Next Article in Journal
Field Evaluations of Two Citrus Interspecific Hybrid Populations Using Desert Lime (Citrus glauca (Lindl.) Burkill) Identify Presumed Resistance and Tolerance to Huanglongbing
Previous Article in Journal
Water and Nitrogen Management Drive Soil Nutrient Dynamics and Microbial–Enzyme Activity in Silage Maize Systems in Northwest China
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Agronomy
Volume 15
Issue 10
10.3390/agronomy15102406
agronomy-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Leaf Spot Disease Caused by Several Pathogenic Species of the Pleosporaceae Family on Agave salmiana and Agave lechuguilla Plants in Mexico, and Their Biocontrol Using the Indigenous Trichoderma asperellum Strain JEAB02

by
José Esteban Aparicio-Burgos
1,
Teresa Romero-Cortes
1,2,
María Magdalena Armendáriz-Ontiveros
1 and
Jaime Alioscha Cuervo-Parra
1,*
1
Escuela Superior de Apan, Universidad Autónoma del Estado de Hidalgo, Carretera Apan-Calpulalpan, Km 8, Chimalpa Tlalayote, Apan 43920, Hidalgo, Mexico
2
Escuela Superior de Ciudad Sahagún, Universidad Autónoma del Estado de Hidalgo, Carretera Ciudad Sahagún-Otumba s/n, Zona Industrial, Cd Sahagún 43990, Hidalgo, Mexico
*
Author to whom correspondence should be addressed.
Agronomy 2025, 15(10), 2406; https://doi.org/10.3390/agronomy15102406
Submission received: 16 September 2025 / Revised: 13 October 2025 / Accepted: 15 October 2025 / Published: 16 October 2025
(This article belongs to the Section Pest and Disease Management)
Browse Figure
Versions Notes

Abstract

The genus Agave (family Asparagaceae) represents the second-most important group of plants in Mexico. Several fungal species have been identified as causal agents of leaf spot disease affecting Agave salmiana and A. lechuguilla, producing necrotic lesions that compromise plant health and productivity. Pathogenicity experiments were conducted under greenhouse conditions, field tests were performed, and in vitro antagonism using Trichoderma asperellum strain JEAB02 against selected pathogenic isolates was evaluated. Phylogenetic analysis of genomic DNA fragments allowed the identification of 26 fungal isolates belonging to Curvularia lunata, C. verruculosa, Bipolaris zeae, Alternaria alternata, Fusarium lactis, Epicoccum sorghinum, Myrmaecium rubricosum, Penicillium diversum, and Aspergillus oryzae. In pathogenicity assays under greenhouse conditions on A. salmiana and A. lechuguilla, treatments T5–T12 exhibited statistically similar levels of disease severity (33.10–37.29%), caused mainly by C. verruculosa, A. alternata, B. zeae, and F. lactis. In field tests, Agave plants inoculated with the selected pathogenic fungi (T4, T5, T7, T8, T10, and T11) showed 21.07–36.73% leaf damage after 75 days. The antagonistic effect of T. asperellum JEAB02 caused complete (100%) growth inhibition of the pathogenic isolate JCPN27 and inhibition levels from 99.81 to 99.98% for isolates JCPN18, JCPN24, JCPN28, JCPN29, JCPN31, and JCPN33, demonstrating its high potential as a biological control agent.

1. Introduction

The genus Agave (family Asparagaceae) is the second-most important group of plants in Mexico for producing non-timber products such as tequila, mezcal, aguamiel, pulque, fibers, and probiotics. The genus comprises about 310 known species, with around 273 found in Mexico, 55% of which are endemic [1,2]. Mexico is regarded as the center of origin and diversification of Agave species, which are mainly found in arid regions [3]. Therefore, they form a group of plants with significant economic importance due to their culinary [4,5], medicinal [6,7], industrial [8,9,10], and ornamental [11] uses, among others. Among all species, the most widely used include Agave tequilana [12], A. americana [13], A. angustifolia [14,15], A. salmiana [16], A. victoria-reginae [11], and A. lechuguilla [10]. Among these, A. salmiana Otto ex Salm-Dick ssp. ferox (Koch) Hochstätter [17], and A. lechuguilla Torr., have a wide distribution in the arid and semi-arid areas of the Llanos de Apan region, which includes the states of Hidalgo, Tlaxcala, Puebla, and the Estado de México [18,19]. Their resources are used by the inhabitants of this region, either directly from wild populations or from commercial plantations [20,21].
However, due to the slow growth of plants in this genus until they reach maturity and can be used [22,23], combined with their low rate of sexual and asexual reproduction [24], the presence of pests [25,26] and fungal diseases such as agave wilt [27], gray spot [28,29], and leaf spot [30,31,32,33,34], as well as growth delays caused by these factors [35], economic losses remain significant. However, despite the economic and ecological importance of Agave species, there is currently no scientific information available in Mexico regarding the economic losses caused by the causal agents of leaf spot disease in A. salmiana and A. lechuguilla, affecting both seedlings and mature plants. Moreover, the information available on control methods remains very limited. To date, only one study has reported the chemical control of Bipolaris zeae using silver nanoparticles in artificially inoculated A. salmiana plants [36]. This highlights the need for further research focused on the development of biological management strategies aimed at the sustainable control of these causal agents. Therefore, to identify which fungal species are responsible for the leaf spot symptoms observed in A. salmiana and A. lechuguilla plants in two municipalities in the state of Hidalgo, the following objectives were set: (i) molecularly characterize the causal agents of the “leaf spots” observed on A. salmiana and A. lechuguilla plants in these municipalities, (ii) evaluate the aggressiveness of the pathogens associated with the leaf spot symptoms in plants grown under both greenhouse and field conditions, and (iii) assess in vitro the effectiveness of a Trichoderma asperellum strain against the pathogenic fungi isolated from Agave plants.
To achieve these objectives, the study was structured in consecutive experimental phases. First, 26 fungal isolates were obtained from symptomatic leaves of Agave salmiana and A. lechuguilla collected in two contrasting environments—one natural (Mineral de la Reforma) and one disturbed (Apan). These isolates were subsequently subjected to morphological and molecular characterization to determine their taxonomic identity. In the second phase, in vitro confrontation assays were conducted using the T. asperellum JEAB02 strain to evaluate its antagonistic potential against the isolated fungi. Finally, pathogenicity tests were performed sequentially under greenhouse conditions (2023) and field conditions (2024), following Koch’s postulates, to identify which isolates were responsible for causing the leaf spot symptoms observed in Agave plants. Additionally, Zea mays plants were included as a comparative host species, since maize is an agriculturally and culturally important crop in Mexico and in the study region. Assessing the pathogenic potential of the isolates on maize provided a relevant reference for understanding cross-host infectivity and ecological adaptability of the fungi within the Helminthosporium complex.

2. Materials and Methods

2.1. The Fungal Strains

Twenty-six pathogenic fungal isolates were obtained from wild and cultivated populations of Agave salmiana and A. lechuguilla in the municipalities of Apan and Mineral de la Reforma, Hidalgo, Mexico, between 2019 and 2023 (Table 1). Based on macroscopic and microscopic morphological characteristics (e.g., ascomata, conidiophores, macroconidia, conidia) observed during cultivation and through comparison with reference fungal isolates, the isolates were identified as belonging to nine genera. Nine strains were assigned to the genus Bipolaris, seven to Curvularia, and two each to Penicillium, Fusarium, Alternaria, and Epicoccum. The remaining two strains were identified as Myrmaecium and Aspergillus.
All fungal isolates were obtained from leaves of sampled maguey plants showing symptoms of leaf spot disease. To process infected tissue, samples were surface sterilized for 1 min in a 3% sodium hypochlorite solution, followed by two rinses in sterile distilled water [37]. Subsequently, samples were allowed to dry and placed in Petri dishes with potato dextrose agar (PDA, BD Bioxon®) in a moist chamber. The fungi were incubated in the dark at 28 °C for one week [38] or until the isolates covered the entire culture surface [30]. Afterwards, each isolate obtained was subcultured on PDA under the same conditions. All fungal strains isolated from maguey leaves exhibiting leaf spot symptoms were maintained on slanted agar tubes at 4 °C and preserved as spore suspensions at −86 °C until further use for molecular characterization.

2.2. Molecular Characterization

Genomic DNA from 26 fungal strains isolated from the leaves of A. salmiana and A. lechuguilla showing leaf spot symptoms (Table 1) was sequenced using the Sanger sequencing method (Eurofins Genomics GmbH, Anzinger, Germany). DNA extraction was performed following the procedure described by Cuervo-Parra et al. [39]. Ribosomal DNA (rDNA) fragments were amplified by PCR as outlined by Romero-Cortes et al. [40], using the primers ITS5 and ITS4B [41]. The amplicons obtained, containing the ITS1/5.8s/ITS2 region, were washed twice with TE buffer (Tris HCl 10 mM, EDTA 1 mM). Subsequently, the PCR products of each strain were analyzed with Chromas 1.45 software (School of Health Science, Griffith University, Gold Coast campus, Southport, Queensland, Australia). The sequences were then subjected to pairwise alignment using ElasticBLAST (v1.4.0) against the GenBank database of the National Center for Biotechnology Information (NCBI) [42].
Amplicon sequences were deposited in the GenBank database. Phylogenetic analysis of the 26 strains isolated in this study, together with 46 related fungal sequences from NCBI, was conducted by constructing a phylogenetic tree using the Neighbor-Joining (NJ) method, the Jukes–Cantor model, with bootstrap support on 100 replicates in MEGA X (v10.2) software [43]. The accessions OL840629 and OL840630 of Irpex latemarginatus, along with OL830229 and OL830230 of Mucor circinelloides, isolated from a textile industry effluent in Mexico, were used as an outgroup for the phylogenetic analysis [41].

2.3. Pathogenicity Tests for Agave Phytopathogens

2.3.1. Greenhouse Experiment Design and Treatments

To confirm which pathogenic fungi isolated from the maguey leaves (Table 1) developed the typical symptoms of leaf spot disease observed in the field on A. salmiana and A. lechuguilla plants, a preliminary experiment was carried out in 2023 at ESAp-UAEH facilities. This experiment aimed to evaluate, under greenhouse conditions, the fungus–plant interaction of one representative isolate per species of the nine fungal species isolated from infected Agave tissues showing leaf spot symptoms, in order to determine whether disease symptoms developed and thereby fulfill Koch’s postulates [44]. For the preliminary experiments, the isolate of each species that exhibited the greatest radial growth after one week of culture in PDA medium was used. The treatments were assigned using a completely randomized design with three replicates per treatment (Table 2).
Disease symptoms in greenhouse-grown A. salmiana and A. lechuguilla were evaluated through systematic weekly visual assessments. Symptoms’ severity was quantified using a standardized 1–9 scale [34,45], which accounted for leaf spots, necrosis, chlorosis, and wilting. Disease incidence was calculated as the percentage of symptomatic plants, with non-inoculated controls included to ensure accurate symptom attribution.
Square plastic pots (11 × 11 × 11 cm; 1 kg capacity) were filled with a substrate consisting of 30 kg of sand, 45 kg of tepetate, and 75 kg of black soil. For each treatment, 15 healthy one-year-old maguey seedlings, obtained from seeds in 2022, each with 7–8 developed leaves, were used. A total of 150 seedlings were used for each of the two Agave species studied. The maguey plants were inoculated with pathogenic fungi isolated from A. salmiana and A. lechuguilla plants, using a 15 mL spore suspension containing equal volumes of conidia from each selected strain: C. lunata, C. verruculosa, B. zeae, A. alternata, F. lactis, E. sorghinum, M. rubricosum, P. diversum, and A. oryzae (Table 2). The suspension was prepared in sterile distilled water adjusted to 5 × 106 conidia/mL and applied directly to the leaves and rhizosphere of each plant, in treatments T3 to T20. In the control treatments (T1 and T2), only sterile distilled water was applied to the leaves and rhizosphere near the base of each A. salmiana and A. lechuguilla plant.
Based on the results of the preliminary greenhouse pathology test and following Koch’s postulates, an experimental design was developed to evaluate the pathogenic effect of the fungal species exhibiting the typical leaf spot symptoms observed under both field and greenhouse conditions.

2.3.2. Field Pathological Tests

To evaluate the impact of the selected pathogenic Agave fungi following Koch’s postulates [44,46], healthy one-year-old rhizome seedlings of A. salmiana and A. lechuguilla obtained from healthy adult plants were planted under field conditions.
The experiment was conducted at the facilities of the Escuela Superior de Apan-UAEH, located at 19°65′48″ N, 98°51′88″ W, and at an elevation of 2488 m above sea level. This site has an average annual temperature of 14.4 °C and a total annual rainfall of 618.2 mm, in the Hidalgo state [47,48]. The total experimental plot was 96 m2 (3 m wide by 32 m long), with treatments arranged in a completely randomized design with three replicates (Table 3). Each treatment consisted of 3 rows, each 3 m long, with 1 m spacing between rows and 0.60 m between plants. A 1 m gap was left between treatments to prevent crossover of results [49].
The maguey rhizome propagules used for the two Agave species were left to rest in the shade for a month before transplanting under field conditions in February 2024. Meanwhile, corn seeds were sown in pots under greenhouse conditions in January to ensure their germination and survival. Subsequently, after 15 days of germination, the seedlings were transplanted to the experimental plot together with the A. salmiana and A. lechuguilla seedlings.
For each treatment, 15 Agave and Zea seedlings with 7–8 fully developed leaves [50,51] were planted under field conditions in three replicates, using a total of 60 seedlings: A. salmiana, A. lechuguilla, and Z. mays. A substrate mixture of 120 kg sand, 120 kg tepetate, and 120 kg black soil was used, with 2 kg of substrate applied per maguey and corn seedling. Agronomic management of the experimental plot from February to July combined conventional practices with the application of chemical and organic fertilizers, following the methodology of Cuervo-Parra et al. [52]. During the dry period from February to April, flood irrigation was employed, whereas during the rainy period from May to July, a rainfed production system was used [53].
The maguey and corn plants were inoculated with each of the evaluated pathogenic fungi following a modified version of the technique described by Mata-Santoyo et al. [54]. For this, a 15 mL suspension containing equal parts of conidia from C. lunata, C. verruculosa, and B. zeae selected strains (Table 3) was prepared in sterile distilled water adjusted to 5 × 106 conidia/mL, directly onto the leaves and rhizosphere of plants in treatments T4 to T12. For the control treatments (T1 to T3), only sterile distilled water was applied to the leaves and rhizosphere near the base of each plant. After a five-day incubation period, evaluations were conducted using the disease incidence scale proposed by Perelló et al. [45] and Romero-Cortes et al. [34]: 1 = plants free of infection; 2 = plants with 1 to 5% leaf spots; 3 = 5 to 12%; 4 = 12 to 20%; 5 = 20 to 35%; 6 = 35 to 45%; 7 = 45 to 60%; 8 = 60 to 80%; and 9 = more than 80% of leaves showing leaf spots. Subsequently, the data were converted to percentages, and an ANOVA analysis was performed using Statistix 10 software [55]. Mean comparison was conducted using the Tukey HSD All-Pairwise test at a significance level of p ≤ 0.05.

2.4. In Vitro Confrontation Tests

Antagonistic Activity of Trichoderma asperellum Strain JEAB02 Against Pathogenic Fungi of Agave

To determine the antagonistic effect of a strain of Trichoderma asperellum, isolated from the leaves and rhizosphere of mangosteen plants [56], confrontation experiments were conducted using dual culture. These involved the Agave fungi (C. lunata, C. verruculosa, B. zeae, A. alternata, E. sorghinum, F. lactis, M. rubricosum, P. diversum, and A. oryzae) and the JEAB02 strain of T. asperellum in 2024, following the methodology proposed by Cuervo-Parra et al. [37]. For the in vitro confrontation test, the strain with the greatest radial growth for each pathogenic fungal species was used, corresponding to the same strains selected for the pathogenicity experiment under greenhouse conditions (Table 2).
Interactions between Trichoderma and each confronted fungal pathogen were measured after one week. Digital images were captured at a fixed distance of 20 cm using an iPhone SE (Model MX9U2LZ/A, Mexico City, Mexico) equipped with iOS 17.2.1, a 12-megapixel single-lens camera, and a wide-angle lens with an ƒ/1.8 aperture. The inhibition percentages of radial growth for each pathogenic fungus exposed to the T. asperellum (JEAB02) strain were reported as the biocontrol index (BCI), calculated using the formula BCI = [T/C] × 100, as described by Cuervo-Parra et al. [37]. Where BCI represents the percentage of inhibition of the radial growth of the pathogenic fungal colony caused by its interaction with the antagonistic Trichoderma fungus, T denotes the area occupied by the Trichoderma colony, and C indicates the total area occupied by both fungi. To calculate the radial growth areas of the BCI, the ImageJ (v1.48q) software was used http://rsbweb.nih.gov/ (accessed on 10 July 2024). Data from the dual culture experiments were analyzed using a completely randomized design with three replicates. Statistical analysis, including analysis of variance (ANOVA) and mean comparisons by the Tukey HSD All-Pairwise test (significance level set at p ≤ 0.05), was conducted using Statistix (v10) software [55].

3. Results and Discussion

3.1. Molecular Characterization

In molecular biology, analysis of the ITS1/5.8s/ITS2 genetic regions is a valuable and straightforward tool for identifying DNA sequences from a wide range of organisms [57]. In this study, the molecular characterization of the ITS regions of the rDNA from 26 new sequences of phytopathogenic fungi isolated from plant tissues of A. salmiana and A. lechuguilla exhibiting disease symptoms was performed. These sequences were compared with related sequences obtained from the GenBank database of the National Center for Biotechnology Information (NCBI).
For the molecular identification of the rDNA region, which included the ITS1/5.8s/ITS2 sequence of each pathogenic fungus isolated in this study, the primers ITS5 and ITS4B were used [41]. Pairwise alignment of DNA fragments from fungal strain was performed using the ElasticBLAST (v1.4.0) program [58], by comparison with sequences of related pathogenic fungal accessions available in the NCBI GenBank database [59]. Similarity percentages for the 26 accessions corresponding to the nine genera of isolated fungi ranged from 99.43 to 100% when compared with related fungal sequences available in GenBank. The length of the amplified rDNA fragments from the fungal strains isolated in this study ranged from 497 to 578 base pairs. These sequences were subsequently deposited in the NCBI GenBank database with the following accession numbers: OR047532-OR047548, OR553111-OR553112, and OR553116-OR553122 (Table 4).
When comparing the ITS regions of the 26 fungal isolates obtained from field-infected maguey plants, the data from this study are consistent with previously reported results for E. sorghinum from Agave angustifolia and A. tequilana plants [64,68,71]; C. lunata [63,72,73]; C. verruculosa [67,74]; F. lactis [61]; M. rubricosum [75]; A. alternata [69,76,77,78]; P. diversum [62]; and B. zeae [34,60,79,80].
Phylogenetic analysis of 26 fungal sequences from maguey plants and 46 related sequences from GenBank was performed, resulting in a phylogenetic tree constructed using the Neighbor-Joining (NJ) method. The tree was rooted with DNA sequences from accessions OL840629 and OL840630 of Irpex latemarginatus, and OL830229 of Mucor circinelloides, which were used as outgroups to root the tree and infer phylogenetic relationships [41]. Analysis of the NJ tree revealed three main clades (I, II, and III). Clade I was represented by sequences from the families Pleosporaceae and Didymellaceae, clade II by sequences from Nectriaceae, Valsariaceae, and Trichocomaceae, and clade III, serving as the external group, included sequences from Irpicaceae and Mucoraceae. All clades were supported by a similarity value of ≥97%.
Among the ITS regions, the ITS2 was the most conserved across all rDNA sequences examined in the multiple sequence alignment in this study. The optimal phylogenetic tree, generated using the NJ method with a total branch length of 1.72543294, is shown. Phylogenetic analysis using the NJ method indicated that the rDNA sequences from the 26 accessions, representing nine pathogenic fungal species isolated from A. salmiana and A. lechuguilla, grouped separately on different branches of the phylogenetic tree (Figure 1).
At the top of the phylogenetic tree, in clade I, were the accessions of the genera Curvularia, Bipolaris, and Alternaria, all belonging to the Pleosporaceae family [81], and supported by a bootstrap value of 100. Within these accessions, those with a bootstrap support value of 92 at the top, included MN688814 (C. pseudobrachyspora), KJ922372 (C. brachyspora), MH858446 (C. lunata var. lunata), KP340066 (C. aeria), KX610322 (C. lunata), MF490823 (C. variabilis), KF018919 (C. hawaiiensis), and MH863648 (C. spicifera), all obtained from different sources and geographical regions [67,73,82,83,84,85,86]. In a sister branch with a bootstrap value of 62, the C. lunata accessions OR047532, OR047533, OR047534, OR047535, and OR047536 reported in this work were grouped with NCBI C. lunata accessions KY100122 and GQ280375, which were isolated from sugarcane and Agave fourcroydes samples [63,72]. In another branch near the C. lunata accessions, with a bootstrap support value of 100, the C. verruculosa accessions OR047537 and OR047538 were grouped with GenBank accessions KJ748697 and MH859788 of C. verruculosa [67,74].
In the upper part of clade I, all accessions of the genus Curvularia grouped together with those of the genus Bipolaris, supported by a bootstrap value of 98 at the node joining these sequences. The top accessions included OR553112 and OR553118 of B. zeae reported in this work, together with MT505867, MT505870, MT645704, and ON630341 of B. zeae from GenBank [34,60,79,80]. The B. zeae accessions OR553111, OR553116, OR553117, and OR553119-OR553122 reported in this work were located adjacent to B. zeae accessions ON630338 and ON630344, which were isolated from A. salmiana plants in Mexico exhibiting leaf spot disease symptoms [34]. Another branch contained the accessions OR047546 and OR047547 of A. alternata isolated from A. lechuguilla plants in this study, along with the accessions KX115418, MN481948, MG255300, MN596827, and MN615420 of A. alternata from GenBank, with a bootstrap value of 100 [65,69,76,77,78]. At the bottom of clade I, along with all Pleosporaceae accessions considered in this analysis and supported by a bootstrap value of 100, are OR047543 and OR047544 of E. sorghinum isolated from A. lechuguilla plants in Mexico, together with GenBank accessions FJ427068, MH497563, OQ346146, and OR388091 of E. sorghinum isolated from A. angustifolia and A. tequilana samples in Mexico [33,64,68,71].
The sequences of OR047541 and OR047542 of the genus Fusarium, isolated from A. salmiana samples in Mexico, clustered at the top of clade II in the phylogenetic tree, together with MT279265 (F. lactis), ON229472 (F. verticillioides), OR426451 (F. fujikuroi; a former synonym of F. verticillioides within the Gibberella fujikuroi species complex [87]), and ON329680 (F. oxysporum) [61,88,89,90], all supported by a bootstrap value of 100 (Figure 1). In addition, OR047545 of M. rubricosum, isolated from A. lechuguilla in this study, was closely aligned with MK041893, MK041894, and OQ975458 of M. rubricosum, which were isolated from A. tequilana and Pinus sp. samples in Mexico [68,75], as well as KP687862 of M. fulvopruinatum from acacia bark in Taiwan [91]. At the bottom of clade II, with a bootstrap value of 97, were the sequences of Penicillium and Aspergillus accessions. OR047539 and OR047540 of P. diversum, isolated from A. salmiana and A. lechuguilla, were located near HM469392 (P. diversum) and KY966028 (P. verrucosum) from Korea and India [62,92], supported by a bootstrap value of 100. OR047548 of A. oryzae, isolated from A. salmiana in this study, aligned with OQ202044 of A. oryzae, isolated in Mexico from A. salmiana [70]. On a nearby branch with a bootstrap value of 100 were OR397993 (A. niger), OK012380 (A. minisclerotigenes), and MT529296 (A. flavus) [93,94,95]. Finally, in clade III, OL840629 and OL840630 of I. latemarginatus and OL830229 and OL830230 of M. circinelloides were positioned as an outgroup [41]. The high bootstrap support values obtained in this phylogenetic analysis corroborate the macroscopic identification and microscopic characterization of each fungal isolate, providing new records of pathogenic fungi associated with maguey plants in Hidalgo, Mexico.

3.2. Pathogenicity Tests

3.2.1. Greenhouse Tests

After completing the pathogenicity experiment under greenhouse conditions, treatments T5–T12 were statistically similar, showing the highest disease severity on A. salmiana and A. lechuguilla, with mean values ranging from 33.10 to 37.29%, caused by the pathogenic fungi C. verruculosa, A. alternata, F. lactis, and B. zeae. The second-highest severity was seen in plants from treatments T3 and T4, with mean values of 24.20 and 25.29%, caused by C. lunata. Treatments T13 and T14, associated with E. sorghinum, showed severity values of 18.24 to 18.26%. The fourth-highest severity was caused by M. rubricosum, with mean values of 11.20 to 11.28% on A. salmiana and A. lechuguilla. The lowest severity, ranging from 7.13 to 7.38% was recorded in treatments T17–T18, associated with P. diversum. Meanwhile, treatments T19 and T20 of A. oryzae, as well as the control treatments T1 and T2, showed no disease symptoms (0.0%) throughout the experimental period. Notably, the treatments that included C. verruculosa, A. alternata, F. lactis, and B. zeae exhibited the highest levels of damage (Table 5).
The symptoms caused by each pathogenic fungus on infected leaves of A. salmiana and A. lechuguilla were as follows. For A. alternata, initial signs were chlorotic marginal spots, especially at the leaf tips, which progressed to tip wilting and ultimately necrosis. These symptoms align with previous reports in Agave tequilana [96,97], a species of significant economic importance in Mexico, particularly in the states of Jalisco, Michoacán, Nayarit, Guanajuato, and Tamaulipas [98]. The isolation of this fungus from A. salmiana and A. lechuguilla in this study is not surprising, as it is an opportunistic pathogen known to infect more than 380 plant species [65,76,77,99]. In the case of F. lactis, the fungus primarily caused root rot and leaf blight, which led to bud wilting and drying of the leaf tips. These symptoms have also been reported in commercial plantations of A. tequilana [96,100,101], Agave potatorum [102], and sweet pepper [103].
In plants infected with E. sorghinum, irregularly shaped lesions were observed, sometimes surrounded by a yellow halo. These lesions became sunken, forming necrotic depressions. The results of this study align with reports for other crops such as rice [104,105,106], cocksfoot grass [107], sugarcane [108], chrysanthemum [109], A. angustifolia [33,64], and A. tequilana [68,71]. In plants infected with M. rubricosum, leaf blisters were observed, which eventually caused wilting and rotting of affected leaves. In some cases, leaf deformation was also observed. These symptoms are consistent with reports for Pinus patula [110,111] and A. tequilana [68,112]. In plants infected with P. diversum, brown spots developed. Over time, these spots caused leaves wilting and stunted growth in affected specimens. Similar symptoms have been reported in several crops of economic importance [113,114,115]. This group of fungi is commonly found on a wide variety of crops and, under favorable humidity and temperature conditions, typically causes the diseases described here, as well as chlorosis and water-soaked tissues that emit a moldy odor [116,117]. Although, A. oryzae is not classified as a direct plant pathogen, it can indirectly affect their plant health as an opportunistic microorganism that thrives under high humidity [118]. No disease symptoms were observed on the leaves and roots of A. salmiana and A. lechuguilla inoculated with this fungus. Instead, improved growth was observed in these plants. These results are consistent with previous studies reporting A. oryzae as a microorganism with potential biotechnological [119] and agricultural [120] applications, and it is classified by the FDA as a food additive within the Generally Recognized As Safe (GRAS) category [121,122].
The fungi C. lunata, C. verruculosa, and B. zeae, which are part of the Helminthosporium complex of pathogenic fungi associated with various crops worldwide [63,82,83,123,124], caused black leaf spots on A. salmiana and A. lechuguilla leaves, consistent with field observations of infected tissues. Additionally, the leaf spot symptoms observed in this study match reports for A. salmiana plants [34,36,125] and A. fourcroydes [72]. C. lunata is recognized as an opportunistic pathogenic microorganism, frequently found in food, decaying plant matter, and soil [126]. It mainly affects cereals [63,82,127,128], cacao [83], and, more recently, maguey plants [125], where it causes leaf spot disease [129]. For C. verruculosa, although specific data on the extent of damage and symptoms in Agave species are not yet available, this fungus is known to infect cultivated grasses such as rice and corn [130,131,132], as well as wild grasses [133,134,135], and legumes [136], leading to significant economic losses. B. zeae is another pathogen responsible for various plant diseases, including seedling death, leaf and stem wilting, and black leaf spots [137]. This pathogen is frequently reported in economically important crops such as grasses [138,139], in polygonaceous plants [140], often cultivated alongside magueys under production systems like metepantle [2,141], and in several wild Poaceae species growing near maguey plantations [142,143,144]. This ecological context may explain the presence of these fungi in the sampled maguey plants, which may have been infected through biotic and abiotic vectors that transported spores of the Helminthosporium complex [145] from infected wild or cultivated grasses to a new host, in this case A. salmiana and A. lechuguilla.

3.2.2. Field Tests

Field evaluations were carried out to confirm the pathogenicity of B. zeae, C. lunata, and C. verruculosa—the fungal species previously identified as causal agents of leaf spot disease under greenhouse conditions (Table 5). Under natural field conditions, disease development was monitored in A. salmiana, A. lechuguilla, and Zea mays plants corresponding to treatments T4–T12 (Table 6). Characteristic leaf spot symptoms appeared 75 days after inoculation in Agave plants and 15 days in maize, with foliar damage ranging from 21.07% to 41.50%. These results confirmed the pathogenic capacity of the three fungi in both hosts and revealed interspecific differences in susceptibility, which are likely associated with structural and physiological adaptations of Agave species to arid environments that delay pathogen penetration and symptom expression.
Under field conditions, plants of the genus Agave show adaptations to Mexico’s arid and semi-arid climates, such as a thick cuticle, wax layers, and complex stomata that help protect them from water loss [31,146]. These adaptations are some of the mechanisms that enable these plants to defend against pathogen attacks. They also explain the long time (75 days) it takes for the three pathogenic fungi to penetrate these biological barriers [34], in contrast to the very short time (15 days) required for the same pathogens to penetrate and cause leaf spot symptoms in the corn plants. Similarly, the tissue of corn leaves, which is formed from thin-walled parenchymal cells with numerous perforations [147], could have facilitated the hyphae of pathogens of the genera Bipolaris and Curvularia evaluated in this study to penetrate and infect the plant tissues of corn in a considerably shorter time (Table 6).
When analyzing results by species, significant differences were observed among treatments T6, T9, and T12, with the highest percentage of foliar damage (41.50%) in corn leaves in treatment T6, which was inoculated with the fungus B. zeae. In second place were corn plants from treatment T9, inoculated with the fungus C. verruculosa, with a leaf damage percentage of 38.08%. In third place were corn plants from treatment T12, with 32.66% leaf damage. These results fall within the damage range reported by other authors for various corn varieties affected by B. setariae [148,149], B. zeicola, B. cynodontis, B. oryzae, B. saccharicola, Curvularia spp. [148], C. lunata [127,150], and C. verruculosa [131].
Damage percentages higher than those reported in this study have been noted for the fungus C. verruculosa, with damage reaching up to 60% in bean plants [136], and for the phytopathogen B. maydis, with damage exceeding 70% in corn crops [148,151]. Therefore, the damage caused by fungi of the Helminthosporium complex in corn crops varies and depends on multiple factors, such as environmental conditions, the susceptibility of the corn variety, and the virulence of the pathogen [127,152,153].
For treatments T4, T7, and T10, significant differences were also observed among the evaluated treatments. The highest percentage of damage was recorded at 36.73% for A. salmiana plants inoculated with the fungus C. verruculosa, followed closely by plants in treatment T4 with 33.27% damage caused by B. zeae, and a lower percentage of 23.67% for maguey plants inoculated with C. lunata. On the other hand, for A. lechuguilla plants in treatments T5 and T11, inoculated with the fungi C. verruculosa and B. zeae, no significant differences were observed, with damage values of 34.07 and 31.73%. For A. lechuguilla plants in treatment T8, the lowest percentage of damage among all treatments was recorded, at 21.07%. Finally, plants in treatments T1, T2, and T3 did not exhibit any disease symptoms throughout the entire duration of the pathogenicity experiment (Table 6).
Information on disease symptoms caused by Bipolaris and Curvularia species in agave leaves is scarce, particularly for maguey [34,125]. Moreover, no reports exist on the pathogenicity of C. verruculosa in agaves. Therefore, our data can only be compared with previous studies on B. zeae, a pathogen of A. salmiana in Hidalgo, Mexico. In this study, B. zeae caused 33.27% foliar damage, which is higher than the 10.92% reported for the JCP N07 strain of B. zeae in A. salmiana [34]. This difference may be explained by the experimental conditions: our work was conducted during the rainy season (lower temperatures, higher humidity), whereas Romero-Cortes et al. [34] performed their study in the dry season (higher temperatures, lower humidity). Such climatic conditions are critical for fungal pathogens to complete their infection cycle [152,154,155]. In particular, reduced moisture decreases nutrient availability and absorption, which lowers fungal colony density [156,157] and delays the development of somatic (mycelium) and reproductive (asci and conidia) structures. Under dry conditions, this delay extended the appearance of disease symptoms in A. salmiana plants to 90 days. By contrast, our results for treatments T4–T12 (inoculated with C. lunata, C. verruculosa, and B. zeae) showed lower tissue damage than the 66.34% reported by [36] for B. zeae on A. salmiana. The latter difference is attributable to methodology: in that study, plants were artificially wounded prior to inoculation, which altered natural infection processes and bypassed the plant’s defense mechanisms [31].
Once the pathogenicity experiment was concluded, a confirmation step was carried out to verify that the leaf spot symptoms observed in maguey (75 days) and corn (15 days) were caused by conidia of the fungi B. zeae, C. lunata, and C. verruculosa. A total of 9 samples of infected tissue from each host species were incubated in PDA medium. The process led to the isolation of pure colonies of the three pathogenic fungi evaluated under field conditions, namely B. zeae, C. lunata, and C. verruculosa. These isolates exhibited the same macroscopic and microscopic morphological characteristics previously reported from maguey, rice, oil palm, and tule plants [34,125,132,134,158]. Subsequently, each isolate was genetically characterized to confirm the identity of each pathogen recovered from the plants of A. salmiana, A. lechuguilla, and Z. mays (Table 7).
The results of pairwise sequence alignment using the NCBI BLAST program for the 9 DNA sequences obtained from infected tissues of A. salmiana, A. lechuguilla, and Z. mays plants showed identity percentages of 99 to 100% with sequences from closely related fungal accessions in the GenBank database. Isolates were then identified to the species level using the ITS1/5.8s/ITS2 rRNA region, and the data were deposited in GenBank (Table 7). Based on molecular characterization and the fulfillment of Koch’s postulates with the fungi isolated from leaf spots on infected maguey and corn tissues, the pathogenicity of B. zeae, C. lunata, and C. verruculosa was confirmed as the causal agents of leaf spot symptoms in the maguey and corn plants analyzed in this work. This is also the first report of these Helminthosporium complex fungi species as pathogenic microorganisms associated with A. salmiana ssp. ferox and A. lechuguilla plants in field conditions.

3.3. In Vitro Antagonism of Trichoderma Against Pathogenic Fungi in Agave

The antagonistic effect of T. asperellum strain JEAB02 on pathogenic fungi isolated from A. salmiana and A. lechuguilla plants in this study was assessed through dual culture confrontation experiments. BCI values were calculated from digital images taken 7 days after confronting all tested pathogenic strains. The interaction of T. asperellum strain JEAB02 in the in vitro antagonism experiment resulted in significantly different levels of inhibition of the Agave phytopathogens tested. The most susceptible isolate was the strain JCPN27, with 100% growth inhibition. It was followed by isolates from strains JCPN18, JCPN24, JCPN28, JCPN29, JCPN31, and JCPN33, which showed growth inhibition percentages ranging from 99.81% to 99.98%. In contrast, isolates from strains JCPN16 and JCPN26 exhibited the lowest inhibition values (Table 8).
Regarding this topic, the ability of fungi to grow rapidly, compete for space and nutrients with antagonistic fungi, and coexist with phytopathogenic microorganisms is an advantageous trait when using them as biological control agents [124,161]. Although the JEAB02 strain of T. asperellum grew quickly in most Petri dishes, the BCI values recorded against the different strains of pathogenic fungi tested ranged from 89.55 to 100%. In this context, studies evaluating the genetic variations observed in isolates of the same species from different geographical regions have reported that the spatiotemporal distribution of a species across a large geographical area could explain the differences in virulence observed between strains of the same fungal species [162,163,164]. This information is particularly relevant when considering that most of the fungal species reported in this work have a global distribution [36,85,112,148,165,166,167,168,169,170,171].
It has been reported that some pathogenic strains of the genus Fusarium, by releasing mycotoxins into the culture medium, can affect genes related to the mycoparasitism process of Trichoderma species [172]. These results align with the lower growth inhibition values recorded in the JCPN16 F. lactis strain evaluated in this study. Although some variation was observed in the biocontrol effect exerted by the T. asperellum strain JEAB02 assessed in this research, most of the pathogenic strains tested showed growth inhibition percentages of 99% or higher, and Trichoderma colonized the surface of the pathogenic strains, which were sporulating on them. According to the classification by Ezziyyani et al. [173], the JEAB02 strain can be considered a strong candidate for biological control of pathogenic fungi that attack Agave plants.
On the other hand, when comparing the results obtained in this study for the interaction of the T. asperellum strain JEAB02 with each pathogenic fungus tested, our data are consistent with reports from other authors on different T. asperellum strains for the pathogenic fungi C. lunata, Bipolaris oryzae, Fusarium oxysporum [174,175], Fusarium sp. [176], A. niger [177], and Colletotrichum spp. [178]. Conversely, some studies have reported lower inhibition rates, ranging from 41.73 and 61.66% for different strains of the Trichoderma genus when confronted with the Curvularia eragrostidis pathogen affecting pineapple crops [179].
Similarly, in studies with other Trichoderma species, Arispe et al. [180] reported antagonistic effects greater than 67.74% for strains of T. harzianum and T. longibrachiatum against several pathogens associated with corn crops, including Alternaria arborescens, Bipolaris shoemakeri, Bipolaris victoriae, Exserohilum sorghinum, Exserohilum longirostratum, Fusarium brevicatenulatum, Penicillium polonicum, Phaeocytostroma ambiguum, and Fusarium equiseti. In addition, strains of T. viride and T. harzianum showed inhibition percentages of 75.04 and 67.83%, respectively, against A. alternata, the causal agent of black point disease in wheat [181]. In another experiment, the biocontrol effect of three antagonistic Trichoderma strains (T. asperellum, T. harzianum, and T. lignorum) was tested individually and in combination against F. oxysporum and Rhizoctonia solani, the pathogens responsible for chili wilt. Disease incidence decreased by 31.35–71.34% under field conditions, with the strongest effect observed when all three antagonistic species were applied together [182]. Since there are no previous reports for most of the species isolated in this study, our results provide the first data on the pathogenicity of these fungal species in A. salmiana and A. lechuguilla plants in Hidalgo, Mexico.

4. Conclusions

Pathogenicity assays supported by Koch’s postulates demonstrated that leaf spot disease in A. salmiana and A. lechuguilla is caused by fungal isolates belonging to the genera Curvularia and Bipolaris, while molecular characterization confirmed the taxonomic identity of these pathogenic species within the group of nine fungal species isolated from field samples. This disease is currently widespread across several municipalities in Hidalgo, Mexico. To date, two Curvularia species (C. lunata and C. verruculosa) and one Bipolaris species (B. zeae) have been identified as responsible for leaf spots on maguey and corn plants. Notably, this study represents the first report of C. lunata and C. verruculosa infecting A. lechuguilla in Mexico. Among the three species, B. zeae is the most prevalent, occurring in the greatest number of municipalities, while C. verruculosa exhibits the highest virulence in the field pathogenicity experiments. Dual culture confrontation assays demonstrated that Trichoderma asperellum strain JEAB02 is an effective biological control agent against these leaf spot pathogens. Consequently, this strain could be employed to manage leaf spot disease in commercial A. salmiana plantations in Mexico.

Author Contributions

Conceptualization, J.A.C.-P. and J.E.A.-B.; methodology, J.A.C.-P.; software, J.A.C.-P.; validation, J.A.C.-P., J.E.A.-B., M.M.A.-O. and T.R.-C.; formal analysis, J.A.C.-P.; investigation, J.A.C.-P., J.E.A.-B. and T.R.-C.; resources, J.A.C.-P., J.E.A.-B. and T.R.-C.; data curation, J.A.C.-P. and J.E.A.-B.; writing—original draft preparation, J.A.C.-P.; writing—review and editing, J.A.C.-P., J.E.A.-B., M.M.A.-O. and T.R.-C.; visualization and supervision, J.A.C.-P. and J.E.A.-B.; project administration, J.A.C.-P. and J.E.A.-B. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Data Availability Statement

Data are contained within the article.

Acknowledgments

All the authors thank the staff of the Escuela Superior de Apan-UAEH for providing all the facilities needed to carry out the field and laboratory experiments. We also appreciate M.M. Armendáriz-Ontiveros’s support in checking and improving the English language of the final manuscript.

Conflicts of Interest

The authors declare no conflicts of interest, and the funders had no role in designing the study, collecting, analyzing, or interpreting data, writing the manuscript, or deciding to publish the results.

References

  1. Castillo, Q.D.; Villarreal, Q.J.A.; Cano, P.A. El género Agave L. bajo cultivo: Taxonomía, distribución y usos. Rev. Cien. For. Mex. 2007, 32, 57–70. [Google Scholar]
  2. Robles, A.M.A.; Cruz, G.A.E. Agave: Importancia y aplicaciones en México. CienciAcierta 2023, 76, 264–288. [Google Scholar]
  3. Servicio Nacional de Inspección y Certificación de Semillas. Agave (Agave spp.), Generalidades de la Red Agaváceas. Gobierno de México. 1 August 2017. Available online: https://www.gob.mx/snics/acciones-y-programas/agave-agave-spp (accessed on 13 December 2024).
  4. García-Mendoza, A.; Cházaro-Basañez, M.J.; Nieto-Sotelo, J.; Sánchez-Teyer, L.; Tapia, E.; Gómez-Leyva, J.; Tamayo-Ordoñez, M.; Narváez-Zapata, J.A.; Rodríguez-Garay, B.; Palomino-Hasbach, G.A.; et al. Panorama del Aprovechamiento de los Agaves en México, 1st ed.; Red Temática Mexicana Aprovechamiento Integral Sustentable y Biotecnología de los Agaves, AGARED: Ciudad de México, México, 2017; pp. 1–49. [Google Scholar]
  5. Onofre-Sánchez, J.; Testón-Franco, N.; Pinón-Vargas, M. La entomofagia y florifagia en el Valle del Mezquital, Hidalgo México, valor cultural y uso alimentario. Sosquua 2022, 4, 9–21. [Google Scholar] [CrossRef]
  6. Rodríguez-Juárez, F.A.; Carrasco-Urbina, H.S.; Hernández-Zapata, A. Pulque: Contenido probiótico y potencial en la industria biotecnológica. RD-ICUAP 2021, 7, 95–110. [Google Scholar] [CrossRef]
  7. Padilla-Camberos, E.; Arrizon, J.; Sandoval, G. Effect of Agave fructan bioconjugates on metabolic syndrome parameters in a murine model. Pharmaceuticals 2023, 16, 412. [Google Scholar] [CrossRef] [PubMed]
  8. Nava-Cruz, N.Y.; Medina-Morales, M.A.; Martínez, J.L.; Rodríguez, R.; Aguilar, C.N. Agave biotecnology: An overview. Crit. Rev. Biotechnol. 2015, 35, 546–559. [Google Scholar] [PubMed]
  9. Gonzáles, M.; Ballesteros, A. Crean Muros Ecológicos con Fibra de Agave y Hielo Seco. 17 August 2015. Available online: https://cuaad.udg.mx/?q=noticia/crean-muros-ecologicos-con-fibra-de-agave-y-hielo-seco (accessed on 7 December 2024).
  10. Carmona, J.E.; Morales-Martínez, T.K.; Mussatto, S.I.; Castillo-Quiroz, D.; Ríos-González, L.J. Propiedades químicas, estructurales y funcionales de la lechuguilla (Agave lechuguilla Torr.). Rev. Mex. Cienc. For. 2017, 8, 100–122. [Google Scholar]
  11. García-Suárez, M.D.; Serrano, H. Agaves de Ornato Asparagaceae. 7 July 2021. Available online: https://tecnoagro.com.mx/no.149/agaves-de-ornato-asparagaceae (accessed on 7 December 2024).
  12. Ceja-Ramírez, R.; González-Eguiarte, D.R.; Ruiz-Corral, J.A.; Rendón-Salcido, L.A.; Flores-Garnica, J.G. Detección de restricciones en la producción de agave azul (Agave tequilana Weber var. azul) mediante percepción remota. Terra Latinoam. 2017, 35, 259–268. [Google Scholar]
  13. Comisión Nacional para el Conocimiento y Uso de la Biodiversidad. Diversidad Biológica Magueys. 25 May 2021. Available online: https://www.biodiversidad.gob.mx/diversidad/alimentos/magueyes/diversidadmagueyes (accessed on 8 December 2024).
  14. Barrientos-Rivera, G.; Esparza-Ibarra, E.L.; Segura-Pacheco, H.R.; Talavera-Mendoza, O.; Sampedro-Rosas, M.L.; Hernández-Castro, E. Caracterización morfológica de Agave angustifolia y su conservación en Guerrero, México. Rev. Mex. Cienc. Agric. 2019, 10, 655–668. [Google Scholar] [CrossRef]
  15. Domínguez-Arista, D.R. Bacanora, mezcal of Sonora: From clandestinity to denomination of origin. Estud. Sociales. Rev. De Aliment. Contemp. Y Desarro. Reg. 2020, 30, 2–23. [Google Scholar]
  16. Servicio de Información Agroalimentaria y Pesquera. Maguey Pulquero: El Estado de Hidalgo Destacó en 2017 con 69.6% de la Producción Nacional. 10 April 2018. Available online: https://www.gob.mx/siap/articulos/maguey-pulquero?idiom=es (accessed on 7 December 2024).
  17. Podda, L.; Meloni, F.; Fenu, G.; Iiriti, G.; Bacchetta, G. The vascular flora of the marine protected area of “Capo Carbonara” (SE-Sardinia). Fl. Medit. 2021, 31, 415–449. [Google Scholar]
  18. Sánchez, J.E.; Torres-Oregón, F.; Pérez-Akaki, P. El mezcal en México: Las tensiones socioculturales con el agroextractivismo. Rev. COPaLa 2020, 9, 143–153. [Google Scholar]
  19. González, L.H.D.; González, U.D.U. Estimación del rendimiento de biomasa y fibra de Agave lechuguilla Torr. en el norte de Zacatecas. Rev. Mex. Cienc. For. 2023, 14, 97–117. [Google Scholar]
  20. Mayorga-Hernández, E.; Rössel-Kipping, D.; Ortiz-Laurel, H.; Quero-Carrillo, A.R.; Amante-Orozco, A. Análisis comparativo en la calidad de fibra de Agave lechuguilla Torr., procesada manual y mecánicamente. Agrociencia 2004, 38, 219–225. [Google Scholar]
  21. Vega, G.M.A.; Álvarez-Rios, G.D.; Figueredo-Urbina, C.J. Sistemas de manejo de agaves pulqueros en el estado de Hidalgo. Pädi Boletín Científico De Cienc. Básicas E Ing. Del ICBI 2023, 10, 92–100. [Google Scholar] [CrossRef]
  22. García-Mendoza, A. Los Agaves de México. Ciencias 2007, 87, 14–23. [Google Scholar]
  23. Guillot, O.D.; Van Der, M.P.; Laguna, L.E.; Rosselló, P.J.A. El género Agave L. en la flora alóctona valenciana. Valencia, España. Monogr. De La Rev. Bouteloua 2009, 3, 94. [Google Scholar]
  24. Domínguez, R.M.S.; González, J.M.L.; González, R.C.; Quiñones, V.C.; Díaz, D.L.; Mireles, O.S.J.; Pérez, M.E. El cultivo in vitro como herramienta para el aprovechamiento, mejoramiento y conservación de especies del género Agave. Investig. Y Cienc. 2008, 41, 53–62. [Google Scholar]
  25. Cuervo-Parra, J.A.; Pérez-España, V.H.; López, P.P.A.; Morales-Ovando, M.A.; Arce-Cervantes, O.; Aparicio-Burgos, J.E.; Romero-Cortes, T. Scyphophorus acupunctatus (Coleoptera: Dryophthoridae): A weevil threatening the production of Agave in Mexico. Fla. Entomol. 2019, 102, 1–9. [Google Scholar]
  26. Servicio Nacional de Sanidad, Inocuidad y Calidad Agroalimentaria (SENASICA). Plagas Reglamentadas del Agave. 10 February 2020. Available online: https://www.gob.mx/senasica/documentos/plagas-reglamentadas-del-agave-110851 (accessed on 12 December 2024).
  27. Quiñones-Aguilar, E.E.; Montoya-Martínez, A.C.; Rincón-Enríquez, G.; López-Pérez, L. Inoculación de bulbilos de Agave tequilana con hongos micorrízicos arbusculares: Efecto en el crecimiento y biocontrol contra Fusarium oxysporum. Cienc. Y Tecnol. Agropecu. 2023, 24, e3043. [Google Scholar] [CrossRef]
  28. Coria-Contreras, J.J.; Mora-Aguilera, G.; Yáñez-Morales, M.J.; Acevedo-Sánchez, G.; Santana-Peñaloza, B.; Mendoza-Ramos, C.; Jiménez-González, L.; Martínez-Bustamante, V.I.; García-Martínez, D.C.; Rubio-Cortés, R. Epidemiología regional aplicada a la caracterización inductive y pronóstico de la mancha gris del agave azul (Cercospora agavicola) en Jalisco, México. Rev. Mex. Fitopatol. 2019, 37, 71–94. [Google Scholar]
  29. Mancera, A.C.P. Detección de Cercospora en Agave a Través del Análisis de Imágenes Multiespectrales. Master’s Thesis, Instituto Tecnológico de Estudios Superiores de Occidente, Tlaquepaque, Jalisco, Mexico, May 2021. [Google Scholar]
  30. Ceballos-Álvarez, A.; Chávez-Díaz, I.F.; Zelaya-Molina, L.X.; Cruz-Cárdenas, C.I.; Mercado-Vargas, T.J.; Reséndiz-Venado, Z. Identificación morfológica de hongos filamentosos aislados de diferentes síntomas asociados a Agave tequilero en los Altos de Jalisco. Cienc. Y Tecnol. Agrop. México 2022, 10, 60–69. [Google Scholar]
  31. Pérez-España, V.H.; Cuervo-Parra, J.A.; Aparicio, B.J.E.; Morales-Ovando, M.A.; Peralta, G.M.; Romero-Cortes, T. Importancia de la capa cuticular durante la colonización del hongo causante de la negrilla en Agave salmiana Otto ex Salm-Dyck ssp. salmiana. Rev. Mex. Cienc. For. 2022, 13, 158–168. [Google Scholar] [CrossRef]
  32. Romero-Cortes, T.; Cuervo-Parra, J.A.; Pérez, E.V.H.; Pescador, R.J.A.; Rangel, C.E. First Report of Leaf Spot of Agave salmiana Caused by Bipolaris zeae in Mexico. 30 May 2022. Available online: https://www.ncbi.nlm.nih.gov/nuccore/ON630338 (accessed on 29 November 2024).
  33. Mora-Aguilera, J.A.; Cabrera-Huerta, E.; Nava-Díaz, C.; Vázquez-López, A.; Camacho-Tapia, M.; Hernández-Catro, E. Primer Informe de Phoma sorghina que Causa Manchas de Hojas Necróticas en Agave-Mezcal (Agave angustifolia) en México. 29 January 2023. Available online: https://www.ncbi.nlm.nih.gov/nuccore/OQ346146 (accessed on 26 May 2025).
  34. Romero-Cortes, T.; Pérez-España, V.H.; Pescador-Rojas, J.A.; Rangel-Cortés, E.; Armendáriz-Ontiveros, M.M.; Cuervo-Parra, J.A. First report of leaf spot disease (“Negrilla”) on Agave salmiana Otto ex Salm-Dyck (ssp. salmiana) plants caused by Bipolaris zeae Zivan in Mexico. Agronomy 2024, 14, 623. [Google Scholar]
  35. Almazán-Morales, A.; Moreno-Godínez, M.E.; Hernández-Castro, E.; Vázquez-Villamar, M.; Mora-Aguilera, J.A.; Cabrera-Huerta, E.; Alvarez-Fitz, P. Phytochemical profile and in vitro activity of Agave angustifolia and A. cupreata extracts against phytopathogenic fungi. Mex. J. Phytopathol. 2022, 40, 169–187. [Google Scholar] [CrossRef]
  36. López, G.I.; Cardozo, M.V.A.; Pérez, E.V.H.; Romero-Cortes, T.; Pescador, R.J.A.; Saucedo, G.M. Nanopartículas de plata como control de Bipolaris zeae Zivan en maguey. Ingenio Y Concienc. Boletín Científico De La Esc. Super. Ciudad Sahagún 2025, 12, 78–84. [Google Scholar]
  37. Cuervo-Parra, J.A.; Pérez, E.V.H.; Zavala-González, E.A.; Peralta-Gil, M.; Aparicio, B.J.E.; Romero-Cortes, T. Trichoderma asperellum strains as potential biological control agents against Fusarium verticillioides and Ustilago maydis in maize. Biocontrol Sci. Technol. 2022, 32, 1–24. [Google Scholar] [CrossRef]
  38. Ramírez-Cariño, H.F.; Guadarrama-Mendoza, P.C.; Sánchez-López, V.; Cuervo-Parra, J.A.; Ramírez-Reyes, T.; Dunlap, C.A.; Valadez-Blanco, R. Biocontrol of Alternaria alternata and Fusarium oxysporum by Trichoderma asperellum and Bacillus paralicheniformis in tomato plants. Antonie Van Leeuwenhoek 2020, 113, 1247–1261. [Google Scholar] [CrossRef]
  39. Cuervo-Parra, J.A.; Sánchez-López, V.; Romero-Cortes, T.; Ramírez-Lepe, M. Hypocrea viridescens ITV43 potential biological control agent for Moniliophthora roreri Cif & Par, Phytophthora megasperma and P. capsici. Afr. J. Microbiol. Res. 2014, 8, 1704–1712. [Google Scholar]
  40. Romero-Cortes, T.; Zavala-González, E.A.; Pérez, E.V.H.; Aparicio-Burgos, J.E.; Cuervo-Parra, J.A. Characterization of Cochliobolus sativus and Pyrenophora teres fungi belonging to the leaf spot complex of barley (Hordeum vulgare) isolated from barley seeds in Mexico. Chil. J. Agric. Anim. Sci. 2021, 37, 277–289. [Google Scholar] [CrossRef]
  41. Juárez-Hernández, J.; Castillo-Hernández, D.; Pérez-Parada, C.; Nava-Galicia, S.; Cuervo-Parra, J.A.; Surian-Cruz, E.; Díaz-Godínez, G.; Sánchez, C.; Bibbins-Martínez, M. Isolation of fungi from a textile industry effluent and the screening of their potential to degrade industrial dyes. J. Fungi 2021, 7, 805. [Google Scholar] [CrossRef]
  42. Johnson, M.; Zaretskaya, I.; Raytselis, Y.; Merezhuk, Y.; McGinnis, S.; Madden, T.L. NCBI BLAST: A better web interface. Nucleic Acids Res. 2008, 36, W5–W9. [Google Scholar] [CrossRef]
  43. Kumar, S.; Stecher, G.; Li, M.; Knyaz, C.; Tamura, K. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol. 2018, 35, 1547–1549. [Google Scholar] [CrossRef]
  44. Reveles, H.R.G.; Maldonado, T.C.; Chávez, R.I.; Muñoz, E.J.J.; Moreno, G.A. Vigencia de los postulados de Koch en la reproducción de Trichinella spiralis en modelo experimental. REDVET 2014, 15, 1–20. [Google Scholar]
  45. Perelló, A.; Sisterna, M.N.; Cortese, P. Scale for appraising the leaf blight of wheat caused by Alternaria triticimaculans. Cereal Res. Commun. 1998, 26, 189–194. [Google Scholar] [CrossRef]
  46. Robledo-D’Angelo, O. Enfermedad de damping-off en plántulas de lechuga: Un modelo didáctico-experimental para la enseñanza de los postulados de Koch. REurEDC 2016, 13, 680–685. [Google Scholar]
  47. Instituto Nacional de Estadística y Geografía (INEGI). Prontuario de Información Geográfica Municipal de los Estados Unidos Mexicanos. Apan, Hidalgo, Clave Geoestadística 13008. 9 December 2009. Available online: https://web.archive.org/web/20170327170244/http://www3.inegi.org.mx/sistemas/mexicocifras/datos-geograficos/13/13008.pdf (accessed on 5 March 2025).
  48. Servició Meteorológico Nacional (SMN) (2010). Estación: 00013002 APAN (DGE) Normales Climatológicas Periodo 1951–2010. 1 December 2010. Available online: https://smn.conagua.gob.mx/es/climatologia/informacion-climatologica/normales-climatologicas-por-estado?estado=hgo (accessed on 5 March 2024).
  49. Romero-Cortes, T.; Tamayo-Rivera, L.; Morales-Ovando, M.A.; Aparicio, B.J.E.; Pérez, E.V.H.; Peralta-Gil, M.; Cuervo-Parra, J.A. Growth and yield of purple Kculli corn plants under different fertilization schemes. J. Fungi 2022, 8, 1–27. [Google Scholar] [CrossRef]
  50. Rodríguez, L.L.; Guevara, H.F.; Ovando, C.J.; Marto, G.J.R.; Ortiz, P.R. Crecimiento e índice de cosecha de variedades locales de maíz (Zea mays L.) en comunidades de la región Frailesca de Chiapas, México. Cult. Trop. 2016, 37, 137–145. [Google Scholar]
  51. Cruz-Vasconcelos, S.T.; Ruiz-Posadas, L.M.; García-Moya, E.; Sandoval-Villa, M.; Cruz-Huerta, N. Growth and CO2 exchange rate of maguey pulquero (Agave salmiana Otto ex Salm-Dyck) obtained by seed. Agrociencia 2020, 54, 911–926. [Google Scholar] [CrossRef]
  52. Cuervo-Parra, J.A.; López-Pérez, P.A.; Aparicio-Burgos, J.E.; Morales-Ovando, M.A.; Romero-Cortes, T. Chalqueño maize (Zea mays L.) yield under different fertilization schemes in the municipality of Apan, Hidalgo, Mexico. Trop. Subtrop. Agroecosystems 2024, 27, 025. [Google Scholar] [CrossRef]
  53. Comisión Nacional del Agua. Reporte del Clima en México. Reporte anual 2020. Available online: https://smn.conagua.gob.mx/tools/DATA/Climatología/Diagnóstico%20Atmosférico/Reporte%20del%20Clima%20en%20México/Anual2020.pdf (accessed on 5 March 2025).
  54. Mata-Santoyo, C.I.; Leyva-Mir, S.G.; Camacho-Tapia, M.; Tovar-Pedraza, J.M.; Huerta-Espino, J.; Villaseñor-Mir, H.E.; García-León, E. Aggressiveness of Bipolaris sorokiniana and Alternaria alternata isolates on wheat cultivars in Mexico. Rev. Mex. Fitopatol. 2018, 36, 432–443. [Google Scholar]
  55. Analytical Software, Statistix 10, 1st ed.; Analytical Software: Tallahassee, FL, USA, 2017; pp. 7–445.
  56. Cuervo-Parra, J.A.; Romero-Cortes, T.; Román, A.R.; Valle, H.J.; Aparicio-Burgos, J.E. Identificación morfológica de hongos aislados de plantas de Garcinia mangostana. Rev. Mex. Cienc. Agric. 2024, 15, e3575. [Google Scholar] [CrossRef]
  57. Mohammadi, A.; Amini, Y. Molecular characterization and identification of Acrostalagmus liteoalbus from Saffron in Iran. Agric. Sci. Dev. 2015, 4, 16–18. [Google Scholar]
  58. Camacho, C.; Boratyn, G.M.; Joukov, V.; Vera, A.R.; Madden, T.L. ElasticBLAST: Accelerating sequence search via cloud computing. BMC Bioinform. 2023, 24, 117. [Google Scholar] [CrossRef]
  59. Zhang, Z.; Schwartz, S.; Wagner, L.; Miller, W. A greedy algorithm for aligning DNA sequences. J. Comput. Biol. 2000, 7, 203–214. [Google Scholar] [CrossRef]
  60. Visagie, C.M.; Bhiya, T.; Kgatle, G. Alternaria Diversity from South Africa. 22 May 2020. Available online: https://www.ncbi.nlm.nih.gov/nuccore/MT505870 (accessed on 25 May 2025).
  61. Camarena-Pozos, D.A.; Flores-Núñez, V.M.; López, M.G.; Partida-Martínez, L.P. Volatiles Emitted by Fungi Associated with the Phyllosphere of Agaves and Cacti are Diverse and able to Promote Plant Growth. 3 April 2020. Available online: https://www.ncbi.nlm.nih.gov/nuccore/MT279265 (accessed on 20 May 2025).
  62. Jang, Y.; Huh, N.; Lee, J.; Lee, J.S.; Kim, G.H.; Kim, J.J. Phylogenetic analysis of major molds inhabiting woods and their discoloration characteristics. Part 2. Genus Penicillium. Holzforschung 2011, 65, 265–270. [Google Scholar] [CrossRef]
  63. Cristobal Alejo, J.; Candelero De La Cruz, J.; Moo Koh, F.A.; Rangel Ortega, A.; Flores Revilla, C. Molecular Identification Phytopathogenic Fungi in Sugar Cane. 5 November 2016. Available online: https://www.ncbi.nlm.nih.gov/nuccore/KY100122 (accessed on 9 May 2025).
  64. Mora-Aguilera, J.A.; Cabrera-Huerta, E.; Nava-Díaz, C.; Hernández-Castro, E. First Report of Epicoccum sorghinum on Agave Mezcal (Agave angustifolia). 19 June 2018. Available online: https://www.ncbi.nlm.nih.gov/nuccore/MH497563 (accessed on 26 May 2025).
  65. Zhang, P.; Qiao, Y.L.; Deng, J.X.; Ma, D.F. First Report of Alternaria alternata Causing leaf spot on Kadsura coccinea in China. Plant Dis. 2020, 104, 3073. [Google Scholar] [CrossRef]
  66. Rodrigues, A.; Mueller, U.G.; Ishak, H.D.; Bacci, M.; Pagnocca, F.C. Ecology of microfungal communities in gardens of fungus-growing ants (Hymenoptera: Formicidae): A year-long survey of three species of attine ants in Central Texas. FEMS Microbiol. Ecol. 2011, 78, 244–255. [Google Scholar] [CrossRef]
  67. Vu, D.; Groenewald, M.; de Vries, M.; Gehrmann, T.; Stielow, B.; Eberhardt, U.; Al-Hatmi, A.; Groenewald, J.Z.; Cardinali, G.; Houbraken, J.; et al. Large-scale generation and analysis of filamentous fungal DNA barcodes boosts coverage for kingdom fungi and reveals thresholds for fungal species and higher taxon delimitation. Stud. Mycol. 2019, 92, 135–154. [Google Scholar] [CrossRef]
  68. Campos-Rivero, G.; Sánchez-Teyer, L.F.; De la Cruz-Arguijo, E.A.; Ramírez-González, M.S.; Larralde-Corona, C.P.; Narváez-Zapata, J.A. Bioprospecting for fungi with potential pathogenic activity on leaves of Agave tequilana Weber var. Azul. J. Phytopathol. 2019, 167, 283–294. [Google Scholar] [CrossRef]
  69. Ma, G.; Wu, X. GenBank Direct Submission. 18 September 2019. Available online: https://www.ncbi.nlm.nih.gov/nuccore/MN481948 (accessed on 26 May 2025).
  70. Martínez-Pérez, A.; Mares-Rodríguez, F.D.; González-Herrera, S.M.; Muñiz-Márquez, D.B.; Rodríguez-Herrera, R.; Rutiaga-Quiñones, O.M. GenBank Direct Submission. 9 January 2023. Available online: https://www.ncbi.nlm.nih.gov/nuccore/OQ202044 (accessed on 26 May 2025).
  71. Valencia, Y.T.; Martin, M.R.; Cruz, L.J.R.; Pérez, B.D.; Magaña, A.A.; Cortes, V.A.; Nexticapan, G.A. Identification and Molecular Characterization Phytopathogenic Fungi in Agave tequilana in Mexico. 2 August 2023. Available online: https://www.ncbi.nlm.nih.gov/nuccore/OR388091 (accessed on 26 May 2025).
  72. Tapia-Tussell, R.; Solis, S.; Pérez-Brito, D. Identification and Molecular Characterization of Ligninolytic Fungi from Yucatan, Mexico. 17 June 2009. Available online: https://www.ncbi.nlm.nih.gov/nuccore/GQ280375.1/ (accessed on 29 November 2024).
  73. Gómez-Cornelio, S.; De la Rosa-García, S.; Partida-Martínez, L.P. Interactions among Dominant Members of a Fungal Community Isolated from Epilithic Biofilms Limestone-Colonizing. 26 July 2016. Available online: https://www.ncbi.nlm.nih.gov/nuccore/KX610322 (accessed on 29 November 2024).
  74. Moo, K.F.A.; Cristóbal, A.J.; Reyes, R.A.; Tun, S.J.M.; Sandoval, L.R.; Ramírez, P.J.A. Activity in vitro of an aqueous extract of Bonellia flammea against fungal phytopathogenic. 15 April 2014. Available online: https://www.ncbi.nlm.nih.gov/nuccore/KJ748697 (accessed on 29 November 2024).
  75. Gutiérrez-Flores, L.M.; López-Reyes, L.; Mauricio-Gutiérrez, A. Control Biológico de Enfermedades Fúngicas en Pinus sp. de Sistemas Forestales de la Sierra Norte de Puebla Como una Alternativa al uso de Plaguicidas. 12 May 2023. Available online: https://www.ncbi.nlm.nih.gov/nuccore/OQ975458 (accessed on 29 May 2025).
  76. Camacho-Tapia, M.; Leyva-Mir, S.G.; Suaste-Dzul, A.P.; Tovar-Pedraza, J.M. First Report of Alternaria Black Spot of Pomegranate Caused by Alternaria alternata in Mexico. 24 October 2017. Available online: https://www.ncbi.nlm.nih.gov/nuccore/1269612204 (accessed on 29 March 2025).
  77. Reyes-Tena, A.; Montoya-Martínez, A.C.; Fernández-Pavía, S.P.; Santillán-Mendoza, R.; Jiménez-Villegas, A.; Rodríguez-Alvarado, G. Leaf Blight of Onion Caused by Stemphylium vesicarium and Alternaria alternata in Mexico. 21 October 2019. Available online: https://www.ncbi.nlm.nih.gov/nuccore/MN596827 (accessed on 29 November 2024).
  78. Romero-Cortes, T.; Pérez, E.V.H.; López, P.P.A.; Rodríguez-Jimenes, G.C.; Robles-Olvera, V.J.; Aparicio, B.J.E.; Cuervo-Parra, J.A. Antifungal activity of vanilla juice and vanillin against Alternaria alternata. J. Food 2019, 17, 375–383. [Google Scholar] [CrossRef]
  79. Visagie, C.M.; Bhiya, T.; Kgatle, G. Alternaria Diversity from South Africa. 22 May 2020. Available online: https://www.ncbi.nlm.nih.gov/nuccore/MT505867 (accessed on 25 May 2025).
  80. Chen, T.; Qi, Y.; Li, C. GenBank Direct Submission. 22 June 2020. Available online: https://www.ncbi.nlm.nih.gov/nuccore/MT645704 (accessed on 29 November 2024).
  81. Manamgoda, D.S.; Cai, L.; McKenzie, E.H.C.; Crous, P.W.; Madrid, H.; Chukeatirote, E.; Shivas, R.G.; Tan, Y.P.; Hyde, K.D. A phylogenetic and taxonomic re-evaluation of the Bipolaris–Cochliobolus–Curvularia Complex. Fungal Divers. 2012, 56, 131–144. [Google Scholar] [CrossRef]
  82. Azizah, K.; Madihan, N.Z.A.; Shahrizim, Z.; Mohd, T.Y.; Nur, A.; Izzati, M.Z. Morphological and molecular characterization of Curvularia and related species associated with leaf spot disease of rice in Peninsular Malaysia. Rend. Lincei Sci. Fis. Nat. 2016, 27, 205–214. [Google Scholar]
  83. Cuervo-Parra, J.A.; Romero-Cortes, T.; Ramírez-Lepe, M. Isolation and Molecular identification of Curvularia lunata/Cochliobolus lunatus causal agent of leaf spot disease of cocoa. In Advances in Science, Biotechnology and Safety of Foods, 1st ed.; Santos, G., García, G.H.S., Nevárez-Moorillón, G.V., Eds.; Asociación Mexicana de Ciencias de los Alimentos: Ciudad de México, Mexico, 2015; pp. 327–334. [Google Scholar]
  84. Manamgoda, D.S.; Rossman, A.Y.; Castlebury, L.A.; Crous, P.W.; Madrid, H.; Chukeatirote, E.; Hyde, K.D. The genus Bipolaris. Stud. Mycol. 2014, 79, 221–288. [Google Scholar] [CrossRef]
  85. Marin-Felix, Y.; Hernández-Restrepo, M.; Crous, P.W. Multi-locus phylogeny of the genus Curvularia and description of ten new species. Mycol. Prog. 2020, 19, 559–588. [Google Scholar] [CrossRef]
  86. Marin-Felix, Y.; Senwanna, C.; Cheewangkoon, R.; Crous, P.W. New species and records of Bipolaris and Curvularia from Thailand. Mycosphere 2017, 8, 1555–1573. [Google Scholar] [CrossRef]
  87. Seifert, K.A.; Aoki, T.; Baayen, R.P.; Brayford, D.; Burgess, L.W.; Chulze, S.; Gams, W.; Geiser, D.; de Gruyter, J.; Leslie, J.F.; et al. The name Fusarium moniliforme should no longer be used. Mycol. Res. 2003, 107, 643–644. [Google Scholar] [CrossRef]
  88. Price, J.L.; Yilmaz, N.; Visagie, C.M. Fungal Diversity Associated with Maize in the Eastern Cape. 11 April 2022. Available online: https://www.ncbi.nlm.nih.gov/nuccore/ON229472 (accessed on 27 May 2025).
  89. Sandoval, M.M.I.; Zavaleta, M.E.; Leyva, M.S.G.; Osnaya, G.M.M.; Soto, R.L.; Nava, D.C. Fungi Associated to Grain and Seed Discoloration and Leaf Spots of Rice in Campeche, Mexico. 24 April 2022. Available online: https://www.ncbi.nlm.nih.gov/nuccore/ON329680 (accessed on 27 May 2025).
  90. Roy, B.; Maitra, D.; Mitra, A.K. GenBank Direct Submission. 11 August 2023. Available online: https://www.ncbi.nlm.nih.gov/nuccore/OR426451 (accessed on 27 May 2025).
  91. Jaklitsch, W.M.; Fournier, J.; Dai, D.Q.; Hyde, K.D.; Voglmayr, H. Valsaria and Valsariales. Fungal Divers. 2015, 73, 159–202. [Google Scholar] [CrossRef]
  92. Ghosh, S.K. GenBank Direct Submission. 11 April 2017. Available online: https://www.ncbi.nlm.nih.gov/nuccore/KY966028 (accessed on 30 November 2024).
  93. Li, C. GenBank Direct Submission. 23 May 2020. Available online: https://www.ncbi.nlm.nih.gov/nuccore/MT529296.1/ (accessed on 30 November 2024).
  94. Omomowo, I.O.; Babalola, O.O. GenBank Direct Submission. 2 September 2021. Available online: https://www.ncbi.nlm.nih.gov/nuccore/OK012380 (accessed on 30 November 2024).
  95. Namakwa, P.; Mumbi, L.; Ouko, A. Investigation of Aflatoxin-Causing Fungal Isolates Obtained from Soil, Seeds and Diseased Plant Samples. 3 August 2023. Available online: https://www.ncbi.nlm.nih.gov/nuccore/OR397993 (accessed on 30 November 2024).
  96. Rubio, C.R. Enfermedades del cultivo de agave. In Conocimiento y Prácticas Agronómicas para la Producción de Agave tequilana Weber en la Zona de Denominación de Origen del Tequila, 1st ed.; Rulfo, V.F.O., Pérez, D.J.F., del Real, L.J.I., Byerly, M.K.F., Eds.; Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Centro de Investigación Regional del Pacífico Centro: Tepatitlán de Morelos, Jalisco, Mexico, 2007; pp. 169–195. [Google Scholar]
  97. Servicio Nacional de Sanidad, Inocuidad y Calidad Agroalimentaria. Medidas de Manejo de Aplicación Contra Alternaria alternata (Fr.) Keissler. 1 January 2023. Available online: https://www.gob.mx/cms/uploads/attachment/file/979054/34._Medidas_de_manejo_Alternaria_alternata.pdf (accessed on 20 June 2025).
  98. AGQLabs. Requerimientos Técnicos para el Cultivo de Agave. 29 January 2024. Available online: https://agqlabs.mx/2024/01/29/cultivo-de-agave/ (accessed on 8 June 2025).
  99. Thomma, B.P. Alternaria spp.: From general saprophyte to specific parasite. Mol. Plant Pathol. 2003, 4, 225–236. [Google Scholar] [CrossRef]
  100. Quezada, S.A.; García, A.C.J.; Florencio, A.J.G.; Hernández, P.S.; Ruiz, G.I.; Bravo, P.D.; Pineda, R.J.M. Marchitez del Agave. Fusarium verticillioides. 2016. Available online: https://www.gob.mx/cms/uploads/attachment/file/244027/Ficha_T_cnica_Fusarium_verticillioides_en_agave_Versi_n_FINAL.pdf (accessed on 10 July 2025).
  101. Quezada, S.A.; García, A.C.J.; Florencio, A.J.G.; Hernández, P.S.; Ruiz, G.I.; Bravo, P.D.; Pineda, R.J.M. Marchitez del Agave. Fusarium oxysporum. 2003. Available online: https://www.gob.mx/cms/uploads/attachment/file/244026/Ficha_T_cnica_Fusarium_oxysporum_en_agave_Versi_n_FINAL.pdf (accessed on 10 July 2025).
  102. Bailón-Ortiz, A.Z.; Amábilis-Sánchez, M.J. Identificación de agentes causales de la pudrición seca de Agave potatorum Zucc en vivero. Rev. Mex. De Agroecosistemas 2024, 2, 11–20. [Google Scholar] [CrossRef]
  103. Van Poucke, K.; Monbaliu, S.; Munaut, F.; Heungens, K.; De Saeger, S.; Van, H.F. Genetic diversity and mycotoxin production of Fusarium lactis species complex isolates from sweet pepper. Int. J. Food Microbiol. 2012, 153, 28–37. [Google Scholar] [CrossRef]
  104. Imran, M.; Khanal, S.; Zhou, X.G.; Antony-Badu, S.; Atiq, M. First report of leaf spot of rice caused by Epicoccum sorghinum in the United States. Plant Dis. 2022, 106, 2758. [Google Scholar] [CrossRef]
  105. Liu, L.M.; Zhao, Y.; Zhang, Y.L.; Wang, L.; Hou, Y.X.; Huang, S.W. First report of leaf spot disease on rice caused by Epicoccum sorghinum in China. Plant Dis. 2020, 104, 2735. [Google Scholar] [CrossRef]
  106. Servicio Nacional de Sanidad, Inocuidad y Calidad Agroalimentaria (SENASICA). Monitor Fitosanitario. 26 August 2022. Available online: https://dj.senasica.gob.mx/Contenido/files/2022/agosto/MonitorFitosanitario26082022_3051d5a2-352e-475d-a38d-d622f8d1e833.pdf (accessed on 10 June 2025).
  107. Kang, Y.; Zhang, J.; Wan, Q.; Xu, T.; Li, C.; Cao, H. First report of leaf brown spot caused by Epicoccum sorghinum on Digitaria sanguinalis in China. Plant Dis. 2019, 103, 1787. [Google Scholar] [CrossRef]
  108. Laurel, N.R.; De Torres, R.L.; Mendoza, J.V.S.; Balendres, M.A.O.; De la Cueva, F.M. Identification of Epicoccum sorghinum and its effect on stalk sugar yield. Sugar Tech. 2021, 23, 1383–1392. [Google Scholar] [CrossRef]
  109. Chen, Q.H.; Li, J.X.; Qi, Y.; Liu, D.H.; Miao, Y.H. First report of leaf spot on white Chrysanthemum (Chrysanthemum morifolium) caused by Epicoccum sorghinum in Hubei province, China. Plant Dis. 2021, 105, 1212. [Google Scholar] [CrossRef]
  110. Gutiérrez-Flores, L.M.; Mauricio-Gutiérrez, A.; Carcaño-Montiel, M.C.; Portillo-Manzano, E.; Gómez-Velázquez, L.; Sánchez-Alonso, P.; López-Reyes, L. Fungi associated with sick trees of Pinus patula in Tetela de Ocampo, Puebla, Mexico. Arch. Phytopathol. Plant Prot. 2020, 53, 591–611. [Google Scholar] [CrossRef]
  111. Gutiérrez, F.L.M. Diagnóstico de la Enfermedad Fúngica de Pinus patula y Propuesta de Biocontrol en Tetela de Ocampo, Puebla. Master’s Thesis, Universidad Autónoma de Puebla, Posgrado En Ciencias Ambientales, Heroica Puebla de Zaragoza, Puebla, Mexico, October 2018. [Google Scholar]
  112. Gutiérrez-Flores, L.M.; López-Reyes, L.; Hipólito-Romero, E.; Torres-Ramírez, E.; Castañeda-Roldán, E.I.; Mauricio-Gutiérrez, A. Biological control perspectives in the pine forest (Pinus spp.), an environmentally friendly alternative to the use of pesticides. Rev. Mex. Fitopatol. 2022, 40, 401–424. [Google Scholar] [CrossRef]
  113. Castro, C.J.; Umaña, R.G. Poblaciones e identificación de los hongos causantes de mohos poscosecha en el pedúnculo de la piña, en dos zonas de Costa Rica. Agron. Costarric. 2015, 39, 61–77. [Google Scholar]
  114. Gipuzkoa. Enfermedad, Penicillium (Penicillium ssp.). 1999. Available online: https://www.gipuzkoa.eus/es/web/sagarrondoak/-/peniciliuma (accessed on 19 May 2025).
  115. Stošić, S.; Ristić, D.; Savković, Z.; Ljaljević, G.M.; Vukojević, J.; Živković, S. Penicillium and Talaromyces species as postharvest Pathogens of pear fruit (Pyrus communis) in Serbia. Plant Dis. 2021, 105, 3510–3521. [Google Scholar] [CrossRef]
  116. Ephytia. Penicillium expansum Link 1809 y Otras Penicillium spp. 6 October 2021. Available online: https://ephytia.inra.fr/es/C/6100/VID-Podredumbre-por-Penicillium (accessed on 20 May 2025).
  117. Plantix. Pudrición de la Mazorca por Penicillium. 2025. Available online: https://plantix.net/es/library/plant-diseases/100053/penicillium-ear-rot/ (accessed on 20 May 2025).
  118. Awuchi, C.G.; Ondari, E.N.; Ogbonna, C.U.; Upadhyay, A.K.; Baran, K.; Okpala, C.O.R.; Korzeniowska, M.; Guiné, R.P.F. Mycotoxins affecting animals, foods, humans, and plants: Types, occurrence, toxicities, action mechanisms, prevention, and detoxification strategies—A revisit. Foods 2021, 10, 1279. [Google Scholar] [CrossRef]
  119. Daba, G.M.; Mostafa, F.A.; Elkhateeb, W.A. The ancient koji mold (Aspergillus oryzae) as a modern biotechnological tool. Bioresour Bioprocess 2021, 8, 1–17. [Google Scholar] [CrossRef]
  120. García-Conde, K.B.; Cerna-Chávez, E.; Ochoa-Fuentes, Y.M.; Velázquez-Guerrero, J.J. Aspergillus oryzae: An opportunity for agriculture. Mex. J. Phytopathol. 2024, 42, 1–18. [Google Scholar] [CrossRef]
  121. Food and Drug Administration. Generally Recognized Safe (GRAS)|FDA. 17 October 2023. Available online: https://www.fda.gov/food/food-ingredients-packaging/generally-recognized-safe-gras (accessed on 19 July 2025).
  122. Gad, S.E. Generally recognized as safe (GRAS). In Encyclopedia of Toxicology, 1st ed.; Wexler, P., Anderson, B.D., Eds.; Gad Consulting Services, Inc., US National Library of Medicine: Bethesda, MD, USA, 2005; Volume 2, pp. 417–420. [Google Scholar]
  123. Cardona, R.; Suleima, G.M. Caracterización y patogenicidad de hongos del complejo Helminthosporium asociados al cultivo del arroz en Venezuela. Bioagro 2008, 20, 141–145. [Google Scholar]
  124. Hernández-Navarro, E.; Agustín-Maravilla, G.A.; Sánchez-Rangel, J.C.; Valadez-Ramírez, P.; Chan-Cupul, W. Evaluación in vitro de fungicidas biológicos contra Curvularia eragrostidis, hongo fitopatógeno del cultivo de piña. Rev. Mex. Fitopatol. 2023, 41, 93–111. [Google Scholar]
  125. Hernández, J.J.; Hernández, D.E.M.; López, V.E.; Álvarez, C.J. Aislamiento e identificación del fitopatógeno causal de viruela o “negrilla” en Agave salmiana de municipios del estado de Hidalgo, México. Sci. Fungorum 2022, 53, e1425. [Google Scholar] [CrossRef]
  126. Moparthi, S.; Kleczewski, N. First report of Curvularia leaf spot on Zea mays caused by Curvularia lunata in North Carolina. Plant Dis. 2023, 107, 2242. [Google Scholar] [CrossRef]
  127. Wang, S.; Lu, Z.; Lang, B.; Wang, X.; Li, Y.; Chen, J. Curvularia lunata and Curvularia leaf spot of maize in China. ACS Omega 2022, 7, 47462–47470. [Google Scholar] [CrossRef]
  128. Zhou, H.K.; Liu, Y.L.; Tang, J.R.; Zhong, F.T.; Li, Y. First report of leaf spot caused by Curvularia lunata on wild rice in China. Plant Dis. 2021, 105, 3300. [Google Scholar] [CrossRef]
  129. Servicio Nacional de Sanidad, Inocuidad y Calidad Agroalimentaria. Ficha Técnica Curvularia lunata, 1st ed.; Secretaría de Agricultura y Desarrollo Rural: Ciudad de México, México, 2018; pp. 1–17. [Google Scholar]
  130. Estrada, G.; Sandoval, I. Patogenicidad de especies de Curvularia en arroz. Fitosanidad 2004, 8, 23–26. [Google Scholar]
  131. Manzar, N.; Kashyap, A.S.; Sharma, P.K.; Srivastava, A.K. First report of leaf spot on maize caused by Curvularia verruculosa in India. Plant Dis. 2024, 108, 793. [Google Scholar] [CrossRef]
  132. Palma, A.A.A.; Ureta, J.C.; Rodríguez, J.D. Caracterización morfológica de hongos fitopatógenos asociados a cuatro cultivares de arroz (Oryza sativa) de Panamá. Investig. Agropecu. 2023, 6, 39–52. [Google Scholar] [CrossRef]
  133. Choi, Y.J.; Lee, J.S.; Lee, H.B.; Kim, H.J. First report of leaf blight caused by Curvularia verruculosa on Zoysiagrass (Zoysia japonica) in Korea. Plant Dis. 2018, 102, 1173. [Google Scholar] [CrossRef]
  134. Fernández-Mendiola, J.A.; Silva-Valenzuela, M.; Zita-Padilla, G.A.; Espadas-Reséndiz, M.; Colección de Hongos Fitopatógenos. AgroUNAM. Facultad de Estudios Superiores Cuautitlán. UNAM. 2020. Available online: https://virtual.cuautitlan.unam.mx/agrounam/Fitopatologia/Catalogo_hongos_fitopatogenos/pdf/6-Curvularia_verruculosa.pdf (accessed on 4 December 2024).
  135. Huang, J.; Zheng, L.; Hsiang, T. First report of leaf spot caused by Curvularia verruculosa on Cynodon sp. in Hubei, China. Plant Pathol. 2025, 54, 253. [Google Scholar] [CrossRef]
  136. Wei, T.; Luo, M.; Zhang, H.; Jia, W.; Zeng, Y.; Jiang, Y. Curvularia verruculosa as new causal pathogen of common vean leaf spot disease in China. Crop Prot. 2022, 162, 106091. [Google Scholar] [CrossRef]
  137. Sisterna, M.; Wolcan, S. Bipolaris zeae on Pennisetum clandestinum in Argentina. Summa Phytopathol. 1990, 16, 184–188. [Google Scholar]
  138. Zibani, A.; Benslimane, H.; Sicora, O.; Marian, M. New Records of Bipolaris Species from Algeria. 10 August 2022. Available online: https://www.ncbi.nlm.nih.gov/nuccore/OP218553 (accessed on 8 July 2025).
  139. Qi, Z. Bipolaris zeae D759 Wheat Isolate. 23 March 2025. Available online: https://www.ncbi.nlm.nih.gov/nuccore/PV368006 (accessed on 8 July 2025).
  140. Chen, T.X.; Qi, Y.J.; Wang, L.H.; Li, C.J. First report of leaf spot disease on Fagopyrum esculentum caused by Bipolaris zeae in China. Plant Dis. 2021, 105, 3301. [Google Scholar] [CrossRef]
  141. Arce-Cervantes, O.; Castañeda-Ovando, A. Diversidad de agaves utilizados para la producción de jarabe de aguamiel en el estado de Hidalgo, México. Polibotánica 2024, 58, 265–290. [Google Scholar]
  142. Liu, Y.; He, P.H.; He, P.; Munir, S.; Wu, Y.; Tang, P.; He, Y.; Kong, B. Bipolaris zeae isolate BSE1 from Axonopus compressus. 22 May 2023. Available online: https://www.ncbi.nlm.nih.gov/nuccore/OR018922.1/ (accessed on 8 July 2025).
  143. Xu, L.; Qian, Y.; Xue, L.; Yu, B.; Li, C. First report of Bipolaris zeae causing leaf spot on Setaria viridis in China. Plant Dis. 2023, 107, 2862. [Google Scholar] [CrossRef]
  144. Xue, L.H.; Liu, Y.; Wu, W.X.; Liang, J.; Zhang, L.; Huang, X.Q.; Johnson, R.D.; Li, C.J.; Wang, J.J.; Hu, J.Q. First report of leaf spot of Hemarthria altissima caused by Bipolaris zeae in China. Plant Dis. 2017, 101, 243. [Google Scholar] [CrossRef]
  145. Sivanesan, A. Graminicolous Species of Bipolaris, Curvularia, Drechslera, Exserohilum and Their Teleomorphs, 10th ed.; Mycol. Pap., CAB International Mycological Institute: Wallingford, UK, 1987; No. 158; pp. 10–267. [Google Scholar]
  146. Espinosa, B.L.A. Generalidades e importancia de los agaves en México. Herb. CICY 2015, 7, 161–164. [Google Scholar]
  147. Prado-Martínez, M.; Anzaldo-Hernández, J.; Becerra-Aguilar, B.; Palacios-Juárez, H.; Vargas-Radillo, J.J.; Rentería-Urquiza, M. Caracterización de hojas de mazorca de maíz y de bagazo de caña para la elaboración de una pulpa celulósica mixta. Madera Y Bosques 2012, 18, 37–51. [Google Scholar] [CrossRef]
  148. Sun, X.; Qi, X.; Wang, W.; Liu, X.; Zhao, H.; Wu, C.; Chang, X.; Zhang, M.; Chen, H.; Gong, G. Etiology and symptoms of maize leaf spot caused by Bipolaris spp. in Sichuan, China. Pathogens 2020, 9, 229. [Google Scholar] [CrossRef]
  149. Xiao, S.Q.; Zhang, D.; Zhao, J.M.; Yuan, M.Y.; Wang, J.H.; Xu, R.D.; Li, G.F.; Xue, C.S. First report of leaf spot of maize (Zea mays) caused by Bipolaris setariae in China. Plant Dis. 2019, 104, 582. [Google Scholar] [CrossRef]
  150. Henrickson, M.; Koehler, A.M. First report of Curvularia lunata causing curvularia leaf spot of corn in Delaware. Plant Dis. 2022, 106, 319. [Google Scholar] [CrossRef]
  151. GBIF Occurrence Download. Available online: https://doi.org/10.15468/c3kkgh (accessed on 9 July 2025).
  152. Almaguer, M.; Rojas, T.I.; Dobal, V.; Bastita, A.; Aira, M.J. Efecto de la temperatura sobre el crecimiento y germinación de conidios en especies de Curvularia y Bipolaris aisladas del aire. Aerobiología 2012, 29, 1–8. [Google Scholar]
  153. Ríos, H.E.N.; Ochoa, F.Y.M.; Cerna, C.E.; Landeros, F.J.; Cepeda, M.S.; Rodríguez, G.R. Fungi associated with the tar spot in maize cultivation in Mexico. Rev. Mex. Cienc. Agric. 2017, 8, 457–462. [Google Scholar]
  154. LA PIÑORRA. La Importancia de la Lluvia para los Hongos. 2025. Available online: https://www.lapinorravinuesa.com/blog/la-importancia-de-la-lluvia-para-los-hongos/ (accessed on 12 July 2025).
  155. Noriega-Cantú, D.H.; Toledo-Aguilar, R.; Vazquez-Ortiz, R.; Alejo-Jaimes, A.; Garrido-Ramírez, E.R.; Pereyda-Hernández, J.; González-Mateos, R. Relationship between spore fluctuations, environmental conditions and severity of calyx spot on roselle (Hibiscus sabdariffa). Mex. J. Phytopathol. 2020, 38, 1–124. [Google Scholar] [CrossRef]
  156. Casadiego, U.M.C. Efecto del Incremento de la Temperatura Sobre Hongos Edáficos en un Bosque Altoandino (Cuenca del río Blanco, Cundinamarca). Bachelor’s Thesis, Pontificia Universidad Javeriana, Bogotá, Colombia, June 2011. [Google Scholar]
  157. Ramos, R.; Meza, V. Efectos de algunos factores meteorológicos sobre la concentración de esporas de hongos en la plaza San Martín de Lima. Ecol. Apl. 2017, 16, 143–149. [Google Scholar] [CrossRef]
  158. Priwiratama, H.; Wiyono, S.; Hendrastuti, H.S.; Wening, S.; Toding, T.E. Identification and characterization of Curvularia, the causal agent of leaf spot disease of oil palm seedlings in Indonesia. J. Saudi Soc. Agric. Sci. 2025, in press. [CrossRef]
  159. Cuervo-Parra, J.A.; Romero-Cortes, T.; Aparicio-Burgos, J.E. Curvularia lunata Strain JCPN21. 25 May 2023. Available online: https://www.ncbi.nlm.nih.gov/nuccore/OR047534 (accessed on 10 July 2025).
  160. Cuervo-Parra, J.A.; Romero-Cortes, T.; Aparicio-Burgos, J.E. Curvularia verruculosa Strain JCPN30. 25 May 2023. Available online: https://www.ncbi.nlm.nih.gov/nuccore/OR047538 (accessed on 10 July 2025).
  161. Gonzalez-Chingaté, E.J.; Lievano, K.S.; Cubillos, D.D. Evaluación de la efectividad de antagonismo de Trichoderma sp. sobre diferentes hongos fitopatógenos presentes en el cultivo de maíz (Zea mays) en condiciones in vitro. Rev. Cienc. Agropec. 2019, 6, 19–34. [Google Scholar] [CrossRef]
  162. Blixt, E.; Olso, Å.; Lindahl, B.; Djurle, A.; Yuen, J. Spatiotemporal variation in the fungal community associated with wheat leaves showing symptoms similar to stagonospora nodorum blotch. Eur. J. Plant Pathol. 2009, 126, 373–386. [Google Scholar] [CrossRef]
  163. Çelik, O.A.; Karakaya, A. Genetic diversity of barley foliar fungal pathogens. Agronomy 2021, 11, 434. [Google Scholar] [CrossRef]
  164. Kaul, N.; Kashyap, P.L.; Kumar, S.; Singh, D.; Pratap, S.G. Genetic diversity and population structure of head blight disease causing fungus Fusarium graminearum in northern wheat belt of India. J. Fungi 2022, 8, 820. [Google Scholar] [CrossRef]
  165. Li, Q.; Li, M.; Jiang, Y.; Wang, S.; Xu, K.; Liang, X.; Niu, J.; Wang, C. Assessing genetic resistance in wheat to black point caused by six species in the Yellow and Huai wheat area of China. Plant Dis. 2020, 104, 3131–3134. [Google Scholar] [CrossRef]
  166. Pérez, R.M.C.J.; Sánchez, H.G.; Martínez, F.R.; Garza, R.J.L.; Espinosa, R.J. Género Aspergillus Características e Importancia. 2021. Available online: https://masam.cuautitlan.unam.mx/mohos_toxigenos_unigras/aspergillus.html (accessed on 5 December 2024).
  167. Pérez, R.M.C.J.; Sánchez, H.G.; Martínez, F.R.; Garza, R.J.L.; Espinosa, R.J. Género Penicillium Características e Importancia. 2021. Available online: https://masam.cuautitlan.unam.mx/mohos_toxigenos_unigras/penicillium.html (accessed on 22 May 2025).
  168. Duke, R.; Fischer, C. m16 Curvularia lunata Scientific Information. January 2022. Available online: https://www.thermofisher.com/phadia/wo/en/resources/allergen-encyclopedia/m16.html (accessed on 19 August 2025).
  169. Servicio Nacional de Sanidad, Inocuidad y Calidad Agroalimentaria. Alternaria alternata Mancha Foliar y Tizón del Amaranto, 1st ed.; Servicio Nacional de Sanidad, Inocuidad y Calidad Agroalimentaria: Ciudad de México, México, 2023; pp. 1–7. [Google Scholar]
  170. Servicio Nacional de Sanidad, Inocuidad y Calidad Agroalimentaria. Fusarium spp. (Hypocreales: Nectriaceae) Podredumbre de Raíces, 1st ed.; Secretario de Agricultura y Desarrollo Rural: Ciudad de México, México, 2020; pp. 1–19. [Google Scholar]
  171. Arunakumar, G.S.; Nisarga, P.M.N.R.; Arya, N.R.; Monika, B.M.; Dolma, C.S.; Anupama, C.; Akhil, S.; Supriya, K.; Supriya, M.; Sruthi, S.; et al. Diversity of fungal pathogens in leaf spot disease of Indian mulberry and its management. Heliyon 2023, 9, e21750. [Google Scholar] [CrossRef]
  172. El_Komy, M.H.; Saleh, A.A.; Eranthodi, A.; Molan, Y.Y. Characterization of novel Trichoderma asperellum isolates to select effective biocontrol agents against tomato fusarium wilt. Plant Pathol. J. 2015, 31, 50–60. [Google Scholar] [CrossRef]
  173. Ezziyyani, M.; Pérez, S.C.; Requena, M.E.; Rubio, L.; Candela, M.E. Biocontrol por Streptomyces rochei –Ziyani–, de la podredumbre del pimiento (Capsicum annuum L.) causada por Phytophthora capsici. An. Biol. 2004, 26, 69–78. [Google Scholar]
  174. Infante, D.; González, N.; Reyes, Y.; Martínez, B. Evaluación de la efectividad de doce cepas de Trichoderma asperellum Samuels sobre tres fitopatógenos en condiciones de campo. Rev. Prot. Veg. 2011, 26, 194–197. [Google Scholar]
  175. Ramírez-Olier, J.; Trujillo-Salazar, J.; Osorio-Echeverri, V.; Jaramillo-Ciro, M.M.; Botero-Botero, L. Antagonismo in vitro de Trichoderma asperellum contra Colletotrichum gloesporioides, Curvularia lunata, y Fusarium oxysporum. Rev. UIS Ing. 2019, 18, 159–166. [Google Scholar] [CrossRef]
  176. Silva-Martínez, K.L.; Allende-Molar, R.R.; Vázquez-Luna, D.; González-Cárdenas, J.C.; Murguía-González, J. Antagonismo in vitro de Trichoderma asperellum contra Fusarium sp. agente causal de gomosis en cítricos. Agro Product. 2016, 9, 20–25. [Google Scholar]
  177. Romero-Cortes, T.; López-Pérez, P.A.; Pérez, E.V.H.; Medina-Toledo, A.K.; Aparicio-Burgos, J.E.; Cuervo-Parra, J.A. Confrontation of Trichoderma asperellum VSL80 against Aspergillus niger via the effect of enzymatic production. Chil. J. Agric. Anim. Sci. 2019, 35, 68–80. [Google Scholar] [CrossRef]
  178. Infante, M.D.; Martínez, C.B. Antagonismo de seis cepas de Trichoderma asperellum Samuels, Lieckfeldt & Nirenberg sobre Colletotrichum spp. Rev. Prot. Veg. 2020, 35, 1–8. [Google Scholar]
  179. Iglesias-Velasco, S.J.; Chan-Cupul, W.; Osuna-Castro, J.A.; Hernández-Ortega, H.A.; Centeno-Leija, S. Antagonismo in vitro, actividad ligninolítica y de hidrolasas de pared celular en la interacción de especies de Trichoderma spp. con Curvularia eragrostidis aislado de piña. Sci. Fungorum 2022, 53, e1430. [Google Scholar]
  180. Arispe, V.J.L.; Sánchez, A.A.; Galindo, C.M.E.; Vázquez, B.M.E.; Oyervides, G.A.; Rodríguez, G.R. Antagonismo de Trichoderma spp. en hongos asociados al daño de Diatraea saccharalis Fabricius (Lepidoptera: Crambidae) en maíz. Bol. Microbiol. 2019, 34, 17–24. [Google Scholar]
  181. Yassin, M.T.; Mostafa, A.A.F.; Al-Askar, A.A. In vitro antagonistic activity of Trichoderma spp. against fungal pathogens causing black point disease of wheat. J. Taibah Univ. Sci. 2022, 16, 57–65. [Google Scholar] [CrossRef]
  182. Gallegos-Morales, G.; Espinoza-Ahumada, C.A.; Figueroa-Reyes, J.; Méndez-Aguilar, R.; Rodríguez-Guerra, R.; Salas-Gómez, A.L.; Peña-Ramos, F.M. Compatibilidad de especies de Trichoderma en la producción y biocontrol de marchitez del chile. Ecosist. Recur. Agropec. 2022, 9, e3066. [Google Scholar] [CrossRef]
Agronomy 15 02406 g001
Figure 1. Neighbor-Joining phylogenetic analysis of pathogenic fungi based on ITS alignment. Numbers at nodes indicate bootstrap values. Three main clades were observed: I (Pleosporaceae, Didymellaceae), II (Nectriaceae, Valsariaceae, Trichocomaceae), and III (Irpicaceae, Mucoraceae), all supported by ≥97% similarity.
Figure 1. Neighbor-Joining phylogenetic analysis of pathogenic fungi based on ITS alignment. Numbers at nodes indicate bootstrap values. Three main clades were observed: I (Pleosporaceae, Didymellaceae), II (Nectriaceae, Valsariaceae, Trichocomaceae), and III (Irpicaceae, Mucoraceae), all supported by ≥97% similarity.
Agronomy 15 02406 g001
Table 1. Geographic origin of the strains isolated from A. salmiana and A. lechuguilla.
Table 1. Geographic origin of the strains isolated from A. salmiana and A. lechuguilla.
StrainsHostSample *SpeciesLocationCoordinatesDate
JCPN09A. salmianaM1Bipolaris zeaeApan19°64′97″ LN, 98°51′93″ LW2019
JCPN10A. salmianaM1Bipolaris zeaeApan19°64′97″ LN, 98°51′93″ LW2020
JCPN11A. salmianaM2Bipolaris zeaeApan19°64′96″ LN, 98°51′91″ LW2020
JCPN12A. salmianaM3Bipolaris zeaeApan19°64′94″ LN, 98°51′88″ LW2020
JCPN13A. salmianaM2Bipolaris zeaeApan19°64′96″ LN, 98°51′91″ LW2021
JCPN14A. salmianaM4Bipolaris zeaeApan19°64′90″ LN, 98°51′84″ LW2021
JCPN7A. salmianaM4Bipolaris zeaeApan19°64′ 90″ LN, 98°51′84″ LW2022
JCPN16A. salmianaM5Fusarium lactisMineral de la Reforma20°07′60″ LN, 98°72′43″ LW2022
JCPN18A. salmianaM6Curvularia lunataMineral de la Reforma20°07′78″ LN, 98°72′00″ LW2022
JCPN19A. lechuguillaM7Curvularia lunataMineral de la Reforma20°07′81″ LN, 98°72′16″ LW2022
JCPN22A. lechuguillaM8Alternaria alternataMineral de la Reforma20°07′67″ LN, 98°72′06″ LW2022
JCPN24A. lechuguillaM9Curvularia verruculosaMineral de la Reforma20°07′51″ LN, 98°72′50″ LW2022
JCPN27A. lechuguillaM10Penicillium diversumMineral de la Reforma20°07′85″ LN, 98°71′57″ LW2022
JCPN28A. lechuguillaM11Epicoccum sorghinumMineral de la Reforma20°07′80″ LN, 98°71′66″ LW2022
JCPN33A. salmianaM12Bipolaris zeaeMineral de la Reforma20°07′93″ LN, 98°71′35″ LW2022
JCPN15A. salmianaM13Fusarium lactisMineral de la Reforma20°07′84″ LN, 98°71′78″ LW2023
JCPN17A. salmianaM14Penicillium diversumMineral de la Reforma20°07′89″ LN, 98°72′18″ LW2023
JCPN20A. lechuguillaM11Epicoccum sorghinumMineral de la Reforma20°07′80″ LN, 98°71′66″ LW2023
JCPN21A. lechuguillaM15Curvularia lunataMineral de la Reforma20°07′78″ LN, 98°72′13″ LW2023
JCPN23A. lechuguillaM16Curvularia lunataMineral de la Reforma20°07′74″ LN, 98°72′09″ LW2023
JCPN25A. lechuguillaM11Curvularia lunataMineral de la Reforma20°07′80″ LN, 98°71′66″ LW2023
JCPN26A. lechuguillaM17Myrmaecium rubricosumMineral de la Reforma20°07′83″ LN, 98°71′57″ LW2023
JCPN29A. lechuguillaM18Alternaria alternataMineral de la Reforma20°07′61″ LN, 98°72′16″ LW2023
JCPN30A. salmianaM19Curvularia verruculosaMineral de la Reforma20°07′53″ LN, 98°72′87″ LW2023
JCPN31A. salmianaM20Aspergillus oryzaeMineral de la Reforma20°07′63″ LN, 98°72′53″ LW2023
JCPN32A. salmianaM21Bipolaris zeaeMineral de la Reforma20°07′85″ LN, 98°71′37″ LW2023
* Percentage of occurrence of fungal isolates by municipality and Agave species. Apan: A. salmiana—B. zeae (100%). Mineral de la Reforma: A. salmiana—B. zeae (18.2%), C. lunata (36.3%), C. verruculosa (9.1%), F. lactis (18.2%), P. diversum (9.1%), A. oryzae (9.1%); A. lechuguilla—C. lunata (12.5%), C. verruculosa (12.5%), A. alternata (25%), P. diversum (12.5%), E. sorghinum (25%), M. rubricosum (12.5%).
Table 2. Experimental design for various pathological experiments evaluated.
Table 2. Experimental design for various pathological experiments evaluated.
KeyTreatment *Inoculation Site
T1A. salmiana controlWithout inoculants
T2A. lechuguilla controlWithout inoculants
T3C. lunata vs. A. salmiana Leaves and rhizosphere
T4C. lunata vs. A. lechuguilla Leaves and rhizosphere
T5C. verruculosa vs. A. salmiana Leaves and rhizosphere
T6C. verruculosa vs. A. lechuguilla Leaves and rhizosphere
T7B. zeae vs. A. salmiana Leaves and rhizosphere
T8B. zeae vs. A. lechuguilla Leaves and rhizosphere
T9A. alternata vs. A. salmiana Leaves and rhizosphere
T10A. alternata vs. A. lechuguilla Leaves and rhizosphere
T11F. lactis vs. A. salmiana Leaves and rhizosphere
T12F. lactis vs. A. lechuguilla Leaves and rhizosphere
T13E. sorghinum vs. A. salmiana Leaves and rhizosphere
T14E. sorghinum vs. A. lechuguilla Leaves and rhizosphere
T15M. rubricosum vs. A. salmiana Leaves and rhizosphere
T16M. rubricosum vs. A. lechuguilla Leaves and rhizosphere
T17P. diversum vs. A. salmiana Leaves and rhizosphere
T18P. diversum vs. A. lechuguilla Leaves and rhizosphere
T19A. oryzae vs. A. salmianaLeaves and rhizosphere
T20A. oryzae vs. A. lechuguilla Leaves and rhizosphere
* For greenhouse experiments, the isolate within each species that exhibited the highest radial growth after 7 days of cultivation was selected. The isolates chosen were: JCPN18 (C. lunata), JCPN24 (C. verruculosa), JCPN33 (B. zeae), JCPN29 (A. alternata), JCPN16 (F. lactis), JCPN28 (E. sorghinum), JCPN26 (M. rubricosum), JCPN27 (P. diversum), and JCPN31 (A. oryzae).
Table 3. Experimental design of field pathological tests.
Table 3. Experimental design of field pathological tests.
KeyTreatment *Inoculation Site ¥
T1A. salmiana controlWithout inoculants
T2A. lechuguilla controlWithout inoculants
T3Z. mays controlWithout inoculants
T4B. zeae vs. A. salmiana Leaves and rhizosphere
T5B. zeae vs. A. lechuguilla Leaves and rhizosphere
T6B. zeae vs. Z. maysLeaves and rhizosphere
T7C. lunata vs. A. salmiana Leaves and rhizosphere
T8C. lunata vs. A. lechuguilla Leaves and rhizosphere
T9C. lunata vs. Z. maysLeaves and rhizosphere
T10C. verruculosa vs. A. salmiana Leaves and rhizosphere
T11C. verruculosa vs. A. lechuguilla Leaves and rhizosphere
T12C. verruculosa vs. Z. maysLeaves and rhizosphere
* Treatments involving B. zeae, C. lunata, and C. verruculosa correspond to the strains JCPN33, JCPN18, and JCPN24, based on their greater growth and pathogenic traits observed during the greenhouse experiment. ¥ In the control treatments T1, T2, and T3, sterilized distilled water was used instead of each pathogenic strain.
Table 4. Fungal strain’s identity based on the ITS 1/5.8s/ITS II region of the rDNA.
Table 4. Fungal strain’s identity based on the ITS 1/5.8s/ITS II region of the rDNA.
StrainGenBank AccessionSequence-Based AssociationPercent IdentityGenBank Closest HitReferences
JCPN7OR553122B. zeae99.63%MT505870Visagie et al. [60]
JCPN09OR553116B. zeae99.61%MT505870Visagie et al. [60]
JCPN10OR553117B. zeae99.60%MT505870Visagie et al. [60]
JCPN11OR553118B. zeae99.43%MT505870Visagie et al. [60]
JCPN12OR553119B. zeae99.60%MT505870Visagie et al. [60]
JCPN13OR553120B. zeae99.60%MT505870Visagie et al. [60]
JCPN14OR553121B. zeae99.63%MT505870Visagie et al. [60]
JCPN15OR047541F. lactis100%MT279265Camarena-Pozos et al. [61]
JCPN16OR047542F. lactis100%MT279265Camarena-Pozos et al. [61]
JCPN17OR047539P. diversum98.88%HM469392Jang et al. [62]
JCPN18OR047532C. lunata100%KY100122Cristóbal et al. [63]
JCPN19OR047533C. lunata99.82%KY100122Cristóbal et al. [63]
JCPN20OR047543E. sorghinum100%MH497563Mora-Aguilera et al. [64]
JCPN21OR047534C. lunata100%KY100122Cristóbal et al. [63]
JCPN22OR047547A. alternata100%MN615420Zhang et al. [65]
JCPN23OR047535C. lunata100%HQ607991Rodrigues et al. [66]
JCPN24OR047537C. verruculosa99.45%MH859788Vu et al. [67]
JCPN25OR047536C. lunata99.82%KY100122Cristóbal et al. [63]
JCPN26OR047545M. rubricosum100%MK041894Campos-Rivero et al. [68]
JCPN27OR047540P. diversum98.91%HM469392Jang et al. [62]
JCPN28OR047544E. sorghinum100%MH497563Mora-Aguilera et al. [64]
JCPN29OR047546A. alternata100%MN481948Ma and Wu [69]
JCPN30OR047538C. verruculosa99.46%MH859788Vu et al. [67]
JCPN31OR047548A. oryzae100%OQ202044Martínez-Pérez et al. [70]
JCPN32OR553111B. zeae99.62%MT505870Visagie et al. [60]
JCPN33OR553112B. zeae100%MT505870Visagie et al. [60]
Table 5. Greenhouse pathogenicity experiment.
Table 5. Greenhouse pathogenicity experiment.
KeyTreatments *Damage Percentage ± SE **
T1A. salmiana control0.00 ± 0.00 f
T2A. lechuguilla control0.00 ± 0.00 f
T3C. lunata vs. A. salmiana 25.29 ± 0.37 b
T4C. lunata vs. A. lechuguilla 24.20 ± 0.18 b
T5C. verruculosa vs. A. salmiana 37.29 ± 1.79 a
T6C. verruculosa vs. A. lechuguilla 36.36 ± 0.32 a
T7B. zeae vs. A. salmiana 34.23 ± 1.27 a
T8B. zeae vs. A. lechuguilla 33.38 ± 0.27 a
T9A. alternata vs. A. salmiana 36.32 ± 0.29 a
T10A. alternata vs. A. lechuguilla 35.64 ± 0.21 a
T11F. lactis vs. A. salmiana 33.10 ± 0.11 a
T12F. lactis vs. A. lechuguilla 34.53 ± 1.66 a
T13E. sorghinum vs. A. salmiana 18.26 ± 0.19 c
T14E. sorghinum vs. A. lechuguilla 18.28 ± 0.22 c
T15M. rubricosum vs. A. salmiana 11.20 ± 0.23 d
T16M. rubricosum vs. A. lechuguilla 11.28 ± 0.08 d
T17P. diversum vs. A. salmiana 7.38 ± 1.21 e
T18P. diversum vs. A. lechuguilla 7.13 ± 0.19 e
T19A. oryzae vs. A. salmiana0.00 ± 0.00 f
T20A. oryzae vs. A. lechuguilla 0.00 ± 0.00 f
* Treatments involving B. zeae, C. lunata, and C. verruculosa correspond to the strains JCPN33, JCPN18, and JCPN24, based on their enhanced growth and pathological traits observed during morphological characterization and greenhouse experiment treatment. ** Means sharing the same letter indicate no significant differences (p < 0.05, ANOVA and Tukey tests); ± Standard error.
Table 6. Field pathogenicity in maguey and maize plants.
Table 6. Field pathogenicity in maguey and maize plants.
KeyTreatments *Damage Percentage ± SE **
T1A. salmiana control0.00 ± 0.00 e
T2A. lechuguilla control0.00 ± 0.00 e
T3Z. mays control0.00 ± 0.00 e
T4B. zeae vs. A. salmiana 33.27 ± 0.41 c
T5B. zeae vs. A. lechuguilla 31.73 ± 0.11 c
T6B. zeae vs. Z. mays41.50 ± 0.94 a
T7C. lunata vs. A. salmiana 23.67 ± 0.61 d
T8C. lunata vs. A. lechuguilla 21.07 ± 0.46 d
T9C. lunata vs. Z. mays32.66 ± 1.88 c
T10C. verruculosa vs. A. salmiana 36.73 ± 0.61 b
T11C. verruculosa vs. A. lechuguilla 34.07 ± 0.30 c
T12C. verruculosa vs. Z. mays38.08 ± 0.35 b
* Treatments involving B. zeae, C. lunata, and C. verruculosa correspond to strains JCPN33, JCPN18, and JCPN24, respectively, based on their enhanced growth and pathogenic traits observed during morphological characterization and greenhouse experiments. ** Means sharing the same letter indicate no significant differences (p < 0.05, ANOVA and Tukey tests); ± Standard error.
Table 7. Molecular identification of the causal agents of leaf spots in maguey and corn.
Table 7. Molecular identification of the causal agents of leaf spots in maguey and corn.
StrainStrain AccessionSpecieHostQuery Cover (%)Accession Closest HitReference
JCPN34PV875568C. lunataA. salmiana100OR047534Cuervo-Parra et al. [159]
JCPN35PV875569C. verruculosaA. salmiana100OR047538Cuervo-Parra et al. [160]
JCPN36PV875570B. zeaeA. salmiana100MT505867Visagie et al. [79]
JCPN37PV875624C. lunataA. lechuguilla100OR047534Cuervo-Parra et al. [159]
JCPN38PV875625C. verruculosaA. lechuguilla100OR047538Cuervo-Parra et al. [160]
JCPN39PV875626B. zeaeA. lechuguilla100MT505867Visagie et al. [79]
JCPN40PV875637C. lunataZ. mays100OR047534Cuervo-Parra et al. [159]
JCPN41PV875638C. verruculosaZ. mays100OR047538Cuervo-Parra et al. [160]
JCPN42PV875639B. zeaeZ. mays99MT505867Visagie et al. [79]
Table 8. Trichoderma asperellum biocontrol index against Agave pathogenic fungi.
Table 8. Trichoderma asperellum biocontrol index against Agave pathogenic fungi.
Pathogenic FungiPathogenic StrainTrichoderma StrainBCI € ± SE *
Penicillium diversumJCPN27JEAB02100 ± 0.0 a
Curvularia lunataJCPN18JEAB0299.98 ± 0.01 b
Bipolaris zeaeJCPN33JEAB0299.89 ± 0.08 b
Curvularia verruculosaJCPN24JEAB0299.86 ± 0.07 b
Alternaria alternataJCPN29JEAB0299.81 ± 0.006 b
Aspergillus oryzaeJCPN31JEAB0299.81 ± 0.007 b
Epicoccum sorghinumJCPN28JEAB0299.81 ± 0.006 b
Fusarium lactisJCPN16JEAB0296.01 ± 0.25 c
Myrmaecium rubricosumJCPN26JEAB0289.55 ± 0.97 d
* Equal letters in the same column indicate that the means did not differ significantly (p < 0.05, ANOVA and Tukey tests) at a confidence level of 95%; ± Standard error. € BCI = Percentage of inhibition of the radial growth of the pathogenic fungal colony due to the antagonistic effect of T. asperellum.
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Aparicio-Burgos, J.E.; Romero-Cortes, T.; Armendáriz-Ontiveros, M.M.; Cuervo-Parra, J.A. Leaf Spot Disease Caused by Several Pathogenic Species of the Pleosporaceae Family on Agave salmiana and Agave lechuguilla Plants in Mexico, and Their Biocontrol Using the Indigenous Trichoderma asperellum Strain JEAB02. Agronomy 2025, 15, 2406. https://doi.org/10.3390/agronomy15102406

AMA Style

Aparicio-Burgos JE, Romero-Cortes T, Armendáriz-Ontiveros MM, Cuervo-Parra JA. Leaf Spot Disease Caused by Several Pathogenic Species of the Pleosporaceae Family on Agave salmiana and Agave lechuguilla Plants in Mexico, and Their Biocontrol Using the Indigenous Trichoderma asperellum Strain JEAB02. Agronomy. 2025; 15(10):2406. https://doi.org/10.3390/agronomy15102406

Chicago/Turabian Style

Aparicio-Burgos, José Esteban, Teresa Romero-Cortes, María Magdalena Armendáriz-Ontiveros, and Jaime Alioscha Cuervo-Parra. 2025. "Leaf Spot Disease Caused by Several Pathogenic Species of the Pleosporaceae Family on Agave salmiana and Agave lechuguilla Plants in Mexico, and Their Biocontrol Using the Indigenous Trichoderma asperellum Strain JEAB02" Agronomy 15, no. 10: 2406. https://doi.org/10.3390/agronomy15102406

APA Style

Aparicio-Burgos, J. E., Romero-Cortes, T., Armendáriz-Ontiveros, M. M., & Cuervo-Parra, J. A. (2025). Leaf Spot Disease Caused by Several Pathogenic Species of the Pleosporaceae Family on Agave salmiana and Agave lechuguilla Plants in Mexico, and Their Biocontrol Using the Indigenous Trichoderma asperellum Strain JEAB02. Agronomy, 15(10), 2406. https://doi.org/10.3390/agronomy15102406

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop