Resistance to thiabendazole in Fusarium species isolated from potato tubers affected by dry rot

Fusarium species obtained from stored potato tubers affected with dry rot were grown on agar containing thiabendazole. All 40 isolates of F. coeruleum and 60 isolates of F. avenaceum tested were sensitive to the fungicide, but 68% of the 85 isolates of F. sulphureum and one isolate of F. culmorum were classified as resistant. When isolates were made from dry rots on tubers that had been treated with thiabendazole during loading into store, all 81 isolates of F. sulphureum were resistant, whereas all the isolates of F. coeruleum (25), F. avenaceum (4) and Phoma foveata (10) were sensitive. Resistance was not found in five isolates of Cylindrocarpon destructans. All the Fusarium spp. were sensitive to imazalil and were pathogenic when inoculated into potato tubers. Resistant and sensitive isolates of F. sulphureum caused rots of similar size.     

Use of quantitative molecular diagnostic assays to investigate fusarium dry rot in potato stocks and soil

ABSTRACT Specific and sensitive quantitative diagnostics, based on real-time (TaqMan) polymerase chain reaction (PCR) and PCR enzyme-linked immunosorbent assay, were developed to detect dry-rot-causing Fusarium spp. (F. avenaceum, F. coeruleum, F. culmorum, and F. sulphureum). Each assay detected Fusarium spp. on potato seed stocks with equal efficiency. Four potato stocks, sampled over two seed generations from Scottish stores, were contaminated with F. avenaceum, F. sulphureum, F. culmorum, F. coeruleum or a combination of species, and there was a general trend towards increased Fusarium spp. contamination in the second generation of seed sampled. F. sulphureum and F. coeruleum caused significantly (P < 0.05) more disease in storage than the other species when disease-free tubers of potato cvs. Spunta and Morene were inoculated at a range of inoculum concentrations (0, 10(4), 10(5), and 10(6) conidia/ml). Increased DNA levels were correlated with increased disease severity between 8 and 12 weeks of storage. The threshold inoculum levels resulting in significant disease development on both cultivars were estimated to be 10(4) conidia/ml for F. sulphureum and 10(5) conidia/ml for F. coeruleum. To study the effect of soil infestation and harvest date on disease incidence, seed tubers of cvs. Morene and Spunta were planted in a field plot artificially infested with the four Fusarium spp. F. culmorum and F. sulphureum were detected in soil taken from these plots at harvest, and F. sulphureum DNA levels increased significantly (P < 0.05) at the final harvest. All four Fusarium spp. were detected in progeny tubers. There was a trend toward higher levels of F. culmorum detected in progeny tubers at the earliest harvest date, and higher levels of F. sulphureum at the final harvest. The use of diagnostic assays to detect fungal storage rot pathogens and implications for disease control strategies are discussed.     

黑龙江省马铃薯干腐病菌种类鉴定及致病性

本研究将采自黑龙江省不同地区的马铃薯干腐病病样进行分离和病原菌纯化,得到27个镰刀菌菌株,通过致病性鉴定,其中的18个菌株具有致病性。运用培养性状和形态特征综合分析的方法,对上述18个菌株进行鉴定,结果显示为6种镰刀菌,分别为拟枝孢镰孢(Fusarium sporotrioides)、茄镰孢(F.solani)、接骨木镰孢(F.sam-bucinum)、拟丝孢镰孢(F.trichothecioides)、燕麦镰孢(F.avenaceum)和茄病镰孢蓝色变种(F.solanivar.coerule-um)。同时对上述6种镰刀菌进行致病性测定,结果表明不同种类镰刀菌致病性不同,以接骨木镰孢、燕麦镰孢和拟丝孢镰孢致病力最强,拟枝孢镰孢致病力最弱。     

Survey of benomyl resistance in Fusavium species on winter wheat in England and Wales in 1986

In 1986, samples from 109 winter wheat crops in England and Wales were examined for benomyl resistance in Fusarium species. Of the 704 isolates obtained, 581 were identified as Fusarium nivale, of which 92.1 % were resistant to benomyl. Of the remaining isolates, 82 were identified as F. avenaceum, 40 as F. culmorum and one as F. poae; none of these isolates were resistant to benomyl. The vast majority of isolates of Fusarium spp. were obtained from superficial nodal or internodal lesions on the stems.     

The incidence of Fusarium spp. in stem bases of selected crops of winter wheat in the Midlands, UK

The incidence of pathogenic Fusarium and Microdochium species in stem bases of winter wheat was investigated in a total of nine crops in 3 years, i.e. 1987, 1988 and 1989. Four Fusarium species were isolated: F. nivale ( = Microdochium nivale ), F. avenaceum , F. culmorum and F. graminearum . The predominant species was F. nivale followed by F. avenaceum and F. culmorum . Isolations of F. graminearum were made only from shoots collected during August 1989. The highest incidence of F. nivale occurred during April 1989 in the cultivar Brock when the fungus was isolated from 65% of the shoots sampled. The highest incidence of F. avenaceum was 60% (August 1988, cv. Slejpner) and F. culmorum 37% (August 1989, cv. Mercia). A delay in the isolation of Fusarium spp. during 1987 was attributed to the low January temperatures, and an upsurge of F. culmorum and F. graminearum during 1989 to the warm dry summer. The incidence of F. nivale fluctuated during the 1988 and 1989 seasons, particularly during spring. The effects of fungicide spray programmes and the growth and development of the wheat crop are discussed as possible contributory factors to this.     

Development of resistance to thiabendazole in Helminthosporium solani (silver scurf) as a result of potato seed tuber treatment

Applying thiabendazole to potato seed tubers affected with silver scurf caused by Helminthosporium solani sensitive to thiabendazole decreased the severity of disease on progeny tubers at harvest, but about 50% of the isolates from these were resistant to the fungicide. The disease was not decreased when samples of the progeny tubers were treated with thiabendazole and planted in the following year, and the incidence of resistant isolates increased. Resistant isolates continued to be present when tubers were planted in the next 2 years without fungicide treatment. Treatment with a mixture of thiabendazole and imazalil also decreased the disease and fewer isolates were resistant than when treated with thiabendazole alone, although the proportion increased after treatment with the mixture in the following year. When seed tubers were infected with thiabendazole-resistant H. solani , silver scurf on progeny tubers was not affected by thiabendazole applied to the seed tubers but was decreased by the mixture of thiabendazole and imazalil. Imazalil was equally effective against H. solani sensitive or resistant to thiabendazole.
Some isolates of H. solani had grey aerial mycelium and of 516 of these isolates obtained in 4 years 29% were resistant to the fungicide. Other isolates produced small, black colonies and their frequency increased with thiabendazole treatment of seed tubers. Of 244 of these isolates, 62% were resistant.     

Agglutination response of the conidia of eight Fusarium species to lectins having different sugar-binding specificities

The agglutinating effect of five lectins having different sugar-binding specificities on the conidia of a variety of strains from eight Fusarium species was examined. Conidia of all the strains of F. solani and F. oxysporum were strongly agglutinated by concanavalin A and Helix pomatia agglutinin; D-mannose (and D-glucose) and N-acetyl-D-galactosamine inhibited the agglutination induced by concanavalin A and Helix pomatia agglutinin, respectively. Conidia of the other species of Fusarlum examined ( F. culmorum, F. sambucinum, F. graminearum, F. avenaceum, F. moniliforme and F. xylarioides ) were not agglutinated by either lectin. No conidia from the Fusarium species were agglutinated by wheat-germ agglutinin, potato lectin and Ulex europaeus agglutinin-I. These results have provided some insight into the chemical nature of the outermost layer of Fusarium conidia and this information may be of taxonomic significance.     

Resistance to thiabendazole in isolates of Polyscytalum pustulans (skin spot) and Fusarium solani var. coeruleum (dry rot) in Scotland

Between 1988 and 1991, isolates of Polyscytalum pustulans were obtained from potato tubers grown in Scotland and tested for sensitivity to thiabendazole (TBZ) on agar. Sensitive isolates did not grow at 1 mg TBZ/1, whereas growth of resistant isolates was unaffected at 1 mg and reduced by 10–30% at 10 mg/1. Resistant isolates were present on all classes of seed potatoes, and were also recovered from the air of stores during grading. Isolates from a stock of seed tubers from Northern Ireland were resistant. All 44 isolates of Fusarium solani var. coeruleum collected in 1991 were sensitive to TBZ.     

Fusarium head blight of cereals in Denmark: species complex and related mycotoxins

Quantitative real-time polymerase chain reaction differentiating 10 Fusarium spp. and Microdochium nivale or M. majus was applied to a total of 396 grain samples of wheat, barley, triticale, oat, and rye sampled across Denmark from 2003 to 2007, along with selected samples of wheat and barley from 1957 to 2000, to determine incidence and abundance of individual Fusarium spp. The mycotoxins deoxynivalenol (DON), nivalenol, zearalenone, T-2, and HT-2 were quantified using liquid chromatography-double mass spectrometry. Major differences in the Fusarium species complex among the five cereals as well as great yearly variation were seen. Fusarium graminearum, F. culmorum, and F. avenaceum were dominant in wheat, with DON as the dominant mycotoxin. F. langsethiae, F. culmorum, and F. avenaceum were dominant in barley and oat, leading to relatively high levels of the mycotoxins T-2 and HT-2. F. graminearum, F. culmorum, and F. avenaceum dominated in triticale and rye. The nontoxigenic M. nivale/majus were present in significant amounts in all cereal species. Wheat and barley samples from 1957 to 1996 exhibited no or very low amounts of F. graminearum, indicating a recent increase of this pathogen. Biomass and mycotoxin data exhibited good correlations between Fusarium spp. and their corresponding mycotoxins under field conditions.     

Incidence of soil-borne plant pathogens isolated from barley and winter wheat, and other crops in the rotation, on Prince Edward Island

Barley and winter wheat were sampled over 3 years to characterize soil-borne organisms involved with cereal crown and root disease complexes. Winter wheat crowns were infected more often by Fusarium avenaceum and F. sambucinum than barley crowns. Bipolaris sorokiniana and F. graminearum were more common in crown tissue of barley underseeded with clover than in winter wheat crowns. In roots, the incidence of F. avenaceum was highest in winter wheat. In soil, populations of Rhizoctonia solani and F. sambucinum were higher in winter wheat than barley underseeded with ryegrass. Stunt nematodes (Tylenchorhynchus spp.) were greatest in winter wheat and barley underseeded with ryegrass. The incidence of F. avenaceum in roots of winter wheat correlated positively with the severity of crown and root rot symptoms. The severity of barley crown and root rot correlated positively with the incidence of R. solani in crowns, F. avenaceum in crowns and roots, B. sorokiniana in roots, and numbers of stunt nematodes in the soil. The incidence of soil-borne organisms was also recorded in annual ryegrass. soybean, potato, pea, and clover which were grown in rotation with barley and winter wheat. The incidence of R. solani and F. avenaceum was highest in clover crown tissue, and F. sambucinum was prevalent in soybean hypocotyls. Soil population levels of R. solani, F. avenaceum , and stunt nematodes were greatest in ryegrass and pea soil just prior to ploughing down or harvesting these crops, respectively.     

马铃薯干腐病优势病原菌Fusarium sambucinum 分子检测技术的建立

马铃薯干腐病是马铃薯最重要的贮藏期病害之一,现已成为马铃薯贮藏期烂窖的重要原因。实现病原菌的快速检测对病害诊断和科学防控具有重要的实践意义。本研究基于镰刀菌翻译延伸因子序列设计了一对检测接骨木镰刀菌Fusarium sambucinum的特异引物Fs-F/Fs-R。该特异引物可从接骨木镰刀菌中获得309bp的特异性扩增片段,而其他种类镰刀菌及马铃薯重要病害病原菌中均无此片段,说明该引物具有专一性。该体系的灵敏度检测结果表明,最低能检测到的接骨木镰刀菌基因组DNA浓度为70pg/μL。该引物也适用于发病马铃薯块茎中接骨木镰刀菌的快速检测。     

Isozyme and RAPD-PCR analyses of Fusarium avenaceum strains from Finland

Differences in isozyme and RAPD-PCR polymorphisms amongst 33 isolates of Fusarium avenaceum were compared using native polyacrylamide gel electrophoresis and agarose gel electrophoresis. The isolates were collected from different regions of Finland. Amongst eight enzymes analysed clear isozyme polymorphism was detected in five enzymes which could be grouped into 20 different electrophoretic phenotypes and three main groups at the similarity level of 70% in unweighted pair group method with arithmetic average (UPGMA) analysis. RAPD-PCR analysis differentiated all F. avenaceum strains from each other. The phenotypes resulting from RAPD-PCR analysis were grouped into five main groups by UPGMA analysis at the similarity level of 55%. These main groups had several similarities with the main groups from isozyme analysis. RAPD-PCR patterns of 16 isolates of Fusarium graminearum F. culmorum F. equiseti F. oxysporum and F. redolens were also studied and strains from each Fusarium species formed individual groups in UPGMA and principal components analyses. Thus, the extent of isozyme and RAPD-PCR polymorphisms found in Fusarium strains potentially provides a method for identifying the fungi both at strain and species level.     

Population Dynamics of Fusarium spp. and Microdochium nivale in Crops and Crop Residues of Winter Wheat

ABSTRACT Naturally occurring populations of Fusarium avenaceum, F. culmorum, F. graminearum, F. poae, and Microdochium nivale were studied in two field experiments from anthesis in June 2003 until harvest in crops of winter wheat, and subsequently during 10 months after harvest until June 2004 on their residues exposed on the soil surface under field conditions. The dynamics of the different pathogens were estimated by quantifying the amount of DNA present in wheat tissues using TaqMan-polymerase chain reaction. While colonization of grain by Fusarium spp. and M. nivale was low, high amounts of DNA of F. avenaceum, F. graminearum, and F. culmorum were found in ear residues, internodes, and nodes of the mature crop. Amounts of DNA of pathogens decreased significantly during the following 10 months in residues of internodes and nodes, but not in residues of stem bases. Knowledge on population dynamics of pathogens will help to develop preventive measures aimed at reduction of inoculum sources of head blight pathogens.     

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.     

Restriction fragment length polymorphism analysis of variation in Fusarium species causing ear blight of cereals

Genetic variation in Fusarium species on wheat was investigated using restriction fragment length polymorphism (RFLP) analysis. Single-spore lines (76) of Fusarium were recovered from 24 ears of wheat in a field plot exhibiting severe symptoms of Fusarium ear blight and identified using classical taxonomic criteria. Four Fusarium species were present, of which F. avenaceum and F. culmorum were predominant with F. lateritium and F. poae present in two ears and one ear, respectively. RFLP analysis using rDNA (pTA71) or total genomic DNA from an F. culmorum isolate clearly distinguished the four species. Genetic fingerprints of the isolates generated using DNA of bacteriophage M13 (which contains a mini-satellite repeat sequence) revealed considerable variation within three of the four species (except F. poae). Generally, only a single clone was recovered from each ear and in all but one case only a single species was obtained from each spikelet. However, in several instances it appeared that more than one clone of a species was present within a single spikelet.     

Fusarium graminearum as a dry rot pathogen of potato in the USA: prevalence,comparison of host isolate aggressiveness and factors affecting aetiology

A 2004–2005 survey of potatoes from stores in the north‐central potato‐producing region of the USA showed that the predominant causes of dry rot were Fusarium graminearum and Fusarium sambucinum. Isolates of F. graminearum originally isolated from potato tubers with dry rot (n = 15), wheat kernels with scab (n = 15), and sugarbeet tap roots with decay (n = 5) were tested for aggressiveness to potato tubers. There were no significant differences in aggressiveness among isolates of F. graminearum, regardless of original host, as measured by their ability to cause dry rot. These findings may have implications for survival of F. graminearum inoculum since potatoes, wheat and sugarbeets are frequently used in crop rotation in the region. Fusarium graminearum required larger wounds for infection of potato tubers than F. sambucinum. Plug‐removal injury, simulating a stolon‐removal injury, resulted in equal incidence of dry rot caused by the two Fusarium species, whereas abrasion and bruising injury were sufficient for infection and dry rot development by F. sambucinum, but not F. graminearum. A change in harvest practices from vine‐killing prior to harvest to mechanical vine‐killing on the day of harvest may be a factor affecting the onset of dry rot caused by F. graminearum, since this process often causes large wounds at the stem end of the tubers when the stolon is forcibly removed.     

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.     

Influence of light-enhanced glycoalkaloids on resistance of potato tubers to Fusarium sulphureum and Fusarium solani var. coeruleum

Increases in resistance to Fusarium sulphureum and Fusartium solani var. coeruleum were determined in seed tubers of cultivars Desiree, Pentland Hawk and Kerr's Pink with light-enhanced glycoalkaloids. Glycoalkaloid concentration in the tubers ranged from 78.0 to 885.9 mg kg⁻¹ fresh weight. Exposure of tubers to light prior to inoculation decreased fungal invasion, the greater reductions generally reflecting longer light exposure periods. In most cases, no direct relationship between resistance and glycoalkaloid concentrations was recorded for F . sulphureum. Surface infection indices and reduced depth of rot caused by F. solani var. coeruleum were closely correlated with tuber glycoalkaloid content. No significant effects on spore length and width were recorded. Exposure to light of seed tubers that are not destined for human or animal consumption could provide a pretreatment to reduce fungal invasion by F. sulphureum and F. solani var. coeruleum .     

Pathogenicity of fungi associated with winter loss and injury in white lupin

Fungi isolated from diseased white lupin plants were tested for pathogenicity in standardized glasshouse tests to assess their potential as causes of plant death in winter in autumn-sown crops. Pleiochaeta setosa and, to a slightly lesser extent, Fusarium avenaceum caused lesions or plant death when inoculated onto hypocotyls of lupin seedlings. Disease was more severe when the hypocotyls were wounded before inoculation. F. avenaceum also caused injury when inoculated onto roots; wounding the roots had no effect. Some isolates of F. solani also caused injury, especially after root inoculation, but isolates of F. oxysporum and Cylindrocarpon destructans were only slightly or non-pathogenic. Isolates of Fusarium and C. destructans , but not of P. setosa , differed in pathogenicity. There were differences in susceptibility to disease among three lupin cultivars.     

Inoculating plant material by jet injection

Both tubers and plants of potato were successfully inoculated with Erwinia carotovora subsp. itroseptica by means of a needle-less medical jet injector. The instrument has also been used to infect potato tubers with Phorna exigua var. foveata, Fusarium solani var. coeruleum and F. sulphureum. It is suggested that high-pressure injection is a potentially useful tool for introducing pathogens into plant materials.