First Report of Alternaria Leaf Spot on Gerbera (Gerbera jamesonii H. Bolux ex J. D. Hook) in Bulgaria

Alternaria leaf spots of gerbera (Gerbera jamesonii H. Bolus ex J. D. Hook) were observed on plants from different greenhouses on commercial plants in Bulgaria. The symptoms of the disease on the leaves were characterized by the development of brown, small, scattered dots, which gradually enlarged and coalesced to form large, oval, circular or irregular, brown to black lesions with concentric rings. Affected plants showed lower vitality, suppressed development and fewer, smaller, distorted in shape flowers. Alternaria isolates, obtained from infected leaf tissues were grown in pure culture and the morphological characteristics of the colony and sporulation apparatus were determined. DNA, extracted from the fungal isolates was subjected to polymerase chain reaction (PCR) with primers ITS1/ITS4, amplifying the internal transcribed spacer (ITS) regions. The resulting products were sequenced and compared for homology with other species in the GeneBank. The isolates showed 94% homology of the ITS region with either Alternaria alternata, A. arborescens, A. tenuissima, A. longipes, A. lini or A. smyrnii. None of the studied isolates was amplified with the A. alternata specific primers AAF2/AAR3, indicating that they are pathogenic varieties of it or belong to another species. Pathogenicity tests on 10 gerbera cultivars revealed that all of them were susceptible to Alternaria leaf spot. Additional tests on nine other crops (Solanum lycopersicum, Calendula officinalis, Capsicum annuum, Celosia argantea, Pelargonium spp., Petunia hybrida, Nicotiana tabacum, Cucurbita moscata and Raphanus sativus var. radicina) and on tomato or pepper fruits, potato tubers and carrot roots also indicated that all tested plant species were potential hosts of the disease. This is the first report of highly virulent isolates of Alternaria spp. in Bulgaria that cause leaf spots on gerbera in greenhouses.     

Cucumis sativus and Ipomoea purpurea: New hosts for Alternaria hydrangeae in China

Alternaria leaf spot is a common disease on various plants worldwide. In this study, an Alternaria species, A. hydrangeae causing leaf spot on Cucumis sativus (cucumber) and Ipomoea purpurea in China was identified based on morphology and multi-locus analysis of the partial ITS, GAPDH, Alt a 1, TEF1, and RPB2 gene regions. The pathogenicity of the present isolates and a representative isolate of A. hydrangeae were assessed on living leaves of C. sativus, I. purpurea and Hydrangea paniculata. Similar symptoms were observed on every plant inoculated with the isolates. The same fungus was re-isolated from inoculated leaves fulfilling Koch's postulates. Morphology of the original and re-isolated isolates showed characteristics similar to A. hydrangeae with some variations. Multi-locus analysis indicated that the present isolate fell into A. hydrangeae clade. This study is the first report of A. hydrangeae as an agent of Alternaria leaf spot in C. sativus and I. purpurea in China, which extends the host range of the fungus.     

Incidence of Alternaria Species Associated with Watermelon Leaf Blight in Korea

Alternaria leaf blight is one of the most common diseases in watermelon worldwide. In Korea, however, the Alternaria species causing the watermelon leaf blight have not been investigated thoroughly. A total of 16 Alternaria isolates was recovered from diseased watermelon leaves with leaf blight symptoms, which were collected from 14 fields in Korea. Analysis of internal transcribed spacer (ITS) region, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and RNA polymerase II second largest subunit (RPB2) were not competent to differentiate the Alternaria isolates. On the contrary, analysis of amplicon size of the histone H3 (HIS3) gene successfully differentiated the isolates into three Alternaria subgroups, and further sequence analysis of them identified three Alternaria spp. Alternaria tenuissima, A. gaisen, and A. alternata. Representative Alternaria isolates from three species induced dark brown leaf spot lesions on detached watermelon leaves, indicating that A. tenuissima, A. gaisen, and A. alternata are all causal agents of Alternaria leaf blight. Our results indicate that the Alternaria species associated watermelon leaf blight in Korea is more complex than reported previously. This is the first report regarding the population structure of Alternaria species causing watermelon leaf blight in Korea.     

Isolation of 12 polymorphic microsatellite loci in the phytopathogenic fungus Alternaria brassicicola

Twelve polymorphic microsatellite markers were isolated from the phytopathogenic fungus Alternaria brassicicola, the causal agent of black spot of crucifers. An enrichment protocol was used to isolate microsatellite loci, which were then analysed in a collection of 46 isolates sampled from seven different countries. The number of alleles detected in 12 loci ranged from two to 10 (mean 3.5). Investigation of cross‐species amplifications showed that the designed primers were specific to A. brassicicola.     

Morphology,phylogeny and pathogenicity of Alternaria species,involved in leaf spot disease of sunflower in northern Iran

Leaf spot disease of sunflower is one of the most important foliar diseases on this crop worldwide. Several fungal groups are known to cause leaf spot disease on sunflower. Species of the genus Alternaria are the most common and serious leaf spot causing fungi on this crop. Leaf spot disease is the most destructive foliar diseases on sunflower in northern Iran; however, the identity of the causal agent remains unknown. The present study was aimed to characterise the identity of the causal agent of the disease by means of morphological and molecular data as well as to evaluate the pathogenicity of the responsible species. For this purpose, a total number of 97 fungal isolates were recovered from sunflower leaves with leaf spot disease symptoms from the sunflower fields in northwestern zone of Iran. All of the isolates were identified as Alternaria alternata based on cultural and morphological characteristics. A subset of isolates was subjected to phylogenetic analysis using sequence data from ITS-rDNA region, gpd and rpb2 genes. Sequence data from ITS-rDNA and gpd did not discriminate A. alternata from the other small-spored Alternaria species. A phylogeny inferred using sequence data from rpb2 gene clustered our isolates in several sub-clades within a single monophyletic clade. Sequence data for the type strain of the other small-spored Alternaria species has to be included in phylogenetic analysis, in order to make sure, whether the observed variations in rpb2 gene sequences are an indication for the population variation in sunflower isolates of A. alternata or define species boundaries among the small-spored Alternaria species. The results of pathogenicity assay on sunflower plants (cultivar Euroflor) under greenhouse condition revealed that A. alternata is pathogenic on sunflower.     

Alternaria alternata f.sp. cucurbitae, the cause of a new leaf spot disease of melon (Cucumis melo)

A leaf spot disease of melon caused by Alternaria alternata f.sp. cucurbitae was recorded for the first time in Crete. Necrotic flecks surrounded by chlorotic halos developed on the cotyledons and the leaves of the middle and the upper part of the plants; the flecks enlarged and coalesced to form lesions of 2 cm or more in diameter with brown fructifications of the pathogen on their surface. Severely affected cotyledons and leaves became chlorotic and died. Of 16 species from eight botanical families that were inoculated, only those of the Cucurbitaceae were susceptible. Of four isolates of A. alternata from tomato, sunflower, pear and cucumber, only the cucumber isolate was pathogenic to melon foliage.     

New species of Alternaria from Marathwada (India)

Summary The paper deals with two new species ofAlternaria, Alternaria nyctanthi onNyctanthes arbortristis Linn. andAlternaria tropaeoli onTropaeolum majus L.     

Aflatoxins in sunflower seeds: Influence of Alternaria alternata on aflatoxin production by Aspergillus parasiticus

The aim of the present work was to determine the influence of Alternaria alternata upon aflatoxin production by Aspergillus parasiticus.A mixture of spores of both strains was inoculated in sunflower seeds at 0,90 a_w, and incubated for 42 days at 28 °C ±1.The cultures were observed and analyzed every 7 days to determine the infection level of the seeds and the production of aflatoxins. Results showed that when the seeds were inoculated only with Aspergillus parasiticus, 100% were infected from the 7th day.When Aspergillus parasiticus and Alternaria alternata were simultaneously inoculated the infection level of the seeds was 100% for Aspergillus parasiticus following 7 days of inoculation and 0% for Alternaria alternata. After the 14th day of inoculation there was no significant difference in the infection percentage of both strains (approximately 80% of each one). As far as toxin production is concerned a remarkable decrease was observed when seeds were inoculated with both strains simultaneously.In accordance to the results, Alternaria alternata would not compete with Aspergillus parasiticus in colonization of seeds but would either degrade the aflatoxins by Aspergillus parasiticus or compete for aflatoxin biosynthesis precursors. Alternaria alternata could also secrete some substance that specifically inhibits aflatoxin synthesis.     

Prevalence of the Alternaria blight of cumin (Cuminum cyminum L.) in Bangladesh: Morphology,phylogeny and pathogenic variation of Alternaria spp.

Cumin (Cuminum cyminum L.) is an important spice crop worldwide and its production is hampered by the infection of Alternaria blight. Cultivation of cumin in Bangladesh is very limited due to the lack of appropriate germplasm and adequate scientific information regarding the prevalence of Alternaria blight. Field trials were conducted with four advanced lines of cumin viz. CN026, CN028, CN031 and CN038 in five agro-ecological zones (AEZ) to know the adaptation possibility of these lines against the incidence and severity of Alternaria blight of cumin in Bangladesh. Among all lines, CN026 was found as the best in germination capacity and other yield parameters in all locations. The incidence and severity of the disease was observed as high as 98% and 88%, respectively, however, out of the five locations, the incidence and severity of the disease was the lowest in Bogura for the line CN026. In an attempt to identify the causal organism of the Alternaria blight of cumin by using molecular tools, a total of twenty three isolates were collected from the plants showing Alternaria blight symptoms from different AEZ in Bangladesh. Based on the molecular analysis, the isolates were identified as Alternaria alternata, A. burnsii, A. gaisen and A. tenuissima. A. alternata was the most prevalent species followed by A. tenuissima. The isolates of the identified species were found to have genetic, morphological and pathogenic variation. An isolate of A. alternata was observed as the most virulent among the isolates. This is the first report of A. alternata causing Alternaria blight disease of cumin in Bangladesh. The findings of this experiment will help in selecting suitable cumin germplasm and designing proper management strategies against Alternaria blight of cumin in Bangladesh.     

Small-spored Alternaria spp. (section Alternaria) are common pathogens on wild tomato species

The wild relatives of modern tomato crops are native to South America. These plants occur in habitats as different as the Andes and the Atacama Desert and are, to some degree, all susceptible to fungal pathogens of the genus Alternaria. Alternaria is a large genus. On tomatoes, several species cause early blight, leaf spots and other diseases. We collected Alternaria-like infection lesions from the leaves of eight wild tomato species from Chile and Peru. Using molecular barcoding markers, we characterized the pathogens. The infection lesions were caused predominantly by small-spored species of Alternaria of the section Alternaria, like A. alternata, but also by Stemphylium spp., Alternaria spp. from the section Ulocladioides and other related species. Morphological observations and an infection assay confirmed this. Comparative genetic diversity analyses show a larger diversity in this wild system than in studies of cultivated Solanum species. As A. alternata has been reported to be an increasing problem in cultivated tomatoes, investigating the evolutionary potential of this pathogen is not only interesting to scientists studying wild plant pathosystems. It could also inform crop protection and breeding programs to be aware of potential epidemics caused by species still confined to South America.     

Characterization,pathogenicity, and fungicide sensitivity of Alternaria isolates associated with preharvest fruit drop in California citrus

Alternaria rot has been recently described as an emerging fungal disease of citrus causing significant damage in California groves. A survey was conducted to determine latent infections on fruits, twigs, and leaves and investigate their seasonal patterns during 2019 and 2020. On fruits, latent infections were more associated with the stem end than with the stylar end, except during spring when a significantly high percentage of flowers (86%) had latent infections. Latent infections on twigs varied markedly between years (28% in 2019 and 9.5% in 2020), while Alternaria spp. were also recovered from citrus leaves. Alternaria isolates collected during the survey were identified based on multigene sequence analysis, confirming that Alternaria alternata and Alternaria arborescens are the two species associated with infections of citrus fruits. Of the 23 isolates, 19 were identified as A. alternata and demonstrated the dominance of this species over A. arborescens. Isolates representing populations of these two species were selected as representative isolates for physiological and morphological studies. A. alternata and A. arborescens showed similar conidial dimensions but differed in the number of conidia produced. Growth rates demonstrated that A. alternata grows faster than A. arborescens at all the temperatures evaluated, except at 25 and 35 °C. The growth patterns were similar for both species. The sporulation rate of the Alternaria isolates was influenced differently by temperature. This parameter also influenced conidial germination and appressorium formation, and no significant differences were observed between Alternaria species. Pathogenicity and aggressiveness tests on detached fruit demonstrated the ability of A. alternata and A. arborescens to cause internal lesions and produce fruit drop in the orchards with no quantitative differences between them (disease severity indexes of 58 and 68%, respectively). The fungicide sensitivity tests showed that DMI fungicides are the most effective fungicides in reducing mycelial growth. The SDHI fungicides had intermediate activity against the mycelial growth but also suppressed spore germination. The spore germination assay suggested that some of the isolates included in this study might have some level of resistance to QoI and SDHI fungicides. The findings of this study provide new information about the pathogens associated with the excessive fruit drop recently observed in some California citrus groves.     

First report of Alternaria leaf spot caused by Alternaria alternata (Fries.) Kiessler on Sonchus oleraceus L. and Convolvulus arvensis L. in Algeria

Alternaria leaf spot caused by Alternaria alternata (Fries.) Kiessler was found on sow thistle (Sonchus oleraceus L.) and field bindweed (Convolvulus arvensis L.) in the experimental station of ENSA (Ecole Nationale Supérieure d'Agronomie) in Algiers, Algeria, during the winter of 2016. Necrotic spots in the form of concentric circles were observed on the leaves of both weeds with disease incidence of approximately 70% and disease severity ranging from 50% to 70%. Fungi were isolated from the infected leaves and identified as A. alternata, based on morphological and molecular analyses (using genetic marker internal transcribed spacer, ITS of rDNA). Pathogenicity tests confirmed that A. alternata is the causing agent of leaf spot disease of sow thistle and field bindweed in accordance with the original symptoms. To the best of our knowledge, this is the first report of sow thistle and field bindweed naturally infected by A. alternata in Algeria.     

Isolation and characterization of polymorphic microsatellite markers for Alternaria alternata

Alternaria alternata is of major significance as a food and feed contaminant and is able to produce a range of mycotoxins that may elicit adverse effects in both animals and humans. We describe the isolation and characterization of five microsatellite markers for studying A. alternata. Marker polymorphism was screened in 64 isolates of A. alternata. The number of alleles per locus ranged from eight to 24, and allelic diversity ranged from 0.425 to 0.882. These markers will be useful in the study of relationships and population genetics amongst isolates of A. alternata.     

An Allele-specific PCR Assay for Detecting Azoxystrobin-resistant Alternaria Isolates from Pistachio in California

Alternaria late blight caused by Alternaria spp. in the alternata, tenuissima and arborescens species‐groups is one of the most common fungal diseases of pistachio in California. A single point mutation resulting in the replacement of a glycine by an alanine at codon 143 (G143A) in the mitochondrial cytochrome b (cyt b) has been found in all azoxystrobin‐resistant isolates of these three species from California pistachio. In this study, a pair of allele‐specific polymerase chain reaction (PCR) primers was developed to detect this point mutation. The allele‐specific PCR assay coupled with a rapid DNA extraction method could detect azoxystrobin‐resistant Alternaria isolates in a few hours. The allele‐specific PCR method was also reliable for detecting azoxystrobin‐resistant Alternaria directly in both laboratory‐inoculated and naturally infected pistachio leaves.     

Phylogenetic and enzymatic variability of Alternaria species isolated from various substrates in Qena governorate of Upper Egypt

Recently, 24 sections were characterised in the genus of Alternaria. In this work, 27 isolates of Alternaria belonging to section Alternaria were isolated from different sources in Qena governorate, Egypt. The collected strains were identified using multi-locus products of internal transcribed spacer (ITS) region, small subunit (SSU), large subunit (LSU) and Alt a1 gene. Based on four loci, the phylogenetic analysis revealed that 26 isolates (96.3% of total isolates) identified as A. alternata and the last one isolate (3.7%) as A. arborescens. The different strains of Alternaria exhibited enzymatic variability ranged from 0.1 ± 0.07–2.3 ± 0.13U/ml for cellulase and 0.6 ± 0.20–3.7 ± 0.47 U/ml (pectinase). Within A. alternata isolates, biochemical properties (Cellulase and pectinase) did not correlate either to phylogenetic analysis or strain origin.     

Analysis of intraspecific and interspecific variation in the genus Alternaria by the use of RAPD-PCR

The RAPD-PCR profiles of 13 phytopathogenic Alternaria species and two closely related outgroups were examined using six different primers. Each species produced a distinct pattern of DNA fragments which were used as a measure of the degree of relatedness between species. A. brassicae isolates of diverse origin showed high levels of similarity but little similarity was noted between other species. The closest interspecific genetic distances were recorded between A. citri, A. alternata and A. longipes. The outgroup genera Embellisia and Stemphylium, which are recognised as distinct, could not be clearly separated using RAPD banding criteria, suggesting a high level of genetic diversity amongst these groups of fungi.     

Alt a 1 from Alternaria interacts with PR5 thaumatin-like proteins

Alt a 1 is a protein found in Alternaria alternata spores related to virulence and pathogenicity and considered to be responsible for chronic asthma in children. We found that spores of Alternaria inoculated on the outer surface of kiwifruits did not develop hyphae. Nevertheless, the expression of Alt a 1 gene was upregulated, and the protein was detected in the pulp where it co-localized with kiwi PR5. Pull-down assays demonstrated experimentally that the two proteins interact in such a way that Alt a 1 inhibits the enzymatic activity of PR5. These results are relevant not only for plant defense, but also for human health as patients with chronic asthma could suffer from an allergic reaction when they eat fruit contaminated with Alternaria.     

Tenuazonic Acid, a Toxin Produced by Alternaria alternata

Fifty-seven of 87 isolates of Alternaria alternata (Fr) Keissler grown on autoclaved, moist corn-rice substrate and fed to rats were lethal. The major toxin produced was isolated and characterized as tenuazonic acid. Twenty of 23 toxigenic Alternaria isolates examined produced tenuazonic acid. No tenuazonic acid could be detected in either of the field samples of sorghum or blackeyed peas, which were heavily invaded by Alternaria.