbaceous test plant as a substitute for woody ornamental cuttings. Pathogen growth in the potting mixture was estimated by measuring the distance over which damping-off of seedlings occurred. Compost from two commercial composting facilities suppressed growth of R. solani in potting mixtures with 20% of the product when the compost was fresh (directly after delivery) or long matured (after 5 to 7 months of additional curing). In contrast, short-matured compost (1 month of additional curing) from the same batches stimulated pathogen growth. In vitro mycelial growth of R. solani on mixtures with mature compost was inhibited by microbial antagonism. Compost-amended potting mixtures responded differentially to the addition of cellulose powder; the effect on suppressiveness depended on curing time and origin of the compost. In long-matured compost, suppressiveness to R. solani was associated with high population densities of cellulolytic and oligotrophic actinomycetes. The ratio of the population density of actinomycetes to that of other bacteria was around 200-fold higher in mature suppressive compost than in conducive compost.     

Infection with Rhizoctonia solani induces defense genes and systemic resistance in potato sprouts grown without light

Rhizoctonia solani is an important soilborne and seedborne fungal pathogen of potato (Solanum tuberosum). The initial infection of sprouts prior to emergence causes lesions and may be lethal to the sprout or sprout tip, which results in initiation and compensatory growth of new sprouts. They emerge successfully and do not suffer significant damage. The mechanism behind this recovery phenomenon is not known. It was hypothesized that infection may induce pathogen defense in sprouts, which was investigated in the present study. Tubers were sprouted in cool and moist conditions in darkness to mimic conditions beneath soil. The basal portion of the sprout was isolated from the apical portion with a soft plastic collar and inoculated with highly virulent R. solani. Induction of defense-related responses was monitored in the apical portion using microarray and quantitative polymerase chain reaction techniques at 48 and 120 h postinoculation (hpi) and by challenge-inoculation with R. solani in two experiments. Differential expression of 122 and 779 genes, including many well-characterized defense-related genes, was detected at 48 and 120 hpi, respectively. The apical portion of the sprout also expressed resistance which inhibited secondary infection of the sprouts. The observed systemic induction of resistance in sprouts upon infection with virulent R. solani provides novel information about pathogen defense in potato before the plant emerges and becomes photosynthetically active. These results advance our understanding of the little studied subject of pathogen defense in subterranean parts of plants.     

Correlation between lipid peroxidation and phenolics content in leaves and roots of sugar beet infected with Rhizoctonia solani

The correlation between intensity of lipid peroxidation and changes in antioxidant capacity of sugar beet plants (cv. ‘Drena’) infected with Rhizoctonia solani Kühn isolate (AG 2-2 IIIB group) was studied. Successful inoculation was confirmed by the presence of infection cushions in a cross section of leaf petioles. On the 7^th day of the experiment, phenylalanine ammonia-lyase (PAL; EC. 4.3.1.5) activity was in negative correlation with intensified lipid peroxidation process in leaves of sugar beet plants (r= –0 .99). Also, in leaves and roots of inoculated sugar beet plants, total flavonoids content (35% and 20%, respectively) and 1,1-diphenyl-2-picrylhydrazyl (DPPH)-scavenging activity (80% and 55%, respectively) were significantly reduced. Necrotic processes resulting from R. solani infection of sugar beet plants was followed by induction of plant phenolics metabolism; however, antioxidant capacity of these plants was reduced.