Leaf spot diseases represent a major constraint to eggplant production under favourable environmental conditions. Therefore, potential pathogenicity of the fungus Cladosporium uredinicola on the leaves of eggplant (Solanum melongena L.), cultivar 'Barcelona F1', was confirmed in this study. Symptomatic leaf tissues were obtained from several greenhouse locations in Basrah governorate, southern Iraq. The first isolation of the fungus was done on potato dextrose agar (PDA) plates, followed by the investigation of morphological and microscopic features. Cladosporium uredinicola fungal growth on the PDA medium was olive-green to dark olivaceous-brown pigmentation and irregular margins, while the microscope conidiophores were light to medium olive colorin colour, smooth to finely roughened, bearing short terminal chains of 2–5 conidia. Pale olivaceous-brown, ellipsoid to oblong in shape, smooth-walled conidia were observed with 0–3 transverse septa. The internal transcribed spacer (ITS) region was targeted for confirming the identification; the ITS sequence was deposited in the National Center for Biotechnology Information (NCBI) GenBank database under accession number PV961371.1. Pathogenicity trials were followed under greenhouse conditions, revealing the pathogenic effect of C. uredinicola on eggplant leaves, showing necrotic lesion symptoms, which initially appeared after five days post-inoculation as small brown, circular to irregular margins, subsequently extended and coalesced into large spots and patches with chlorosis symptoms and ended with leaf abscission. The fungal pathogen was re-isolated from diseased leaves to confirm Koch's postulates. The current study represents the first report of C. uredinicola as a potential pathogen on eggplant leaves in Iraq.
In 2024 fall, symptoms of leaf blade and petiole spots were observed on strawberry plants 'Rociera FNM' and 'Marisma FNM'. In 450 randomly selected plants from 3 nurseries of Segovia province (Spain) 3.5% of incidence was recorded. Lesions on the leaf were reddish-brown with yellowish halos and became necrotic. Dark brown lesions with defined edges were noted on petioles. Affected tissue sections were excised, rinsed with 0.1% Tween 20, submerged in 70% ethanol (20 s), followed by 1% NaOCl (60 s), and then placed onto potato dextrose agar (Condalab, Madrid, Spain) with 50 mg/l of streptomycin sulphate. Plates were incubated at 28°C under darkness. From 6 crowns and 2 petioles 8 isolates (Di 3 to Di 10) were obtained from different pycnidia via single conidia isolation. Colonies had petal-shaped margins. Hyphae were initially hyaline, later developing an orange coloration in the center of the colony, and black conidiomata were visible two weeks post-incubation. Conidiomata were ellipsoidal to globose. Single-celled conidia were brown, ellipsoidal with slightly mucronate apices, narrowly truncated bases and smooth walls. This morphology was consistent with Coniella sp. as described by Álvarez et al. (2016). Isolates were stored in 50% glycerol solution at -80ºC. Genomic DNA was extracted from the 8 selected isolates. The internal transcribed spacer (ITS) region, translation elongation factor 1-alpha (TEF1), and large subunit ribosomal RNA (LSU) were amplified using the primer pairs ITS1/ITS4 (White et al. 1990), EF728/EF986 (Carbone and Kohn 1999), and LR0R/LR7 (Chethana et al. 2017), respectively. Sequence alignments performed in MEGA v11.0 (Tamura et al. 2021) showed that all isolates were identical across the three analyzed loci. Sequences of Di6 and Di10 isolates were deposited in GenBank. BLASTn analyses revealed that ITS (PX663172, PX663173), TEF1 (PX666006, PX666007), and LSU (PX663628, PX663629) sequences showed 100% identity with C. fragariae strain CBS 198.18 for ITS and 99% and 100% identity with strain CBS 167.84 for TEF1 and LSU, respectively. A concatenated phylogenetic tree was constructed with representative Coniella species and outgroup sequences from NCBI GenBank (Chethana et al., 2017) and confirmed the identity. Pathogenicity test was conducted using isolates Di6 and Di10. Inoculation with each isolate was performed by spraying 25 mL of a conidial suspension (5 x 104 conidia/mL) onto 5 plants of 'Rociera FNM'. Control plants were sprayed with sterile water. The assay was carried out twice. After inoculation, plants were covered with a plastic bag and kept in darkness for two days. Growth chamber was maintained at 25±2°C with 12-h photoperiod. Identical symptoms to the initially observed appeared on inoculated plants 6 days post-inoculation. Both isolates exhibited comparable severity (proportion of symptomatic leaf area) 20 days after inoculation. An assay was also performed on 'Marismas FNM', giving the same results. Fungal colonies re-isolated from the lesions had the same morphology and ITS, TEF-1 and LSU sequences of the inoculated isolates, thereby fulfilling Koch's postulates. In Switzerland and Great Britain C. fragariae has previously been isolated from strawberry plants (Rigotti et al. 2003; Jones and Baker, 2007). To our knowledge, this is the first report of C. fragariae on strawberry in Spain. Monitoring of the pathogen is essential since it can spread through asymptomatic plants into strawberry’s nurseries and fields.
Pepper (Capsicum annuum L.) is one of the most economically important cash crops in China. In July 2024, leaf spot was observed on pepper cv. ‘Sujiao 5’ in a 1-ha pepper plantation in Hangzhou (30°17′N, 118°52′E), Zhejiang Province. Among eight greenhouses surveyed, 30 plants per greenhouse were inspected. Disease incidence ranged from 10% to 20%. Early symptoms were round, grayish-white lesions. As the disease advanced, lesions expanded into circular spots with grayish-white centers and brown margins, and a grayish-brown mold layer emerged on the lesions. Thirty diseased leaves were randomly sampled from twenty infected plants for pathogen isolation. Under a stereomicroscope, a single conidium from the mold layer on diseased leaves was transferred onto potato dextrose agar (PDA) medium using a sterilized inoculating needle and incubated at 26°C for 7 days. Twenty single-conidium isolates were successfully obtained. After 15 days on PDA, colonies were grayish-green, velvety, with grayish-white margins. On synthetic nutrient-poor agar (SNA), colonies were light grayish-green with sparse mycelium. The conidiophores were solitary, erect, light brown, and septate. Ramoconidia were light brown, cylindrical to obovoid, aseptate, and measured 6.0-15.3 × 2.5-4.3 μm (n = 30). Conidia were light brown, obovoid to limoniform, forming branched chains with one to four conidia per branch, and measured 3.3-5.7 × 2.3-3.3 μm (n = 30). The morphological features were consistent with Cladosporium sp. (Bensch et al. 2015). All 20 single-conidium isolates shared identical colony morphology and conidial characteristics, so three representative isolates were randomly selected for sequencing. The primer pairs ITS1/4 (White et al., 1990), EF728/EF986 (Carbone and Kohn, 1999), and ACT512/ACT783 (Carbone and Kohn, 1999) were used to amplify the partial fragments of the internal transcribed spacers (ITS, GenBank: PZ437871-PZ437873), translation elongation factor 1-α gene (tef1, GenBank: PZ445478-PZ445480), and actin gene (act, GenBank: PZ445475-PZ445477). The sequences of the three isolates were identical at all three loci. BLASTn analysis showed 99% identity with Cladosporium sp. (ITS: OQ629129; tef1: HM148442; act: MT154165). The phylogeny suggests that three isolates were integrated into the Cladosporium tenuissimum clade (ML bootstrap support values/Bayesian posterior probabilities = 93/1). Based on morphology and phylogeny, the isolates were identified as C. tenuissimum. Since all isolates were morphologically and molecularly identical, isolate LJYB-LA1 was selected as a representative strain for pathogenicity testing. The isolate was cultured on PDA at 26°C in continuous darkness for 14 days. Conidia were rinsed with sterile water containing 0.1% Tween-80 and adjusted to 2 × 10
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conidia/mL using a hemocytometer. Two-month-old pepper plants cv. ‘Sujiao 5’ were each sprayed with 200 mL of conidial suspension; the negative control plants were sprayed with 200 mL of sterile water. The plants were incubated in a greenhouse at 26°C and 85% relative humidity for 48 h. The experiment was repeated three times with five plants per replicate. After 10 days, inoculated plants developed symptoms similar to those observed in the field, whereas control plants remained asymptomatic. The species C. tenuissimum was reisolated from symptomatic tissues and confirmed by morphology and molecular data, confirming Koch’s postulates. The species C. tenuissimum has been reported to infect Hydrangea paniculata in China (Li et al. 2021) and Phaseolus vulgaris in China (Gao et al. 2024). To our knowledge, this is the first report of C. tenuissimum causing leaf spot on C. annuum in China.
In the multilocus phylogenetic tree constructed using maximum likelihood (ML) analysis of concatenated ITS-TEF1-RPB2-ALT a1 sequences, isolate HF1 clustered with reference A. alternata strains with 95% bootstrap support, fulfilling Koch’s postulates.
Habofanoe Fosa, W. Swart, N. Muzhinji et al.· Plant Disease· 0 citations
To the authors' knowledge, this is the first report of P. ramorum occurring on C. siliqua globally and the first confirmation of Koch’s postulates for this host-pathogen combination.
Dinh-Hieu Pham, T. Pastalka, Huiying Liu et al.· Plant Disease· 0 citations
Based on morphology and multilocus analyses, the pathogen was identified as C. eragrostidis, and the obtained sequences were deposited in GenBank placed the three isolates within the C. eragrostidis clade.
Xiang Lu, Chumiao Chen, Yunzhi Bai et al.· Plant Disease· 0 citations
The findings provide a theoretical foundation for accurate disease diagnosis, epidemiological surveillance, and the development of effective chemical control strategies for rubber plantations.
Zhiying Cai, Lili He, Li-Ming Dai et al.· Plants· 0 citations
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