A fungitoxic substance extracted from tulips and its possible role as a protectant against disease

A specialized strain ofFusarium oxysporum is the cause of a disease important in tulip bulb cultivation. From observations in the field it was concluded that the fungus is not able to penetrate into the tulip bulb during the greater part of the growing season. A protecting barrier was demonstrated to be present in the white skin. From extracts of this white skin (and from other parts of the tulip plant) a fungitoxic substance was isolated, which was identified as α-methylene butyrolactone. A possible precursor of this lactone has been isolated which, under certain conditions, yields the lactone in vitro. The implications of the presence of the fungitoxic substance in extracts made from different parts of the plant in relation to the susceptibility toFusarium are discussed.     

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.     

Influence of soil temperature on field infection of tulip bulbs by Fusarium oxysporum

Soil temperature is an important factor in field infection of tulip bulbs byF. oxysporum in both autumn and late spring. Three types of infection and the associated symptoms are discussed in relation to temperature.

1. The infection through the browning tunic at any place in the outer scale, which occurs during the last few weeks before harvest, being most common in the Netherlands.
2. A basal rot is thought to be favoured by a relatively high temperature after planting in combination with a warm spring, conditions which sometimes also cause premature death of plants; these symptoms are attributed to infection of the root plate immediately after planting.
3. After a warm spring period several weeks before normal lifting time brown spots may be present in the otherwise white tunic, frequently accompanied by an infection of the scale below the spot.

It is suggested that a high inoculum level built up in the decaying scales of the planted bulb under warm conditions may weaken the tunic barrier by local reduction of the tuliposid concentration in the white tunic, which compound is a precursor of the fungitoxic tulipalin. A semi-quantitative assay for estimation of the number of fungus propagules in the old scales is described.     

Cytological karyotyping of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> by the germ tube burst method (GTBM)

Fusarium oxysporum is an ascomycete fungus including plant pathogenic and nonpathogenic strains. Genome analyses have indicated that the karyotype of F. oxysporum is diverse among isolates. Here we used the germ tube burst method (GTBM), a more reliable method than conventional cytology or pulsed field gel electrophoretis, to karyotype isolates of F. oxysporum ff. spp. lycopersici and conglutinans and nonpathogenic F. oxysporum. In this first application of GTBM for F. oxysporum, pathogenic isolates were found to have more chromosomes than in nonpathogenic isolates. We also used a ribosomal DNA probe and fluorescence in situ hybridization (FISH) to analyze chromosome structure.     

Inactivation of soil-borne plant pathogens during small-scale composting of crop residues

Samples of heavily infested crop residues were incorporated in static compost heaps (2.5–4.6 m³) of the Indore type. Temperature increased to 50–70°C within 6 days depending on the type of crop residues used and the location within the heap. The heat phase (>40 °C) lasted 2–3 weeks and was followed by a c. 5-months maturation phase (<40 °C). Among the 17 pathogens tested, onlyOlpidium brassicae and one of the four formae speciales ofFusarium oxysporum that were tested survived composting, but also their inoculum was greatly reduced.Survival during specific phases of composting was studied by incorporation and retrieval of samples at various stages of the process.F. oxysporum f. sp.melonis was completely inactivated andO. brassicae andPlasmodiophora brassicae were almost completely inactivated during the short heat phase. The three pathogens survived the long-lasting maturation phase without loss of viability. Heat evolved during composting was found to be the most important factor involved with sanitation of crop residues. The possible involvement of fungitoxic conversion products and microbial antagonism is discussed.     

Five plant families support natural sporulation of <Emphasis Type="Italic">Cronartium ribicola</Emphasis> and <Emphasis Type="Italic">C. flaccidum</Emphasis> in Finland

Fusarium is one of the most destructive fungal genera whose members cause many diseases on plants, animals, and humans. Moreover, many Fusarium species secrete mycotoxins (e.g. trichothecenes and fumonisins) that are toxic to humans and animals. Fusarium isolates from date palm trees showing disease symptoms, e.g. chlorosis, necrosis and whitening, were collected from seven regions across Saudi Arabia. After single-sporing, the fungal strains were morphologically characterized. To confirm the identity of morphologically characterized Fusarium strains, three nuclear loci, two partial genes of translation elongation factor 1 α (tef1α) and β-tubulin (tub2), and the rDNA-ITS region, were amplified and sequenced. Of the 70 Fusarium strains, 70 % were identified as F. proliferatum that were recovered from six regions across Saudi Arabia. Fusarium solani (13 %), as well as one strain each of the following species: F. brachygibbosum, F. oxysporum, and F. verticillioides were also recovered. In addition, five Fusarium-like strains were recognized as Sarocladium kiliense by DNA-based data. The preliminary in vitro pathogenicity results showed that F. proliferatum had the highest colonization abilities on date palm leaflets, followed by F. solani. Although F. oxysporum f. sp. albedinis is the most serious date palm pathogen, F. proliferatum and F. solani are becoming serious pathogens and efforts should be made to restrict and control them. In addition, the potential toxin risks of strains belonging to F. proliferatum should be evaluated.     

Unraveling the multilevel aspects of least explored plant beneficial <Emphasis Type="Italic">Trichoderma saturnisporum</Emphasis> isolate GITX-Panog (C)

Species of Trichoderma offer an eco-friendly and economical solution to the management of plant pathogens in agricultural crops. The secretion and spectrum of different bioactive molecules by Trichoderma spp. is known to follow adaptive and species/strain behavior, depending upon the edaphic conditions. In present study, GITX-Panog (C), an antagonistic isolate of Trichoderma saturnisporum against Fusarium oxysporum was screened for the production of different glycosyl hydrolases and secondary metabolites. The evaluation of antagonistic isolates for lytic enzymes and secondary metabolites revealed their significance as plant probiotics. Profiling of lytic enzymes such as chitinases and β-glucanases secreted by T. saturnisporum GITX-Panog (C) in the presence of autoclaved mycelium of F. oxysporum and colloidal chitin or laminarin showed increased enzyme activities. Production of bioactive secondary metabolites such as siderophore, peptaibols, 6-pentyl-α-pyrone, and the known potential of these compounds in mineral acquisition and suppression of plant F. oxysporum, revealed the potential significance of Trichoderma saturnisporum isolate GITX-Panog C in agricultural application against biotic and abiotic stresses. The antagonistic isolate also showed significant increases in fresh biomass and seed production in Arabidopsis thaliana in the greenhouse compared to controls. Thus, the current study highlights the multiple attributes of an antagonistic isolate of T. saturnisporum for future agricultural applications as a plant probiotic agent.     

Check-list for scientific names of common parasitic fungi. Series 3b: Fungi on bulbs: Amaryllidaceae and Iridaceae

This list is a continuation of Series 3a (Neth. J. Pl. Path. 94 (1988), Supplement 1), an account of the nomenclature of common parasitic fungi on bulbs as used in official publications of the Netherlands Society of Plant Pathology and the Netherlands Ministry of Agriculture and Fisheries. The selected names include one new species,Curvularia gladioli Boerema & Hamers, one new pathogenic form,Fusarium oxysporum f.sp.croci Boerema & Hamers, and one new combination,Sclerotium narcissi (Sacc.) Boerema & Hamers.     

Special issue “Deepen knowledge in plant pathology for innovative agroecology”

This study aimed to determine the genetic variability of isolates of rotting of pineapple fruitlet core in Brazil on the states of Paraiba, Pernambuco and Rio Grande do Norte, based on phylogenetic analysis of the RPB2 gene, morphocultural markers and aggressiveness of the isolates. The phylogenetic reconstruction of maximum parsimony and bayesian inference of the isolates were performed. Twenty-seven phylogenetic lineages were characterized with morphocultural markers on potato dextrose agar and synthetic nutrient-poor agar. The aggressiveness of these lineages were characterized in leaves and fruits of pineapple ‘Pérola’ cultivar. The Phylogenetic reconstruction showed close relationship between isolates of this study and phylogenetic lineages of F. guttiforme, F. ananatum and F. oxysporum by analysis of RPB2 gene. Phylogenetic lineages of this study shared significant morphocultural markers similar to those described for this species. Overall, the lineages related with Fusarium oxysporum Species Complex were more aggressive to the fruits of the Fusarium lineages related with Fusarium fujikuroi Species Complex. It is possible that F. ananatum and F. guttiforme or other lineages closely related to these species or F. oxysporum are present in the fields causing pineapple rotting fruitlet core in fruits of pineapple ‘Pérola’ in Brazil. The used markers determined high genetic variability in isolates analyzed in leaves and pineapple fruit ‘Pérola’ in the states of Paraíba, Pernambuco and Rio Grande do Norte of Brazil and the pathogenic lineages analyzed were better adapted to the fruits than to the detached leaves in pineapple ‘Pérola’ analyzed.     

<Emphasis Type="Italic">Fusarium oxysporum</Emphasis> and <Emphasis Type="Italic">Fusarium avenaceum</Emphasis> associated with yield-decline of pyrethrum in Australia

Fusarium wilt, one of the destructive diseases of cucumber can be effectively controlled by using biocontrol agents such as Trichoderma harzianum. However, the mechanisms controlling T. harzianum-induced enhanced resistance remain largely unknown in cucumber plants. Here we screened the potent T. harzianum isolate TH58 that could effectively control F. oxysporum (FO). Glasshouse efficacy trials also showed that TH58 decreased disease incidence by 69.7 %. FO induced ROS over accumulation, while TH58 inoculation suppressed ROS over accumulation and improved root cell viability under F. oxysporum infection. TH58 inoculation could reverse the FO-induced cell division block and regulate the proportional distribution of nuclear DNA content through inducing 2C fraction. Moreover, the expression levels of cell cycle-related genes such as CDKA, CDKB, CycA, CycB, CycD3;1 and CycD3;2 in TH58 - pre-inoculated seedlings were up-regulated compared with those infected with FO alone. Taken together, these results suggest that T. harzianum improved plant resistance against Fusarium wilt disease via alterations in nuclear DNA content and cell cycle-related genes expression that might maintain a lower ROS accumulation and higher root cell viability in cucumber seedlings.     

<Emphasis Type="Italic">In vivo</Emphasis> evaluation of essential oils and biocontrol agents combined with heat treatments on basil cv Genovese Gigante seeds against <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">basilici</Emphasis>

Seed treatments with essential oils (from savory and thyme) and biocontrol agents (Pseudomonas spp. and Fusarium oxysporum) have been evaluated in vivo after dry hot air treatments against Fusarium oxysporum f. sp. basilici on basil seeds. The savory and thyme essential oils showed a significant pathogen control activity because of their innate antifungal activity and because of the seed application method, but the dry hot pre-treatment did not show any obvious effect on the performance of the essential oil treatments. The dry heat treatment improved the Pseudomonas seed dressing effect against F.oxysporum f. sp. basilici, and showed important reductions in plant infection and the disease index on the treated seed plants, without any negative effect on seed germination. However, the pathogen control provided by the heat treatments combined with the application of the biocontrol agents never reached the same performance as the chemical treatments considered as the reference. Thus, short dry heat treatments on basil seeds have been shown to be a valid but complementary seed disinfection method against Fusarium wilt.     

Identification and characterization of pathogens associated with root rot of winter peas grown under organic management in Germany

Root rots are limiting factor for pea production worldwide. This disease is caused by a pathogen complex and the role of single pathogens is unclear. This study aimed at identifying pathogens involved in a root rot of organically grown field pea in Germany, and establishing their importance in the disease complex. The potential of yard waste compost to suppress the diseased was also studied. Average disease severity index was similar in 2010 and 2011 (DI of 4.56 to 4.59, respectively) but it increased in 2012 to DI 5.8. Peyronellaea pinodella was most frequently isolated pathogen, with isolation frequency from 86%, 73% and 86% in 2010, 2011 and 2012, respectively. In addition, Didymella pinodes, Fusarium solani f. sp. pisi, F. oxysporum f. sp. pisi and F. avenaceum were the main fungi recovered from pea roots. In pathogenicity test all of the tested pathogens caused weak symptoms on the pigmented winter variety EFB33 and moderate to severe symptoms on the white flowering summer variety Santana. F. avenaceum was the most aggressive pathogen on Santana with DI of 7.4 followed by P. pinodella with DI of 5.7. The high aggressiveness combined with the wide host range highlights the possibility of F. avenaceum emerging as potential risk for organic crop rotation. High levels of resistance of EFB33 against all pathogens shows the potential of this variety to serve as a resource in further research for identification and development of new sources of resistance against root rot diseases of pea.     

Panama disease of banana occurred in Miyakojima Island,Okinawa, Japan

In 2016, in Miyakojima, Okinawa, Japan, banana plants (Musa?×?paradisiaca) ‘Shima-banana’ developed yellowing and wilt associated with vascular discoloration of the pseudostems. Fusarium oxysporum, identified based on morphological characters, was frequently isolated from the vascular tissue of the infected plant and reproduced the original symptoms on ‘Shima-banana’ after drench inoculation with a spore suspension. Thereby, we determined that the disease is Panama disease caused by F. oxysporum. This is the first official report of Panama disease (Panama-byo in Japanese) of banana in Japan.     

In vitro and field efficacy of three fungicides against <Emphasis Type="Italic">Fusarium</Emphasis> bulb rot of garlic

Fusarium proliferatum has been identified as the main causal agent of bulb rot of garlic (Allium sativum L.). This disease occurs after the drying process and can rot almost 30 % of the bulbs. Few studies are available regarding the effectiveness of chemical treatments to reduce F. proliferatum incidence in garlic. The efficacy of three commercial fungicides of different chemical groups to reduce seven strains of F. proliferatum mycelial growth was tested in vitro. These three fungicides were also evaluated by foliar spreading of aqueous suspension in a field crop. Fluopyram 20 % + tebuconazole 20 % and tebuconazole 50 % + trifloxystrobin 50 % were highly effective at reducing mycelial growth in F. proliferatum with EC₅₀ values <2 ppm. In general, the effectiveness of the fungicides was enhanced with increasing dosage. Our results indicate that the fungicides evaluated in this study may lead to a risk of resistance appearing in F. proliferatum at low concentrations and this risk is maintained at higher doses for the fungicide dimethomorph 7.2 % + pyraclostrobin 4 %. Although several of the fungicides affected in vitro mycelial growth of F. proliferatum, as a part of an strategy to measure the efficacy of resistance management it is necessary to monitor the ongoing efficacy of fungicides under commercial conditions. All fungicidal treatments tested in field application failed to control garlic bulb rot during storage.     

Environmental effects on growth and sporulation of <Emphasis Type="Italic">Fusarium</Emphasis> spp. causing internal fruit rot in bell pepper

Internal fruit rot in bell pepper (Capsicum annuum L.) is mainly caused by members of the Fusarium lactis species complex (FLASC) and to a lesser extent by Fusarium oxysporum and Fusarium proliferatum. Despite the importance of the disease, there is hardly no information about growth, sporulation and germination dynamics of FLASC. In order to understand the dominance of FLASC as main pathogen of internal fruit rot, the effects of temperature (5 °C – 35 °C), water activity (a_w 0.76–0.96), pH (pH 3 - pH 9) and oxygen concentration (2.5% - 20%) on growth and sporulation of all three Fusarium species were compared. In addition, germination kinetics were also investigated. FLASC showed optimal mycelium growth and sporulation in the narrow range of 25 °C, while both other strains were also tolerant for higher temperatures to 30 °C. FLASC was also characterized by a broad pH optimum from pH 3–7 while F. oxysporum (pH 4–7) and F. proliferatum (pH 5–8) were more demanding concerning pH. In addition, optimal sporulation occurred in the acid region for FLASC (pH 3) whilst neutral and alkaline pH were more favourable for the other species. Germination kinetics revealed that FLASC did not benefit from an earlier and/or faster germination process. A thorough understanding of the growth characteristics and dominance of FLASC as main pathogen for internal fruit rot is inevitable to develop sustainable control measures for the disease.     

Characterization of <Emphasis Type="Italic">Trichoderma</Emphasis> species isolated in Ecuador and their antagonistic activities against phytopathogenic fungi from Ecuador and Japan

Native Trichoderma spp. were isolated from agricultural fields in several regions of Ecuador. These isolates were characterized via morphological observation as well as molecular phylogenetic analysis based on DNA sequences of the rDNA internal transcribed spacer region, elongation factor-1α gene and RNA polymerase subunit II gene. Fifteen native Trichoderma spp. were identified as T. harzianum, T. asperellum, T. virens and T. reesei. Some of these strains showed strong antagonistic activities against several important pathogens in Ecuador, such as Fusarium oxysporum f. sp. cubense (Panama disease) and Mycosphaerella fijiensis (black Sigatoka) on banana, as well as Moniliophthora roreri (frosty pod rot) and Moniliophthora perniciosa (witches’ broom disease) on cacao. The isolates also showed inhibitory effects on in vitro colony growth tests against Japanese isolates of Fusarium oxysporum f. sp. lycopersici, Alternaria alternata and Rosellinia necatrix. The native Trichoderma strains characterized here are potential biocontrol agents against important pathogens of banana and cacao in Ecuador.     

Identity and pathogenicity of <Emphasis Type="Italic">Fusarium</Emphasis> species associated with crown rot on wheat (<Emphasis Type="Italic">Triticum</Emphasis> spp.) in Turkey

An extensive survey was carried out to collect Fusarium species colonizing the lower stems (crowns) of bread wheat (Triticum aestivum L.) and durum wheat (T. durum Desf.) from different wheat growing regions of Turkey in summer 2013. Samples were collected from 200 fields representing the major wheat cultivation areas in Turkey, and fungi were isolated from symptomatic crowns. The isolates were identified to species level by sequencing the translation elongation factor 1-alpha (TEF1-α) gene region using primers ef1 and ef2. A total of 339 isolates representing 17 Fusarium species were isolated. The isolates were identified as F. culmorum, F. pseudograminearum, F. graminearum, F. equiseti, F. acuminatum, F. brachygibbosum, F. hostae, F. redolens, F. avenaceum, F. oxysporum, F. torulosum, F. proliferatum, F. flocciferum, F. solani, F. incarnatum, F. tricinctum and F. reticulatum. Fusarium equiseti was the most commonly isolated species, accounting for 36% of the total Fusarium species isolated. Among the damaging species, F. culmorum was the predominant species being isolated from 13.6% of sites surveyed while F. pseudograminearum and F. graminearum were isolated only from 1% and 0.5% of surveyed sites, respectively. Six out of the 17 Fusarium species tested for pathogenicity caused crown rot with different levels of severity. Fusarium culmorum, F. pseudograminearum and F. graminearum caused severe crown rot disease on durum wheat. Fusarium avenaceum and F. hostae were weakly to moderately virulent. Fusarium redolens was weakly virulent. However, F. oxysporum, F. equiseti, F. solani, F. incarnatum, F. reticulatum, F. flocciferum, F. tricinctum, F. brachygibbosum, F. torulosum, F. acuminatum and F. proliferatum were non-pathogenic. The result of this study reveal the existence of a wide range of Fusarium species associated with crown rot of wheat in Turkey.     

Diversity of <Emphasis Type="Italic">Fusarium</Emphasis> species associated with root rot of sugar beet in China

Sugar beet is widely grown throughout the world and represents the second largest crop used to produce sugar. Root rot in sugar beet, caused by Fusarium, significantly reduces yield, juice purity, and sugar concentration. Here, 307 Fusarium isolates were collected from sugar beet roots exhibiting typical root rot symptoms in eight provinces or autonomous regions of China from 2009 to 2012. Based on morphological characteristics and sequence data of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) and the translation elongation factor 1α (EF-1α), Fusarium oxysporum (38.4%) was identified as the most prevalent species, followed by F. solani (20.9%), and F. equiseti (18.9%). These three species were widely distributed in all eight of the provinces and autonomous regions. F. tricinctum (5.9%), F. brachygibbosum (4.6%), F. redolens (3.3%), F. proliferatum (3.3%), F. graminearum (2.3%), F. verticillioides (1.6%), F. nygamai (0.7%), and F. culmorum (0.3%) were less frequently obtained. Of the 307 Fusarium isolates, 117 representing different species and geographic locations were demonstrated to cause tip rot and vascular discoloration in sugar beet roots, with disease incidence ranging from 84.2 to 100.0% and disease index ranging from 41.94 to 75.83. This is the first detailed report of Fusarium species, in particular F. tricinctum, F. brachygibbosum, F. redolens, F. proliferatum, F. nygamai, and F. culmorum, causing sugar beet root rot in China.     

A putative RNA silencing component protein FoQde-2 is involved in virulence of the tomato wilt fungus <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lycopersici</Emphasis>

RNA silencing pathways in filamentous fungi are composed of multiple component proteins and known to be involved in vegetative growth, virulence or sexual reproduction. We found that the tomato wilt fungus, Fusarium oxysporum f. sp. lycopersici (Fol), carries four homologues genes of Qde-2, an argonaute protein gene and one of the main component protein genes in Neurospora crassa. Gene targeting revealed that FoQde-2, one of the Qde-2 homologues in Fol, is involved in virulence to tomato but not in vegetative growth.     

Interaction of root colonizing biocontrol agents demonstrates the antagonistic effect against <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lycopersici</Emphasis> on tomato

Plant growth promoting Bacillus subtilis MSS9 and Bacillus licheniformis MSS14 were isolated from the tomato rhizosphere. These isolates were capable of inhibiting the fungal pathogen, Fusarium oxysporum f. sp. lycopersici causing fusarium wilt in tomato, tested by dual culture method and by mycolytic enzyme production. The isolates have the capacity to form biofilm on the microtitre plate. Scanning electron microscopy revealed good colonization capacity of Bacillus licheniformis MSS14 on tomato plant root as compared to Bacillus subtilis MSS9, pot experiments were also analyzed to study the effects of both rhizobacterial cultures on pathogen development and plant growth. It was observed that MSS14 reduces the incidence of Fusarium oxysporum f. sp. lycopersici in tomato and there was significant increase in vegetative parameters like root length, shoot length, plant wet weight, dry weight and chlorophyll content after which indicates that the root colonization property of the culture MSS14 helps in enhancing the biocontrol capacity against pathogen than that of MSS9.