Influence of the fungal hyperparasite Trichoderma harzianum on the growth of Epichloë typhina,an agent of choke disease in grasses

Journal of Plant Diseases and Protection - In its sexual stage, the fungus Epichloë typhina (Ascomycetes: Clavicipitaceae) is a pathogen that causes choke disease in many grass species. It...     

An isolate of Epichloë festucae,an endophytic fungus of temperate grasses,has growth inhibitory activity against selected grass pathogens

The symbiotic association of epichloae endophytes (Epichloë/Neotyphodium species) with temperate grasses of the subfamily Pooideae is known to enhance plant host tolerance to abiotic and biotic stresses. While the protection of the host plant from insect herbivory by epichloae endophytes is well characterized, the mechanism by which they protect their host against grass pathogens is largely unknown. Here, we assessed a geographically diverse collection of 14 Epichloë festucae isolates for in vitro antifungal activity against 8 grass pathogens. Isolate E437 of E. festucae, which had the broadest antifungal spectrum, inhibited growth of Drechslera erythrospila, D. siccans, D. dictyoides, Colletotrichum graminicola and Bipolaris sorokiniana. As shown with confocal microscopy, the endophyte reduced hyphal tip growth and differentiation of the pathogen, but did not cause any lysis. The isolate produced a thermostable, low-molecular-weight antifungal compound in culture. Disease symptoms caused by D. erythrospila on perennial ryegrass plants infected with E437 were reduced, suggesting the antifungal compound produced by E. festucae E437 isolate could be involved in the protection of the host plant.     

Enkele gegevens omtrent sclerospora in Indonesië

Samenvatting De gevoelige periode van de maisplant voor infectie doorSclerospora maydis (Rac.) Butler werd bepaald, alsmede de incubatietijd (grafieken 1 en 2). Behalve deze soort komt sinds 1947 op Noord CelebesSclerospora philippinensis Weston voor, die daar aanzienlijke schade aan mais toebrengt. De verschillen van deze schimmel metSclerospora sacchari zijn slechts gering en een nader onderzoek dienaangaande is dan ook zeer gewenst.De eerste oögoniën-vorm vanSclerospora in Indonesië werd gevonden en wel vanSclerospora northii Weston op eenErianthus spec. op het eiland Soemba.Summary The nomenclature ofSclerospora maydis (Rac.) Butler is discussed. The susceptible period of the host and the time of incubation of this fungus (graphs 1 and 2) were determined.Since 1947 a species ofSclerospora has caused considerable damage on maize in North Celebes. It is stated to beSclerospora philippinensis Weston. The differences between this fungus andSclerospora sacchari T. Miyake are very slight. AsSclerospora sacchari is still unknown in Indonesia, it is very important for the culture of sugarcane on Java to investigate by means of inoculation trials, ifSclerospora philippinensis can attack sugarcane. Sclerospora northii Weston has been found onErianthus spec. on the isle of Sumba; it is the only knownSclerospora species in Indonesia which forms oögonia.Vroeger verbonden aan het Instituut voor Plantenziekten te Bogor, Indonesia, thans aan het Inst. v. Plantenziektenkundig Onderzoek (I.P.O.) te Wageningen.     

De gomziekte der Amygdaleeën in vergelijking met den boomkanker

Ohne Zusammenfassung     

Development of a qPCR test to detect and quantify Elsinoë piri in unsprayed and organic apple orchards and assessment of apple cultivar susceptibility to the disease

Recently, Elsinoë piri has become an important pathogen in unsprayed and organic apple orchards. The increasing demand for organic apples as well as the limited use of fungicides imposed by European legislation has turned this pathogen, which has been known for over a century in Europe, into a resurgent phytosanitary threat. It also represents a new trait to be taken into account in apple breeding programmes. As E. piri is extremely difficult to isolate and causes symptoms that can be confused with those caused by other pathogens, a quantitative PCR (qPCR) test has been developed based on the internal transcribed spacer (ITS) region of the rDNA gene. This test, which displays a low limit of detection, allows the early detection of the fungus in apple leaves. As a quantitative method, it is a promising tool for breeding purposes as it can be used on leaves to assess the level of quantitative resistance of apple cultivars to the disease. The molecular test is also useful to gain knowledge on the epidemiology of this re-emerging fungal disease. Spore trapping conducted in 2015, 2016 and 2020 using Rotorod devices and the qPCR test showed that airborne inoculum is released at the end of the growing season. Tests were also carried out to demonstrate that the pathogen was detected in buds during winter. A comparison of 18 E. piri isolates based on the sequencing of five genome regions highlighted a high genotypic diversity. All these isolates were detected with the qPCR test developed in this study.     

Cellulase in the Host–parasite System Phaseolus vulgaris (L.)–Uromyces appendiculatus [Pers.] Link

Activity of cellulase and xylanase in the intercellular washing fluid (IWF) of bean plants (Phaseolus vulgaris, cultivar Fori) was monitored during infection with bean rust (Uromyces appendiculatus). In infected plants, cellulase activity could be detected at 2 days after inoculation and reached its maximum between 7 and 8 days after inoculation. The enzyme activity was not detected in healthy controls. The cellulase had a pH optimum at pH 5.5 and a temperature optimum at 30°C. Complete inactivation of cellulase occurred after heating to 50°C for 30min. In non-denaturing polyacrylamide gradient gels, the enzyme exhibited four bands (molecular masses approximately 70, 95, 120, 170kDa). After isoelectric focusing, eight cellulase isoforms with pI values pI 4.6–4.8; 5–5.1; 5.4; 5.5; 5.9; 6; 6.5; 7 appeared. Two dimensional electrophoresis yielded 13 cellulase isoforms. Unlike cellulase, low levels of xylanase were detected in healthy controls. The activity of this hydrolase did not increase due to rust infection.     

Characterization of Populations of Rhizoctonia solani in Paddy Rice Fields in Côte d'Ivoire

ABSTRACT Isolates of Rhizoctonia solani were obtained from plant and soil samples that had been systematically collected in a field experiment in C?te d'Ivoire to study the diversity of the pathogen and the influence of three different rice rotations on the pathogen population. Characterization by morphology, anastomosis testing, pathogenicity testing, and restriction fragment length polymorphisms (RFLPs) of AT-rich DNA (AT-DNA) showed that there were no differences in isolates from different experimental plots, suggesting that the soil as well as the plant population of the fungus was indistinguishable throughout the experiment and was not influenced by crop rotation. Analysis of AT-DNA showed that the isolates obtained from plant material and one from soil shared a distinct banding pattern, identical with the AT-DNA RFLP obtained for the reference strain of anastomosis group 1 (AG-1). The remaining soil isolates produced a consistent RFLP pattern that was distinct from that of the plant isolates. Morphological characterization of isolates produced two major clusters consisting of the same groups of isolates as found by AT-DNA RFLP. Diversity in morphological characters was much higher in plant than in soil isolates and indicated that the population might consist of several clones. Anastomosis testing revealed that soil as well as plant isolates were able to fuse with the tester strain of AG-1. Significant differences in disease severity were observed between the two groups of isolates in pathogenicity tests on rice plants, with plant isolates being distinctively more virulent.     

Epidemiology of flavescence dorée in vineyards in northwestern Italy

ABSTRACT A serious outbreak of flavescence dorée (FD) was reported in Piemonte, northwestern Italy, in 1998, and since then, the disease has compromised the economy of this traditional wine-growing area, even following the application of compulsory insecticide treatments to control Scaphoideus titanus, the vector of the causal phytoplasma. Affected vines show severe symptoms, varying according to the cultivar, and are rogued to reduce disease spread. Following winter and pruning, a previously affected vine may appear symptomless and free of phytoplasmas in its aerial as well as its root system, even by nested-polymerase chain reaction assays. Such plants are considered to be "recovered". Since 1998 homogenous data on the incidence of newly infected, healthy, or recovered plants productivity, presence of vectors, and treatment schedules have been collected in seven severely affected vineyards of southern Piemonte for 5 years (1999 to 2003). Infectivity and recovery rates were also calculated each year. From 1999 to 2003, the average number of healthy plants decreased and the numbers of recovered plants and those with symptoms increased. Productivity of recovered vines, although lower than that of healthy ones, was always higher than that of vines with symptoms and was not influenced by the time elapsed from date of recovery. The relationships between the ln-transformed number of vectors trapped in the vineyards the previous year and the infection and the recovery rates were fitted by an exponential (R(2) = 0.95) and an asymptotic (R(2) = 0.93) model, respectively.     

Occurrence of boscalid resistance in cucumber powdery mildew in Japan and molecular characterization of the iron–sulfur protein of succinate dehydrogenase of the causal fungus

A total of 74 mass isolates of cucumber powdery mildew fungus (Podosphaera xanthii) were collected from commercial greenhouses with a history of boscalid use in Ibaraki Prefecture, Japan, and tested in a leaf disk assay for their sensitivity to boscalid. The mildew development of 40 of 74 isolates and five sensitive reference isolates on the disks was completely suppressed at 5 μg boscalid/ml. The minimum inhibitory concentrations (MIC) for the remaining 34 isolates were 50 μg/ml or higher, and 21 of these isolates also grew well at 500 μg/ml. Six single-spore isolates were resistant to boscalid with MIC values higher than 500 μg/ml; four of these were moderately resistant (MR), and two were very highly resistant (VHR) isolates. The growth of MR isolates was almost completely suppressed at 500 μg/ml, whereas two VHR isolates grew vigorously at 500 μg/ml. In foliar inoculation tests of potted cucumber plants, the efficacy of boscalid (500 μg/ml) against both MR and VHR isolates was very low. Partial DNA fragment of the iron–sulphur protein subunit (SdhB) gene of succinate dehydrogenase was PCR-amplified from five sensitive and five resistant isolates and directly sequenced, revealing that VHR isolates possess a substitution from a highly conserved histidine (CAT) to tyrosine (TAT) in a third cysteine-rich center of a putative SdhB, whereas MR isolates so far have not been found to have such substitution in SdhB.     

Induction of β-1,3-glucanase in Seedlings of Pearl Millet in Response to Infection by Sclerospora graminicola

Differential resistance of pearl millet cultivars to downy mildew disease was correlated with the levels of -1,3-glucanase in their seeds. Higher activity of the enzyme in highly resistant cultivars and lower activity in the highly susceptible ones suggested the possible use of -1,3-glucanase as a biochemical marker for screening pearl millet cultivars for downy mildew disease. Inoculation of seedlings with the downy mildew pathogen Sclerospora graminicola resulted in increased enzyme levels in resistant cultivars. Mesocotyl and shoot regions of seedlings recorded higher levels of enzyme than the root. Isoelectric focusing revealed four basic isoforms with pI 9.6, 9.0, 8.9 and 8.2 and two acidic isoforms with pI 4.9 and 6.2 of -1,3-glucanase in pearl millet. The pI 9.6 isoform was a major isoform of the enzyme in the pearl millet seedlings with a probable developmental function. Isoforms pI 6.2 and pI 8.2 appeared to be involved in resistance and pI 4.9 isoform seemed to be involved in pathogenesis of pearl millet-downy mildew.     

Sources of inoculum and reappearance of spot blotch of wheat in rice–wheat cropping

A study was undertaken to examine the main source of inoculum of Bipolaris sorokiniana responsible for its reappearance in rice–wheat cropping regions of eastern India. Soil samples were collected at monthly intervals during April–October in the years 2000 and 2001 from fields having rice–wheat cropping. Bipolaris sorokiniana conidia were isolated and their viability was found to decline sharply with the onset of flooding in the month of August. In contrast to 82% in April, viability was 4% and <1% in August and September, respectively. Viable conidia were multiplied in the laboratory and inoculated on to susceptible cv. Sonalika under controlled conditions for test of pathogenicity. Appearance of symptoms typical to spot blotch were recorded after 7 days. Twenty-two different species (weeds and grasses) normally found to be associated with rice–wheat fields were tested for the presence of B. sorokiniana to evaluate their possible role as alternative hosts. Only three species, i.e. Setaria glauca, Echinochloa colonum and Pennisetum typhoids, were found to naturally harbour B. sorokiniana. Isolates from these hosts were tested for pathogenicity and also for their possible spread to wheat. When reisolated from these hosts, the pathogen did not infect wheat. Seeds of 25 different wheat genotypes were tested for B. sorokiniana infection. All genotypes were infected and the incidence of infection varied from 26% to 86%. Five isolates of wheat and one isolate from each of the three species (S. glauca, E. colonum and P. typhoids) were subjected to RAPD analysis. Two broad clusters were formed, suggesting that the wheat isolates were different from the isolates originating from other hosts. The results indicate that seeds are the most important source of inoculum for the reappearance of spot blotch of wheat in rice-wheat cropping systems in eastern India.     

Characterisation of benzimidazole resistance of Cercospora beticola in Serbia using PCR-based detection of resistance-associated mutations of the β-tubulin gene

A survey to detect and characterise benzimidazole resistance within populations of Cercospora beticola in Serbia was performed. From 52 field isolates collected from sugar beet and beet root, only eight were found to be benzimidazole-sensitive based on the inhibition of mycelial growth by discriminatory concentrations of carbendazim and thiophanate-methyl. Sensitivity tests revealed the presence of three resistant phenotypes among the tested isolates: high-resistance (HR), low-resistance (LR) and moderate-resistance (MR). The benzimidazole resistant isolates were characterised based on the DNA sequence of the β-tubulin gene and temperature sensitivity. The HR isolates showed no temperature sensitivity regardless of carbendazim concentration, whereas the LR and MR isolates were sensitive at lower temperatures. Analysis of the β-tubulin gene sequence revealed two amino acid replacements in the benzimidazole-resistant isolates of C. beticola. One was a glutamic acid to alanine change at position 198 (codon GAG to GCG) that was identified in HR isolates; this mutation has previously been reported to be associated with the development of benzimidazole resistance in C. beticola. The second replacement was a novel point mutation of phenylalanine (TTC) to tyrosine (TAC) at position 167, identified in low and moderate benzimidazole-resistant isolates, sharing a single LR/MR β-tubulin genotype. A diagnostic PCR-RFLP assay utilising a BsaI restriction site present in the benzimidazole sensitive and LR/MR genotypes but absent in the HR genotype was developed for the routine detection of high resistance. A mutation-specific PCR assay was developed for the diagnosis of LR/MR genotype based on a mutation from T to A at codon 167, which is unique to this genotype.     

Evidence of the Presence of Two Serotypes of Rice Yellow Mottle Sobemovirus in Côte d'Ivoire

Serological variability of isolates of rice yellow mottle virus (RYMV) collected in Côte d'Ivoire was assessed by immunological tests with polyclonal and monoclonal antibodies (MAbs). Two serotypes (named S1 and S2) were distinguished. The S1 isolates had common epitopes which were absent in S2 isolates, whereas they lacked epitopes shared by S2 isolates. There was no evidence of S1 and S2 mixtures, although S1 and S2 isolates were sometimes found in nearby sites. Serotype S2 was more prevalent in Côte d'Ivoire than S1, and was in a large majority in the centre and the south of the country. By contrast, S1 occurred more widely in the north. S1 isolates were also found in neighbouring countries at the north of Côte d'Ivoire. In tests with monoclonal antibodies, three additional serotypes were found, one in West-Africa and two in East-Africa. Using the primers developed against an S2 isolate from Côte d'Ivoire, all S2 but not the S1 isolates were transcribed and amplified by RT-PCR, and another set of primers was developed to amplify S1 isolates. S1 and S2 have different biological properties, and competition between isolates of the two strains was apparent resulting in S2 dominance over S1. This was assessed using S1 and S2 strain specific MAbs, and it occurred whatever the pattern of inoculation or the rice variety tested. Differences in pathogenicity and virus titre did not account for strain competition, as there was no relation between symptom severity, virus content and serotype of the isolates in Oryza sativa indica cultivars.     

Seasonal progression of leaf rust in ‘Niagara Rosada’ grapevine in a biannual crop system in Brazil

The soil-borne fungus Fusarium oxysporum can cause both Fusarium yellows and Fusarium root rot diseases with severe yield losses in cultivated sugar beet. These two diseases cause similar foliar symptoms but different root response and have been proposed to be caused by two distinct F. oxysporum formae speciales. Fusarium yellows, caused by F. oxysporum f. sp. betae, presents vascular discoloration, whereas Fusarium root rot, due to F. oxysporum f. sp. radicis-betae, appears as black rot visible on the root surface. The aim of this work was to study the host-pathogen interaction between sugar beet lines and isolates originally characterized as Fusarium oxysporum f. sp. betae. Eight susceptible sugar beet lines, selected by the USDA-ARS (US) and UNIPD (University of Padova, Italy) breeding programs, were inoculated with three different isolates of F. oxysporum f. sp. betae, the causal agent of Fusarium yellows, representing different genetic groups. All inoculated lines developed symptoms, but severity, expressed as area under the disease progress curve (AUDPC), differed significantly (P < 0.05) among lines. Two lines from UNIPD, 6 and 9, were the most susceptible to the disease, whereas the other lines showed similar levels. The three isolates of F. oxysporum f. sp. betae differed significantly (P < 0.05) in disease severity. Five weeks after inoculation the plants were harvested and roots examined. Surprisingly, severe root rot was observed in the susceptible UNIPD lines when inoculated with all three isolates, while this symptom was never observed in the USDA germplasm. The development of this disease symptom obviously depends on the plant genotype.