Effect of coinoculation by Verticillium dahliae and Colletotrichum coccodes on disease symptoms and fungal colonization in four potato cultivars

Interactions between Verticillium dahliae and Colletotrichum coccodes , major causal agents of potato early dying (PED) syndrome, were studied in four potato cultivars that differ in their susceptibility to these pathogens . Aseptic plantlets of Nicola, Desiree, Alpha and Cara were inoculated with identical concentrations of each pathogen or with a mixture of the pathogens, and grown for 4 weeks in a monitored growth chamber. Coinoculation of Nicola with both pathogens caused more severe foliar disease symptoms and crown rot and greater C. coccodes colonization, than inoculation with each pathogen separately. Significant reductions in weight and height were also observed in plants coinoculated with both pathogens, as compared with plants inoculated with each pathogen separately or noninoculated plants. In Desiree, more roots were covered with C. coccodes sclerotia and disease symptoms were significantly more severe in plants inoculated with both pathogens together. However, plant weight and height were similar to those of plants inoculated with C. coccodes only. In Alpha, disease symptoms and levels of sclerotia in the roots were not affected by simultaneous inoculation with both pathogens. Weight and height of all plants were similar, whether inoculated with each pathogen separately or with both pathogens together. In Cara, plants inoculated with the mixture or either pathogen alone were smaller than the noninoculated control. Disease symptoms and occurrence of sclerotia were similar in plants inoculated with the combination and with a single pathogen. Compared with the effects of inoculation with either pathogen, simultaneous inoculation with both pathogens can, in some cultivars, increase the incidence of PED syndrome and thus severely decrease yields.     

Resistance of potato genotypes to common and netted scab-causing species of Streptomyces

Common and netted scabs are two disfiguring bacterial diseases of potato tubers, caused by various groups of Streptomyces species. Common scab, caused primarily by Streptomyces scabies and Streptomyces europaeiscabiei , is characterized by more or less deep pustules on the tuber surface, while symptoms of netted scab, caused mainly by Streptomyces reticuliscabiei , are superficial, corky alterations of the tuber periderm. Some isolates of S. europaeiscabiei are able to induce both common and netted scab symptoms, and therefore constitute a third pathogenicity group. Like most bacterial diseases, potato scabs would be best controlled by using resistant cultivars. Repeated experiments with soil artificially infested with isolates of three species representative of the three pathogenicity groups showed the level and stability of cultivar resistance, as well as the existence of a range of aggressiveness among different isolates. The distribution of scab severity indexes recorded on a collection of 16 potato cultivars and 27 breeding clones grown in soil infested with common scab-inducing isolates was continuous, suggesting isolate nonspecific quantitative resistance. Least susceptible cultivars were Nicola, BF15, Sirtéma, and Charlotte, while Urgenta, Désirée, Ondine and Bintje were very susceptible. The same genotypes proved either highly susceptible (e.g. cvs Bintje, Désirée or Carmine) or highly resistant (e.g. cvs Charlotte, Sirtéma, Monalisa, BF15 or Belle de Fontenay) to isolates forming netted scab symptoms, suggesting isolate-specific qualitative resistance. The ability was confirmed of some isolates of S. europaeiscabiei to induce one or the other type of symptoms depending on cultivar and soil temperature.     

Development of symptoms caused by Erwinia carotovora ssp. atroseptica under field conditions and their effects on the yield of individual potato plants

Infection of seed tubers by pectinolytic Erwinia species can lead to the development of various symptoms during vegetative growth of potato crops, including non-emergence of plants, chlorosis, wilting, haulm desiccation and typical blackleg. The relationships between types of symptoms and yield are poorly documented, and are investigated by following the development of symptoms in potato plants grown under field conditions from seed tubers artificially inoculated with E. carotovora ssp. atroseptica ( Eca ) , and measuring the yield of each plant. Symptoms were classified into five main types (non-emergence, wilting/chlorosis, blackleg, haulm desiccation and plant death). Each plant was scored for types of symptom on four successive dates; plants without visible symptoms were scored as healthy. The method of inoculation and inoculum concentration proved major factors for the subsequent development of symptoms. Disease development was more severe after vacuum infiltration of bacteria into seed tubers than after shaking tubers in contaminated sand. Disease usually progressed from chlorosis and/or wilting to partial or total desiccation on a given plant. Yield losses varied according to symptom type, but the relationship between symptoms recorded and yield also depended on scoring dates. The data suggest that the beginning of tuber growth might be the most suitable stage for predicting yield losses from symptom observations. In both cultivars studied (Bintje, highly susceptible, and Désirée, moderately resistant), the yield of symptomless plants growing from inoculated seed tubers was significantly less than that of control plants, indicating that the presence of bacteria on the seed tuber was detrimental, even in the absence of visible symptoms. Differences in symptom expression in the field between cultivars matched the level of visible infection of tubers at harvest, as Bintje tubers showed a higher incidence of rot than Désirée tubers.     

Differential stimulation of germination of sclerotia of Sclerotium cepivorum by cultivars of onion and its effect on white rot disease

Differences were found in the ability of onion cultivars to stimulate the germination of sclerotia of Sclerotium cepivorum under UK conditions. The differences between European and Egyptian cultivars were particularly marked. Differences also occurred in the development of white rot disease in European and Egyptian cultivars. These differences were related to the pattern of growth of the cultivars and not to levels or types of flavour and odour compounds. The effect of the pattern of growth of plants on germination of sclerotia was confirmed in experiments with different types of planting material. Germination beneath plants grown from sets was particularly rapid, but very slow beneath plants produced by direct seeding. An experiment with plants grown under different levels of sulphate nutrition showed that low levels of flavour and odour compounds were associated with a weak capacity to stimulate the germination of sclerotia. No evidence was found to suggest that control of white rot disease by selection for growth features or for differences in flavour and odour characteristics would be a practicable proposition for the foreseeable future.     

Growth in vitro and infectivity of Colletotrichum coccodes on potato tubers at different temperatures

To investigate the ability of black dot symptoms to develop on infected potato tubers during storage, the growth of Colletotrichum coccodes was followed in vitro on malt agar at temperatures ranging from 5–27°C, and in vivo on artificially infected potato tubers kept at 5, 10 and 15°C. In vitro , 13 isolates from different geographical origins grew at all temperatures tested; growth started with a delay of 10 days at 5°C and of 4 days at 10°C, and was fastest at 27°C. All isolates had similar growth patterns and produced conidia and sclerotia at all temperatures. Minitubers were successfully infected at 5, 10 and 15°C by depositing either a mycelial plug or a drop of conidial suspension on the tuber surface. Sclerotia were observed after 7 days at the point of inoculation. Symptoms extended in all cases, although more slowly at 5 and 10 than at 15°C. Latent infections were detected in up to 21% of tubers without black dot symptoms at harvest. These results show that latent infections by C. coccodes are probably quite frequent, and that the pathogen is able to develop at low temperatures in controlled conditions. This suggests that black dot symptoms can increase during storage if stores are not adequately managed.     

Production, survival and infectivity of oospores of Phytophthora infestans

The formation of oospores of Phytophthora infestans was studied in tomato and potato crops and volunteer plants under field conditions, and in laboratory tests with leaf discs of potato cultivars differing in their level of race-nonspecific resistance. Oospores were readily detected in blight-affected tomato leaflets and fruits, and in leaflets of field crops and volunteer potato plants. Oospores extracted from blighted potato leaflets yielded 13 oospore-derived progeny. Oospores were also produced following inoculation of leaf discs of eight potato cultivars expressing different levels of race-nonspecific resistance with a mixture of sporangia of A1 and A2 isolates. The highest numbers of oospores were produced in cvs Bintje (susceptible) and Pimpernel (resistant), and the lowest in Nicola (intermediate resistance). The relationship between lesions per leaflet and oospore incidence, affected by varying A1 : A2 ratios, was explored using a simple mathematical model, and validated by comparing actual oospore production in leaflets with multiple lesions of the race-nonspecific-resistant potato clone Lan 22-21 with the predictions generated by the model. Survival of oospores was investigated after their incorporation in either a sandy or a light clay soil in buried clay pots exposed to the local weather conditions. Over 6 years these soils were regularly assessed for their infection potential using floating leaflets in a spore-baiting bioassay. Sandy and clay soils contaminated with oospores remained infectious for 48 and 34 months, respectively, when flooded. Infections of floating potato leaflets occurred within 84–92 h and ceased after 11 days. Soil samples remained infective if dried and re-flooded on two, but not more, occasions.     

Effects of temperature and light on non-race-specific resistance of potato leaflets to late blight

Effects of temperature and illumination on colonization by Phytophthora infestans of detached leaflets of five potato cultivars differing in field resistance to blight were investigated using an ELISA system to quantify the pathogen. Leaflets of cvs Teena and Shelagh, and in one experiment cv. Brodick, were more resistant to colonization when infected leaflets were incubated at 10°C than at 20°C, but temperature conditions before inoculation had little effect. Both photoperiod and light intensity during illumination of intact plants before inoculation interacted with genotype to determine subsequent colonization of infected leaflets incubated in darkness. Leaflets from plants of cv. Teena were more resistant to colonization after exposure to low, rather than to high, light intensities but photoperiod had no apparent effect. Leaflets of cv. Shelagh grown in a 20 h day were more resistant than those grown in a 10 h day but light intensity had no effect. Leaflets of cv. Brodick were more resistant after a 20 h day than a 10 h day and after exposure to low, rather than to high, light intensities. Leaflets of cv. Bintje were extensively colonized and those of cv. Torridon remained relatively resistant to colonization, irrespective of temperature and lighting conditions. There was a high level of unexplained variation in all the experiments.     

Susceptibility of cultivars of garlic to Sclerotium cepivorum

Five garlic cultivars, including one claimed to be resistant to Allium white rot, were similar in their effects on the germination of sclerotia of Sclerotium cepivorum. There was no evidence of resistance to the pathogen in any of the cultivars when tested under field conditions.     

Distribution and fungicide sensitivity of Colletotrichum coccodes in French potato-producing areas

Black dot, caused by Colletotrichum coccodes , is of potential concern to potato production in France as part of the tuber-blemishing disease complex. The lack of information about the actual distribution of the pathogen in potato-producing areas led to a survey of the occurrence of the disease. Black dot symptoms were observed on roots, stems and/or tubers of the 82 potato cultivars examined in 1994. A baiting bioassay, using cuttings of potato cultivars Bintje and Urgenta, revealed the presence of the pathogen in all 37 soil samples tested, which had been collected throughout the main French potato growing areas. In vitro , growth of five C. coccodes isolates recovered from diseased potatoes grown in western and southern France was severely affected by imazalil, tolchlofos-methyl and, to a lesser extent, mancozeb and thiabendazole. Conversely, iprodione, flutolanil and pencycuron were ineffective in reducing the growth of these isolates. These data indicate that C. coccodes is widespread in French potato cropping areas, that currently popular cultivars are susceptible to the disease, and that at least some of the fungicides commonly applied to seed tubers are ineffective against the pathogen. A better diagnosis of the disease, but also the insensitivity of the pathogen to several chemicals frequently used on seed tubers for controlling black or silver scurfs, might thus provide explanations for the apparent increase in black dot occurrence in recent years.     

Effects of salinity and its interactions with disease incidence on potatoes grown in hot climates

The interaction of two diseases — Verticillium wilt, caused byVerticillium dahliae, and early blight, caused byAlternaria solani — with salinity was investigated. The use of a system of two overlapping overhead irrigation lines enabled results to be obtained under a continuous gradient of salinity. Three cultivars were investigated, namely, Cara, Désirée and Nicola. The results illustrate the increase in symptom expression of the two diseases, as well as in associated characters such as height and maturity, in the presence of salt. Differences existed between cultivars in their response to the diseases in the presence of varying levels of salt.     

Optimizing conditions for growth and sporulation of Pyrenophora semeniperda

The effects of photoperiod, light quality, wounding and sealing of culture plates on in vitro growth and sporulation of Pyrenophora semeniperda were examined to define conditions conducive to the abundant and rapid production of conidia. For maximum growth and sporulation, the leaf-spotting and seedborne plant pathogen required an alternating light/dark sequence. Fewer conidia were produced under conditions of constant illumination or constant darkness. Growth was enhanced by light of wavelengths longer than 500 nm, while sporulation was enhanced by light of wavelengths shorter than 500 nm. Sporulation was enhanced under conditions of alternating temperature with a diurnal photoperiod and by mycelial wounding. Sealing of Petri plates had an inhibitory effect on sporulation. Significantly more conidia of higher quality formed when unsealed cultures, wounded after 7 days, were exposed to 23°C during the light phase and 19°C during darkness. Under these optimal conditions, conidial numbers were increased by 800% or more compared with unwounded cultures grown at constant 25°C in 12 h alternating cool-white light.     

Colonization of roots, stolons, tubers and stems of various potato (Solanum tuberosum) cultivars by the black-dot fungus Colletotrichum coccodes

In an attempt to better understand the importance of tuber-borne inoculum in black dot development, several potato cultivars were inoculated with various Colletotrichum coccodes isolates. Symptoms developed first on underground organs (starting 2 weeks after inoculation on roots, and later on stolons and tubers) of inoculated plants; stem infections developed only after 7–10 weeks, depending on the cultivar. Infection with C. coccodes resulted in a reduction in numbers of stolons and tubers in cv. Bintje, but not in the later maturing cv. Roseval. Significant isolate by cultivar interactions were detected from the analysis of root symptoms after inoculation of three potato cultivars (Bintje, Spunta and Desiree) with five C. coccodes isolates. Such an interaction was also detected for stolon/tuber symptoms at the latest scoring date (98 days after inoculation), but not at earlier dates (58, 70 and 84 days after inoculation). These results suggest that protocols based on root colonization might be used for investigating cultivar response to black dot and pathogenicity of C. coccodes isolates, and that some specificity exists in the reaction of potato genotypes to this pathogenic fungus.     

Pathogen resistance in aerial tubers of potato cultivars

Increases in resistance of aerial tubers (cultivars Désirée, Epicure and King Edward) against nine pathogenic potato fungi were determined in vitro using freeze-dried powder derived from subterranean and aerial tubers as a culture medium, and in vivo by inoculating both tuber types with the storage pathogens Fusarium sulphureum, F . solani var. coeruleum, Phoma exigua var. foveata and Polyscytalum pustulans . Significantly reduced growth rates and spore and conidiophore numbers were recorded when pathogens were cultured on freeze-dried powder derived from aerial tubers. No effects on spore size of F . solani var. coeruleum were recorded but spore and conidiophore lengths of remaining pathogens were reduced and width increased, except for Colletotrichum coccodes , where the diameter of acervuli increased, and P . exigua var. foveata , where spore length and width were reduced. Reduced disease incidence, severity and spore numbers were recorded on aerial compared with subterranean tubers inoculated in vivo with the four storage pathogens. Spore length of P . pustulans was reduced and spore width increased, while spore length and width of P . exigua var. foveata were reduced on aerial compared with subterranean tubers inoculated with these pathogens. No effects on spore size of Fusarium spp. were recorded. Concentrations of glycoalkaloids, chlorogenic acid, calcium and magnesium were higher in aerial than in subterranean tuber tissue, which may have contributed to increased resistance and effects on size and morphology of fungal spores. The implications of using aerial tubers as an alternative technique for the production of seed potatoes and their importance to the potato industry are discussed.     

Changes in the content of steroidal glycoalkaloids in potato tubers grown in the field and in the greenhouse under different conditions of light, temperature and daylength

Differences in the content of steroid glycoalkaloids (SGA) were noted in the peels of potato tubers cultivars Alpha, Cara, Désirée and LT-7. Cultivar LT-7 had always the highest SGA content, Désirée was intermediate and Alpha and Cara had the lowest SGA content, regardless of growing conditions, tuber ageing and tuber treatments. Tubers grown during the summer developed the highest SGA content as compared to those grown in the spring and winter seasons, respectively. Tubers grown during winter showed an increase in SGA toward the end of the storage period. Similar results were obtained with tubers of LT-7 grown in a greenhouse under cool temperatures in combination with short days (which imitate the winter conditions in Israel). A decrease in SGA content was observed in tuber slices treated with a fungal cell-wall preparation fromPhytophthora citrophthora. This decrease was much stronger in the pith than in the peel, indicating a possible role for SGA in the defence mechanism because peels are the first part of the tuber to be invaded by pathogens. The above results were observed in tubers of all cultivars regardless of the growing season or tuber ageing.     

Effect of light duration and growth season on verticillium wilt in potato

The tolerance response to Verticillium in potato is affected by external abiotic and biotic factors. Short-day photoperiod enhanced disease level in the susceptible cvs. ‘Nicola’ and ‘Maris Bard’, as well as in the tolerant cvs. ‘Cara’ and ‘Désirée‘. This was observed under field conditions, where plant reaction in the spring was compared with that in autumn. In some cases the disease was more pronounced in autumn, with its shorter days, than in spring, when the days are longer. The effect was measured by disease symptom severity, fungus colonization in the plants, plant height and final yield. This disease enhancement was more pronounced under controlled growth room conditions, where the only different parameter was illumination. A short photoperiod of 8 h increased the disease level relative to 16 h of light.     

Weed suppression ability of three winter wheat varieties at different row spacing under organic farming conditions

For three seasons, six field trials were conducted in winter wheat ( Triticum aestivum ) at different sites in North Rhine-Westphalia, Germany. The aim was to evaluate the performance of three varieties, which differ in their shading behaviour, in suppressing weeds under organic farming (Ökologischer Landbau) conditions and grown at three row spacings. The experimental factors were cultivar, row width and row orientation. Key parameters of crop and weed development, including ground cover and biomass, were assessed at different growth stages and analysed with anova . Growth of the site-specific weed vegetation was significantly affected in five of six trials. Weed growth was reduced by cvs Astron and Pegassos compared with cv. Greif, and was lower at 12 cm compared with 24 cm row spacing. No effect of row orientation (East-West, North-South) on weed growth was observed. The more competitive cultivars Astron and Pegassos were taller than the less competitive cv. Greif and had higher ground cover and light interception, presumably induced by planophile leaf inclination and partly, in the case of cv. Astron, because of higher leaf area index (LAI). Narrow row spacing (12 cm), resulting in a more even spatial plant distribution, increased crop ground cover, LAI, dry matter and light interception. In two trials, the variety with planophile leaf inclination performed better at wider row spacing, suggesting that planophile wheat cultivars may be advantageous in wider row stands.     

Effect of environmental factors and Sclerotium cepivorum isolate on sclerotial degradation and biological control of white rot by Trichoderma

Laboratory assays demonstrated that two isolates of Trichoderma viride and one isolate of Trichoderma pseudokoningii degraded up to 80% of sclerotia of four isolates of Sclerotium cepivorum in a silty clay soil, and also degraded up to 60% of sclerotia in three other soil types. Relationships were defined between the degree of sclerotial degradation by the two T. viride isolates in the silty clay soil and both temperature and soil water potential. Sclerotia were degraded between 10 and 25°C at −0·00012 MPa, but there was little activity of T. viride at 5°C or at −4 MPa. Degradation of S. cepivorum sclerotia also occurred in the absence of Trichoderma at soil water potentials approaching saturation . Experiments using onion seedling bioassays showed that the efficacy of Trichoderma isolates for the control of white rot using the same selection of soils and S. cepivorum isolates was variable, but that there was significant disease control overall. The importance of environmental factors and pathogen isolate in relation to effective biological control of white rot is discussed.     

Opportunities for the microbial control of Allium white rot1

The microbial control of Allium white rot (Sclerotium cepivorum) has shown promise experimentally but has yet to be used in commerce. Allium crops are grown from dry or fluiddrilled seed at a range of seed rates, from bare root and module-grown transplants and from sets and cloves. The period of susceptibility to infection may be prolonged or short in cool and hot climates, respectively. Primary infection of the Allium host results from the growth of hyphae from germinating sclerotia to the roots; secondary infection results from hyphal growth between plants. Sclerotial populations in the soil are often low, hence sclerotia are widely dispersed and therefore inaccessible to microbial control. New sclerotia form on host stem tissue, rarely on roots, rendering them accessible to microbial control. Hyphal parasites may therefore be effective at reducing primary and secondary infection and the formation of new sclerotia. Parasites of sclerotia may restrict the survival of new sclerotia and hence reduce primary infection in future, but not the current season's crops. The opportunities and problems that the variety of growing techniques and conditions of Allium crops present for microbial control of white rot will be reviewed.     

Black dot (Colletotrichum coccodes): an increasingly important disease of potato

In recent years black dot ( Colletotrichum coccodes ) has become an economically important disease problem in potato ( Solanum tuberosum ). It is characterized by silvery lesions on the tuber surface which result in a deterioration in skin quality. In addition to causing tuber blemish symptoms, C. coccodes also causes symptoms on stems and foliage, resulting in crop losses in some countries, and is implicated as a factor in the potato early dying disease complex. In the past, the incidence and severity of black dot might have been underestimated, as tuber symptoms were often mistaken for silver scurf ( Helminthosporium solani ). Inoculum of C. coccodes can be both seed tuber- and soilborne, and disease control is difficult as there are few chemical control methods and little resistance in commercial cultivars. Cultural control options offer the only potential means to control this disease at present. Current developments in rapid and specific PCR-based detection methods are being used to address epidemiological questions.     

生菜软腐和菌核病拮抗菌贝莱斯芽胞杆菌BPC6鉴定与防效

软腐病和菌核病是生菜生产中两大毁灭性病害。为有效控制这2种病害，利用平板稀释和对峙法，从生菜根际土壤中分离到一株拮抗菌BPC6，其菌悬液、无菌滤液对软腐病菌Pectobacterium carotovorum和菌核病菌Sclerotinia sclerotiorum的生长均具有抑制效果，该菌株也能抑制其他14种植物病原菌的生长。形态学、16S rDNA及基因组序列分析将其鉴定为贝莱斯芽胞杆菌Bacillus velezensis。BPC6能产生纤维素酶和蛋白酶，不能产生几丁质酶和嗜铁素。比浊法分析的生长曲线显示，BPC6能在1%～10% NaCl及pH 5.0～9.0条件下生长。通过分析不同浓度BPC6对软腐病菌和菌核病菌侵染生菜离体叶片的影响，明确了菌株BPC6适宜用量为1.4×10⁹～2.8×10⁹ CFU/mL。菌株BPC6对生菜软腐病和菌核病盆栽防效分别为44.09%和53.58%，对菌核病大田防效为77.41%。本研究表明BPC6是一株对生菜软腐病和菌核病具有拮抗作用的潜在生防菌。