Moist Chamber Research Articles

First Report in China of Leaf Blight on Spatholobus suberectus Dunn Caused by Diaporthe tulliensis.

Spatholobus suberectus Dunn is an important Chinese medicinal plant, which is the only officially recognized authentic source of the Chinese herbal medicine Spatholobi caulis in the China Pharmacopeia (Qin et al. 2024). In November 2023, a serious leaf disease was observed in the S. suberectus planting base in Nanning city (108.22E, 22.51N), Guangxi, China, with an incidence rate of 80%. Initially the symptoms appeared in the diseased leaves tips and was irregular in shape. The necrosis displayed discoloration of the gray color with a deep brown margin and this gradually expanded through into the lamina. To isolate the causal agent, 10 symptomatic leaves were collected and small segments (5×5 mm) of infected tissues were cut, and then they were surface sterilized (30s in 75% EtOH, 2 min in 2.5% NaOCl, and rinsed three times in sterilized distilled water), placed on potato dextrose agar (PDA) and incubated at 28℃ for 5 days. The hyphal tips were transferred to PDA to obtain pure cultures. 36 isolates were obtained, 23 of them had similar morphology, which were isolated from all the symptomatic leaves and were considered pathogens, and named YY-3-S. After 7 d, the colonies were white, fluffy, with white aerial mycelium. Sporulation was induced using pine needle medium (Su et al. 2012). After 30 days, the globose pycnidia developed, which were superficial to semi-immersed on the pine needle medium, and they appeared to be orange to black in color, with pale yellow conidial droplets. The alpha conidia spores were 3.53 to 6.85×1.75 to 3.70 μm (n = 50). They were unicellular, hyaline, aseptate and ellipsoidal to fusiform. These morphological features were consistent with Diaporthe sp (Sun et al. 2021). For further molecular identification, isolate YY-3-S underwent sequencing of the internal transcribed spacer (ITS), beta-tubulin (TUB2), and translation elongation factor 1-alpha (TEF1) sections (Huang et al. 2021). Obtained sequences of ITS, TUB2 and TEF1 (Genebank accessions nos. PP838571, PP865050 and PP865051) displayed a 99% similarity to Diaporthe tulliensis (Genebank accessions nos. OP163562, KR936132, OR662168, respectively). Phylogenetic analysis was based on a combination of ITS, TUB2 and TEF1 sequence data using MEGA 11 software to construct a phylogenetic tree with Neighbor-Joining. In the phylogenetic tree, the combined sequences attributed YY-3-S to D. tulliensis. The pathogenicity test was conducted on 1-year-old S. suberectus seedlings. We sprayed the mixed suspension of spores and mycelia on the unwounded leaves, while control seedlings received sterile water. 3 leaves per plant and 5 plants per treatment were selected for assessment. All seedings were placed in pots in a moist chamber at 28°C and 90% humidity. After 5 d, browning was observed on all the inoculated leaves tips. 14 d after inoculation, necrotic lesions expanded outward and their symptoms like those in the field, while control plants remained healthy. D. tulliensis was reisolated from the inoculated leaves, fulfilling the Koch's postulates. D. tulliensis has been reported to necrotic spots and leaf blight on Rambutan (Serrato-Diaz et al. 2023), leaf spot on bodhi tree (Li et al. 2022), stem canker on Jasmin (Hsu et al. 2023). To our knowledge, this is the first report of D. tulliensis causing leaf blight on S. suberectus in China. It is important to ensure timely management of the newly emerging disease.

The Occurrence of Myxomycete Communities in Taxodium distichum Forests: Influence of Substrates and Seasons

Myxomycete communities in Taxodium distichum forests and the influence of substrates and seasons on myxomycete diversity were studied. In total, 80 substrates (40 living barks and 40 litters) were collected from the sample site in winter, spring, summer and autumn of 2023, followed by moist chamber cultures. A total of 733 records representing 34 myxomycete species in 21 genera were obtained. Arcyria cinerea and six other species were abundant (A). Six species were common (C), six species were occasional (O), and fifteen species were rare (R). Species diversity (H’ = 2.04 ± 0.20) of corticolous communities was significantly higher than that (H’ = 1.49 ± 0.31) of litter-inhabiting communities (p < 0.05). Nonmetric Multidimensional Scaling (NMDS) analyses showed significant differences between these two communities (p < 0.01). Species diversity was slightly higher in summer and autumn compared to winter and spring. NMDS analyses indicated no significant differences among seasonal communities. Redundancy Analysis showed that the corticolous species were closely related to wind speed (p < 0.05), while litter-inhabiting species were mainly determined by temperature (p < 0.05). The study revealed that substrates and seasons influenced myxomycete communities in T. distichum forests, with the communities being particularly affected by the substrates in which they live.

Community structure and assembly of myxomycetes in northern Chinese forests under geographic barriers

ABSTRACT The study of myxomycete biogeography has a long-standing history and has consistently drawn scholarly interest. Nevertheless, studies focusing specifically on the spatial and temporal distribution patterns of myxomycete diversity are relatively limited, with even fewer investigating the mechanisms driving the generation and maintenance of myxomycete diversity. Therefore, this study selected two geographically distant sampling sites within northern Chinese forests to investigate myxomycete species composition, community structure, environmental drivers, and assembly patterns under geographic barriers. We established plots in the Altai Mountains (ALE) and the Greater Khingan Mountains (GKM), gathered bark and litter, and conducted 80-day moist chamber cultures of myxomycetes. Additionally, myxomycete specimens were collected in the field simultaneously to supplement the data set. This study collected 541 myxomycete specimens belonging to 73 species from 28 genera, spanning 12 families and eight orders. The ALE and the GKM had 20 identical species, accounting for 27% of the total species. Myxomycetes from both regions exhibited abundant occurrence 18 days after cultivation, with the quantity on bark substrates notably higher than on litter. Arcyria pomiformis and Comatricha elegans were the most common species in moist chamber cultures. Mantel test outcomes revealed that environmental factors had no significant impact on myxomycete community similarity between the two areas, aligning with findings from the neutral community model analysis, indicating a predominant influence of stochastic processes on myxomycete community structure in moist chamber cultures. This study represents the first application of a quantitative framework to analyze myxomycete community assembly cultivated in moist chambers.

Impact of nursing interventions on the prevention of ocular surface disorders in critical care patients: A systematic review.

Intensive care unit (ICU) patients are at an increased risk of ocular surface injuries because of various factors such as reduced tear production and impaired protective mechanisms. Despite the significance of ocular care in ICU settings, there is a lack of consensus on effective interventions, leading to inadequate prevention of ocular surface disease (OSD). This systematic review aimed to assess the effectiveness of nursing eye care in preventing OSD in ICU patients. Secondary objectives included identifying primary risk factors for ocular injuries and examining the most effective preventive methods. A systematic review following PRISMA guidelines was conducted, encompassing a literature search, article selection, quality assessment and data synthesis. Studies meeting inclusion criteria were observational studies and clinical trials, focusing on adults admitted to ICUs under sedation and receiving mechanical ventilation. Of 3545 initially identified articles, 12 studies met inclusion criteria. These studies involved a total of 1853 participants. Various interventions were assessed, including saline rinsing, lubricating drops, gel lubricants, occlusion with polyethylene dressing, passive blinking and eyelid closure with tape. Moist chamber occlusion every 6 h combined with gel lubrication emerged as the most effective method in preventing OSD. Gel lubrication along with moist chamber occlusion proved to be the most effective strategy in preventing ocular injuries in ICU patients. Conversely, the routine use of physiological saline was associated with increased severity of corneal lesions. Properly defined protocols and well-trained nursing teams are crucial for reducing ocular injuries in ICU settings. The findings underscore the importance of implementing evidence-based eye care protocols in ICUs, emphasizing the use of gel lubrication and ocular surface protection to mitigate the risk of OSD. This highlights the need for comprehensive training programmes for ICU nursing staff to ensure optimal ocular care delivery.

First report of Colletotrichum queenslandicum associated with chilli anthracnose in Mauritius.

Globally, chilli (Capsicum annuum L.) is one of the most economically important and widely cultivated crop which elicits ethnomedicinal and nutritional potential as well as enhancing the taste and aroma of foods (Ayob et al., 2022; Kiran et al., 2020). Anthracnose disease is regarded as a prime constraint in chilli production, leading to enormous losses in tropical and subtropical countries. In September 2022, chilli fruit displaying sunken, shriveled and dark bown to black lesion with abundant acervuli on the surface was obtained from Flacq, Mauritius. From the symptomatic tissue, small pieces of the diseased tissue were excised, surface-disinfected using 1% sodium hypochlorite, twice rinsed using sterilized distilled water, air-dried and plated on PDA. After 7 days of incubation at room temperature, white to greyish white colony with dense white cottony aerial mycelium was recovered. Out of two isolates, CHF and CH10, the latter was considered for morphological and molecular characterization. The observed conidia (n=30) were unicellular, straight, cylindrical with rounded ends and slight constriction near the centre and had average length and width of 20.5 µm and 6 µm, respectively. For growth rate measurement of the isolate, two 5×5 mm of fungal agar plugs were taken from growing edge of colony, inoculated at centre of individual PDA plate and incubated at room temperature with a natural light/dark cycle. The diameter of the cultures were measured perpendicularly for a period of 7 days and the growth rate was calculated as 7-day average of daily growth (mm day-1). The growth rate of the fungal isolate (CH10) was 13.5 mm day-1 on PDA. Based on the morphological characters, the isolate was classified within the C. gloeosporioides species complex. For precise identification of the isolate, DNA was extracted from fungal mycelium using traditional DNA isolation methods (Ranghoo and Hyde, 2000), followed by PCR amplification and DNA sequencing using primer pairs ITS4/ITS5 (White et al., 1990), GDF/GDR and T1/Bt2b (Gan et al., 2016), respectively. ITS gene sequence (600 bp) confirmed that the isolate was Colletotrichum, with 99.83% similarity to KR704204 while GADPH (277 bp), TUB2 (733 bp) and ApMat (801 bp) gene sequences showed 99.64 to 100% similarity to C. queenslandicum with GenBank reference sequences, KT372374, KU221378 and MG674932 respectively. The gene sequences of isolate CH10 were deposited in GenBank database under the following accession numbers OR681557 (ITS), OR233734 (GADPH), OR475575 (TUB2) and PP622748 (ApMat). Koch's postulates were confirmed by spraying disease-free chilli plants with 10µL of conidial suspension (1 × 106 spores/ml) prepared from 7 days old colony of isolate CH10. Healthy chilli plants inoculated with sterile distilled water served as a negative control experiment. The plants were grown in pots in a moist chamber at 25˚C. After 5 days post-inoculation, anthracnose symptoms were developed on test plants while the control plant remained asymptomatic. The original isolate was successfully recovered from the test fruits, thus fulfilling Koch's postulates. The experiment was repeated thrice and revealed the same results. To the best of our knowledge, this is the first record of C. queenslandicum in Mauritius and is the first time to report anthracnose of chilli caused by this fungus. Colletotrichum queenslandicum has previously been reported in Europe, Mexico, US, Puerto Rico, Australia, Fiji, Brazil, Indonesia and China. Furthermore, the latter was associated with papaya, avocado, cashew, coffee, Persian lime, Licania tomentosa, white mangrove, lychee, mango, Nephelium lappaceum, olive, passionfruit, Dracaena cambodiana and Syzygium australe (Câmara and Vieira, 2022; Shidiq et al., 2024; Wang et al., 2022). This study will allow local farmers training and extension facilities to increase awareness among farmers about this disease-causing agent and allow them to take necessary measures for building up chilli crops resilience against this new and emerging pathogen in Mauritius.

First Report of Bacterial Leaf Spot Disease on Sesame (Sesamum indicum L.) Caused by Pseudomonas syringae pv. sesami in North Carolina.

In July 2022, dark brown to black, angular, water-soaked lesions were observed on sesame leaves (Sesamum indicum L.) in a research plot established to assess yield potential for eight varieties at the North Carolina (NC) Sandhills Research Station (Chavez 2023). Symptoms were indicative of a bacterial leaf spot (BLS). At early flowering stage, leaf spots were present on scattered plants; varieties ES108, SS3301, and ES201 exhibited up to 75% disease prevalence, with lower frequency in ES103, S39, S4302, S3251, and S3276. Symptomatic leaves from 3-4 plants were collected on four different dates from July through September. A section of symptomatic tissue was excised and macerated in sterile deionized water (SDW). A 10 µL aliquot was streaked onto SPA medium (15 g sucrose, 5.0 g proteose peptone, 0.50 g MgSO4 7H2O, 0.25 g K2HPO4, 15 g agar per liter of SDW) and incubated at 28ºC. After 72 h, numerous, smooth, white-cream colored, convex-shaped, colonies were individually isolated. Five randomly selected isolates from the different collection dates, designated as AHP108-AHP111 and AHP116, were genotyped. The 16S rRNA, gyrB, rpoD, and gapA genes were sequenced (Heuer et al. 1997; Hwang et al. 2005) and deposited to NCBI (GenBank Accessions: P213467- PP213470; OQ628040-OQ628042; PP214983-PP214994; and PP255798). These five isolates shared 100% sequence identity for gyrB and rpoD. AHP108-AHP111 shared 100% sequence identity for 16S rRNA and gapA, with 99.7% and 90.8% identity, respectively, for AHP116. A phylogenetic tree was inferred from a maximum-likelihood analysis of concatenated gyrB, rpoD, and gapA sequences of the five isolates and the top 11 hts from a blastn search of the NCBI nucleotide database. Those hits included closely related sequences from Pseudomonas syringae pv. sesami type strains ICMP 763T and ICMP 7459T. Based on this phylogenetic analysis AHP108-AHP111 and AHP116 are P. syringae pv. sesami. Recent genomic analysis suggests this pathovar is part of P. amygdali (Gomila et al. 2017), but an official name change has not been proposed. Each of the five isolates were infiltrated into leaves of sesame varieties ES108, ES103, and S327, consistently resulting in similar symptoms. Thus, strain AHP116, as a representative, was used to fulfill Koch's postulates using five, 30-day-old potted sesame plants (var. S3301). Plants were spray-inoculated with a bacterial suspension of ~108 CFU/ml until runoff; plants were incubated in moist chambers 24 h pre and post inoculation at 28ºC with 80% relative humidity and a 12 h photoperiod. At 13 days post inoculation, symptoms resembling those on plants at the Sandhills Research Stations in 2022 were evident. Reisolated bacteria were confirmed to be AHP116 through 16S rRNA and gyrB amplification and sequencing. No symptoms were observed on the five water-inoculated plants. BLS of sesame has been reported in Asia and is thought to be seedborne (Firdous et al. 2009; Prathuangwong and Yowabutra 1997). To our knowledge, this is the first report of P. syringae pv. sesami causing BLS on sesame in North Carolina. Sesame cultivation in the state increased from approximately 2,000 acres in 2022 to 13,000 acres in 2023 and there is interest in cultivating sesame as a rotational and alternative crop because it requires minimal input costs. Potential outbreaks of BLS in this warm, humid region could negatively affect sesame production, where little is known about the economic impact of the disease.

Myxobiota of the İskenderun Gulf (Mediterranean Sea/Türkiye) and its Environment

This study aims to determine the myxobiota of the İskenderun Gulf and its environment. This study was carried out on samples collected from 9 different stations in the İskenderun Gulf district in 2019-2022. Myxomycetes samples were collected from leaves, tree bark, and decayed and non-rotting plant materials. This aim is to detect both the myxobiota of this region and the halophytic myxomycetes species. The collected samples have developed myxomycete sporophores by using the moist chamber technique. In addition, myxomycetes which were developed in their natural environment were collected. As a result of the field and laboratory studies, 161 samples from different localities were collected and processed and 111 myxomycete samples were obtained from these samples. 41 species belonging to 6 ordo, 12 families, and 20 genera have been defined. 3 species were obtained only from the natural environment, 30 species were obtained only from moist chamber culture, and 8 species were obtained from both moist chamber culture and natural area. Arcyria cinerea, Didymium difforme, D. squamulosum, and Physarum pusillum were determined as abundant. While the number of species is rare in the seaside regions, it was observed that the further away from the sea, the number of species increased, especially in forest areas. A. cinerea, D. difforme, and D. dubium are the most common species on the beach and near the seaside. While cosmopolitan species were observed in abundance, Physarida members were found to be common in this results area. This study has contributed to the myxobiota of Türkiye.

First Report of Neofabraea actinidiae causing a Cranberry Fruit Rot in Oregon.

Cranberry (Vaccinium macrocarpon, L.) is a commercial small fruit that is native to North America. Oregon ranks fourth in cranberry production in the U.S.A. with 1052 Ha of cranberry beds and annual production of 23,590 metric tonnes (USDA NASS 2021). Cranberry fruit rot diseases are caused by a complex of 15 fungal pathogens belonging to 10 genera (Polashock et al. 2017). In fruit rot surveys of 'Stevens' cranberry beds in Coos and Curry Counties, Oregon, berries were collected before harvest and sorted by symptoms (e.g. softening, shriveling, or discoloration). Cranberries with field rot symptoms were surface-disinfested and lesion margin tissue placed on V8 agar. Cultures were incubated at room temperature and outgrowing fungi were transferred twice onto fresh V8 agar to obtain single isolates. Storage rots that developed on asymptomatic cranberries held at 4°C for 8 weeks were processed similarly. Since 2017, we periodically isolated Neofabraea actinidiae, which is not a member of the cranberry fruit rot complex, from rotted cranberries (Polashock et al. 2017). In 2022, N. actinidiae was isolated from 22% of 45 cranberries collected from an organically managed farm and developed storage rot and from 6% of 50 storage-rot cranberries from three conventionally managed farms. Symptoms associated with N. actinidiae on 'Stevens' cranberries often include softening and brown tissue surrounded by a yellow-colored ring. On V8 agar, N. actinidiae grew as compact white circular colonies with dense aerial hyphae near the center and accompanied by a red pigment in the agar. Pink-colored mucoidal irregular conidiomata often developed on the colony after 3 weeks. Conidia were hyaline, aseptate, and ellipsoidal to fusiform ranging from 7.5 to 12.6 µm long X 3.5 to 5.6 µm wide (n=100). Genomic DNA was extracted from N. actinidiae isolates from cranberries in 2017 and 2022. β-tubulin and the ITS 5/4 region were amplified and sequenced with primers Bt-T2m-Up/Bt-LEV-Lo1 and ITS5/ITS4 using conditions of de Jong et al. (2001) and White et al. (1990), respectively. Sequences were deposited in NCBI Genbank (Accessions: OR606303, OR606305, OR606306, & OR606309 for ITS; OR610314, OR610316, OR610317, & OR610320 for β-tubulin). BlastN analysis of β-tubulin (695 bp) and ITS (489-490 bp) had a 99.6 to 99.8% and 99.8 to 100% identity, respectively, to Neofabraea actinidiae (CBS 121403) (Accession numbers: KR859285 β-tubulin and KR859079 ITS). Phylogenetic analysis of concatenated sequences using Tamura-Nei neighbor-joining (Tamura et al. 2004) confirmed identity of cranberry isolates as N. actinidiae. Koch's Postulates were confirmed with four N. actinidiae isolates from cranberry. Agar or hyphal plugs were placed in a 3 mm wound on the side of six surface-disinfested, asymptomatic berries and incubated in a moist chamber at 20°C for 15 days or 4°C for 55 days. Similar symptoms developed on each berry inoculated with N. actinidiae, but not agar alone. The fungus was recovered from symptomatic tissues and identity confirmed by colony morphology. N. actinidiae causes a ripe rot and storage rot in kiwifruit and is one of the species causing Bull's Eye Rot of pome fruits (Tyson et al. 2019). Cryptosporiopsis actinidiae (anamorph) was isolated from cranberries roots in British Columbia, CA, but considered unlikely to be the causal agent of dieback disease of cranberry vines (Sabaratnam et al. 2009). We have demonstrated that N. actinidiae causes a cranberry fruit rot in beds and in storage. Its prevalence, associated fruit rot symptoms, and disease incidence will continue to be monitored.

First report of Colletotrichum incanum causing leaf spots on common bean in Europe (Slovenia).

A Phaseolus vulgaris L. leaf showing necrotic spots was collected in an experimental bean field in central Slovenia in August 2021. The field contained diverse common bean lines sourced from genebank collections, with each line represented by 10 plants. While symptomatic leaves were seen across various lines, the reported species derived exclusively from a Huasca Huallaga Colorado plant (single-seed descent, USDA accession PI153714, doi: 10.18730/H7P9N), a Peruvian landrace. After incubating the leaf for 2 d at ambient temperature in a moist chamber, setose acervuli developed producing curved, distally tapering and proximately truncated conidia. Single-spore cultures developed equally-shaped conidia measuring 14.5-21.5 (avg. 18.5) × 3-4 (avg. 3.5) μm (n=60) on corn meal agar when mounted in lactic acid. Obtained morphological characters and sequences of the partial actin (GenBank accession, OR208162), beta-tubulin (OR208164), and histone 3 (OR208165) gene identified the isolate as Colletotrichum incanum H.-C. Yang, J.S. Haudenshield & G.L. Hartman. Sequences were identical to those from CBS 133485 (= NRRL 62592, IL6A), ex-type strain of C. incanum (KC110823, KC110814, and KC110796). Partial sequences of the chitin synthase (CHS) gene (OR208163), not available for the ex-type strain, was identical to sequences of other C. incanum strains reported from China (KP145539, ON189040, and OQ613679-OQ613686) or differed in two nucleotide positions (OL471268 and OL471269). The strain from Slovenia was deposited in the CBS biobanks of the Westerdijk Fungal Biodiversity Institute (Utrecht, The Netherlands) as CBS 150848. Pathogenicity of the strain was tested by spraying ca. 3×105 conidia as a watery spore suspension onto each leaf of 6 greenhouse-grown and 3 wk-old common bean plantlets (cv. KIS Amand). Nonsterile commercial substrate (Potgrond H, AGRO-FertiCrop) was used and natural light conditions at ambient temperatures (18-23°C) applied. Sterile water was sprayed on 6, equally grown negative control plants. Treated plants showed small brownish spots after 3 wks similar to those described by Yang et al. (2014) on soybean. Setose acervuli formed within 5 days after detached leaves were incubated in moist chambers. No acervuli formed on negative control plants. Conidia re-isolated from these acervuli and obtained cultures were morphologically identical to originally obtained conidia and cultures and those used for performing the pathogenicity test. Anthracnose is an important disease of common bean attributed to various races of C. lindemuthianum (Sacc. & Magnus) Briosi & Cavara (Nunes et al. 2021). Reporting an additional agent potentially able to cause diseases in common bean and so far not known to occur in Europe is of high relevance as the various genetic bean lines used in Europe may show alternative susceptibility levels to it. However, symptoms caused by C. incanum seem to be less severe as those caused by C. lindemuthianum and the species belongs to the C. spaethianum species complex, whose members have so far not been considered as pathogens of economic importance (Talhinhas & Baroncelli 2021). Yang et al. (2014) based C. incanum on isolates from soybean petioles (USA) and associated it with common bean by re-identifying strain ATCC 64682 obtained by Tu (1990) in Canada. Database queries revealed that it was encountered also on sugar beet (USA; Hanson et al. 2023) and on various crop hosts in China (e.g., chili; Diao et al. 2017), but not in Europe. The work was funded by the Ministry of Agriculture, Forestry and Food and conducted as part of research programs P4-0072 and P4-0431, financed by the Slovenian Research and Innovation Agency ARIS, and the Horizon 2020 project INCREASE funded by the European Union.

Myxomycetes of Subantarctic Macquarie Island

An expedition to subantarctic Macquarie Island was carried out during the period of late January to early May of 1995 with the objective of documenting myxomycete biodiversity. Collecting on the island yielded 412 field collections, with another 14 collections were obtained with the use of the moist chamber culture technique. A total of twenty-six species representing 13 genera was recorded. This is the largest set of data yet available on the myxomycetes associated with any high-latitude area of the Southern Hemisphere. Prior to the expedition, only a single species was known from Macquarie Island. The purpose of this paper is to describe this expedition from the viewpoint of the author.

Evaluation of elite rice lines for resistance to Kenya blast fungus (Magnapothe Oryzae)

Blast resistance tends to often break down, these necessitate search-resistant genes. The screen house experiment was conducted in 2019 and 2020. A total of 56 rice genotypes, (Elite lines, monogenic lines, and local), were screened against ten Kenya isolates of Mangnaporthe oryzae. The establishment was by direct seeding of previously sprouted seeds. A completely Randomized Design (CRD) with two replicates was adopted. Inoculation was done 21 days after planting. Plants were maintained in the moist chamber (26–28 0C) for 48 hrs, then moved into an incubation chamber (25 ◦C ± 2). Disease assessment commenced 48 hours after inoculation until 21 days to full infection. Disease evaluations were performed according to the Standard Evaluation System of IRRI. Polymerase Chain Reactions (PCR) were carried out, blast resistant loci were identified using varied genetic markers that co-segregate with specific resistant loci Analysis was implemented in R version 3.3.2 (R Core Team, 2016). BLUPs were computed, in a mixed model to estimate the random effect of the genotypes. The data was subjected to analysis of variance and means separated by Tukey’s test at P < 0.05. Cluster analyses were performed in JMP software version 11.2 for Windows. Seven genotypes (IR12A311, IR10M210, IR74, 1R02A127, IR09A130, R66 and IR10N230) were more susceptible, IR13N152 and IR14F711 were more resistant to all Isolates. The monogenic lines IRBLsh-B and IRBLkh-K3 which carry blast-resistant genes, were susceptible to at least seven Kenya isolates. The local checks (Basmati 370, BAS 270 (pi9), BW196, and BAS370 (pi9) were highly susceptible. The genotype IR12F711 was consistently resistant to all Kenya Isolates.

An inexpensive moist chamber culture technique for finding microbiota on live tree bark.

Traditional moist chamber cultures (MCs) prepared in aseptic laboratory environments using sterile Petri dishes are commonly used to quantify the microbiota of rough-bark tree species and woody vines. MCs are typically expensive and may be difficult to make, so a less expensive option made from easily available supplies was developed. These cost-friendly MCs were compared with standard laboratory methods to demonstrate their efficacy. Modified MCs were made using inexpensive, store-bought supplies; compared to a standard laboratory setting, the modified MCs are shown to be less expensive with a faster setup time and larger size that facilitates a variety of tree and woody vine species. MC use resulted in the discovery of new species of fungi and myxomycetes with associated locality records. We provide detailed instructions for creating modified MCs, as well as a list of myxomycete species and their associated bark characteristics, pH values, and water-holding capacity. This new, low-cost MC technique makes the study of microbiota more inclusive and accessible for those in research laboratories, classrooms, and homes, including both amateurs and professionals. MCs are easy to prepare, versatile, and applicable for many areas of botany and the biological sciences, potentially allowing exploration into unexplored areas in urban ecosystems.

First Report of Fusarium proliferatum Causing Basal Rot and Dry Rot on Onion Bulbs (Allium cepa) in Spain.

Spain is the second largest onion-producing country in Europe with 1,465,430 tons and an export value of 584 million € in 2021 (MAPA 2022). In summer 2022 rot bulb symptoms were observed in five commercial fields and during the storage of cultivars 'Orlenda', 'Veleta', 'Mallory', 'Citation' and 'Pantano' from La Roda in Albacete (Castilla-La Mancha, Spain). Approximately 20% of sampled bulbs (113 bulbs analyzed) were affected with dry scales showing brown to dark brown rot on the top and basal plate of the onion bulbs. Occasionally, white to light pink fungal mycelium was observed between rotten scales and the plate basal. Sections of dry scales (5-10 mm) of the apical and basal plate were cut and placed on potato dextrose agar (PDA) and Komada medium (Komada 1975). From 5-day-old cultures typical white to light pink mycelium with microconidia in chains formed on polyphialides and macroconidia resembling Fusarium proliferatum (Nelson et al. 1983). To confirm the pathogen identity, partial translation elongation factor 1-alpha (TEF1) and RNA polymerase II subunit 2 (RPB2) genes were amplified and sequenced using primers reported in O´Donnell et al. (1998) and Samuels et al. (2002) for TEF1 and Liu et al. (1999) for RPB2. In BLAST analyses, the sequences showed 100% identity to the corresponding region of F. proliferatum (KP964908 and JF740801). Sequences were submitted to GenBank, and registered accession numbers are OR061014-16 for TEF1 and OR061017-19 for RPB2. Pathogenicity tests were conducted by inoculating healthy onion bulbs (five replicates per treatment) on the apical and basal plate by placing a 7-day old mycelial plug (10 mm diameter) from PDA cultures. Two onion cultivars ('Pandero' by Nunhems USA and 'Mallory' by Bejo The Netherlands) were inoculated separately with three isolates (PRO1, PRO9, PRO12). Control bulbs were inoculated with sterile PDA. The experiment was carried out twice. All bulbs were placed in a moist chamber and incubated at 25°C in the dark. After 15 days, bulbs inoculated with mycelial plugs showed similar symptoms to those of the original diseased bulbs. Browning dry rot was observed on the apical and basal plate of bulbs. When bulbs were cut longitudinally inner progressing rot was observed. Control bulbs remained symptomless. In both experiments, F. proliferatum was successfully re-isolated and morphologically confirmed from symptomatic bulbs to fulfill Koch's postulates. These results confirmed that isolates PRO1, PRO9 and PRO12 were the pathogen causing basal and dry rot on onion bulbs. This pathogen has recently been identified in China on Allium cepa L. var. agrogatum (Liu et al. 2022) and Idaho on onion (Beck et al. 2020) and could become a serious threat to onion production in Spain, reducing the quality and yield of onion.

Spread of dangerous walnut diseases in plantations of different ages in the Western Forest Steppe of Ukraine

Goal. To investigate the development of the phytopathogenic complex of walnut in the Western Forest Steppe of Ukraine.
 Methods. The research was carried out during 2021—2022 in the agrocenosis of the walnut garden of the Ukrainian Research Plant Quarantine Station of the Institute of Plant Protection of the National Academy of Sciences. For a detailed account of the disease, twenty trees were examined from four sides of the crown and 25 accounting organs (leaves, shoots, fruits) were analyzed and the intensity of their damage was determined according to the scale. Diagnosis of diseases was carried out by selecting affected tissues and placing them in a moist chamber for three days for subsequent identification of the type of pathogen by microscopy and determination of systematically important morphological features of the fungus. Isolation of fungi from living plants was carried out by transferring mycelia or spores from their surface to a new environment.
 Results. A laboratory study of plant material with various symptoms of damage showed the dominance of phytopathogenic fungi: brown spotting Marssonina juglandis (Lieh.) Magn., white spotting Microstroma juglandis Sacc. and bacteriosis Xanthomonas juglandis Sacc. When surveying old tree plantations (more than 40 years old), the frequency of brown spotting of walnut Marssonina juglandis (Lieh.) Magn. accounted for 68.7%, bacterial burn (Xanthomonas juglandis Sacc.) — 31.3%. In stands of 6-year-old trees, the frequency of brown spotting Marssonina juglandis (Lieh.) Magn. was 72.5% and bacterial burn Xanthomonas juglandis Sacc. — 27.5%. When inspecting the plantations in the nursery (trees 2—3 years old), the frequency of brown spotting of the walnut Marssonina juglandis (Lieh.) Magn. accounted for 80.3% of bacterial burn Xanthomonas juglandis Sacc. — 12.1%, white spot Microstroma juglandis Sacc. — 7.6%.
 Conclusions. During the examination of walnut plantations (in trees of different ages), it was found that the high incidence of diseases, in particular, brown spotting Marssonina juglandis (Lieh.) Magn. was noted in nurseries, where on the day of the last record on susceptible forms of culture, damage by brown spot was 100.0%, while the development of the disease was 47.7%.

First Report of Colletotrichum tropicale Causing Anthracnose on Pitaya (Hylocereus costaricensis) in Brazil.

Pitaya, Hylocereus costaricensis, is a species of the Cactaceae family and originated in the Americas (Ortiz & Livera, 1995). It has been cultivated in Brazil and has shown a great potential for fruit production and is currently present in several markets (Faleiro et al. 2021). In July 2018, infected plants of pitaya with symptoms of anthracnose were obtained from an orchard in Fortaleza, Ceará Brazil, (3°44'24.5"S 38°34'30.8"W), with 50% disease incidence. The symptoms observed consisted of well-defined and depressed stains, that initially appeared as reddish-orange spots and were surrounded by a border of dark-brown color. As the lesion progressed, the center became light brown or whitish in color, with black dots appearing later. Four cladodes were collected with anthracnose symptoms. The pathogen was isolated from symptomatic cladodes, which were surface disinfected with 1% v/v NaClO and 70% v/v ethanol, rinsed with sterile distilled water, transferred onto potato dextrose agar (PDA) medium and incubated under a light/dark (12h/12h) photoperiod. Two isolates were recovered from the lesions on cladodes. Pure cultures were obtained from single conidia produced on colonies grown on PDA medium, using an inoculation needle under a microscope. Colonies on PDA exhibited white aerial mycelia with an orange conidial mass. The colonies were light grey in the front and light orange in the reverse of the plate. Morphological features suggested that the isolates had the same characteristics as previously described for Colletotrichum spp. (Weir et al., 2012). In order to identify the species of the isolates, the genomic DNA of UFCM 0684 and UFCM 0685 isolates was extracted using the CTAB method and the ITS region, TUB2, ACT, GS, GAPDH gene fragments were amplified. PCR products were sequenced and the resulting sequences were submitted to phylogenetic analyses based on maximum likelihood for the combination of the genes. The isolates grouped within Colletotrichum tropicale with 99% bootstrap support. The sequences obtained in this study were deposited in GenBank as ACT (accession no. OL799311, OL799312), TUB2 (OL799313; OL799314), GAPDH (OL799315, OL799316), GS (OL799317; OL799318) and ITS (OL799319; OL799320). After that, the UFCM 0685 isolate was selected to study for further characterization. Conidia (n = 50) were 13.7 (length) × 4.7 μm (width) in average, hyaline, aseptate and cylindrical. To complete Koch's postulates, pathogenicity tests were performed in moist chamber for one week at 25°C with 80% relative humidity on a 12 h fluorescent light/dark photoperiod. The cladodes were wounded using a sterilized needle and inoculated with 10 µl of a conidial suspension (1 × 106 conidia/ml) on three cladodes with five wounds each. The same number of uninoculated cladode was used as control. The experiment was performed twice. Two weeks later, all inoculated cladodes showed necrotic symptoms, which were similar to the symptoms previously observed in the field. The uninoculated cladode remained symptomless. The fungus was reisolated from the inoculated cladode and its morphological characteristics were similar to the original isolate. Colletotrichum tropicale has been reported to cause anthracnose on H. costaricensis in Mexico (Nunez-Garcia et al. 2023), H. undatus, H. monocanthus and H. megalanthus (Evallo et al. 2022). For the best of our knowledge, this is the first report of anthracnose caused by C. tropicale in H. costaricensis in Brazil.

First Report of Colletotrichum gloeosporioides Causing Anthracnose on Peanut in Chongqing, China.

In July 2022, dieback and discoloration were detected on infected stems of peanut in Qijiang District of Chongqing (106.56°E,29.41°N), China, with an incidence up to 5%. These peanut stems had disease symptoms typical of anthracnose with irregular gray-brown spots with dark brown edges, sunken, and necrotic. High temperature and high humidity were favorable for the growth of the pathogen. To isolate the pathogen, we collected 10 typical infected peanuts and cut one piece from each of symptomatic stems, surface sterilized with 0.5% NaClO for 1 min, and 75% ethanol for 30 s, then rinsed three times with sterile distilled water and dried on sterilized filter paper. These pieces were incubated on potato dextrose agar (PDA) at 25°C in the dark. Pure cultures were obtained from hyphal tips of each colony. It was found that isolates with the same colony morphology were isolated from each infected stem. A representative isolate (L7) was used for morphological characterization, molecular analysis, phylogenetic analysis, and pathogenicity tests. The colonies appeared white to gray, with white margins and aerial hyphae, and the reverse of the colonies was gray to brown. Conidia were cylindrical, aseptate, with obtuse to slightly rounded ends, 13.4 to 18.8 × 4.2 to 5.8 μm (n=50). Morphological characteristics were generally consistent with those of Colletotrichum gloeosporioides species complex (Cannon et al., 2012). For molecular identification, genomic DNA was extracted using a CTAB method and partial sequences of β-tubulin (TUB2), actin (ACT) genes, chitin synthase (CHS) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were amplified and sequenced using primers T1/T2, ACT-512F/ACT-783R, CHS-79F/CHS-345R, and GDF1/GDR1, respectively (Damm et al., 2012; Dowling et al., 2020). Using the BLAST, TUB2, ACT, CHS and GAPDH gene sequences (GenBank accession No. OR714793, OP168707, OP168708 and OR714794, respectively) were100% (429 bp out of 429 bp), 99.22% (256 bp out of 258 bp), 99.64% (276 bp out of 277 bp) and 100% (253 bp out of 253 bp) identical to C. gloeosporioides CBS:112999 (JQ005587, JQ005500, JQ005326, and JQ005239), respectively. Using Neighbor-Joining algorithm, phylogenetic analysis was conducted based on the concatenated sequences of published TUB2, ACT, CHS and GAPDH genes. The identified isolate (L7) was closely related to C. gloeosporioides. To evaluate pathogenicity, the stems of ten peanut (Zhonghua12) seedlings (2 weeks) were wounded with a sterile toothpick and mycelial plugs (5 mm in diameter) or 20 μl of conidial suspension (105/ml) were inoculated. Non-colonized agar plugs or 20 μl of sterile distilled water were treated as control. After inoculation, the peanuts were kept in a moist chamber at 28°C with 80% humidity in the dark for 24 h, and subsequently transferred to the moist chamber with 12 h light and darkness cycle for 6 days, similar symptoms were observed on all inoculated peanuts. Controls remained asymptomatic. C. gloeosporioides was reisolated from the diseased stems and confirmed using morphological features and sequence analysis of TUB2, ACT, CHS and GAPDH. Anthracnose caused by C. truncatum and C. fructicola has been reported on peanut leaves in China (Gong et al., 2023; Yu et al., 2019). To our knowledge, this is the first report of anthracnose on peanut stem caused by C. gloeosporioides in Chongqing. Our report will provide crucial information for studying on epidemiology and management of this disease.

First report of canker caused by Cytospora sorbicola on Sweet Cherry (Prunus avium) in Chile.

Chile leads cherry exports in the southern hemisphere with a total of 415.315 t exported in the 2022 to 2023 season (IQonsulting, 2023). Cytospora canker, produced by Cytospora spp., causes destructive infections and limit the productivity of sweet cherry orchards (Luo et al. 2019). This study was focused on isolating Cytospora strains to identify and characterize the species present in sweet cherry. During the period 2019-2022, ten samples of stem or branch presenting canker, dieback, gummosis or dead buds, were collected from sweet cherry cultivars 'Skeena', 'Lapins', 'Santina', 'Sweetheart', and 'Regina', in the regions Ñuble and O'Higgins, Chile. Five mm pieces from the necrotic wood margins of the samples were rinsed with sterile deionized water, placed on potato dextrose agar (PDA, Difco) and incubated at 20±2 ºC for 5 days. One isolate was recovered from each sample, resulting in ten Cytospora-like strains. Single hyphal tips were transferred onto PDA plates and all isolates were deposited in the Chilean Collection of Microbial Genetic Resources (CChRGM). Colonies grown on PDA reached 89 mm in diameter in 10 d at 25 °C, showing irregular margin, lacking aerial mycelium, initially off-white to cream that turned greenish gray in the center, which darkens with age. After 20 days of culturing on pine needle agar (Chen et al. 2015), isolates produced conidiomata pycnidial, semi-immersed, black, and subglobose (362)445-555(681)×(357)528-700(1053) µm (n=10), generating amber slimy conidia masses; Conidiophores were phialidic, cylindrical, aseptate, hyaline (6.77)9-10.04(12.88)×(0.82)1.1-1.28(1.99) µm (n = 30); conidia were abundant, allantoid, hyaline to light brown, aseptate (3.39)4.28-4.57(5.36)×(0.69)0.96-1.09(1.47) µm (n = 30) (Supplementary Figure 1). No sexual morph was observed. With the exception of the strain RGM 3390, all the isolates shared morphological characters to the descriptions of Cytospora sorbicola Norphanph., Bulgakov, T. C. Wen & K. D. Hyde (Norphanphoun et al. 2017). Isolates were identified at species level, by sequencing DNA regions described by Pan et al. (2020): ITS1-5.8S-ITS2, LSU; act, tef-1α, and tub2 with the exception of the RBP2, because this region could not be amplified in seven out of ten isolates. The consensus tree included the concatenated sequences of the ten isolates and those of reference Cytospora species reported by Ilyukhin et al. (2023) using a maximum likelihood analysis with the tool IQ-TREE webserver. MLSA confirmed the taxonomic affiliation of nine of the isolates with C. sorbicola and one isolate with Cytospora sp. (RGM 3390), that might represent a novel species (Supplementary Figure 2). The isolates RGM 3399 and RGM 3400, were selected randomly for pathogenicity tests. Inoculations were performed on 2-year-old sweet cherry cv. 'Lapins' grow in pots in a greenhouse at 26±6°C. Seven plants per isolate were cut to about 6-cm length from the main stem, and inoculated onto fresh cuts with 5-mm mycelium PDA plugs of 5-d-old culture and wrapped in moist sterile cotton and parafilm to keep moisture. Six plants were inoculated with non-colonized PDA agar plugs as control. The average canker length 3 months after inoculation was 3.1 and 0.8 cm, for RGM 3389 and RGM 3400, respectively (Supplementary Figure 1). Symptomatic twigs were incubated in moist chambers at 20±2 ºC for 10 d, resulting in the re-isolation of Cytospora strains that produced pycnidia and conidia structures in agreement with C. sorbicola. Both strains were reidentified to fulfill Koch's postulates, control twigs remained asymptomatic and no fungus was isolated from these twigs. This is the first report of C. sorbicola causing canker on sweet cherry in Chile. Our findings suggest that this species could be the most recurrent in cherry in central Chile, coinciding with it found in California where C. sorbicola has been described as the main causal agent of Cytospora canker of stone fruits in California (Lawrence et al. 2018).

Didymium arenosum, a myxomycete new to science from the confluence of deserts in northwestern China.

A new myxomycete species, Didymium arenosum, was described based on morphological evidence and phylogenetic analyses. The species was discovered in the arid region at the confluence of the Badain Jaran desert and Tengger desert on the leaves of Betula platyphylla and was cultivated in a moist chamber culture. Morphologically, the species is distinguished by the greenish-yellow calcium carbonate crystals on the surface and the spores covered with small warts, some of which are connected into a short line. A phylogenetic analysis of D. arenosum strongly supports its classification as a separate clade. The spore to spore agar culture of D. arenosum requires 23 days, and this study provides a detailed description of its life cycle.

First Report of Anthracnose Caused by Colletotrichum nymphaeae on Strawberry Fruits in Central Argentina.

Strawberry (Fragaria x ananassa) production in Argentina extends to around 1700 hectares. Coronda city, located in Santa Fe province, is an important strawberry producer due to ideal agroecological conditions for culture and a high specialization for production. In November 2021, anthracnose symptoms were observed on strawberries cvs. 'San Andreas' and 'Splendor' in Coronda (31°58'S, 60°55'W), central Argentina. During these years, the incidence of the disease reached 40% of the production. Symptoms included 2-3 mm circular to irregular dark brown spots which enlarged rapidly and became sunken. Under high humidity conditions, concentric rings of pinhead-size salmon-colored acervuli developed on the lesions. The causal agent was isolated by touching acervuli with a sterile needle and monosporic cultures were obtained on PDA after 10 days at 25°C, with a 12-h light period. Colonies were white to gray on the top and orange on the underside, where concentric rings of salmon acervuli were clearly distinguished. The width and length of one hundred conidia were examined in three isolates (CF1, CF2, and CF3), ranging from 3.27 to 5.53 μm (avg.= 4.3 μm), and from 10.37 to 19.52 μm (avg.= 14.27 μm), respectively. The conidia were hyaline, smooth-walled, aseptate, and cylindric-clavate with one end round and one end acute. These morphological characteristics correspond to species belonging to the C. acutatum complex (Damm et al. 2012; Liu et al. 2022). To accurately identify the species, DNA was extracted from isolates, and ß-tubulin (TUB2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and histone (HIS3) genes were partially amplified and sequenced (Vieira et al. 2020). TUB2, GAPDH, and HIS3 sequences presented a 100% of identity with species of Colletotrichum nymphaeae. The nucleotide sequences were deposited in GenBank (OR271556-OR271558, TUB2; OR271559-OR271561, GAPDH; and OR271562-OR271564, HIS3). Multilocus phylogenetic analyses performed with reference sequences (Damm et al. 2012) showed that the three isolates clustered with C. nymphaeae, in accordance with BLAST results. To confirm pathogenicity, each isolate was inoculated in eight detached fruits of the cultivar from which it was originally obtained. Two drops of 10 μl of conidial suspension (1x105 conidia per ml) were deposited in non-wounded areas on fruits previously disinfested with 1% sodium hypochlorite solution for 1 min and rinsed twice with sterile distilled water. Drops of sterile water were deposited in eight fruits as control. Pathogenicity tests were repeated twice. Fruits were kept in moist chamber (80+5% relative humidity) at 25°C for ten days. First symptoms appeared 4 days after inoculation. After that, all of the isolates produced symptoms identical to those previously described, whereas the controls remain symptomless. The pathogen was re-isolated from lesions, and identified as C. nymphaeae by morphological characteristics and based on the TUB2 sequences, as previously described. Strawberry anthracnose in Argentina was previously associated with Colletotrichum acutatum, C. gloeosporioides and C. fragariae species based on morphological characteristics (Ramallo et al. 2000; Monaco et al. 2000) but molecular identification was not performed until today. To our knowledge, this is the first report of C. nymphaeae causing anthracnose on strawberry in Argentina. This accurate identification will help to develop more efficient management strategies.

First Report of Colletotrichum chrysophilum Causing Anthracnose on Nance (Byrsonima crassifolia) in Mexico.

Nance fruit [Byrsonima crassifolia (L.) HBK] is a native crop widely distributed in Mexico and South America (Medina-Torres et al. 2018). It has been reported that nance is a good source of active compounds with anti-inflammatory, neuropharmacological and antioxidant effects. In 2022, the annual production of nance fruit in Mexico was of 7,713.13 tons and average yield of 5.64 t/ha with economic value of 51,952.66 million pesos (SIAP, 2022). This production generated significant economic income for the communities at a local, regional, and national level. In January 2023, irregular necrotic spots were observed on leaves and fruit of nance in an orchard of 50 nance trees located in San Sebastián Nopalera (16°54'52.73"N; 97°47'50.35"W), Oaxaca, Mexico. The incidence of the disease ranged from 50 to 60% of the trees. Infected fruit first showed dark-brown lesions with defined borders that coalesced to form large necrotic area. Isolates were purified by single spore isolation method (Choi et al. 1999).Colletotrichum strains were grown in PDA medium and five monoconidial isolates were obtained. A representative sample was selected (CNC-NP3) and deposited in the Culture Collection of Phytopathogenic Fungi of Plant Pathology Laboratory of the CIIDIR-Oaxaca of the Instituto Politécnico Nacional. Colony on PDA was white with sparse aerial mycelium, and the center was dark grey with abundant acervuli. Conidia (n = 100) were hyaline, aseptate, cylindrical with rounded apex, 13.5 to 15.2 × 4.3 to 5.1 μm. Appressoria (n = 20) were terminal or lateral, obovoid to clavate and some with slightly lobed, 9.9 to 11.6 × 5.3 to 6.6 μm. Based on the morphology, the isolate was identified as belonging to the Colletotrichum gloeosporioides species complex (Jayawardena et al. 2016). The representative isolate CNC-NP3 was identified by multilocus phylogenetic analysis using nucleotide sequences of internal transcribed spacer (ITS), actin (ACT), β-tubulin (TUB2), and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) (Jayawardena et al. 2016). The sequences were deposited in GenBank (accessions nos. OQ861102 (ITS), OQ870548 (ACT), OQ870549 (TUB2), OQ870550 (GAPDH). The phylogenetic analysis was carried out by Maximum likelihood method using concatenated sequences of ITS, ACT, TUB2 and GAPDH genes (Kozlov et al. 2019). The multilocus phylogenetic analysis revelated clearly the isolate CNC-NP3 as Colletotrichum chrysophilum. To confirm pathogenicity of CNC-NP3, 30 healthy fruits were inoculated. Fifteen disinfected nance with wounds and fifteen nance without wounds were inoculate with 10 µL of conidial suspension (1×105 spores/mL) from 7-day old culture. And controls were inoculated using sterile distilled water. Fruits were placed in a moist chamber covered with plastic bag at 25 °C for 48 h to maintain high humidity. After 4 days the inoculation sites development symptoms that were identical to those initially observed in the field, whereas the control group remained symptomless. The pathogenicity test was performed twice, with the same results. The pathogen was re-isolated from the lesion to fulfill Koch's postulates. Currently, Colletotrichum chrysophilum has been reported causing anthracnose disease in several crops: apple in New York (Khodadadi et al. 2020), papaya in Mexico (Pacheco- Esteva et al.2022), Blueberry (Soares et al. 2022) and banana in Brazil (Astolfi et al. 2022). To our knowledge, this is the first report of anthracnose in Byrsonima crassifolia caused by Colletotrichum chrysophilum in Oaxaca, Mexico.