A comparison of controlled environment and field trials for detection of resistance in cereal cultivars to root rot caused by Rhizoctonia solani

A screening test was developed to study the effect of different quantities of inoculum of Rhizoctonia solani on root rot and growth of cultivars of wheat, barley, oats, rye and triticale in a controlled environment and the field in two seasons. The ranking of relative susceptibility to disease of cultivars within a cereal differed between the controlled environment and field experiments, and between seasons in the field. The ranking of relative susceptibility between cereals also differed between experiments. In the controlled-environment experiment, wheat had the highest disease rating, followed by oats, barley, triticale and rye, but in the field barley had the highest rating followed by wheat and then oats. The overall differences between the cereals were small.     

Identification of Brassica accessions with enhanced resistance to Verticillium longisporum under controlled and field conditions

Journal of Plant Diseases and Protection - Verticillium longisporum (VL) is a soil-borne vascular fungal pathogen with host-specificity to cruciferous plants such as oilseed rape, threatening its...     

抗玉米纹枯病材料的鉴定及抗性遗传研究

 通过1997~2000年抗性鉴定,获得CML270为高抗玉米纹枯病且抗性稳定的抗性材料。该自交系在不同年份,对不同地理来源的玉米纹枯病强致病菌丝融合群AG 1 IA表现高抗(病情指数10左右),抗性表现明显优于国内玉米骨干自交系478和48-2。抗性遗传初步分析表明,玉米纹枯病抗性表现为质量-数量性状,抗性遗传受主效基因控制,同时受微效多基因修饰,最少基因数目4~7对。玉米纹枯病抗原的获得为最终解决玉米纹枯病危害提供了技术和材料支撑。     

Effect of growth media, storage environment, soil temperature and delivery to soil on binucleate Rhizoctonia AG-G for protection of potato from Rhizoctonia canker

Binucleate Rhizoctonia (BNR) isolates propagated for 20 days at 24°C on oat kernels and for 30 days on vermiculite amended with potato broth were recovered from an average of 62% of whole kernels, 100% of chopped kernels and 71 % of vermiculite particles within the cultures, respectively. Viability of BNR isolates 232-CG and JF-3S4-3 was higher when stored at 5 than at 24°C, and was slightly affected by the vacuum used to reduce the O₂ level. After 17 weeks of storage at 5°C in air, BNR isolates 232-CG and JF-3S4-3 maintained similar viability (75% viability on whole oat kernels and 100% viability on chopped oat kernels), but in vermiculite amended with potato broth, viability of isolate 232-CG remained at 100% while that of JF-3S4-3 was 28%. In the glasshouse, BNR isolates 232-CG and JF-3S4-3 protected potato plants from Rhizoctonia canker caused by R. solani in soil maintained at 11, 17 and 23°C. Protection from Rhizoctonia canker was greater when BNR was delivered to soil than when placed on seed pieces. BNR-colonized-whole oat kernels placed in soil (15 g m of row) gave the greatest protection from Rhizoctonia canker in all experiments. In two field experiments in soil naturally infested with R. solani AG-3. the amount of BNR-colonized oat kernels was reduced from 15 g/m of row to 1-9 g m of row without affecting protection of potato plants from Rhizoctonia canker.     

Evaluation of apricot (Prunus armeniaca L.) resistance to Apple chlorotic leaf spot virus in controlled greenhouse conditions

Apple chlorotic leaf spot virus (ACLSV) seems to be the causal agent of apricot viruela disease. This disease has become an important problem for apricot production in Spain, mainly affecting the ‘Búlida’ cultivar, although no information is available about the behaviour of other cultivars with regards to ACLSV. In this study, the behaviour of 29 apricot cultivars against ACLSV (Apr 62 isolate) was evaluated under controlled conditions in an insect-proof greenhouse. Three different rootstocks, ‘GF305’ peach, ‘Real Fino’ apricot and ‘Adesoto’ plum, were first inoculated by grafting ACLSV-infected bark and were later grafted with the apricot cultivar to be evaluated. Apricot cultivars were evaluated during three cycles of study. ACLSV was asymptomatic on the leaves of all cultivars and rootstocks, so level of susceptibility or resistance was determined by virus detection through RT-PCR. ‘GF305’ rootstock showed a greater susceptibility level than ‘Real Fino’ and ‘Adesoto’. Most of the cultivars were susceptible to ACLSV with different levels of susceptibility, and only ‘Bergeron’ and ‘Mauricio’ were resistant.     

Selection of bacterial antagonists for the biological control of Rhizoctonia solani in oilseed rape (Brassica napus)

Spore-forming bacteria isolated from oilseed rape (OSR) roots were tested for biological control activity against Rhizoctonia solani damping-off of OSR, using a simple slurry coat formulation. From a total of 239 strains which were tested, Bacillus subtilis 205 was selected as the most promising biocontrol agent (BCA). A known isolate of B. subtilis (NCIMB 12376), which had previously shown activity against R. solani in cotton, was used as a standard comparison throughout. In an in vivo screening, pre-emergence damping-off disease control by B. subtilis 205 was excellent, with plant stands being almost as high as non-diseased controls. However, post-emergence survival was less good, although the surviving plants exhibited significantly reduced hypocotyl rot. Growth-room trials which assessed the effect of ambient temperature on the biological control efficacy of B. subtilis strains 12376 and 205 showed that isolate 205 was more effective at lower temperatures. Disease control by isolate 205 at 15°C/10°C (day/night) suggests that this bacterium could operate under autumn sowing field conditions. Further investigations also indicated that the addition of iron (III) to the BCA formulation may enhance disease control by both bacilli. The potential of bacterial BCAs for the control of seedling diseases is discussed.     

宁夏马铃薯主栽品种对黑痣病的抗性鉴定

为明确宁夏马铃薯主栽品种对黑痣病的抗性水平,利用田间和室内接种相结合的方法对供试马铃薯品种的抗病性进行评价。室内接种试验表明,供试的20份马铃薯材料中没有免疫及高抗品种,接种量与抗性水平呈负相关,其中‘陇薯7号’,‘黑美人’,‘青薯9号’在接种量为5~15g水平上表现为中抗。通过聚类分析,接种量为5g和10g时,以欧式距离13作为最佳聚类分割点,供试品种可分为3个类群。田间接种综合评价分析表明,不同品种间抗性存在显著差异,通过聚类分析,以欧式距离13作为最佳聚类分割点,试验品种可分为4个高感品种、11个中感品种和5个中抗品种。     

Screening resistance against Sclerotinia sclerotiorum in Brassica crops with use of detached stem assay under controlled environment

Sclerotinia sclerotiorum is a major pathogen that infects stem tissue, causing yield loss and poor seed quality in rapeseed (Brassica napus). Here we report a reliable detached stem inoculation method for evaluating resistance under controlled environment. Two sets of Brassica materials were employed, including 17 genotypes from five Brassica crops in experiment 1, and 71 F₂ lines derived from a cross between susceptible and resistant lines of B. oleracea in experiment 2. High correlations were detected between stem and branch for lesion length in both experiments and between stem and sections of stem in experiment 1. Although the lesion length of detached stem inoculation under controlled environment was positively correlated with that of toothpick inoculation in the field, the variation of lesion length in the detached stem inoculation was lower than that in toothpick inoculation in experiment 2. Moreover, no significant association was detected between lesion length and the diameter of stem or branch. These findings suggest that the detached stem inoculation under controlled conditions is a large-scale, flexible and reliable method of screening for resistance against S. sclerotiorum in Brassica crops. The application of detached stem inoculation is discussed in resistance breeding programs against S. sclerotiorum in Brassica crops.     

Biological diversity of Rhizoctonia solani (AG-3) in a northern potato-cultivation environment in Finland

A total of 119 isolates of Rhizoctonia were collected from stem canker lesions, stolon and root lesions, hymenia on stems, or from black scurf on tubers of potato plants ( Solanum tuberosum ) in Finland (latitudes 60–67°N). All isolates except three belonged to anastomosis group 3 (AG-3) of R. solani , as determined by phylogenetic analysis of the internal transcribed spacer sequences (ITS1 and ITS2) of ribosomal RNA (rRNA) genes. Sensitivity of the 119 isolates to the fungicide flutolanil was tested in vitro (EC₅₀ values 0·14–0·75  µ g active ingredient mL⁻¹). The isolates also varied considerably in growth rate (5·1–14·8 mm day⁻¹). The severity of disease caused by 99 isolates was determined based on the proportion of potato sprouts affected by lesions, discoloration or death, which was c . 1–60%. Only two isolates that were able to cause severe symptoms showed particularly low sensitivity to the fungicide and rapid growth rate. One isolate each of anastomosis groups AG-2-1 and AG-5 and an unknown, binucleate Rhizoctonia sp. were detected. The AG-5 isolate and the binucleate isolate caused mild symptoms on potato sprouts, whereas the AG-2-1 isolate was not pathogenic. Taken together, AG-3 of R. solani was the predominant causal agent of the stem canker and black scurf diseases of potato in Finland and showed considerable variability in disease severity, fungicide sensitivity and growth rate in vitro .     

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.     

水稻纹枯病菌菌核上的真菌和细菌种类及其对水稻纹枯病菌生长的影响

 A total of 136 fungal and 86 bacterial isolates were isolated from the sclerotia of Rhizoctonia solani,collected from three different sources.These fungal isolates were identified as the following 12 genera:Alternaria,Aspergillus,Cladosporium,Coniothyrium,Curvularia,Gliocladium,Fusarium,Metarhizium,Peni-cillium,Phoma,Phytophthora and Trichoderma.Some isolates of Trichoderma and Gliocladium were found to inhibit the growth of R.solani strongly in vitro.Among the 86 bacterial isolates,20 of them could inhibit the growth of R.solani,whereas 5 isolates were found to promote the growth of mycelia and the formation of sclerotia of R.solani.These bacterial isolates with antagonistic actives to R.solani were identified as Bacillus and Pseudomonas.     

Relationships between stem canker, stolon canker, black scurf (Rhizoctonia solani) and yield of potato (Solanum tuberosum) under different agronomic conditions

Relationships between diseases caused by Rhizoctonia solani on different parts of potato plants ( Solanum tuberosum ) at different stages of crop growth were investigated under selected agronomic conditions. The effects of different densities of tuber-borne inoculum, date of planting, irrigation, size of seed tubers and their interactions on the incidence of stem and stolon canker during crop growth, the incidence and severity of black scurf and the yield of progeny tubers at harvest were quantified in a multifactorial experiment. Differences in the incidence of stem canker, stolon canker and black scurf were dominated by the effect of density of inoculum on seed tubers at planting. Highly positive correlations between the disease variables indicated a close relationship between the incidence of disease at each stage of crop growth although the degree of association between variables measured at an early growth stage and those measured at progressively later stages of crop growth weakened as the time interval increased. Total yield of progeny tubers was not affected by the density of tuber-borne inoculum although there was a shift in the size distribution, with a decrease in the yield of main-sized tubers and an increase in the yield of baker- and oversized tubers at the higher density of inoculum. Of the remaining factors, the effect of season tended to be more pronounced than any of the agronomic treatments although the use of irrigation and later dates of planting did influence the incidence of infection to a limited extent.     

Characterization of isolates of binucleate Rhizoctonia spp. associated with strawberry black root rot complex using fatty acid methyl ester (FAME) profiles

Black root rot is an important disease of strawberry caused by a complex of fungi that includes species of Rhizoctonia. In this study, a modified MIDI method (Microbial Identification System) was investigated for its utility to differentiate isolates of the three different anastomosis groups (AGs) of binucleate Rhizoctonia spp., associated with strawberry black root rot complex representing AG-A, AG-G, and AG-I. A total of 11 fatty acids were detected, and the FAME profiles for isolates of the three different AGs of Rhizoctonia spp. varied quantitatively and qualitatively. Moreover, the modified MIDI method will be a useful discriminatory tool for fungal identification and classification of the AGs of binucleate Rhizoctonia spp. associated with strawberry black root rot complex.     

水稻品种抗纹枯病田间鉴定方法改进

田间试验表明,插大穴(基本苗30株,穴左右)诱发与常规穴(6株,穴左右)插植人工接茵秆法鉴定水稻品种对纹枯病的抗病性结果基本一致,以插大穴诱发鉴定发病更为充分,从而提出了水稻纹枯病田问新的鉴定方法,且该方法省工、节支.     

Effects of granular nematicides on growth and microbial antagonism to Rhizoctonia solani

Effects of nematicides on growth and microbial antagonism toRhizoctonia solani were investigated as part of a study on the mechanisms involved in the increased incidence of this pathogen in nematicide-treated potato crops.Ethoprophos inhibited mycelial growth ofR. solani on potato dextrose agar (PDA), Czapek Dox agar (CDA) and on water agar (WA). Aldicarb stimulated its growth on PDA up to 14% but not on CDA and WA. Oxamyl inhibited mycelial growth on CDA and WA, but not on PDA.Ethoprophos and aldicarb stimulated development of the mycoparasiteVerticillium biguttatum on cultures ofR. solani. The effect was dependent on the medium on which the host fungus was grown. ForRhizoctonia cultures on PDA, growth of the mycoparasite was highly promoted by aldicarb and to a lesser extent by ethoprophos. WhenR. solani was grown on CDA, the development of the mycoparasite was not affected by aldicarb, slightly stimulated by ethoprophos and slightly inhibited by oxamyl. On water agar, its development on the host mycelium was not affected.In field trials on sandy soil, nematicides encouragedV. biguttatum probably by increased availability of substrate (i.e.Rhizoctonia mycelium) perhaps through reduced activity of the mycophagous fauna.Soil fungistasis was increased by ethoprophos and to a lesser extent by aldicarb at very high doses. At normal field rates, no effects can be expected on fungistasis. So the increased stem and stolon infection of potatoes in nematicide-treated fields was not caused by a direct effect of the nematicides on growth ofR. solani or by suppressing the microbial antagonism.Samenvatting De invloed van granulaire nematiciden op de groei vanRhizoctonia solani en op het microbiële antagonisme tegen deze schimmel werd onderzocht in het kader van een studie over de mechanismen die een rol spelen bij de toename van de aantasting in een met nematiciden behandeld aardappelgewas.Ethoprofos remde de myceliumgroei vanR. solani op aardappeldextrose agar (PDA), Czapek Dox agar (CDA) en op wateragar (WA). Aldicarb stimuleerde op PDA de groei met maximaal 14%. Op CDA en WA werd geen effect van aldicarb waargenomen. Oxamyl veroorzaakte groeiremming op CDA en WA, maar niet op PDA.Ethoprofos en aldicarb stimuleerden de ontwikkeling van de mycoparasietVerticillium biguttatum op cultures vanR. solani. De mate van groeistimulering was afhankelijk van de voedingsbodem waarop de waard,R. solani, werd gekweekt. De groei vanV. biguttatum werd sterk gestimuleerd door aldicarb en in geringere mate door ethoprofos, wanneer de waard gekweekt werd op PDA. Aldicarb had geen effect op de mycoparasiet wanneerR. solani op CDA gekweekt werd, terwijl ethoprofos de groei wel iets stimuleerde en oxamyl een gering remmend effect had. Op WA werd geen effect van de nematiciden op het mycoparasitisme vastgesteld.In veldproeven op zandgrond stimuleerden de nematiciden het voorkomen vanV. biguttatum op de stolonen. Het effect werd waarschijnlijk veroorzaakt door een verhoogde substraat beschikbaarheid (d.w.z. mycelium vanR. solani). De verhoogde beschikbaarheid van dit mycelium kan samenhangen met een door nematiciden gereduceerde activiteit van de fungivore bodemfauna.De bodemfungistase werd verhoogd door ethoprofos en, in geringere mate, door aldicarb bij hogere doseringen. Bij de in de praktijk aanbevolen doseringen kan echter geen effect op de fungistase verwacht worden. De toename in stengel- en stolonaantasting van aardappelen, geteeld in met granulaire nematiciden behandelde percelen, kon niet worden toegeschreven aan een direct effect van de nematiciden op de groei vanR. solani of aan een vermindering van het microbiële antagonisme.     

Antagonistic potential of Trichoderma spp. against Rhizoctonia solani and use of M13 microsatellite-primed PCR to evaluate the antagonist genetic variation

Journal of Plant Diseases and Protection - Rhizoctonia solani is the most important pathogen involved in cotton seedling disease in Egypt. Variations in the antagonistic activity of 20 isolates...     

Medicago truncatula as a model host for studying legume infecting Rhizoctonia solani and identification of a locus affecting resistance to root canker

The broad‐host‐range necrotizing fungal pathogen Rhizoctonia solani is responsible for economically significant diseases to crops as diverse as wheat, maize, barley, canola, sugar beet, potato, soyabean, bean, lupin and alfalfa. Germplasm screens in many of the crop hosts have not identified strong genetic resistance which, together with the lack of effective control, mean the pathogen remains a substantial problem for agriculture in many parts of the world. Following the establishment of a robust inoculation assay, a germplasm collection of the model legume Medicago truncatula was screened with various legume‐infecting isolates of R. solani. While some significant differences in susceptibility/resistance were detected between some lines, in the majority of cases M. truncatula was susceptible to R. solani. Comparison of a legume‐ and cereal‐infecting AG8 isolate with a legume‐specific AG11 isolate revealed no difference in pathogenicity between the two isolates when infecting M. truncatula. The most significant differences in susceptibility occurred with an AG6 isolate, which caused root canker. This included significant differences between the moderate resistance of the M. truncatula reference genotype A17 and the high susceptibility of line A20. The analysis of a recombinant inbred line population derived from A17 and A20 revealed a single locus contributing to the resistance in A17. Interestingly, the locus only affected the development of post‐emergent (late) symptoms, such as necrosis of cotyledons at 11 days after inoculation and root‐ and above‐ground‐weights, but not pre‐emergent seedling damping off. These findings pave the way for further studies to dissect the genetic and molecular mechanisms of resistance.     

Molecular characterization and pathogenicity of binucleate Rhizoctonia AG-F associated to the watermelon vine decline in Italy

In recent years, watermelon (Citrullus lanatus) has been subjected to significant losses due to vine decline in Sicily (southern Italy). During a survey conducted in 2009, the predominant fungal species associated with root rot and vine decline were Rhizoctonia spp. The most isolates were characterized as binucleate Rhizoctonia AG-F through morphological observation, nuclear condition, anastomosis tests and sequence homology of rDNA-ITS. Occasionally, R. solani was found. The pathogenicity of binucleate Rhizoctonia and virulences of different isolates were tested in growth chamber on watermelon seedlings. All isolates were pathogenic on watermelon seedlings and showed statistically significant differences on the disease incidence and severity among them. To our knowledge, this is the first report worldwide of the occurrence of pathogenic binucleate Rhizoctonia responsible for root rot and associated with watermelon vine decline.     

水稻纹枯病菌拮抗菌CZB40的筛选、鉴定及其发酵条件优化

为获得对水稻纹枯病有生防效果的拮抗放线菌,以水稻纹枯病菌(Rhizoctonia solani)为指示菌,从湖南郴州天鹅山风景区土样中分离筛选出5株拮抗放线菌株,其中菌株CZB40无菌滤液的5倍稀释液对指示菌菌丝生长抑制率高达90.45%,能使菌核菌丝萌发指数降低56.30%,而且能显著抑制菌核的形成。经形态观察、培养性状、生理生化鉴定,并结合16SrDNA序列分析,将CZB40菌株鉴定为极暗黄链霉菌(Streptomyces fulvissimus)。抗菌谱研究表明,该菌株对稻瘟病菌(Magnaporthe grisea)、桃褐腐病菌(Monilinia fructicola)、辣椒疫霉病菌(Phytophthora capsici)、草莓灰霉病菌(Botrytis cinerea)和辣椒白绢病菌(Sclerotium rolfsii)都有拮抗作用,抑菌率达27.94%~85.71%。通过单因子变量法和均匀设计法对菌株CZB40进行发酵条件研究。结果表明,其最佳的发酵条件为:玉米粉25g/L、酵母粉5g/L、MgSO40.2g/L、NaCl 0.8g/L、K2HPO40.4g、水1 000mL、29℃、pH 6.0。经优化后,菌株CZB40无菌滤液的10倍稀释液对水稻纹枯菌丝生长抑制率最高,达到94.85%,比优化前明显提高。该研究结果为水稻纹枯病的生物防治提供科学依据。