Characterisation of Rhizoctonia solani Anastomosis Groups Causing Bottom Rot in Field-Grown Lettuce in Germany

Bottom rot caused by Rhizoctonia solani is an increasing problem in field-grown lettuce in Germany. During the growing seasons of 1999 and 2000, 95 isolates of R. solani from lettuce plants with bottom rot symptoms were collected from eight locations. The isolates were characterised using hyphal anastomosis, pectic zymograms and morphological characteristics. Ninety-three isolates were identified as anastomosis group (AG) 1-IB, one as AG 1-IC and one as AG 2-1. Optimum hyphal growth was measured over a temperature range of 20–30 °C with an optimum at 25 °C. Aggressiveness of the AG 1-IB isolates varied from weak to strong when tested on detached lettuce leaves. The pathogenic potential of six AG 1-IB isolates was determined on 14 plant species in comparison with lettuce under conditions favourable for the fungus. Radish, broccoli, kohlrabi, spinach and millet seedlings were as severely infected as lettuce seedlings. The same isolates caused little symptoms on maize, tomato and onion. Knowledge about the host range of AGs of R. solani are important for planning an effective crop rotation as part of a control management system.     

Root Rot of Miniature Roses Caused by Pythium helicoides

Miniature roses growing in an ebb-and-flow watering system developed dieback during the summer growing season of 1996 in Gifu Prefecture. The main diagnostic symptoms were chlorosis of leaf followed by blight, and a brown, water-soaked root rot followed by dieback. Pythium isolates were recovered from the rotted root. The isolates form proliferous ellipsoidal papillate sporangia, spherical smooth oogonia, elongate antheridia, and aplerotic oospores. The optimum temperature for hyphal growth was 35°C with a growth rate of 34 mm/24 hr. Optimum temperature of zoospore formation (25-30°C) was lower than that of mycelial growth, and zoospores were produced even at 10°C. The isolates were identified as P. helicoides on the basis of these characteristics. In pathogenicity tests disease severity was highest at the highest tested temperature (35°C) at which the disease naturally occurred in summer. Four days after inoculation, the leaves turned yellow and the roots had a water-soaked rot, followed by leaf blight and root dieback after 7 days. The disease transmission test showed that diseased plants were found throughout the bench after 10 days. Received 4 July 2001/ Accepted in revised form 10 October 2001     

Nonpathogenic Binucleate Rhizoctonia spp. and Benzothiadiazole Protect Cotton Seedlings Against Rhizoctonia Damping-Off and Alternaria Leaf Spot in Cotton

ABSTRACT Recent reports have shown induction of resistance to Rhizoctonia root rot using nonpathogenic strains of binucleate Rhizoctonia spp. (np-BNR). This study evaluates the biocontrol ability of several np-BNR isolates against root and foliar diseases of cotton in greenhouse trials, provides evidence for induced systemic resistance (ISR) as a mechanism in this biocontrol, and compares the disease control provided by np-BNR with that provided by the chemical inducer benzothiadiazole (BTH). Pretreatment of cotton seedlings with np-BNR isolates provided good protection against pre- and post-emergence damping-off caused by a virulent strain of Rhizoctonia solani (AG-4). Seedling stand of protected cotton was significantly higher (P < 0.05) than that of nonprotected seedlings. Several np-BNR isolates significantly reduced disease severity. The combination of BTH and np-BNR provided significant protection against seedling rot and leaf spot in cotton; however, the degree of disease reduction was comparable to that obtained with np-BNR treatment alone. Significant reduction in leaf spot symptoms caused by Alternaria macrospora occurred on cotyledons pretreated with np-BNR or sprayed with BTH, and the np- BNR-treated seedlings had significantly less leaf spot than BTH-treated seedlings. The results demonstrate that np-BNR isolates can protect cotton from infections caused by both root and leaf pathogens and that disease control was superior to that observed with a chemical inducer.     

Characterization of Phytophthora spp. Causing Outbreaks of Citrus Brown Rot in Florida

ABSTRACT Epidemics of citrus brown rot from 1994 to 1997 in the south-central and east-coast citrus areas of Florida were characterized and the causal Phytophthora spp. identified. Two species of Phytophthora, P. palmivora and P. nicotianae, were consistently associated with brown rot. Epidemics caused by P. palmivora appeared to be initiated on immature fruit dropped on the orchard floor. The soilborne fungus infected and sporulated on these fruit and was then disseminated to fruit above 1 m in the canopy. In contrast, infection by P. nicotianae, the common cause of root rot, was confined to the lowest 1 m of the canopy. Fruit infected by P. palmivora produced large amounts of ellipsoidal sporangia available for splash dispersal, whereas those infected by P. nicotianae produced far fewer spherical sporangia. Isolates from brown rot epidemics were compared with P. nicotianae from citrus in Florida and Texas, P. citrophthora in California, P. palmivora, and selected Phytophthora spp. from other hosts. Brown rot symptoms produced by the different pathogenic citrus isolates on inoculated fruit were indistinguishable. Morphology, mating behavior, and isozyme patterns of brown rot isolates from 1988 to 1997 matched P. palmivora from citrus roots, other host plants, and other locations, but were different from characterized isolates of P. citrophthora in California and P. nicotianae in Florida and Texas. Cellulose acetate electrophoresis of the isozyme glucose-6-phosphate isomerase rapidly identified the causal citrus pathogen from infected fruit and soil isolation plates. Although P. palmivora is an aggressive pathogen of citrus roots, bark, and fruit, populations in orchard soils were low compared with P. nicotianae.     

Etiology and Population Genetics of Colletotrichum spp. Causing Crown and Fruit Rot of Strawberry

ABSTRACT Isolates of Colletotrichum spp. from diseased strawberry fruit and crowns were evaluated to determine their genetic diversity and the etiology of the diseases. Isolates were identified to species using polymerase chain reaction primers for a ribosomal internal transcribed spacer region and their pathogenicity was evaluated in bioassays. Isolates were scored for variation at 40 putative genetic loci with random amplified polymorphic DNA and microsatellite markers. Only C. acutatum was recovered from diseased fruit. Nearly all isolates from crowns were C. gloeosporioides. In crown bioassays, only isolates of C. gloeosporioides from strawberry caused collapse and death of plants. A dendrogram generated from the genetic analysis identified several primary lineages. One lineage included isolates of C. acutatum from fruit and was characterized by low diversity. Another lineage included isolates of C. gloeosporioides from crowns and was highly polymorphic. The isolates from strawberry formed distinctive clusters separate from citrus isolates. Evaluation of linkage disequilibrium among polymorphic loci in isolates of C. gloeosporioides from crowns revealed a low level of disequilibrium as would be expected in sexually recombining populations. These results suggest that epidemics of crown rot are caused by Glomerella cingulata (anamorph C. gloeosporioides) and that epidemics of fruit rot are caused by C. acutatum.     

防治人参锈腐病木霉菌的筛选及防治效果

人参锈腐病是人参的主要病害,应用生物防治技术对人参质量安全保证和人参的可持续种植具有重要意义。通过离体对峙拮抗筛选试验,从种植人参土壤和其他土壤中分离获得的107株木霉菌中筛选出了5株拮抗效果较好的菌株。应用这些筛选到的菌株的厚垣孢子处理人参苗床,人参的出苗率不受影响。播种后1年,5株木霉处理病情指数都在10以下,对照达到40.69,差异极显著,5株木霉防治效果都在75%以上,木霉菌株间防效差异不显著。播种后2年,5株木霉处理病情指数在13左右,对照达到46.84,差异极显著,5株木霉防治效果在67%以上,木霉菌株间防效差异不显著。     

Anastomosis Groups, Pathogenicity and Sensitivity to Fungicides of Rhizoctonia solani Isolates Collected on Potato Crops in France

A collection of 241 isolates of Rhizoctonia solani obtained from potato plants grown in different areas in France was characterized for anastomosis grouping, symptomatology on tubers of different cultivars and sensitivity to three fungicides. Most isolates collected belonged to (anastomosis groups (AGs)) AG 3, but 2% and 4% of the isolates were AG 5 and AG 2-1. AG 3 and AG 2-1 isolates were mostly obtained from sclerotia on tubers, but all AG 5, some AG 3 and some AG 2-1 isolates were recovered from superficial tuber alterations, like deformations, corky or scabby lesions. Sclerotia were formed on tubers produced by healthy stem cuttings grown in soil artificially infested with AG 3, but not on tubers grown in soil infested with either AG 5 or AG 2-1. No variation in susceptibility to sclerotial formation was observed among five potato cultivars. In all cases, a large proportion of tubers showed superficial corky lesions, often associated with deformations. The proportion of tubers with lesions and deformations was highest in soil infested with AG 2-1 and significantly lower on cv. Samba in all treatments. All isolates were highly sensitive to flutolanil, iprodione and pencycuron, except the AG 5 isolates, moderately sensitive to pencycuron. These results show that, although AG 3 is the most common R. solani group on potato in France, AG 5 and AG 2-1 may be present. Isolates differed for pathogenicity. In vitro sensitivity to fungicides varied among AGs.     

Fusarium Redolens f.sp asparagi, Causal Agent of Asparagus Root Rot, Crown Rot and Spear Rot

Two Fusarium species, F. oxysporum f.sp. asparagi and F. proliferatum, are known to be involved in the root and crown rot complex of asparagus. We have investigated reports on the involvement of F. redolens, a third species, which until recently was considered conspecific with F. oxysporum because of morphological similarities. RFLP analysis of the rDNA internal transcribed spacer region and AFLP fingerprinting identified eight strains from asparagus unambiguously as F. redolens. Four of these were tested and found to be pathogenic to asparagus either in this study (two strains) or in a previous one in which they were classified as F. oxysporum (three strains). Disease symptoms and disease development were the same as with F. oxysporum f.sp. asparagi and F. proliferatum. Present data and literature reports identify F. redolens as a host-specific pathogen involved in root, crown and spear rot of asparagus. The pathogen is formally classified as F. redolens Wollenw. f.sp. asparagi Baayen.     

Comparison of Soil Receptivity to Thielaviopsis basicola, Aphanomyces euteiches, and Fusarium solani f. sp. pisi Causing Root Rot in Pea

ABSTRACT Soil receptivity as a quantifiable characteristic ranging from conduciveness to suppressiveness to soilborne pea pathogens Thielaviopsis basicola and Aphanomyces euteiches was determined by analysis of differences in disease response curves obtained by artificial introduction of inoculum into natural field soil samples. Several parameters, including maximum root rot severity, the area under the health index curve, scores on the first axis of a principal component analysis (PCA) on dose responses, and Weibull model fitting were used to describe the disease responses. In all cases, the Weibull model gave satisfactory fits. PCA yielded a first axis that comprised 86% of the variance found when using Weibull predicted responses for T. basicola and 74% of the variance found for A. euteiches. This PCA axis essentially represented the average increase in disease severity due to the addition of increasing doses of inoculum to the soil. The Weibull scale parameter B, which represents the amount of inoculum necessary to increase root rot severity by 63% with respect to the level caused by pathogens naturally present in the soil, is another means of quantifying the receptivity of soils to these plant pathogens. Weibull parameter B, maximum root rot severity, the areaunder the health index curve, and the scores on the first PCA axis were strongly correlated for each of the pathogens tested individually. To compare the extent and behavior of soil receptivity responses to different pathogens, Weibull parameters B and C (slope at dose B) were chosen because of their universal definition, in contrast to PCA scores. Comparison of the average levels of Weibull parameters B and C indicated significant differences between the pathogens. Yet, no significant similarity in the ranking of the soils was found for the three pathogens, demonstrating that individual soils may interact with different pathogens in totally different ways. In general, soils were suppressive to T. basicola but conducive to A. euteiches, whereas their response to Fusarium solani f. sp. pisi ranged from conducive to suppressive. Therefore, risk assessment of soils prior to planting may require different strategies for each pathogen. Bioassays with soil samples taken before the last pea crop in 1987 and 1991 revealed a significant increase in the natural inoculum potential of soils that mainly was accounted for by A. euteiches and Pythium spp. These results strongly indicate that A. euteiches must be considered one of the most threatening pathogens to pea crops in the Netherlands.     

Efficacy of Hypovirulent Binucleate Rhizoctonia sp. to Control Banded Leaf and Sheath Blight in Corn

Water agar, artificially infested soil and leaf sheath inoculation methods were used to assess the suitable time of application, varietal host response and persistence of Rhv7, a hypovirulent, binucleate Rhizoctonia collected from soil in the Philippines, to effectively control virulent Rhizoctonia solani AG1-IA isolate RS35 on corn. With the water agar method, prior inoculation (2 to 3 days) with the biocontrol agent is essential to achieve maximum control of the pathogen. Disease protection was increased from 23 to 70% as the pre-incubation time of Rhv7 prior to the challenge inoculation with virulent isolate was lengthened from 0 to 3 days. Disease severity and incidence of banded leaf and sheath blight (BLSB) on the three corn hybrids were also suppressed in artificially infested soil. In leaf sheath inoculation, the suppressive ability of Rhv7 against BLSB on corn persisted during lesion expansion. This suppression was expressed as slower disease progress in plants with Rhv7 than in plants without Rhv7. Early and timely brace root formation that detached infected sheaths, also aided in reducing the number of diseased plants in Rhv7-treated plots at the final observation. Mycelial growth activity of RS35 was reduced when corn plants were pre-inoculated with Rhv7 before challenge even if there was no contact detected between Rhv7 and RS35, which suggests that Rhv7 protects corn against BLSB by induced resistance. The hypovirulent, binucleate Rhizoctonia Rhv7 strain effectively controlled R. solani AG1-IA isolate RS-35 in corn. Received 30 August 1990/ Accepted in revised form 25 October 1999     

Systemic Induction of Peroxidases, 1,3-beta-Glucanases, Chitinases, and Resistance in Bean Plants by Binucleate Rhizoctonia Species

ABSTRACT Inoculation of bean hypocotyls with a nonpathogenic binucleate Rhizoctonia (BNR) species induced systemic resistance and protection of the roots and cotyledons to later challenge with the root rot pathogen Rhizoctonia solani or the anthracnose pathogen Colletotrichum lindemuthianum. Bean seedlings that were treated with BNR 48 h prior to their challenge with R. solani or C. lindemuthianum had few necrotic lesions and reduced disease severity as compared with seedlings not treated with BNR. Treatment with BNR 48 h prior to their challenge also elicited a significant and systemic increase in all cellular fractions of peroxidases, 1,3-beta-glucanases, and chitinases compared with the diseased and control plants. Compared with control plants, total peroxidases and glucanases increased twofold and eightfold, respectively, in all protected bean tissues. BNR 232-CG could not be recovered from the challenged hypocotyls or cotyledons, indicating that there was no contact between the inducer and the pathogen. Both the 1,3-beta-glucanases and the peroxidases were positively correlated with induced resistance.     

Characterization of Mycorrhizal Isolates of Rhizoctonia solani from an Orchid, Including AG-12, a New Anastomosis Group

ABSTRACT Isolates of Rhizoctonia solani collected from mycorrhizal orchid (Pterostylis acuminata) plants and adjacent leaf litter were characterized. Of 23 selected isolates, 20 were members of a new anastomosis group (AG-12) and the rest were members of AG-6. There were no bridging anastomosis reactions observed between AG-12 and other AGs of R. solani. Among the 20 isolates of AG-12 evaluated, 18 vegetatively compatible populations were detected, indicating diversity within the AG. Mature cultures were dark brown, as were mature sclerotia. Some cultures produced alternating dark- and light-colored concentric rings, with sclerotia forming in the darker rings. Most cultures were appressed to the agar surface. In tests run to characterize pathogenic potential, selected mycorrhizal isolates of AG-12 and AG-6 did little damage to potato and barley seedlings, moderate damage to head lettuce seedlings, and more extensive damage to seedlings of cauliflower and radish. Isolates of AG-12 have not been observed to fruit in nature, and all attempts to induce formation of the teleomorph (Thanatephorus cucumeris) in the laboratory by selected isolates of AG-12 failed.     

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.     

铁皮石斛镰刀菌根腐病病原菌的鉴定及其对链霉菌发酵液的敏感性分析

根腐病是危害浙江省铁皮石斛的重要病害之一,鉴定根腐病的病原菌是有效防治该病害的前提条件.为进一步明确前期经柯赫法则验证为病原菌的镰刀菌(编号为GF-14)分类地位,本研究通过形态学鉴定,并结合内源转录间隔区(rDNA-ITS)、翻译延伸因子(TEF-1α)和RNA聚合酶基因(RPB1和RPB2)等序列的同源性比对及系统...     

Root and Collar Rot of Milkwort Caused by Cylindrocladium pauciramosum, a New Record for Europe

A new disease of milkwort (Polygala myrtifolia) was observed on several commercial nurseries in southern Italy. Diseased plants showed wilting, stunting, chlorosis or loss of foliage, and rotting of the basal stem as well as the crown and roots. A Cylindrocladium species was consistently found associated with crown, basal stem, and root lesions. The etiology of this disease was proved on milkwort, by fulfilling Koch's postulates. Two hundred Cylindrocladium isolates were collected from the most important Sicilian and Calabrian ornamental production areas from different host plants. Isolates were identified as Cylindrocladium pauciramosum (teleomorph Calonectria pauciramosa) on the basis of their obpyriform to broadly ellipsoidal terminal vesicles, conidiophore branching pattern, conidium morphology, as well as mating type studies with tester strains of C. pauciramosum for selected isolates. This is the first record of this pathogen from Europe and it is the first report of C. pauciramosum on milkwort.     

芽孢杆菌BH1防治大豆根腐病的效果及机制

芽孢杆菌BH1分离于大豆根际，在温室试验中表现出较好的防治效果。田间试验对尖孢镰刀菌引起的大豆根腐病防效达56．1％，增产7．6％。该菌株发酵无菌滤液对尖孢镰刀菌F3、茄孢镰刀菌W1和镰刀菌Sl孢子萌发和菌丝生长有抑制作用；对热处理较稳定。对其抗利福平标记株B-R^ -4与大豆根系的亲和性研究表明，BH1可定殖于无菌土和病土中生长的大豆根际，且动态变化趋势相似，但在无菌土中数量略高于病土。组织印记法表明：B-R^ -4可在整个大豆根系生长。扫描电镜观察发现，BH1主要存在于根表凹陷处，在根尖或侧根处则很少。透射电镜观察，根细胞间隙内未检测到BH1。在大豆根际的定殖试验说明，B-R^ -4与大豆根系有很好的亲和性，对营养和空间位点的竞争是其拈抗病原菌的主要机制。     

芽孢杆菌BH_1防治大豆根腐病的效果及机制

芽孢杆菌BH1分离于大豆根际,在温室试验中表现出较好的防治效果。田间试验对尖孢镰刀菌引起的大豆根腐病防效达56.1%,增产7.6%。该菌株发酵无菌滤液对尖孢镰刀菌F3、茄孢镰刀菌W1和镰刀菌S1孢子萌发和菌丝生长有抑制作用;对热处理较稳定。对其抗利福平标记株B R+ 4与大豆根系的亲和性研究表明,BH1可定殖于无菌土和病土中生长的大豆根际,且动态变化趋势相似,但在无菌土中数量略高于病土。组织印记法表明:B R+ 4可在整个大豆根系生长。扫描电镜观察发现,BH1主要存在于根表凹陷处,在根尖或侧根处则很少。透射电镜观察,根细胞间隙内未检测到BH1。在大豆根际的定殖试验说明,B R+ 4与大豆根系有很好的亲和性,对营养和空间位点的竞争是其拮抗病原菌的主要机制。     

Characterization, Genetic Structure, and Pathogenicity of Rhizoctonia spp. Associated with Rice Sheath Diseases in India

ABSTRACT Isolates of Rhizoctonia spp. were obtained from rice in India during 2000-2003. Characterization by conventional techniques and polymerase chain reaction showed that from 110 isolates, 99 were R. solani and 11 were R. oryzae-sativae. Of 99 isolates identified as R. solani, 96 were AG1-IA, 1 was AG1-IB, and 2 were AG1-IC. Amplified fragment length polymorphism (AFLP) analyzes were used to determine genetic relationships in Rhizoctonia pathogen populations collected from different geographic regions. Cluster analysis based on the AFLP data separated isolates belonging to the three different intraspecific groups of R. solani AG1 and differentiated R. solani from R. oryzae-sativae. Analysis of molecular variance (AMOVA) revealed that geographic region was the dominant factor determining population structure of R. solani AG1-1A; host cultivar had no significant effect. Pathogenicity tests on Oryza sativa cv. Zenith revealed that isolates of R. solani AG1-1A and AG1-1B were more virulent than R. solani AG1-IC and R. oryzae-sativae isolates.     

Hyphal Anastomosis Reactions, rDNA-Internal Transcribed Spacer Sequences, and Virulence Levels Among Subsets of Rhizoctonia solani Anastomosis Group-2 (AG-2) and AG-BI

ABSTRACT Hyphal anastomosis reactions, rDNA-internal transcribed spacer (ITS) sequences, and virulence of isolates representing Rhizoctonia solani AG-BI and six subsets of anastomosis group (AG)-2 (-2-1, -2-2 IIIB, -2-2 IV, -2-2 LP, -2-3, and -2-4) were compared. AG-2-4 is a subset described for the first time in this report. Anastomosis reactions within AG-BI and the listed subsets of AG-2 were generally strong but, between subsets, ranged from strong to a very weak "bridging" -type reaction. Anastomosis reaction alone generally did not provide adequate evidence for placement of an isolate into a subset of AG-2. Anastomosis reactions between AG-BI and the original subsets of AG-2 (-2-1 and -2-2) are very strong; for this reason, we propose that it be included as a subset of AG-2 (designation AG-2 BI). Subsets -2-3 and -2-4 show very weak bridging-type anastomosis reactions with all other subsets of AG-2 and thus may be candidates for independent AG status. Grouping within AG-2 based on rDNA-ITS sequences was consistent with the abovementioned subsets. However, grouping based on virulence as measured herein does not conform to established grouping patterns within AG-2 and does not seem useful as a group-defining criterion. A broad range of damage was observed among members of the most virulent subsets (-2-1, -2-2 IIIB, -2-2 IV, and -2-4), whereas other subsets (-2 BI, -2-2 LP, and -2-3) were similar to one another in causing a minimal level of damage. Group-specific primer pairs for each of the seven subsets of AG-2 were designed based on the abovementioned rDNA-ITS sequences. Primer pairs proved dependable and subset specific in polymerase chain reaction amplifications of purified genomic DNA from 109 isolates of R. solani and two isolates of binucleate Rhizoctonia. These primers will provide a simple and useful method for subset-specific characterization within AG-2 if further critical evaluations confirm their specificity.