Strawberry Anthracnose Research Articles

CHARACTERIZATION OF COLLETOTRICHUM ACUTATUM, THE CAUSAL SPECIES OF ANTHRACNOSE OF STRAWBERRY AND OTHER HOSTS IN BULGARIA

Strawberry anthracnose was first reported in Bulgaria in 2002. Since then, the disease has reached epidemic levels in production fields. More than one-hundred Colletotrichum cultures were isolated from different strawberry cultivars, pepper and tomato fruits, and the weed species Cirsium arveuse that were cultivated in close proximity with infected strawberry field crops. The Colletotrichum isolates were identified and characterized by classical morphological criteria and by various molecular methods. Morphological characterization identified all the tested isolates as C. acutatum, which was further confirmed by species-specific polymerase chain reaction (PCR) amplification using the universal ITS4 and CaInt2 primers. No PCR products were amplified using C. gloeosporoides specific primers. Arbitrarily-primed PCR determined that all the studied isolates were uniform, regardless of origin of plant species, indicating that they belong to an asexually reproducing population. Sequence analysis of the complete internal transcribed spacer (ITS) region confirmed that the populations belonged to the species C. acutatum. Pathogenicity tests using representative isolates caused cross infections on the respective alternative hosts. These findings emphasize that various crops including weeds may serve as a potential inoculum source for C. acutatum.

Identification of the Fungal Pathogen that Causes Strawberry Anthracnose in Bangladesh and Evaluation of In Vitro Fungicide Activity

This study was conducted to identify the Colletotrichum species causing anthracnose disease of strawberry in Balgladesh and to evaluate in vitro activity of commercial fungicides it. Based on morphological and cultural characteristics, all 22 isolates were identified as Colletotrichum gloeosporioides. They developed white or glittery colonies with grey to dark grey reverse colony colors and they produced cylindrical conidia. The efficacy of five commercial fungicides, Bavistin DF, Dithane M-45, Sulcox 50 WP, Corzim 50 WP and Rovral 50 WP, were tested against the fungus. Bavistin inhibited radial growth completely and was followed in efficacy by Dithane M-45. In Bavistin DF treated media, the fungus did not produce conidia. The percent inhibition of radial growth of the fungus was increased with the increasing concentrations of fungicide.

Development of protocols for detection of Colletotrichum acutatum and monitoring of strawberry anthracnose using real‐time PCR

Real‐time PCR (TaqMan®) assays were developed for the specific detection and discrimination of Colletotrichum spp., C. acutatum and C. gloeosporioides causing anthracnose in strawberry using the most divergent area of the internal transcribed spacers (ITS1 and ITS2) and 5·8S ribosomal RNA (rRNA) gene region. The specificity of the new assays was tested using DNA from six species of Colletotrichum and nine fungal species commonly found associated with strawberry material, and additionally by comparing the sequences with those from databases using a blast search. The sequences only showed identity with homologous sequences from the desired target organisms. The new assays were 10–100 times more sensitive than conventional PCR methods previously published for the diagnosis of strawberry anthracnose. When real‐time PCR was compared with ELISA methods, PCR improved the sensitivity of the identification by obtaining positive results for samples of strawberry plant material that tested negative with ELISA. The development of C. acutatum was monitored using artificially infected strawberry crowns from two strawberry cultivars (Camarosa and Ventana) and a real‐time PCR assay specific for this species between January and June 2006. The amount of C. acutatum detected using real‐time PCR varied significantly by month (P < 0·001), but not by cultivar (P = 0·394). The new assays were shown to be useful tools for rapid detection and identification of these pathogens and to allow rapid and accurate assessment of the casual agents of anthracnose in strawberry.

Nitrogen Fertilizer Affects the Severity of Anthracnose Crown Rot Disease of Greenhouse Grown Strawberries

The influence of nitrogen, phosphorus, and potassium on the severity of anthracnose crown rot was evaluated in three greenhouse studies. Strawberry plants were fertilized three times weekly with a modified Hoagland's Nutrient Solution containing the treatments and inoculated eight weeks after treatment applications began with a conidial suspension of the causal pathogen, Colletotrichum fragariae. Disease severity was rated 30 days later on a scale of 0 (no symptoms) to 6 (plant dead). In the first study, the effect of N, P, and K levels was evaluated using 16 treatments: eight N levels with either low P and K or high P and K. Disease severity ratings and percent foliar N increased as N level increased but were not influenced by the level of P and K. In two other experiments, seven N sources were each evaluated at three levels. Plants receiving 160 ppm N had higher disease severity ratings than plants receiving 0 or 40 ppm N. Among plants receiving 160 ppm N, those treated with Ca(NO3)2 had the least disease. When N fertilizer is applied to strawberry plants as Ca(NO3)2, anthracnose crown rot severity should be less severe than when N is applied in ammonium forms. Accepted for publication 13 April 2009. Published 9 June 2009.

Antagonism of Bacillus spp. Isolated from Marine Biofilms Against Terrestrial Phytopathogenic Fungi

We aimed at determining the antagonistic behavior of bacteria derived from marine biofilms against terrestrial phytopathogenic fungi. Some bacteria closely related to Bacillus mojavensis (three isolates) and Bacillus firmus (one isolate) displayed antagonistic activity against Colletotrichum gloeosporioides ATCC 42374, selected as first screen organism. The four isolates were further quantitatively tested against C. gloeosporioides, Colletotrichum fragariae, and Fusarium oxysporum on two culture media, potato dextrose agar (PDA) and a marine medium-based agar [yeast extract agar (YEA)] at different times of growth of the antagonists (early, co-inoculation with the pathogen and late). Overall antagonistic assays showed differential susceptibility among the pathogens as a function of the type of culture media and time of colonization (P < 0.05). In general, higher suppressive activities were recorded for assays performed on YEA than on PDA; and also when the antagonists were allowed to grow 24 h earlier than the pathogen. F. oxysporum was the most resistant fungus while the most sensitive was C. gloeosporioides ATCC 42374. Significant differences in antagonistic activity (P < 0.05) were found between the different isolates. In general, Bacillus sp. MC3B-22 displayed a greater antagonistic effect than the commercial biocontrol strain Bacillus subtilis G03 (Kodiak). Further incubation studies and scanning electronic microscopy revealed that Bacillus sp. MC3B-22 was able to colonize, multiply, and inhibit C. gloeosporioides ATCC 42374 when tested in a mango leaf assay, showing its potential for fungal biocontrol. Additional studies are required to definitively identify the active isolates and to determine their mode of antifungal action, safety, and biocompatibility.

Colletotrichum fragariae Is a Pathogen on Hosts Other Than Strawberry.

Evidence that Colletotrichum fragariae causes disease on hosts other than strawberry is limited. In the fall of 2006, fungal isolates from silver date palm with leaf spot symptoms and from cyclamen with leaf spot and stem rot symptoms were identified as C. fragariae. After confirming the pathogenicity of the isolates on their host of origin, a representative isolate from each host was compared to C. fragariae and to C. gloeosporioides/Glomerella cingulata isolates from strawberry. Date palm and cyclamen isolates bore conidia on setae, and conidia were tapered and in the size range of C. fragariae reference isolates. Sequence data from the combined internal transcribed spacer (ITS) regions 1 and 2 and the gene for the 5.8 ribosomal RNA from the cyclamen and date palm isolates matched the sequence for C. fragariae reference isolates. Based on these characteristics, it was concluded that the C. fragariae species designation was correct for both isolates. However, the date palm isolate was a weak pathogen on strawberry compared with other isolates and had a distinct AT-rich DNA banding pattern. The ability of the cyclamen isolate to cause crown rot on strawberry was comparable with the strawberry reference isolates, and the AT-rich DNA banding pattern of the cyclamen isolate was identical to the C. fragariae isolates from strawberry. The results indicate that C. fragariae is a pathogen on hosts other than strawberry and that there is more diversity among C. fragariae isolates than previously reported.

Management of strawberry anthracnose by choice of irrigation system, mulching material and host resistance

We studied, by means of field experiments, the combined effects of irrigation system, mulching material and genetic resistance on the incidence of strawberry anthracnose caused by Colletotrichum acutatum. All trials involved artificial inoculation with fruits colonized by the pathogen. One experiment examined the effects of drip irrigation, overhead sprinkler irrigation, and localized low-pressure tape irrigation, and the effects of grass (Brachiaria sp.) and plastic (polyethylene) mulches, in a split plot, randomized complete block design with four replicates. Lower disease incidence, indicating lower pathogen dispersal, was verified with low-pressure tape and drip irrigation. Flower blight and fruit rot incidence in drip-irrigated plots was minimal (ca. 2% of the disease levels in the plots otherwise irrigated). Grass mulch reduced flower blight in the sprinkler system (up to 80%), but had no significant effect when disease levels were already low due to use of other irrigation systems. A second experiment compared the effects of grass mulch, pine (Pinus elliotti) mulch, and plastic mulch with overhead sprinkler irrigation on strawberry cultivars partially resistant (‘Dover’) and susceptible (‘Campinas’) to the disease in a factorial completely randomized design (CRD). Both organic mulches reduced flower blight and diseased fruit incidence by up to 76% compared to the plastic mulch treatment, especially in the early stages of the epidemics. ‘Dover’ had 70% lower incidence of flower blight than ‘Campinas’, and the effects of organic mulches were more pronounced in ‘Campinas’. A third experiment, with all three irrigation systems combined with inoculated and non-inoculated plots, estimated yield effects due to disease and irrigation system. All plots were covered with plastic mulch in a CRD with six treatments. This experiment clearly confirmed the higher flower blight and fruit rot incidences when sprinkler irrigation was used. In inoculated treatments, fruit yield was significantly reduced in tape-irrigated plots, and more so in sprinkler-irrigated plots. In drip-irrigated plots yield was reduced by only 1%. Overall, our results confirm the importance of cultural and genetic factors as valuable means of strawberry anthracnose management. Adoption of localized irrigation systems, the use of organic mulches and choice of a partially resistant cultivar significantly reduced disease levels in field plots. The importance of water splash for C. acutatum dispersal and its dependency on the soil mulch characteristics are the likely causes for the results observed.

First Report of Anthracnose Fruit Rot Caused by Colletotrichum acutatum on Strawberry in Korea

Symptoms typical of anthracnose fruit rot; sunken, dark brown lesions on maturing fruits, were found in a commercial field of strawberry (Fragaria × ananassa) cv. Cal Giant in Yangyang County, Korea in May 2007. Masses of conidia were produced in acervuli in the center of lesions. The fungus was isolated on potato dextrose agar (PDA). Colonies grown on PDA were pale to mouse gray and became dark green to black in reverse. Conidia were formed in orange-to-salmon pink masses in the center of the culture. The average size of conidia on PDA was 15.2 × 4.6 μm, and they were hyaline, straight, cylindrical, with pointed ends, and aseptate (1). The fungus did not form an ascigerous stage in culture. Mycelial growth rate was 7.5 mm per day at 25°C on PDA. The identity of two isolates was confirmed as Colletotrichum acutatum J.H. Simmonds by PCR amplification using species-specific primers TBCA and TB5 (2), resulting in a characteristic 330-bp band on agarose gel. Morphological characters were in accordance with previous reports on C. acutatum. A pathogenicity test was conducted with five healthy plants of cvs. Cal Giant, Maehyang, Seolhyang, Kumhyang, Akihime, and Redpearl. After fruits and flowers were sprayed with a conidia suspension (105 conidia per ml), the plants were maintained at 10 to 25°C and 100% relative humidity in a greenhouse. As a control, five healthy plants were sprayed with sterile distilled water and incubated under the same conditions. Dark brown, water-soaked spots appeared on mature fruits of all cultivars after 5 days, and lesions on green fruits appeared on individual achenes. Flowers developed dark lesions, dried out, and died. No symptoms were found on the control plants. After the pathogen was reisolated from fruits and flowers lesions, the morphological characters developed in culture as described above. To our knowledge, this is the first report of C. acutatum causing strawberry anthracnose in Korea.

Chemical Composition and Antifungal Activity of Angelica sinensis Essential Oil Against three Colletotrichum Species

Chemical fungicides are an important component in disease management for most crops. As part of a program to discover natural product-based fungicides, several sensitive assay systems have been developed for the evaluation of naturally occurring antifungal agents. In this study, we focused on the discovery and evaluation of new natural product-based fungicides from Angelica sinensis (Dong Quai) for use in control of strawberry anthracnose caused by Colletotrichum species. A. sinensis oil was evaluated for antifungal activity against three Colletotrichum species using direct-bioautography assay and a strawberry detached leaf assay using C. fragariae. Chemical composition and fingerprinting were conducted by gas chromatography and gas chromatography-mass spectrometry. Major components were identified as ( Z) -ligustilide (43.1%), ( E)-3-butylidene phthalide (14.5%), ( Z)-β-ocimene (12.9%), and apiole (11.2%).

Isolation and Identification of Antifungal Fatty Acids from the Basidiomycete Gomphus floccosus

Bioautography of extracts of the fruiting bodies of the basidiomycete Gomphus floccosus (Schw.) Singer indicated the presence of fungitoxic compounds in the ethyl acetate fraction against the plant pathogens Colletotrichum fragariae, Colletotrichum gloeosporioides, and Colletotrichum acutatum. Bioassay-guided fractionation of this fraction resulted in the isolation of the bioactive fatty acids (9 S,10 E,12 Z)-9-hydroxy-10,12-octadecadienoic acid (1), (9 E,11 Z)-13-oxo-9,11-octadecadienoic acid (2), and (10 E,12 E)-9-oxo-10,12-octadecadienoic acid (3). These three oxylipins were further evaluated for activity against a greater range of fungal plant pathogens (C. fragariae, C. gloeosporioides, C. acutatum, Botrytis cinerea, Fusarium oxysporum, Phomopsis obscurans, and Phomopsis viticola) in in vitro dose-response studies. Phomopis species were the most sensitive fungi to these compounds. At 120 h of treatment, the IC50 values for compounds 1, 2, and 3 for P. obscurans were 1.0, 4.5, and 23 microM, respectively, as compared to 1.1 microM for the captan positive control.

Development and Optimization of a Real-Time Detection Assay for Xanthomonas fragariae in Strawberry Crown Tissue with Receiver Operating Characteristic Curve Analysis

Angular leaf spot of strawberry is caused by the bacterium Xanthomonas fragariae. The disease is transmitted primarily through systemically infected nursery stock. This creates problems for nurseries wishing to export plants to Europe because of quarantine restrictions. Currently, field inspections for symptoms are used to certify plants free of X. fragariae, but visual inspection is not useful for detecting plants infected systemically. To detect systemic infections, polymerase chain reaction (PCR) is the desired tool because of its sensitivity, specificity, and ease of use. In this study, we developed three sets of real-time PCR primers and probes and determined optimal reaction conditions for use of these primers for the detection of the bacterium X. fragariae in strawberry crown tissue. Real time detection proved to be both more sensitive and specific than standard PCR. Moreover, the detection of X. fragariae in crown tissue extract was possible with real-time PCR but not with standard PCR which is a significant improvement over standard PCR. The information on sensitivity and specificity of the primer sets was used to evaluate the performance of these primers with receiver operating characteristic (ROC) curve analysis under different tolerances. The results of this analysis can be used to provide guidance on threshold selection to manage disease below unacceptable levels. The results of this research may be useful to regulators and inspectors who must certify that plants meet European and Mediterranean Plant Protection Organization standards.

Epidemiology and Pathology of Strawberry Anthracnose: A North American Perspective

Three Colletotrichum species—Colletotrichum acutatum J.H. Simmonds (teleomorph Glomerella acutata J.C. Guerber &amp; J.C. Correll), Colletotrichum fragariae A.N. Brooks, and Colletotrichum gloeosporioides (Penz.) Penz. &amp; Sacc. in Penz. [teleomorph Glomerella cingulata (Stoneman) Spauld. &amp; H. Schrenk]—are major pathogens of strawberry (Fragaria ×ananassa). Strawberry anthracnose crown rot has been a destructive disease in commercial strawberry fields in the southeastern United States since the 1930s. The causal fungus, C. fragariae, may infect all aboveground plant parts; however, the disease is most severe when the fungus infects the crown, causing crown rot, wilt, and death. Colletotrichum gloeosporioides was responsible for an epidemic of anthracnose crown rot in strawberry nurseries in Arkansas and North Carolina in the late 1970s. The anthracnose fruit rot pathogen, C. acutatum, was first reported in 1986 on strawberry in the United States. Since the 1980s, increased losses due to anthracnose fruit and crown rots in the United States may be related to changes in cultivars and to widespread use of annual plasticulture production rather than the matted-row production system. Anthracnose investigations in the United States have concentrated on its epidemiology and differences among the three causal Colletotrichum spp. in their cultural, morphological, and molecular characteristics; their infection processes; and their pathogenicity. Results from these studies have resulted in a better understanding of the diseases and have led to better disease control. Strawberries grown in soils with high nitrogen levels are more susceptible to anthracnose than are those grown in soils with lower nitrogen levels or those amended with calcium nitrate. Anthracnose is spread more rapidly in fields that have overhead irrigation and plastic mulch than in fields where drip irrigation and straw mulch are used. Fungicide efficacy has been determined in in-vitro, greenhouse, and field studies, and pathogen resistance to some fungicides has been detected. Anthracnose-resistant cultivars are a major objective of most strawberry breeding programs in the southern United States.

Occurrence of Soilborne Diseases and Root Knot Nematodes in Strawberry Plants Grown on Compacted Rice Straw Bales Compared with Naturally Infested Soils

The present investigation deals with a possible use of rice straw bales as soilless cultiva- tion medium, thus escaping the problems inherent in the natural soils and avoiding a serious pollution when disposed about 5°C million tons of rice straw annually by burning. Strawberry fruits of good quality and quantity were harvested from plants cultivated on compacted rice straw bales in compari- son with the control plots under natural soil conditions. A higher temperature of 2 to 5 in comparison to natural soil conditions favours all physiological activities including the absorption of nutrients by roots and thus stimulating the vegetative and the generative growth of strawberry plants. The pH values around the roots in straw bales ranged from 5.5 to 6.5, while values obtained around the root system in natural soil ranged from 7.5 to 8.5. So, growing strawberry on rice straw bales helps avoid and overcome the problem of alkalinity and salinity in the rhizosphere. This is very important, as strawberries are very sensitive to salinity. Fruit rot diseases reached 0.8% on rice straw bales while on the control plots these were about 52% of fruits were infected with fungi. Cultivating strawberry on rice straw bales keeps the fruits away from contacting the soil and thus limits the possibility of injec- tion by soilborne fungi. The occurrence of damping-off, root rot, crown rot and root knot nematodes in strawberry plants grown on rice straw bales reached 4.0, 0.85, 0.35 and 0.0%, respectively. However, the corresponding figures for strawberry plants grown in natural soil under the same conditions were 27.0, 16.15, 11.70 and 13.20%, respectively, 135 days after planting. Isolation from strawberry plants grown in natural soil showing symptoms of crown rot and black root rot yielded several fungi identi- fied as Phytophthora cactorum, Colletotrichum fragariae, Pythium ultimum, Rhizoctonia solani and Fusarium oxysporum. The wilt symptoms observed on few strawberry plants on rice straw bales might be attrib-

Occurrence of fungicides resistant isolates of Glomerella cingulata causing the strawberry anthracnose in Nara Prefecture

Occurrence of fungicides resistant isolates of Glomerella cingulata causing the strawberry anthracnose in Nara Prefecture

Evaluation of Elite Native Strawberry Germplasm for Resistance to Anthracnose Crown Rot Disease Caused by Colletotrichum Species

Anthracnose crown rot of cultivated strawberry (Fragaria ×ananassa Duchesne ex Rozier) has been a major disease problem in the strawberry producing regions of the southeastern United States since the early 1970s. Chemical controls are often inadequate, but use of resistant cultivars is seen as a credible option for managing this disease. Only a small portion of Fragaria L. germplasm has been screened for resistance to anthracnose crown rot. A core subset of the Fragaria collection maintained at the U.S. Department of Agriculture National Clonal Repository in Corvallis, OR, has been constructed to contain an elite group of native F. virginiana Mill. and F. chiloensis (L.) Mill. This collection, referred to as the “core collection,” has been characterized for many horticultural traits, including reactions to several common foliar diseases, resistance to black root rot (causal organisms unknown), and resistance to northern root-knot nematode (Meloidogyne hapla Chitwood) and root-lesion nematode [Pratylenchus penetrans (Cobb) Filipjev &amp; Shuurmans Stekhoven]. Our objective was to evaluate the core collection for resistance to a selection of isolates of three Colletotrichum Corda species known to cause strawberry anthracnose, Colletotrichum fragariae A.N. Brooks, Colletotrichum gloeosporioides (Penz.) Penz. &amp; Sacc. in Penz. [teleomorph Glomerella cingulata (Stoneman) Spauld. &amp; H. Schrenk], and Colletotrichum acutatum J.H. Simmonds (teleomorph Glomerella acutata J.C. Guerber &amp; J.C. Correll). No Fragaria subspecies or geomorph was more resistant than any other; rather, individual genotypes within these groups were identified as sources from which resistance can be obtained. Collecting germplasm in areas of intense disease pressure may not be as beneficial as one might assume, at least where anthracnose crown rot disease is concerned.

Molecular Characterization and Diagnosis of QoI Resistance in Cucumber and Eggplant Fungal Pathogens

ABSTRACT The molecular mechanism of QoI fungicide resistance was studied using isolates of cucumber Corynespora leaf spot fungus (Corynespora cassiicola) and the eggplant leaf mold (Mycovellosiella nattrassii). In both pathogens, a mutation at position 143 from glycine to alanine (G143A) was detected in the cytochrome b gene that encodes for the fungicide-targeted protein. Moreover, the nucleotide sequence at amino acid position 143 was converted from GGT or GGA in sensitive (wild-type) to GCT or GCA in resistant (mutant-type) isolates. The methods of polymerase chain reaction restriction fragment length polymorphism commonly used for QoI resistance monitoring were employed successfully, leading to the amplified gene fragment from resistant isolates being cut with the restriction enzyme ItaI. However, heteroplasmy (the coexistence of wild-type and mutated alleles) was found when the resistant isolates of C. cassiicola, M. nattrassii, and Colletotrichum gloeosporioides (strawberry anthracnose fungus) were subcultured in the presence or absence of QoI fungicides. QoI resistance of cucumber powdery and downy mildew isolates persisted for a few years following the removal of the selection pressure imposed by the fungicide under both laboratory and commercial greenhouse conditions. The proportion of mutated sequences in cytochrome b gene decreased over time in the pathogen population. The protective efficacy of the full dose of azoxystrobin decreased when the populations of powdery and downy mildews contained resistant isolates at 10%. Using FMBIO, a fluorescence bio-imaging analyzer, the mutant allele from the QoI-resistant isolates could be detected at the level of 1%, whereas the detection sensitivity of ethidium-bromide-stained gels was approximately 10 times lower.

Isolation and pathogenicity of Colletotrichum spp. causing anthracnose of strawberry in south west Spain

Anthracnose, caused by Colletotrichum spp., is a major disease of cultivated strawberry, Fragaria × ananassa. This study identifies the Colletotrichum spp. which causes strawberry anthracnose in the southwest of Spain. A survey of the region was carried out, and the strains isolated were identified as C. acutatum by using the polymerase chain reaction (PCR) with genus and species-specific primers, demonstrating that this species is currently the causal agent of strawberry anthracnose in the studied region. The pathogenicity of C. acutatum and C. gloeosporioides strains was evaluated on two principal strawberry cultivars (cvs Camarosa and Ventana) under field conditions, the latter being more pathogenic than the former.

Antifungal Activity of Strawberry Fruit Volatile Compounds against Colletotrichum acutatum

Eight volatile products characterizing strawberry aroma, which is generated from the oxidative degradation of linoleic and linolenic acids by a lipoxygenase (LOX) pathway, were examined because of their antifungal activity against Colletotrichum acutatum, one of the causal agents of strawberry anthracnose. In this study, the effects of aldehydes, alcohols, and esters on mycelial growth and conidia development were evaluated. (E)-Hex-2-enal was found to be the best inhibitor of mycelial growth [MID (minimum inhibitory doses)=33.65 microL L(-1)] and of spore germination (MID=6.76 microL L(-1)), while hexyl acetate was the least effective of all volatile compounds tested (MID=6441.89 microL L(-1) for mycelial growth and MID=1351.35 microL L(-1) for spore germination). Furthermore, the antifungal activity of (E)-hex-2-enal on susceptibility of strawberry fruits to C. acutatum was also confirmed. The presence of these molecules in jars containing strawberry fruits inoculated with a suspension of spores inhibited the fungus growth and prevented the appearance of symptoms. Moreover, a study of the effects of (E)-hex-2-enal on conidial cells of C. acutatum was evaluated by transmission electron microscopy. This volatile compound altered the structures of the cell wall and plasma membrane, causing disorganization and lysis of organelles and, eventually, cell death.

Anthracnose: A new strawberry disease in Serbia and its control by fungicides

Anthracnose is a destructive disease of strawberry fruits in warm and continental climate. During 2004, in the vicinity of Valjevo, there were severe losses in two strawberry plantations due to fruit anthracnose. Two fungal isolates, GG-6A and GG-JUP were recovered from strawberry stolons and fruits showing severe anthracnose symptoms. Based on morphological and pathological characteristics, and PCR analyses with specific primers of reference species, isolate GG-6A was identified as Colletotrichum gloeosporioides, and GG-JUP isolate as C. acutatum. This is the first identification of C. acutatum in strawberry in Serbia. In order to control strawberry anthracnose, five fungicides and their combinations were applied four times during the flowering. The best fruit protection was achieved by fungicides Metiram + piraclostrobin (Cabrio top), Captan FL and Fludioksinil + ciprodinil (Swich). Less effective were Benomil (Benlate) and Krezoksim-metil (Stroby). Pathogen is transmitted by planting material, so phytosanitary measures are extremely important in preventing the disease.

Two subpopulations of Colletotrichum acutatum are responsible for anthracnose in strawberry and leatherleaf fern in Costa Rica

Strawberry, Fragaria × ananassa, and leatherleaf fern, Rumohra adiantiformis, are two important crops in Costa Rica. One of the most severe diseases affecting these crops is anthracnose, caused by members of the fungal genus, Colletotrichum (teleomorph; Glomerella). Eighty single-spore isolates from strawberry and leatherleaf fern were identified as Colletotrichum acutatum by species-specific PCR, and were further characterised by Universally Primed PCR (UP-PCR) fingerprinting analysis, and sequence analysis of the ribosomal internal transcribed spacer (ITS) region. Morphological differences, genotypic variation revealed by UP-PCR fingerprinting analysis, and a single sequence polymorphism within the ITS2 region were found between the isolates from strawberry and leatherleaf fern, respectively. The UPGMA cluster analysis of the fingerprints clearly separated the isolates derived from strawberry and leatherleaf fern into two different clusters. Pathogenicity assays on detached strawberry fruits confirmed the apparent difference between the two groups of isolates. It is therefore suggested that the pathogens responsible for strawberry anthracnose fruit rot and leatherleaf fern anthracnose in Costa Rica, belong to two distinct subpopulations of C. acutatum.