Colletotrichum-induced anthracnose is a major disease of avocado (Persea americana), causing significant losses in production worldwide. In 2023, a survey of avocado fruits in markets within the K & imath;z & imath;ltepe district of Mardin, T & uuml;rkiye, revealed symptoms of anthracnose, characterized by brown-black lesions and internal necrosis. To identify the causal agent, 20 symptomatic avocado fruits were collected and fungal isolates resembling Colletotrichum species were obtained. Morphological characterization, multilocus sequence analysis of the internal transcribed spacer (ITS), actin (act), the apn2-Mat1-2 intergenic spacer (ApMat), and calmodulin (cal) loci, and pathogenicity testing were performed for only two representative isolates (Pa01 and Pa02). These analyses identified the pathogen as Colletotrichum perseae. The isolates readily formed ascomata on all culture media tested, suggesting a high capacity for sexual reproduction. Pathogenicity tests confirmed that C. perseae caused characteristic dark, sunken anthracnose lesions on avocado fruits. This is the first report of C. perseae causing anthracnose in avocados in T & uuml;rkiye, expanding the known geographical distribution of this pathogen. The findings highlight the need for effective disease management strategies and further research to understand the pathogen's lifecycle, environmental conditions favoring infection, and potential for seedborne transmission.
Kiwifruit (Actinidia deliciosa) production in northeastern T & uuml;rkiye is facing an emerging threat from Armillaria root rot. Surveys conducted in August 2021 and 2022 revealed that 10.5%-17.5% of the 200-500 vines within each of 35 kiwifruit orchards in the Per & scedil;embe, G & uuml;lyal & imath; and Alt & imath;nordu districts of Ordu province exhibited symptoms of Armillaria root rot, including complete defoliation. Diseased plants exhibited diagnostic signs on their root collars and woody roots, including white mycelial fans, extensive wood rot and rhizomorphs. A single Armillaria isolate was obtained from each of the 35 symptomatic orchards. Molecular identification, employing DNA sequencing of the large subunit (LSU) region of rDNA, the translation elongation factor subunit 1-alpha (TEF1) gene and the second largest RNA polymerase II B-subunit (RPB2) gene, identified 20 isolates as Armillaria gallica and 15 as A. mellea. Phylogenetic analyses, based on TEF1 alignments, further supported the species identification with a 100% bootstrap value. To examine genetic diversity, start codon targeted (SCoT) marker 13 was used, successfully differentiating the two Armillaria species and revealing three distinct clades. Clades I and II comprised 15 and 5 isolates of A. gallica, respectively, while Clade III contained all 15 A. mellea isolates. Pathogenicity testing on kiwifruit seedlings (cv. Hayward) revealed the ability of both A. gallica and A. mellea isolates to induce disease. Rhizomorph-producing isolates in kiwifruit seedlings displayed high virulence, inducing severe leaf necrosis and seedling mortality, while isolates lacking these structures were non-pathogenic. This study identifies the first global instance of A. gallica causing kiwifruit root rot. This finding, along with the identification of A. mellea as a causal agent in the country, highlights the emerging threat of Armillaria root rot to kiwifruit production in the region.
The primary objective of this study was to characterize lactic acid bacteria (LAB) strains derived from sourdough for possible utilization as functional starters to produce sourdough and various cereal-based fermented foods. A total of 350 autochthonous LAB strains were isolated from 65 Type I sourdough samples and characterized using six random amplified polymorphic DNA (RAPD) primers at intra- and interspecific levels. Species identification of selected strains representing distinct clusters from RAPD analysis was performed based on the 16S rRNA region. The LAB strains were identified as Companilactobacillus crustorum (n = 135), Levilactobacillus brevis (n = 125), Latilactobacillus curvatus (n = 40), Companilactobacillus paralimentarius (n = 32), and Lactiplantibacillus plantarum (n = 18). A total of 66 LAB strains were selected for technological characterization along with two commercial strains. The characterization involved acidity development, EPS production potential, leavening activity, and growth abilities under harsh conditions. Principle component analysis (PCA) identified 2 Lp. plantarum and 14 Lev. brevis strains as the most relevant technologically. Among them, Lp. plantarum L35.1 and Lev. brevis L37.1 were resistant to tetracycline. Evaluation of probiotic characteristics (survival in pH 2.5 and bile presence, auto aggregation capacity, hydrophobic activity, antioxidant activity, antimicrobial activity) by PCA identified four strains with relevance to Lactobacillus rhamnosus GG (LGG), which were further selected for in vitro digestion assays. Lactiplantibacillus plantarum L7.8, Lev. brevis L55.1, and L62.2 demonstrated similar viability indices to LGG, along with increased auto aggregation capacity and antioxidant activity. These strains are promising as candidate starters for producing sourdough and sourdough-related fermented food products.
In the southeastern province of Mardin in Turkey, a severe and novel fungal infection affected all chickpea plant parts, resulting in blight symptoms on stem, petiole, branch, and leaf, defoliation, seed and root rot, and death. Neoscytalidium dimidiatum was identified as the agent responsible for this new blight and root rot using sequence analysis of the ITS, tef1, and tub2 loci as well as morphological data. The pathogen was found in all surveyed districts and fields, with varying incidences of blight and root rot, blight being nearly twice (40%) as common as root rot (21%), and root-rotted and blighted plants co-occurring in the majority of instances. All 92 N. dimidiatum isolates from various tissues induced necrotic lesions on the inoculated plants, indicating they were pathogenic for chickpea. Conidia exhibited germination across a temperature range, with increasing temperatures positively influencing germination rates, and mycelial growth was significantly influenced by temperature, with the optimal growth temperature observed to be 35 degrees C. The response of 25 Turkish chickpea cultivars and three other genotypes when subjected to inoculation with Ciar 12 and Ciar 78 isolates, representative of phylogenetic clusters, was evaluated based on the severity of blight and root rot. The majority of cultivars and genotypes displayed high susceptibility and suffered mortality when exposed to either soil or spray inoculation with each isolate. Among the assessed cultivars and genotypes, cagatay had the lowest severity of root rot. This study is the first to report a natural infection of chickpea plants by N. dimidiatum. Under global warming, this may be detrimental to chickpea cultivation and habitat in southeastern Turkey, which is the origin of chickpeas.
Fungal species associated with crown and root rot diseases in wheat have been extensively studied in many parts of the world. However, no reports on the relative importance and distribution of pathogens associated with wheat crown and root rot in Kyrgyzstan have been published. Hence, fungal species associated with wheat crown/root rot were surveyed in three main wheat production regions in northern Kyrgyzstan. Fungal species were isolated on 1/5 strength potato-dextrose agar amended with streptomycin (0.1 g/L) and chloramphenicol (0.05 g/L). A total of 598 fungal isolates from symptomatic tissues were identified using morphological features of the cultures and conidia, as well as sequence analysis of the nuclear ribosomal internal transcribed spacer (ITS) region, the translation elongation factor 1α (TEF1), and the RNA polymerase II beta subunit (RPB2) genes. The percentage of fields from which each fungus was isolated and their relative percentage isolation levels were determined. Bipolaris sorokiniana, the causal agent of common root rot, was the most prevalent pathogenic species isolated, being isolated from 86.67% of the fields surveyed at a frequency of isolation of 40.64%. Fusarium spp. accounted for 53.01% of all isolates and consisted of 12 different species. The most common Fusarium species identified was Fusarium acuminatum, which was isolated from 70% of the sites surveyed with an isolation frequency of 21.57%, followed by Fusarium culmorum, Fusarium nygamai, Fusarium oxysporum, and Fusarium equiseti, all of which had a field incidence of more than 23%. Inoculation tests with 44 isolates representing 17 species on the susceptible Triticum aestivum cv. Seri 82 revealed that Fusarium pseudograminearum and F. culmorum isolates were equally the most virulent pathogens. The widespread distribution of moderately virulent B. sorokiniana appears to be a serious threat to wheat culture, limiting yield and quality. With the exception of F. culmorum, the remaining Fusarium species did not pose a significant threat to wheat production in the surveyed areas because common species, such as F. acuminatum, F. nygamai, F. oxysporum, and F. equiseti, were non-pathogenic but infrequent species, such as Fusarium redolens, Fusarium algeriense, and F. pseudograminearum, were highly or moderately virulent. Curvularia inaequalis, which was found in three different fields, was mildly virulent. The remaining Fusarium species, Fusarium solani, Fusarium proliferatum, Fusarium burgessii, and Fusarium tricinctum, as well as Microdochium bolleyi, Microdochium nivale, and Macrophomina phaseolina, were non-pathogenic and considered to be secondary colonizers. The implications of these findings are discussed.
Kazakhstan is the fourteenth largest wheat producer in the world. Despite this fact, there has not been a comprehensive survey of wheat root and crown rot. A quantitative survey was conducted for the purpose of establishing the distribution of fungi associated with root and crown rot on wheat (Triticum spp.). During the 2019 growing season, samples were taken from the affected plants’ roots and stem bases. A total of 1221 fungal isolates were acquired from 65 sites across the central (Karagandy region), eastern (East Kazakhstan region), and southeastern (Almaty region) parts of the country and identified using morphological and molecular tools. The internal transcribed spacer (ITS), translation elongation factor 1-alpha (EF1-α), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sequences were successfully used to identify the species of fungal isolates. It was found that Bipolaris sorokiniana (44.80%) and Fusarium acuminatum (20.39%) were the most predominant fungal species isolated, which were present in 86.15 and 66.15% of the fields surveyed, respectively, followed by F. equiseti (10.16%), Curvularia spicifera (7.62%), F. culmorum (4.75%), F. oxysporum (4.10%), F. redolens (2.38%), Rhizoctonia solani AG2-1 (1.06%), Nigrospora oryzae (0.98%), C. inaequalis (0.90%), F. pseudograminearum (0.74%), F. flocciferum (0.74%), Macrophomina phaseolina (0.66%), F. cf. incarnatum (0.33%), Fusarium sp. (0.25%), and F. torulosum (0.16%). A total of 74 isolates representing 16 species were tested via inoculation tests on the susceptible Triticum aestivum cv. Seri 82 and the results revealed that F. culmorum and F. pseudograminearum, B. sorokiniana, Fusarium sp., R. solani, F. redolens, C. spicifera, C. inaequalis, and N. oryzae were virulent, whereas others were non-pathogenic. The findings of this investigation demonstrate the presence of a diverse spectrum of pathogenic fungal species relevant to wheat crown and root rot in Kazakhstan. To the best of our knowledge, this is the first report of F. pseudograminearum, Fusarium sp., C. spicifera, and C. inaequalis as pathogens on wheat in Kazakhstan.
In 2020, a canker disease with dieback of branches and decline of various fig trees in Şanlıurfa and Mardin provinces of Turkey was observed. The causal pathogen was identified as Neoscytalidium dimidiatum based on morphological characteristics and phylogenetic analysis of ITS, tef1, and tub2 loci. Koch’s postulates were confirmed by successful re-isolation of N. dimidiatum only from plants inoculated with the pathogen. This is the first report of N. dimidiatum associated with dieback and canker of common fig in Turkey.
Fusarium crown rot of wheat is an economically important disease that leads to significant yield and quality losses, especially in many arid and semi-arid wheat-growing areas worldwide. In June 2020, winter wheat (Triticum aestivum L.) plants exhibiting crown rot symptoms were identified in a commercial field located in the Tokbay location (43.033719°N, 74.325623°E), Chuy Province, Kyrgyzstan. The diseased plants were stunted and had brown discoloration on internodes of the stem bases and roots. Disease incidence was about 3%. A total of 10 plants were sampled at the ripening stage from the field to identify the causal agent. Symptomatic tissues were excised, surface disinfected with 1% NaOCl, rinsed three times with distilled water, and placed on one-fifth strength potato dextrose agar (PDA) followed by incubation at 23°C in the dark for 5 days. A total of 8 Fusarium isolates were recovered from tissues and purified by the hyphal tips method onto fresh PDA and Spezieller-Nährstoffarmer agar (SNA) plates (Leslie and Summerell 2006). Sequence analysis of the translation elongation factor 1α (TEF1) and the RNA polymerase II beta subunit (RPB2) genes were performed with primers EF1 and EF2 (O'Donnell et al. 1998), and 5f2 (Reeb et al. 2004) and 7cr (Liu et al. 1999), respectively. The sequences of three isolates showed 100% identities with the corresponding sequences of the strain NRRL 66652 of Fusarium algeriense Laraba & O'Donnell (TEF1: MF120515 and RPB2: MF120504), and the sequences of a representative isolate (KyrFa01) were deposited in GenBank (TEF1: OM135603 and RPB2: OM135604). On PDA, fungal colonies were initially yellowish-white but gradually turned yellowish-brown. Ellipsoidal microconidia produced in false heads on monophialides were usually aseptate (8.30 ± 1.17 μm, n = 50) and occasionally one-septate (21.89 ± 2.01 μm, n = 50). Sporodochial macroconidia were mostly 3-4 septate measuring 43.41 ± 2.83 μm (n = 50), slightly curved and formed generally on monophialides on SNA. No chlamydospores formation was detected after 15 days on SNA or PDA. Morphological characteristics described above were consistent with the morphology of F. algeriense, as reported by Laraba et al. (2017). To confirm pathogenicity, seeds of wheat cultivar Seri 82, Fusarium crown rot susceptible, were treated in 1% NaOCl for 2 min, rinsed twice, and placed in plates containing a piece of sterile filter paper saturated with water to induce germination for 3 days. Five pregerminated seeds were placed on the soil surface for each 9-cm-diameter pot, which was filled with a sterile potting mix containing peat, vermiculite, and soil (1:1:1 by v/v/v). A 1-cm-diameter mycelial plug taken from the margin of actively growing colonies (PDA) of the representative isolate KyrFa01 was contacted with each seed, and then seeds were covered with the same potting mix. The seeds in control pots were treated with sterile PDA plugs. The experiment was conducted in a growth chamber in a completely randomized design with five replicated pots at 23°C with a 12-h photoperiod. Disease assessment was made after 4 weeks of fungal inoculation. The isolate KyrFa01 induced discoloration on the crown and root tissues of inoculated plants similar to those observed in the field-grown plants, whereas no symptoms were observed on plants grown in the control pots. The pathogen was successfully reisolated from the symptomatic tissues, confirming Koch's postulates. To the best of our knowledge, this is the first report of crown rot caused by F. algeriense on wheat in Kyrgyzstan. Fusarium algeriense was firstly described within the Fusarium burgessii species complex by Laraba et al. (2017) as a crown rot pathogen of wheat in Algeria. The pathogen was secondly reported from wheat-growing areas in Azerbaijan (Özer et al. 2020a) and thirdly from Kyrgyzstan in this report. Özer et al. (2020b) confirmed the coexistence of this pathogen with other Fusarium species. The result warrants the need to further investigate the potential of this species in the Fusarium crown rot complex of wheat.
In August 2021, jimson weed (Datura stramonium) plants growing as weeds in potato fields in Bolu province, Turkey, exhibited leaf spots with dark concentric rings. Sunken and lens-shaped lesions with a light center were also frequently observed on petioles, branches, and stems. Based on morphological characteristics and phylogenetic analysis of the transcription elongation factor 1-α, RNA polymerase second largest subunit, and glyceraldehyde-3- phosphate dehydrogenase loci, the causal agent was identified as Alternaria crassa. The pathogen was successfully re-isolated from inoculated jimson weed plants in the pathogenicity assay, proving Koch’s postulates. Alternaria crassa caused necrotic lesions on potato plants, similar to those of early blight, confirming them as an alternative host of the pathogen. This is apparently the first report of leaf spot caused by A. crassa on jimson weed in Turkey.
Fusarium crown rot, caused by several species within the genus, is a major constraint that results in significant losses in wheat production worldwide. In June 2019, diseased wheat plants with typical symptoms of crown rot, including discoloration on the first two or three internodes of the stem just above the soil line and stunted, dry rotted, and discolored roots were collected in several bread wheat fields during the maturity stage in Almaty, East Kazakhstan, and Karaganda Regions of Kazakhstan. For each field, approximately twenty tillers were randomly sampled. Symptomatic tissues were surface sterilized in 1% NaClO for 2 min, rinsed with sterile distilled water three times, air-dried in a laminar flow hood, and then transferred to Petri dishes containing one-fifth strength potato dextrose agar (PDA). After incubating in the dark at 23°C for 5 days, 79 single-spore isolates showing cultural and microscopic characteristics of Fusarium were obtained on PDA and Spezieller-Nährstoffarmer agar (SNA). Colonies were initially white but later produced a beige to pink diffusible pigment in PDA. Microconidia that formed on aerial monophialides were hyaline, 0 to 1 septum, oval- to kidney-shaped, and measured 4.3 to 10.3 × 1.9 to 3.4 µm (average 7.8 × 2.6 µm), whilst macroconidia were straight to slightly curved, 3 to 5 septate, and measured 18.7 to 38.8 × 2.9 to 6.6 µm (average 29.9 × 4.7 µm), with foot-shaped basal cells on SNA. Chlamydospores were present on PDA. Sequence analysis based on portions of translation elongation factor 1α (TEF1) and the nuclear ribosomal internal transcribed spacer region (ITS rDNA) loci with primers EF1/EF2 (O'Donnell et al. 1998) and ITS1/ITS4 (White et al. 1990) identified 29 of the 79 isolates as Fusarium redolens Wollenw. The sequences of the five representative isolates with 99.85% of similarity to those of F. redolens strains available in GenBank e.g., ITS (MT435063) and TEF1 (GU250584). The TEF1 (accession nos. MW403914-MW403918) and ITS rDNA (accession nos. MW397138-MW397142) sequences of the isolates were deposited in GenBank. The morphological features are consistent with the described features of F. redolens (Leslie and Summerell 2006). To confirm pathogenicity of the five isolates, five pre-germinated seeds of wheat cultivar Seri 82 were placed in a 9-cm-diameter pot filled with a sterile potting mix containing equal volumes of peat, vermiculite, and soil. An approximately 1-cm-diameter 7-day-old mycelial plug of each isolate was individually placed in contact with the seeds. Seeds were covered with the same potting mix, and then the pots were maintained for four weeks in a growth chamber at 23°C with a 12-h photoperiod. The experiment was conducted twice with three replicate 15-cm pots with 5 plants per pot. Controls were inoculated with sterile agar plugs using the same procedure. After four weeks, all the inoculated plants showed stunted growth with brown discoloration in most parts of the crown and roots, whereas no symptoms were observed in the control plants. The mean severity of the disease for each isolate was between 2.1 and 2.7 according to the scale of 1 to 5 described by Gebremariam et al. (2015). The pathogen was reisolated from crowns of diseased plants, but not from asymptomatic control tissues, and identified morphologically based on the methods described above, fulfilling Koch's postulates. Although several morphological features are shared by F. oxysporum and F. redolens, Baayen et al. (2001) showed that these species could be easily distinguished using molecular data. The pathogen was previously reported as F. redolens associated with crown rot of wheat in Turkey (Gebremariam et al. 2015) and Saskatchewan, Canada (Taheri et al. 2011). The presence of F. redolens causing crown rot is confirmed in the six wheat fields surveyed in Kazakhstan, for the first time. This pathogen may pose a risk for wheat production, and further studies needed to determine the impact on the crop in Kazakhstan.
In June 2020, many plants exhibited symptoms of root rot and foliar blight in the experimental field of common sage in Şanlıurfa province, Turkey. The pathogen was identified as Neoscytalidium novaehollandiae based on morphological characteristics and phylogenetic analysis of partial sequence of the transcription elongation factor 1-α gene and the internal transcribed spacer of rDNA. Koch’s postulates were fulfilled by successful re-isolation of the pathogen from inoculated plants in the pathogenicity assay. To our best knowledge, this is the first report of N. novaehollandiae causing root rot and foliar blight of common sage worldwide.