Journal of Fungi

35 pages, 36488 KiB

Open AccessArticle

Revealing Brownish Mycena Diversity in China: New Discoveries and Taxonomic Insights

by Renxiu Wei, Yupeng Ge, Liangliang Qi, Menghui Han, Hui Zeng, Yaping Hu, Li Zou, Xianhao Cheng, Xiaoming Wu and Qin Na

J. Fungi 2024, 10(6), 439; https://doi.org/10.3390/jof10060439 - 20 Jun 2024

Cited by 2 | Viewed by 860

Abstract

Within the genus Mycena, species exhibiting brownish basidiomata present considerable challenges in identification due to similar coloration. This study underscores the significance of pileipellis types and cheilocystidia characteristics as critical in delimiting brownish Mycena species. To clarify the principal taxonomic characters and [...] Read more.

Within the genus Mycena, species exhibiting brownish basidiomata present considerable challenges in identification due to similar coloration. This study underscores the significance of pileipellis types and cheilocystidia characteristics as critical in delimiting brownish Mycena species. To clarify the principal taxonomic characters and their utility in distinguishing between brownish Mycena species, a morphological taxonomy and phylogenetic analysis were performed. Five new species from China were introduced and characterized through a comprehensive morphological anatomy and phylogenetic substantiation: M. campanulatihemisphaerica sp. nov., M. digitifurcata sp. nov., M. kunyuensis sp. nov., M. limitis sp. nov., and M. oryzifluens sp. nov. Discussions of these taxa are supplemented with morphological illustrations. The phylogenetic relationships were inferred using Bayesian Inference and Maximum Likelihood methods based on sequences from the internal transcribed spacer and the large subunit regions of nuclear ribosomal RNA. With the addition of these five new species, the worldwide count of brownish Mycena increases to 94, and a key to the 29 known species of brownish Mycena from China is presented. Full article

(This article belongs to the Special Issue Taxonomy, Systematics and Evolution of Forestry Fungi, 2nd Edition)

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16 pages, 3002 KiB

Open AccessArticle

Volatile Semiochemicals Emitted by Beauveria bassiana Modulate Larval Feeding Behavior and Food Choice Preference in Spodoptera frugiperda (Lepidoptera: Noctuidae)

by Arturo Ramírez-Ordorica, Sandra Goretti Adame-Garnica, Hilda Eréndira Ramos-Aboites, Robert Winkler and Lourdes Macías-Rodríguez

J. Fungi 2024, 10(6), 438; https://doi.org/10.3390/jof10060438 - 20 Jun 2024

Cited by 1 | Viewed by 1262

Abstract

Beauveria bassiana is an entomopathogenic fungus that parasitizes and kills insects. The role of volatile organic compounds (VOCs) emitted by B. bassiana acting as semiochemicals during its interaction with lepidopterans is poorly explored. Here, we studied the effect of VOCs from B. bassiana [...] Read more.

Beauveria bassiana is an entomopathogenic fungus that parasitizes and kills insects. The role of volatile organic compounds (VOCs) emitted by B. bassiana acting as semiochemicals during its interaction with lepidopterans is poorly explored. Here, we studied the effect of VOCs from B. bassiana and 3-methylbutanol (as a single compound) on the feeding behavior of L2 larvae of Spodoptera frugiperda in sorghum plants. Additionally, we assessed whether fungal VOCs induce chemical modifications in the plants that affect larval food preferences. Metabolomic profiling of plant tissues was performed by mass spectrometry and bioassays in a dual-choice olfactometer. The results showed that the larval feeding behavior was affected by the B. bassiana strain AI2, showing that the insect response is strain-specific. Furthermore, 80 µg of 3-methylbutanol affected the number of bites. The larval feeding choice was dependent on the background context. Fragment spectra and a matching precursor ion mass of 165.882 m/z enabled the putative identification of 4-coumaric acid in sorghum leaves exposed to fungal VOCs, which may be associated with larval deterrent responses. These results provide valuable insights into the bipartite interaction of B. bassiana with lepidopterans through VOC emission, with the plant as a mediator of the interaction. Full article

(This article belongs to the Collection Entomopathogenic and Nematophagous Fungi)

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Experimental strategy used to investigate the effect of volatiles emitted by Beauveria bassiana on the larval feeding behavior (A) and (B) food choice preference in Spodoptera frugiperda. Full article ">Figure 2
Spodoptera frugiperda larval feeding behavior in response to volatiles emitted by the AI2 (A–C) and AS5 (D–F) strains of Beauveria bassiana. Panels (A,D) correspond to the feeding area (mm2) (removed tissue after 12 h of consumption). Panels (B,E) show the percentage of herbivory (missing leaf area with respect to the total leaf area). Panels (C,F) correspond to the number of bites per leaf inflicted by larvae. Control refers to treatments with uninoculated PDA culture medium. n = 114–183, glm with gamma distribution, α = 0.05. Full article ">Figure 3
Larval dual-choice olfactometry bioassays in response to pairs of odor sources. (A) The bioassays utilized mycelium from the strains AI2 and AS5 cultured in PDA culture medium. n = 40–63. (B) Bioassays were performed on sorghum foliage unexposed and exposed to VOCs from the strains AI2 and AS5. n = 30–46. The bars indicate the frequency of insects that selected either odor source. Pie charts show proportions of responding (black) and not responding (white) individuals. Statistical significance according to Chi-square two-sides test (* p < 0.05, *** p < 0.001) at α = 0.05. Full article ">Figure 4
Larval feeding behavior of L2 larvae of Spodoptera frugiperda on Sorghum bicolor foliage exposed to 8 and 80 µg of 3-methylbutanol. (A) Feeding area (mm2) (removed tissue after 12 h of consumption). (B) Percentage of herbivory (missing leaf area with respect to the total leaf area). (C) Number of bites per leaf inflicted by larva. n = 114–183, analyzed with glm with gamma distribution, α = 0.05. Different letters indicate statiscally significant differences, ns means not statiscally significant. (D) Olfactometry dual-choice bioassay of L2 larvae exposed to 8 μg of 3-methylbutanol compared to water and artificial diet. The bars indicate the frequency of insects that selected either odor source. Pie charts show proportions of responding (black) and not responding (white) individuals. Statistical significance according to Chi-square two-sides test (*** p < 0.001) at α = 0.05. Full article ">Figure 5
(A) Representative chromatogram of the chemical composition of sorghum foliage exposed to VOCs emitted by B. bassiana. Insets correspond to the abundance of the ion signals 165.882, 353.347, and 437.402 m/z. Green, red, blue, and black colors indicate VOCs AS5, VOCs AI2, 3-methylbutanol, and control treatments, respectively. (B) Heatmap of the normalized signal intensities of the 116 ions obtained from the metabolic profiles. Hierarchical clustering with Euclidean distance. Full article ">Figure 6
(A) Biomarker ions obtained by random forest model for differentiating between the Sorghum bicolor foliage treatments, including exposure to VOCs AI2, VOCs AS5, and 8 µg of 3-methylbutanol. Random forest with 500 trees. (B) Principal component analysis from the chemical profiles of sorghum plants differentially treated with the fungal VOCs. PERMANOVA test, α = 0.05. (C) Mean intensities from the ion signals with the highest Gini index, n = 6. Different letters indicate statiscally significant differences by one-way ANOVA, Tukey post hoc, α = 0.05. Full article ">

17 pages, 10950 KiB

Open AccessArticle

Role of Flavohemoglobins in the Development and Aflatoxin Biosynthesis of Aspergillus flavus

by Xiaoling Zhou, Dongyue Chen, Min Yu, Yuan Jiao and Fang Tao

J. Fungi 2024, 10(6), 437; https://doi.org/10.3390/jof10060437 - 19 Jun 2024

Viewed by 863

Abstract

Aspergillus flavus is notorious for contaminating food with its secondary metabolite—highly carcinogenic aflatoxins. In this study, we found that exogenous nitric oxide (NO) donor could influence aflatoxin production in A. flavus. Flavohemoglobins (FHbs) are vital functional units in maintaining nitric oxide (NO) [...] Read more.

Aspergillus flavus is notorious for contaminating food with its secondary metabolite—highly carcinogenic aflatoxins. In this study, we found that exogenous nitric oxide (NO) donor could influence aflatoxin production in A. flavus. Flavohemoglobins (FHbs) are vital functional units in maintaining nitric oxide (NO) homeostasis and are crucial for normal cell function. To investigate whether endogenous NO changes affect aflatoxin biosynthesis, two FHbs, FHbA and FHbB, were identified in this study. FHbA was confirmed as the main protein to maintain NO homeostasis, as its absence led to a significant increase in intracellular NO levels and heightened sensitivity to SNP stress. Dramatically, FHbA deletion retarded aflatoxin production. In addition, FHbA played important roles in mycelial growth, conidial germination, and sclerotial development, and response to oxidative stress and high-temperature stress. Although FHbB did not significantly impact the cellular NO level, it was also involved in sclerotial development, aflatoxin synthesis, and stress response. Our findings provide a new perspective for studying the regulatory mechanism of the development and secondary mechanism in A. flavus. Full article

(This article belongs to the Special Issue Mycotoxins and Toxigenic Fungi 2nd Edition)

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20 pages, 4267 KiB

Open AccessArticle

Comparative Bioremediation of Tetradecane, Cyclohexanone and Cyclohexane by Filamentous Fungi from Polluted Habitats in Kazakhstan

by Mariam Gaid, Wiebke Jentzsch, Hannah Beermann, Anne Reinhard, Mareike Meister, Ramza Berzhanova, Togzhan Mukasheva, Tim Urich and Annett Mikolasch

J. Fungi 2024, 10(6), 436; https://doi.org/10.3390/jof10060436 - 19 Jun 2024

Cited by 1 | Viewed by 958

Abstract

Studying the fates of oil components and their interactions with ecological systems is essential for developing comprehensive management strategies and enhancing restoration following oil spill incidents. The potential expansion of Kazakhstan’s role in the global oil market necessitates the existence of land-specific studies [...] Read more.

Studying the fates of oil components and their interactions with ecological systems is essential for developing comprehensive management strategies and enhancing restoration following oil spill incidents. The potential expansion of Kazakhstan’s role in the global oil market necessitates the existence of land-specific studies that contribute to the field of bioremediation. In this study, a set of experiments was designed to assess the growth and biodegradation capacities of eight fungal strains sourced from Kazakhstan soil when exposed to the hydrocarbon substrates from which they were initially isolated. The strains were identified as Aspergillus sp. SBUG-M1743, Penicillium javanicum SBUG-M1744, SBUG-M1770, Trichoderma harzianum SBUG-M1750 and Fusarium oxysporum SBUG-1746, SBUG-M1748, SBUG-M1768 and SBUG-M1769 using the internal transcribed spacer (ITS) region. Furthermore, microscopic and macroscopic evaluations agreed with the sequence-based identification. Aspergillus sp. SBUG-M1743 and P. javanicum SBUG-M1744 displayed remarkable biodegradation capabilities in the presence of tetradecane with up to a 9-fold biomass increase in the static cultures. T. harzianum SBUG-M1750 exhibited poor growth, which was a consequence of its low efficiency of tetradecane degradation. Monocarboxylic acids were the main degradation products by SBUG-M1743, SBUG-M1744, SBUG-M1750, and SBUG-M1770 indicating the monoterminal degradation pathway through β-oxidation, while the additional detection of dicarboxylic acid in SBUG-M1768 and SBUG-M1769 cultures was indicative of the fungus’ ability to undertake both monoterminal and diterminal degradation pathways. F. oxysporum SBUG-M1746 and SBUG-M1748 in the presence of cyclohexanone showed a doubling of the biomass with the ability to degrade the substrate almost completely in shake cultures. F. oxysporum SBUG-M1746 was also able to degrade cyclohexane completely and excreted all possible metabolites of the degradation pathway. Understanding the degradation potential of these fungal isolates to different hydrocarbon substrates will help in developing effective bioremediation strategies tailored to local conditions. Full article

(This article belongs to the Special Issue Bioremediation of Contaminated Soil by Fungi)

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15 pages, 2673 KiB

Open AccessArticle

Fauna and Ecology of Macromycetes (Basidiomycota) in the Arctic Tree and Shrub Ecosystems of Central Siberia

by Sergey Sergeevich Kulakov, Andrey Ivanovich Tatarintsev, Denis Aleksandrovich Demidko and Natalia Pavlovna Khizhniak

J. Fungi 2024, 10(6), 435; https://doi.org/10.3390/jof10060435 - 19 Jun 2024

Viewed by 821

Abstract

The research was aimed at studying the taxonomic diversity, habitat specialization, and trophic characteristics of mycobiota, including Basidiomycota, in the northern ecosystems of the Krasnoyarsk Krai (Central Siberia) near Norilsk. Larch forests and woodlands in the Siberian permafrost zone are distinctive and Basidiomycota, [...] Read more.

The research was aimed at studying the taxonomic diversity, habitat specialization, and trophic characteristics of mycobiota, including Basidiomycota, in the northern ecosystems of the Krasnoyarsk Krai (Central Siberia) near Norilsk. Larch forests and woodlands in the Siberian permafrost zone are distinctive and Basidiomycota, as a component of these ecosystems, plays an essential role in their functioning. Currently, there is a paucity of information about this group in Arctic ecosystems, both in terms of floristic and ecological aspects. Seventy species of macromycetes belonging to different trophic groups were discovered and identified. Only 15% of species occur regularly, while most species are found rarely or only once. The identified species belong to 44 genera, 25 families, and 8 orders, which are included in the class Agaricomycetes. The leading families in terms of the number of species are Russulaceae, Polyporaceae, Tricholomataceae, Suillaceae, Strophariaceae, and Cortinariaceae. Mycorrhizal fungi and wood decay fungi dominate the structure of mycobiota of the study area (the total share is 71%). The rest of the species (29%) are fungal decomposers inhabiting plant litter, the forest floor, and humus. The largest number of species occur in forest ecosystems, which are dominated by mycorrhizal and wood decay fungi (up to 70%), which are trophically associated with woody plants and debris. The fungal decomposers inhabiting plant litter, the forest floor, and humus dominate (about 80%) in the species composition of tundra, where, in the absence of woody substrate, wood decay fungi have not been found at all. The species richness of tree and shrub Arctic ecosystems is low, yet the taxonomical and ecological structure of Basidiomycota is similar to that observed in taiga and temperate forests. These data permit a more comprehensive description of the biodiversity of the Arctic and may prove useful in studying biological processes in these ecosystems. Full article

(This article belongs to the Special Issue Macromycetes: Diversity and Biotechnological Potential)

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17 pages, 2909 KiB

Open AccessArticle

Card9 Broadly Regulates Host Immunity against Experimental Pulmonary Cryptococcus neoformans 52D Infection

by Isabelle Angers, Wided Akik, Annie Beauchamp, Irah L. King, Larry C. Lands and Salman T. Qureshi

J. Fungi 2024, 10(6), 434; https://doi.org/10.3390/jof10060434 - 19 Jun 2024

Viewed by 968

Abstract

The ubiquitous soil-associated fungus Cryptococcus neoformans causes pneumonia that may progress to fatal meningitis. Recognition of fungal cell walls by C-type lectin receptors (CLRs) has been shown to trigger the host immune response. Caspase recruitment domain-containing protein 9 (Card9) is an intracellular adaptor [...] Read more.

The ubiquitous soil-associated fungus Cryptococcus neoformans causes pneumonia that may progress to fatal meningitis. Recognition of fungal cell walls by C-type lectin receptors (CLRs) has been shown to trigger the host immune response. Caspase recruitment domain-containing protein 9 (Card9) is an intracellular adaptor that is downstream of several CLRs. Experimental studies have implicated Card9 in host resistance against C. neoformans; however, the mechanisms that are associated with susceptibility to progressive infection are not well defined. To further characterize the role of Card9 in cryptococcal infection, Card9^em1Sq mutant mice that lack exon 2 of the Card9 gene on the Balb/c genetic background were created using CRISPR-Cas9 genome editing technology and intratracheally infected with C. neoformans 52D. Card9^em1Sq mice had significantly higher lung and brain fungal burdens and shorter survival after C. neoformans 52D infection. Susceptibility of Card9^em1Sq mice was associated with lower pulmonary cytokine and chemokine production, as well as reduced numbers of CD4⁺ lymphocytes, neutrophils, monocytes, and dendritic cells in the lungs. Histological analysis and intracellular cytokine staining of CD4⁺ T cells demonstrated a Th2 pattern of immunity in Card9^em1Sq mice. These findings demonstrate that Card9 broadly regulates the host inflammatory and immune response to experimental pulmonary infection with a moderately virulent strain of C. neoformans. Full article

(This article belongs to the Special Issue New Perspectives on Cryptococcus and Cryptococcosis)

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Card9 signaling is required for survival and control of fungal burden after infection with Cryptococcus neoformans 52D. Wild-type (WT) and Card9em1Sq mice were infected intratracheally with 104 CFU of C. neoformans strain 52D. (A) Mice were observed for up to 36 days for survival analysis (n = 16 mice/strain, using a log-rank test). (B,C) Fungal burden in the lung and brain at serial time intervals was determined by plating tissue homogenates on Sabouraud dextrose agar. CFU data are shown as mean ± SEM and combine at least two independent experiments (n = 9–16 mice /strain/time point). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001. Full article ">Figure 2
Histological analysis of C. neoformans pulmonary infection. Wild-type (WT) and Card9em1Sq mice were infected intratracheally with 104 CFU of C. neoformans 52D. Lungs were harvested at day 21 post-infection; perfused with phosphate-buffered saline; embedded in paraffin; and stained with (A,B) hematoxylin–eosin (H&E), (C,D) periodic acid–Schiff (PAS) or (E,F) mucicarmine. Rectangular boxes correspond to magnified regions (Balb/c; (A,C,E) and Card9em1Sq; (B,D,F)). Arrows identify findings that are detailed in the text of the manuscript. Full article ">Figure 3
Pulmonary inflammatory mediator expression after C. neoformans infection. Wild-type (WT) and Card9em1Sq whole lung proteins were collected at 14 days post infection with 104 CFU of C. neoformans strain 52D. Milliplex and ELISA (IFN-γ) were performed to determine the level of (A) pro-inflammatory immune mediators, (B) chemokines, (C) Type 3 and (D) Type 2 cytokines. Mediators with levels below detectable levels are not shown. Data is shown as mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01, and *** p ≤ 0.001. Full article ">Figure 4
Pulmonary T-lymphocyte recruitment following C. neoformans infection. Lungs of Card9em1Sq mice display fewer CD4+ and CD8+ T lymphocytes during the adaptive phase of immunity after Cryptococcus neoformans 52D infection. Total number of (A) CD3+CD4+ T lymphocytes and (B) CD3+CD8+ T lymphocytes in the lungs at 0, 14, 21, and 28 days post-infection. (C) Representative flow plots and (D) percentages of IFN-γ+CD4+, IL-17A+CD4+ and IL-13+CD4+ T cells. (E) percentages of IFN-γ+CD8+, IL-17A+CD8+ and IL-13+CD8+ T cells. Data are shown as mean ± SEM (n = 5–7 mice/strain/time point). * p ≤ 0.05 and ** p ≤ 0.01. Full article ">Figure 5
Pulmonary myeloid cell recruitment following C. neoformans infection. Lungs of Card9em1Sq mice have decreased monocytes, dendritic cells, and neutrophil recruitment and increased alveolar macrophages and eosinophil recruitment to the lungs after Cryptococcus neoformans 52D infection. (A) Absolute numbers of total CD45+ cells in the lungs at 0,14, 21 and 28 days post-infection. Total number of (B) alveolar macrophages (CD45+CD11c+F4/80+SiglecF+), (C) monocytes (CD45+SiglecF−CD11b+Ly-6G−Ly-6Chi), (D) macrophages (CD45+SiglecF−CD11b+Ly-6G−Ly-6Clo-intCD11c− F4/80+) (E) dendritic cells (CD45+SiglecF−CD11b+Ly-6G−Ly-6Clo-intCD11c+) (F) neutrophils (CD45+SiglecF−CD11b+Ly-6G+Ly-6Chi) (G) NK cells (CD45+SiglecF−CD11b−NK1.1+), and (H) eosinophils (CD45+CD11c−F4/80int-hi) in the lungs at 14, 21 and 28 days post-infection. Data are shown as mean ± SEM (n = 5–7 mice/strain/time point). * p ≤ 0.05 and ** p ≤ 0.01. Full article ">

17 pages, 1504 KiB

Open AccessArticle

Metabolic Engineering of Saccharomyces cerevisiae for Production of Canthaxanthin, Zeaxanthin, and Astaxanthin

by Peerada Promdonkoy, Akaraphol Watcharawipas, Suriyaporn Bubphasawan, Kitisak Sansatchanon, Nattida Suwanakitti, Kanokarn Kocharin and Weerawat Runguphan

J. Fungi 2024, 10(6), 433; https://doi.org/10.3390/jof10060433 - 18 Jun 2024

Viewed by 979

Abstract

The sustainable production of natural compounds is increasingly important in today’s industrial landscape. This study investigates the metabolic engineering of Saccharomyces cerevisiae for the efficient biosynthesis of valuable carotenoids: canthaxanthin, zeaxanthin, and astaxanthin. Utilizing a tailored parental yeast strain, Sp_Bc, we optimized the [...] Read more.

The sustainable production of natural compounds is increasingly important in today’s industrial landscape. This study investigates the metabolic engineering of Saccharomyces cerevisiae for the efficient biosynthesis of valuable carotenoids: canthaxanthin, zeaxanthin, and astaxanthin. Utilizing a tailored parental yeast strain, Sp_Bc, we optimized the carotenoid pathway by screening and identifying CrtW and CrtZ enzymatic variants. The CrtW variant from Bradyrhizobium sp. achieved a canthaxanthin titer of 425.1 ± 69.1 µg/L, while the CrtZ variant from Pantoea ananatis achieved a zeaxanthin titer of 70.5 ± 10.8 µg/L. Additionally, we optimized carotenoid production by exploring enzyme fusion strategies for all three studied carotenoids and organelle compartmentalization specifically for enhancing astaxanthin synthesis. We further improved carotenoid production by integrating the optimal gene constructs into the yeast genome and deleting the GAL80 gene, enabling the use of sucrose as a carbon source. The engineered strain Sp_Bc-Can001 ∆gal80 was evaluated in a 5 L bioreactor fermentation, achieving a notable canthaxanthin titer of 60.36 ± 1.51 mg/L using sucrose. This research conclusively establishes S. cerevisiae as a viable platform for efficient carotenoid biosynthesis and, for the first time in this yeast system, illustrates sucrose’s viability as a carbon source for canthaxanthin production. These findings pave the way for sustainable, cost-effective carotenoid production at an industrial scale. Full article

(This article belongs to the Special Issue New Perspectives on Industrial Yeasts)

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A schematic representation of engineered carotenoid biosynthesis in S. cerevisiae. The pathway initiates with the conversion of β-carotene into zeaxanthin through the enzymatic action of β-carotene hydroxylase (CrtZ). An alternative route involves β-carotene ketolase (CrtW) catalyzing the formation of canthaxanthin from β-carotene. The subsequent transformation of zeaxanthin to astaxanthin is facilitated by β-carotene ketolase, and conversely, astaxanthin can also be synthesized from canthaxanthin through the hydroxylation process mediated by β-carotene hydroxylase. Full article ">Figure 2
Carotenoid production and fermentation metrics in engineered yeast. (a) Canthaxanthin concentration achieved by genetically modified yeast strains; (b) zeaxanthin concentration measured in the same set of strains; (c) optical density (OD600) indicating cellular growth; (d) remaining concentration of galactose after a 72 h fermentation period. Engineered yeast strains were incubated in SCG-Ura medium with an initial mixture of 0.2% glucose and 1.8% galactose at 30 °C, with shaking at 250 rpm. Galactose levels were quantified using HPLC at the end of the fermentation process. Statistically significant differences are marked with ‘**’ (p < 0.05) using a two-tail, unpaired, heteroscedastic Student’s t-test. Full article ">Figure 3
The effects of enzyme fusion on canthaxanthin (a), zeaxanthin (b), and astaxanthin (c) titers. Carotenoids were measured after a 72 h fermentation period. LF stands for the long GGGGSGGPGS linker, while GS stands for the short GS linker. The orientation of the fused enzymes is presented from the N-terminus to the C-terminus. Engineered yeast strains were incubated in SCG-Ura medium with an initial mixture of 0.2% glucose and 1.8% galactose at 30 °C, with shaking at 250 rpm. Experiments were conducted in triplicate, and values are presented as the mean ± standard deviation. Statistically significant differences are marked with ‘***’ (p < 0.01) of ‘**’ (p < 0.05) using a two-tail, unpaired, heteroscedastic Student’s t-test. Full article ">Figure 4
The effects of enzyme compartmentalization on astaxanthin titer. GS stands for the short GS linker. The orientation of the fused enzymes is presented from the N-terminus to the C-terminus. Carotenoids were measured after a 72 h fermentation period. Engineered yeast strains were incubated in SCG-Ura medium with 0.2% glucose and 1.8% galactose at 30 °C, with shaking at 250 rpm. Experiments were conducted in triplicate, and values are presented as the mean ± standard deviation. N.D. means not detected. Statistically significant differences are marked with ‘***’ (p < 0.01) of ‘**’ (p < 0.05) using a two-tail, unpaired, heteroscedastic Student’s t-test. Full article ">Figure 5
Carotenoid production in 50 mL conical tubes across varying sucrose-to-galactose ratios. The fermentation of strains Sp_Bc-Can001 ∆gal80, Sp_Bc-Zea001 ∆gal80, and Sp_Bc-Asta002 ∆gal80 in 50 mL conical tubes using media with different sucrose-to-galactose ratios. Four sucrose-to-galactose ratios were tested: Media A, 100% sucrose; Media B, 2:1 ratio of sucrose to galactose; Media C, 1:1 ratio; and Media D, 1:2 ratio, maintaining a total concentration of 2%. (a) Canthaxanthin production in strain Sp_Bc-Can001 ∆gal80. (b) Zeaxanthin production in strain Sp_Bc-Zea001 ∆gal80. (c) Astaxanthin production in strain Sp_Bc-Asta002 ∆gal80. The experiments were conducted in triplicate, with results presented as the mean ± standard deviation. Statistically significant differences are marked with ‘***’ (p < 0.01) of ‘**’ (p < 0.05) using a two-tail, unpaired, heteroscedastic Student’s t-test. Full article ">Figure 6
The shake-flask fermentation of strain Sp_Bc Can001 ∆gal80 in yeast medium containing either 2% sucrose as the sole carbon source (YPS) or a 2% mixed carbon source (sucrose and galactose at a 1:2 ratio) (YPSG). (a) Canthaxanthin production (mg/L). (b) Biomass accumulation as dry cell weight (DCW, in g/L). The experiments were conducted in triplicate, and values are presented as the mean ± standard deviation. Statistically significant differences are marked with ‘***’ (p < 0.01) of ‘**’ (p < 0.05) using a two-tail, unpaired, heteroscedastic Student’s t-test. Full article ">Figure 7
The fed-batch fermentation of strain Sp_Bc Can001 ∆gal80 in a 5 L fermenter using sucrose as the sole carbon source. (a) The fermentation profile including canthaxanthin titer, biomass as dry cell weight (DCW), ethanol production, glycerol production, and residual sugars (sucrose, fructose, and glucose) over time. Data points represent the averages and standard deviations from duplicate runs. The fermentation began with a 2 L batch phase using a basal salt medium supplemented with 20 g/L sucrose. The fed-batch stage started at the 10th hour, with periodic sucrose additions at 5 h intervals across 20 cycles, cumulatively reaching 120 g/L of sucrose. (b) The visual progression of the bioreactor culture over time, with an intensifying red/orange hue reflecting the increasing canthaxanthin concentration, providing a visual confirmation of successful carotenoid biosynthesis. Full article ">

9 pages, 1272 KiB

Open AccessCommunication

Rapid Molecular Diagnosis of Sporotrichosis Directly from Biological Samples from a Reference Center in Brazil

by Amanda Gabriela da Silva, Arthur Felipe Cavalcanti de Matos, Bruna Rodrigues de Sousa, Claudia Elise Ferraz, Raul Leal Faria Luiz, Rejane Pereira Neves, Reginaldo Gonçalves de Lima-Neto and Manoel Marques Evangelista Oliveira

J. Fungi 2024, 10(6), 432; https://doi.org/10.3390/jof10060432 - 18 Jun 2024

Viewed by 894

Abstract

The gold standard diagnosis of sporotrichosis is the isolation of Sporothrix sp. in culture media, but this is a time-consuming test that is susceptible to contamination and can be affected by the fungal load. Molecular methods such as nested PCR are gaining more [...] Read more.

The gold standard diagnosis of sporotrichosis is the isolation of Sporothrix sp. in culture media, but this is a time-consuming test that is susceptible to contamination and can be affected by the fungal load. Molecular methods such as nested PCR are gaining more ground in the management of several infections as they are tools for the rapid and accurate identification of microorganisms from pure cultures or directly from biological samples. This study aimed to apply a nested PCR molecular protocol for the rapid detection of Sporothrix spp. directly from clinical samples. Thirteen samples—six from skin biopsies, five from skin exudates, and two from conjunctival secretions—were obtained from patients diagnosed with sporotrichosis due to S. brasiliensis. Calmodulin gene sequencing identified all the isolates as S. brasiliensis. Nested PCR was able to detect all the Sporothrix sensu lato directly from clinical samples as well as the CBS 120339 reference strain. The nested PCR protocol stands out as a diagnostic alternative, as it allows the identification of Sporothrix spp. directly from clinical samples without the need for fungal isolation. Full article

(This article belongs to the Section Fungal Pathogenesis and Disease Control)

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13 pages, 13230 KiB

Open AccessArticle

The Cysteine Protease CfAtg4 Interacts with CfAtg8 to Govern the Growth, Autophagy and Pathogenicity of Colletotrichum fructicola

by Shufeng Guo and Shengpei Zhang

J. Fungi 2024, 10(6), 431; https://doi.org/10.3390/jof10060431 - 18 Jun 2024

Viewed by 720

Abstract

Camellia oleifera is a native woody oil plant in southern China and is infected with anthracnose wherever it is grown. We previously identified Colletotrichum fructicola as the major causal agent of anthracnose on C. oleifera and found that CfAtg8 regulates the pathogenicity and [...] Read more.

Camellia oleifera is a native woody oil plant in southern China and is infected with anthracnose wherever it is grown. We previously identified Colletotrichum fructicola as the major causal agent of anthracnose on C. oleifera and found that CfAtg8 regulates the pathogenicity and development of C. fructicola. Here, we revealed that CfAtg4 interacts with CfAtg8, contributing to the formation of autophagosomes. The CfAtg8^1–160 allele, which only contains 1–160 amino acids of the CfAtg8, partially recovered the autophagosome numbers and autophagy defects of the ΔCfatg4 mutant. Consequently, these recoveries resulted in the restoration of the defects of the ΔCfatg4 mutant in growth and responses to different external stresses, albeit to an extent. Importantly, we illustrated the critical roles of CfAtg8^1–160 in appressoria formation, and pathogenicity. Collectively, our findings provide new insights into the importance of the interaction between CfAtg8 and CfAtg4 in the growth, autophagy and pathogenicity of the phytopathogenic fungi. Full article

(This article belongs to the Special Issue Plant Fungal Diseases and Crop Protection)

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CfAtg4 interacts with CfAtg8. (A) The Y2H assays between CfAtg4 and CfAtg8. The pGADT7 and pGBKT7 vectors fused the related genes and were co-transformed into the yeast strain AH109. Leu: leucine; Trp: tryptophan; and His: histidine. (B) The schematic of the interaction between CfAtg4 and CfAtg8. Full article ">Figure 2
The alignment of Atg8 proteins among different species. The Atg8 proteins were aligned by the BioEdit v7.0 software and an asterisk indicates the conserved glycine of Atg8 proteins among fungi. The related species names are as follows: C. fructicola, C. gloeosporioides, C. siamense, S. cerevisiae, M. oryzae, A. oryzae, A. nidulans, U. maydis, F. graminearum, N. crassa, R. norvegicus and A. thaliana. Full article ">Figure 3
CfAtg81–160 partially restores the growth and asexual reproduction of ΔCfatg4. (A) The growth of WT, ΔCfatg4, ΔCfatg4/CfAtg81–160, and ΔCfatg4/CfATG4 strains in CM and MM medium for 3 days. (B) The colony diameters were measured and statistically analyzed. (C) Statistical analysis of conidia. Asterisks mean the difference is significant (**, p < 0.01; *, 0.01 ≤ p < 0.05). The experiments were performed with 3 biological replications. The data were analyzed by Student’s t-test, and the error bars indicate standard deviation (SD). Full article ">Figure 4
CfAtg81–160 partially restores the resistance of ΔCfatg4 to rapamycin. (A) The WT, ΔCfatg4, ΔCfatg4/CfAtg81–160, and ΔCfatg4/CfATG4 strains were incubated on CM plates with 50 nM of rapamycin at 28 °C for 3 days. (B) Statistical analysis of growth inhibition rates of the strains to rapamycin stress. The data were analyzed by Student’s t-test, and the error bars indicate SD. Asterisks indicate the difference is significant (**, p < 0.01). Full article ">Figure 5
CfAtg81–160 is important for autophagy. (A) Micrographs of GFP-CfAtg8-labeled autophagosomes in the WT, ΔCfatg4, and ΔCfatg4/CfAtg81–160. (B) Statistical analysis of autophagosome number in ΔCfatg4 and ΔCfatg4/CfAtg81–160 after being induced for 0 h, 2 h, and 5 h in MM-N liquid medium. The data were analyzed by Student’s t-test, and the error bars indicate SD. Asterisks indicate the difference is significant (**, p < 0.01; *, 0.01 ≤ p < 0.05) (C) Immunoblot analysis of GFP-CfAtg8 proteolysis. The upper and lower lanes point to the intact GFP-Atg8 (46 kDa) and free GFP (26 kDa), respectively. Full article ">Figure 6
CfAtg81–160 partially restores the pathogenicity of ΔCfatg4. (A) The WT, ΔCfatg4, ΔCfatg4/CfAtg81–160 and ΔCfatg4/CfATG4 strains were inoculated on wounded C. oleifera leaves. (B) The disease spot areas of strains on wounded C. oleifera leaves were measured by ImageJ, and the lesion ratios of the related strains to WT were statistically analyzed. (C) The strains were inoculated on wounded apples and photographed. (D) Statistical analysis of the related strains to WT on apples. CK: compared control. The data were analyzed by Student’s t-test, and the error bars indicate SD. Asterisks indicate the difference is significant (**, p < 0.01; *, 0.01 ≤ p < 0.05). Full article ">Figure 7
CfAtg81–160 partially restores conidia germination and appressoria formation in ΔCfatg4. (A) The conidia of the WT, ΔCfatg4, ΔCfatg4/CfAtg81–160, and ΔCfatg4/CfATG4 strains were cultured on hydrophobic glass for 12 h and 24 h, and the appressoria formation was observed. (B) Conidia germination and appressoria formation were statistically analyzed. The data were analyzed by Student’s t-test and the error bars indicate SD. Asterisks indicate the difference is significant (**, p < 0.01; *, 0.01 ≤ p < 0.05). Arrows represent appressoria. Bar = 5 μm. Full article ">Figure 8
CfAtg81–160 helps ΔCfatg4 respond to external environmental stresses. (A) The strains of the WT, ΔCfatg4, ΔCfatg4/CfAtg81–160, and ΔCfatg4/CfATG4 were cultured on CM and CM plus 1 M of NaCl, 1 M of KCl, 0.1% SDS, and 2.5 mM of DTT for 3 days. (B) The inhibition rate of strains to stresses was statistically analyzed against an untreated control. The data were analyzed by Student’s t-test, and the error bars indicate SD. Asterisks indicate the difference is significant (**, p < 0.01; *, 0.01 ≤ p < 0.05). Full article ">

12 pages, 8293 KiB

Open AccessArticle

The Near-Gapless Penicillium fuscoglaucum Genome Enables the Discovery of Lifestyle Features as an Emerging Post-Harvest Phytopathogen

by Dianiris Luciano-Rosario, Wayne M. Jurick II and Christopher Gottschalk

J. Fungi 2024, 10(6), 430; https://doi.org/10.3390/jof10060430 - 18 Jun 2024

Viewed by 924

Abstract

Penicillium spp. occupy many diverse biological niches that include plant pathogens, opportunistic human pathogens, saprophytes, indoor air contaminants, and those selected specifically for industrial applications to produce secondary metabolites and lifesaving antibiotics. Recent phylogenetic studies have established Penicillium fuscoglaucum as a synonym for [...] Read more.

Penicillium spp. occupy many diverse biological niches that include plant pathogens, opportunistic human pathogens, saprophytes, indoor air contaminants, and those selected specifically for industrial applications to produce secondary metabolites and lifesaving antibiotics. Recent phylogenetic studies have established Penicillium fuscoglaucum as a synonym for Penicillium commune, which is an indoor air contaminant and toxin producer and can infect apple fruit during storage. During routine culturing on selective media in the lab, we obtained an isolate of P. fuscoglaucum Pf_T2 and sequenced its genome. The Pf_T2 genome is far superior to available genomic resources for the species. Our assembly exhibits a length of 35.1 Mb, a BUSCO score of 97.9% complete, and consists of five scaffolds/contigs representing the four expected chromosomes. It was determined that the Pf_T2 genome was colinear with a type specimen P. fuscoglaucum and contained a lineage-specific, intact cyclopiazonic acid (CPA) gene cluster. For comparison, a highly virulent postharvest apple pathogen, P. expansum strain TDL 12.1, was included and showed a similar growth pattern in culture to our Pf_T2 isolate but was far more aggressive in apple fruit than P. fuscoglaucum. The genome of Pf_T2 serves as a major improvement over existing resources, has superior annotation, and can inform forthcoming omics-based work and functional genetic studies to probe secondary metabolite production and disparities in aggressiveness during apple fruit decay. Full article

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FastANI phylogenetic tree and heatmap of different Penicillium type specimen reference genomes, including the Pf_T2 genome marked by a red colored star. Full article ">Figure 2
BUSCO phylogeny of nineteen different Penicillium type specimen reference genomes with the P. fuscoglaucum Pf_T2 genome marked by a red star. Each genome is also indicated with corresponding genus and species name followed by isolate name. Full article ">Figure 3
Collinearity with whole genome alignments between Pf_T2 assembly and the NCBI reference genome FM041 for P. fuscoglaucum. Full article ">Figure 4
P. fuscoglaucum Pf_T2 isolate and P. expansum TDL 12.1 growth and virulence. (A) Growing in culture on glucose minimal medium. (B) Lesion diameter in “Honeycrisp” apple fruit over time. Error bars denote standard deviation of the mean. Asterisks denote significant differences between strains *** p < 0.001. Full article ">Figure 5
Cyclopiazonic acid (CPA) biosynthesis cluster in Penicillium camemberti species complex that includes P. fuscoglaucum. (A) CPA cluster in P. camemberti var. “camemberti” reference genome FM013 recreated from Ropars et al. 2020 [<a href="#B7-jof-10-00430" class="html-bibr">7</a>]. The cluster contains 8 loci with 6 CpA genes: Reg (transcriptional regulator/C6 transcription factor), CpaT (multi-function substrate transporter), CpaM (hypothetical protein), CpaH (Cytochrome P450), CpaO (FAD oxidase), CpaD (dimethylallyl synthase), CpaA (Non-ribosomal polyketide synthase), and Arrestin-like gene. (B) CPA Cluster in P. fuscoglaucum isolate Pf_T2 isolate contains 7 of the 8 genes present in P. camemberti gene cluster and is missing cpaM (hypothetical protein) and contains an extra transcription factor. TF (transcriptional regulator C6 transcription factor), TF (Gal4 transcriptional regulator), MFS (multi-function substrate transporter), P450 (Cytochrome P450 monoxygenase), FAD (FAD oxidase), TDAT (Tryptophan Dimethylallyl transferase), PKS (polyketide synthase), and Arrestin-like protein. Full article ">

8 pages, 778 KiB

Open AccessBrief Report

Coccidioidomycosis in Immunocompromised at a Non-Endemic Referral Center in Mexico

by Carla M. Román-Montes, Lisset Seoane-Hernández, Rommel Flores-Miranda, Andrea Carolina Tello-Mercado, Andrea Rangel-Cordero, Rosa Areli Martínez-Gamboa, José Sifuentes-Osornio, Alfredo Ponce-de-León and Fernanda González-Lara

J. Fungi 2024, 10(6), 429; https://doi.org/10.3390/jof10060429 - 18 Jun 2024

Viewed by 829

Abstract

The incidence and distribution of coccidioidomycosis are increasing. Information scarcity is evident in Mexico, particularly in non-endemic zones and specific populations. We compared the treatment and outcomes for patients with isolated pulmonary infections and those with disseminated coccidioidomycosis, including mortality rates within six [...] Read more.

The incidence and distribution of coccidioidomycosis are increasing. Information scarcity is evident in Mexico, particularly in non-endemic zones and specific populations. We compared the treatment and outcomes for patients with isolated pulmonary infections and those with disseminated coccidioidomycosis, including mortality rates within six weeks of diagnosis. Of 31 CM cases, 71% were male and 55% were disseminated. For 42% of patients, there was no evidence of having lived in or visited an endemic region. All patients had at least one comorbidity, and 58% had pharmacologic immunosuppressants. The general mortality rate was 30%; without differences between disseminated and localized disease. In our research, we describe a CM with a high frequency of disseminated disease without specific risk factors and non-significant mortality. Exposure to endemic regions was not found in a considerable number of subjects. We consider diverse reasons for why this may be, such as climate change or migration. Full article

(This article belongs to the Special Issue Dimorphic Fungal Pathogen Coccidioides and Coccidioidomycosis)

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24 pages, 2170 KiB

Open AccessArticle

Colonization and Biodegradation Potential of Fungal Communities on Immersed Polystyrene vs. Biodegradable Plastics: A Time Series Study in a Marina Environment

by Aurélie Philippe, Marie Salaun, Maxence Quemener, Cyril Noël, Kévin Tallec, Camille Lacroix, Emmanuel Coton and Gaëtan Burgaud

J. Fungi 2024, 10(6), 428; https://doi.org/10.3390/jof10060428 - 18 Jun 2024

Viewed by 1087

Abstract

Plastic pollution of the ocean is a major environmental threat. In this context, a better understanding of the microorganisms able to colonize and potentially degrade these pollutants is of interest. This study explores the colonization and biodegradation potential of fungal communities on foamed [...] Read more.

Plastic pollution of the ocean is a major environmental threat. In this context, a better understanding of the microorganisms able to colonize and potentially degrade these pollutants is of interest. This study explores the colonization and biodegradation potential of fungal communities on foamed polystyrene and alternatives biodegradable plastics immersed in a marina environment over time, using the Brest marina (France) as a model site. The methodology involved a combination of high-throughput 18S rRNA gene amplicon sequencing to investigate fungal taxa associated with plastics compared to the surrounding seawater, and a culture-dependent approach to isolate environmentally relevant fungi to further assess their capabilities to utilize polymers as carbon sources. Metabarcoding results highlighted the significant diversity of fungal communities associated with both foamed polystyrene and biodegradable plastics, revealing a dynamic colonization process influenced by the type of polymer and immersion time. Notably, the research suggests a potential for certain fungal species to utilize polymers as a carbon source, emphasizing the need for further exploration of fungal biodegradation potential and mechanisms. Full article

(This article belongs to the Section Environmental and Ecological Interactions of Fungi)

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Principal Coordinate Analysis (PCoA) based on the 18S dataset, showing the relationship between the variables “polymers” (A) and “immersion time” (B). The PCoA vas generated using Bray–Curtis dissimilarity as the distance measure, with each point representing a sample. Ellipses connect groups of samples that were subjected to the same conditions. Full article ">Figure 2
Bubble plot representing the proportions of phyla or genus, based on the 18S dataset, according to the “Polymers” (A,B) and “Type of plastic” (C,D) variables. Full article ">Figure 3
Bubble plot representing the proportions of phyla (A) or genus (B), based on the 18S dataset, according to the ‘Time’ variable. Full article ">Figure 4
Bubble plot showing the proportions of isolates per isolation medium (BH+PVC, +PS, +PHBV, +PET, +PE, +PCL) from the initial collection (A), or stratified and weighted sampling (B) to select representative isolates from this collection. Full article ">Figure 5
UpSet plot representing the most promising isolates utilizing either a single polymer (light grey circles) or several polymers (dark grey circles) as carbon sources. Full article ">

13 pages, 4364 KiB

Open AccessArticle

Arbuscular Mycorrhizal Fungi Regulate Lipid and Amino Acid Metabolic Pathways to Promote the Growth of Poncirus trifoliata (L.) Raf

by Yihao Kang, Gratien Twagirayezu, Jie Xu, Yunying Wen, Pengxiang Shang, Juan Song, Qian Wang, Xianliang Li, Shengqiu Liu, Tingsu Chen, Tong Cheng and Jinlian Zhang

J. Fungi 2024, 10(6), 427; https://doi.org/10.3390/jof10060427 - 18 Jun 2024

Viewed by 850

Abstract

Arbuscular mycorrhizal (AM) fungi can enhance the uptake of soil nutrients and water by citrus, promoting its growth. However, the specific mechanisms underlying the action of AM fungi in promoting the growth of citrus were not fully elucidated. This study aimed to explore [...] Read more.

Arbuscular mycorrhizal (AM) fungi can enhance the uptake of soil nutrients and water by citrus, promoting its growth. However, the specific mechanisms underlying the action of AM fungi in promoting the growth of citrus were not fully elucidated. This study aimed to explore the role of AM fungi Funneliformis mosseae in the regulatory mechanisms of P. trifoliata growth. Pot experiments combined with non-targeted metabolomics methods were used to observe the growth process and changes in metabolic products of P. trifoliata under the conditions of F. mosseae inoculation. The results showed that F. mosseae could form an excellent symbiotic relationship with P. trifoliata, thereby enhancing the utilization of soil nutrients and significantly promoting its growth. Compared with the control, the plant height, stem diameter, number of leaves, and aboveground and underground dry weight in the F. mosseae inoculation significantly increased by 2.57, 1.29, 1.57, 4.25, and 2.78 times, respectively. Moreover, the root system results confirmed that F. mosseae could substantially promote the growth of P. trifoliata. Meanwhile, the metabolomics data indicated that 361 differential metabolites and 56 metabolic pathways were identified in the roots of P. trifoliata and were inoculated with F. mosseae. This study revealed that the inoculated F. mosseae could participate in ABC transporters by upregulating their participation, glycerophospholipid metabolism, aminoacyl tRNA biosynthesis, tryptophan metabolism and metabolites from five metabolic pathways of benzoxazinoid biosynthesis [mainly enriched in lipid (39.50%) and amino acid-related metabolic pathways] to promote the growth of P. trifoliata. Full article

(This article belongs to the Section Fungi in Agriculture and Biotechnology)

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The growth and scanning of the root system of P. trifoliata inoculated without (a–c) and with AM fungi (d–f). Full article ">Figure 2
Plant height (a), stem diameter (b), and number of leaves (c) of orange P. trifoliata in the CK and AMF treatments at different times after inoculation. All data are presented as the mean ± standard deviation (n = 6). * represents a significant difference between treatments (p < 0.05). Full article ">Figure 3
Top dry weight (a), root dry weight (b), root diameter (c), root length (d), root volume, (e) and root surface area (f) in CK and AMF treatment. The labels of CK and AMF represent without (control) and with AM fungi (F. mosseae). All data are presented as the mean ± standard deviation (n = 6). * represents a significant difference between treatments (p < 0.05). Full article ">Figure 4
A pie chart of the classification of metabolites in the root samples of P. trifoliata is at the superclass level. Different colors in the pie chart represent different HMDB classifications; the area means the relative proportion of metabolites. Full article ">Figure 5
(a) Volcano map for the positive and negative ion merging mode. Up represents significantly upregulated metabolites, down represents significantly downregulated metabolites, and nosing represents metabolites with no differences; (b) pie chart of differential metabolite classification at the superclass level, where different colors represent different HMDB classifications, and the area means the relative proportion of metabolites in the classification. Full article ">Figure 6
Pathways for differential metabolites between CK and AMF treatment. Full article ">Figure 7
Heat map of correlation between the differential metabolites of roots and the growth indexes of P. trifoliata in CK and AMF treatment. ‘**’ represents a significant correlation of 0.01 level, whereas ‘*’ represents a significant correlation of 0.05 level. Full article ">

17 pages, 788 KiB

Open AccessReview

Inbred Mouse Models in Cryptococcus neoformans Research

by Minna Ding and Kirsten Nielsen

J. Fungi 2024, 10(6), 426; https://doi.org/10.3390/jof10060426 - 17 Jun 2024

Viewed by 846

Abstract

Animal models are frequently used as surrogates to understand human disease. In the fungal pathogen Cryptococcus species complex, several variations of a mouse model of disease were developed that recapitulate different aspects of human disease. These mouse models have been implemented using various [...] Read more.

Animal models are frequently used as surrogates to understand human disease. In the fungal pathogen Cryptococcus species complex, several variations of a mouse model of disease were developed that recapitulate different aspects of human disease. These mouse models have been implemented using various inbred and outbred mouse backgrounds, many of which have genetic differences that can influence host response and disease outcome. In this review, we will discuss the most commonly used inbred mouse backgrounds in C. neoformans infection models. Full article

(This article belongs to the Special Issue Cryptococcus Infections and Pathogenesis)

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21 pages, 1908 KiB

Open AccessArticle

The Diversity and Floristic Analysis of Rust Diseases in the Sanjiangyuan Forest Plants

by Qi Xu and Luchao Bai

J. Fungi 2024, 10(6), 425; https://doi.org/10.3390/jof10060425 - 16 Jun 2024

Viewed by 756

Abstract

Between 2020 and 2023, rust fungus specimens were collected from the primary forested regions of the Sanjiangyuan area in Qinghai Province, resulting in over 300 samples. A taxonomic and phylogenetic study of the rust fungi from these forests was conducted using morphological and [...] Read more.

Between 2020 and 2023, rust fungus specimens were collected from the primary forested regions of the Sanjiangyuan area in Qinghai Province, resulting in over 300 samples. A taxonomic and phylogenetic study of the rust fungi from these forests was conducted using morphological and molecular biological techniques. The investigation identified rust fungi from 7 families, 12 genera, 56 species and varieties, including 10 new host records, 1 new record for China, and 2 novel species. The host plants involved belonged to 26 families, 48 genera, and 78 species. Pucciniaceae and Coleosporiaceae were the dominant families, with the genera Puccinia, Melampsora, and Gymnosporangium being prevalent. The rust fungi in the Sanjiangyuan forests showed a biogeographical affinity with the North Temperate Zone. Floristic comparisons revealed a higher similarity with rust fungi from Inner Mongolia, Gansu, and Tibet and a lower similarity with those from Hainan. An analysis of the life forms of rust fungus host plants indicated that herbaceous plants were the most common, followed by shrubs and trees. In different regions of Sanjiangyuan, rust fungi were found as follows: Golog Prefecture with 6 families, 9 genera, and 28 species; Yushu Prefecture with 5 families, 8 genera, and 31 species; Huangnan Prefecture with 5 families, 9 genera, and 26 species; and Hainan Prefecture with 4 families, 5 genera, and 10 species. The families Pucciniaceae, Melampsoraceae, and Coleosporiaceae were common across all four regions. Moreover, the families Rosaceae, Asteraceae, Ranunculaceae, Salicaceae, and Caprifoliaceae were shared among the host plants in these regions. Full article

(This article belongs to the Special Issue Rust Fungi)

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12 pages, 1412 KiB

Open AccessArticle

Secondary Metabolites from Marine-Derived Fungus Penicillium rubens BTBU20213035

by Xiuli Xu, Yifei Dong, Jinpeng Yang, Long Wang, Linlin Ma, Fuhang Song and Xiaoli Ma

J. Fungi 2024, 10(6), 424; https://doi.org/10.3390/jof10060424 - 16 Jun 2024

Cited by 2 | Viewed by 936

Abstract

Two new polyketide derivatives, penirubenones A and B (1 and 2), and two naturally rare amino-bis-tetrahydrofuran derivatives, penirubenamides A and B (3 and 4), together with nine known compounds (5–13) were isolated from the marine-derived [...] Read more.

Two new polyketide derivatives, penirubenones A and B (1 and 2), and two naturally rare amino-bis-tetrahydrofuran derivatives, penirubenamides A and B (3 and 4), together with nine known compounds (5–13) were isolated from the marine-derived fungus Penicillium rubens BTBU20213035. The structures were identified by HRESIMS and 1D and 2D NMR analyses, and their absolute configurations were determined by a comparison of experimental and calculated electronic circular dichroism (ECD) spectroscopy and ¹³C NMR data. We found that 6 exhibited antibacterial activity against Staphylococcus aureus, with an MIC value of 3.125 μg/mL, and 1 and 2 showed synergistic antifungal activity against Candida albicans at 12.5 and 50 μg/mL with 0.0625 μg/mL rapamycin. Full article

(This article belongs to the Special Issue The Gift of Marine Fungi: Abundant Secondary Metabolites)

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22 pages, 11983 KiB

Open AccessArticle

Morphological and Phylogenetic Analyses Reveal Three New Species of Entomopathogenic Fungi Belonging to Clavicipitaceae (Hypocreales, Ascomycota)

by Zhi-Qin Wang, Jin-Mei Ma, Zhi-Li Yang, Jing Zhao, Zhi-Yong Yu, Jian-Hong Li and Hong Yu

J. Fungi 2024, 10(6), 423; https://doi.org/10.3390/jof10060423 - 14 Jun 2024

Viewed by 956

Abstract

This study aims to report three new species of Conoideocrella and Moelleriella from Yunnan Province, Southwestern China. Species of Conoideocrella and Moelleriella parasitize scale insects (Coccidae and Lecaniidae, Hemiptera) and whiteflies (Aleyrodidae, Hemiptera). Based on the phylogenetic analyses of the three-gene nrLSU, tef-1α [...] Read more.

This study aims to report three new species of Conoideocrella and Moelleriella from Yunnan Province, Southwestern China. Species of Conoideocrella and Moelleriella parasitize scale insects (Coccidae and Lecaniidae, Hemiptera) and whiteflies (Aleyrodidae, Hemiptera). Based on the phylogenetic analyses of the three-gene nrLSU, tef-1α, and rpb1, it showed one new record species (Conoideocrella tenuis) and one new species (Conoideocrella fenshuilingensis sp. nov.) in the genus Conoideocrella, and two new species, i.e., Moelleriella longzhuensis sp. nov. and Moelleriella jinuoana sp. nov. in the genus Moelleriella. The three new species were each clustered into separate clades that distinguished themselves from one another. All of them were distinguishable from their allied species based on their morphology. Morphological descriptions, illustrations, and comparisons of the allied taxa of the four species are provided in the present paper. In addition, calculations of intraspecific and interspecific genetic distances were performed for Moelleriella and Conoideocrella. Full article

(This article belongs to the Special Issue Ascomycota: Diversity, Taxonomy and Phylogeny, 2nd Edition)

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Phylogenetic relationships of 14 genera in Clavicipitaceae are based on the maximum likelihood (ML) and the Bayesian inference (BI) analyses using nrLSU, tef-1α, and rpb1 sequences. Statistical support values greater than 70% are shown at the nodes for the BI posterior probabilities/the ML bootstrap proportions. The new taxa are highlighted in bold and T for ex-type material. Full article ">Figure 2
Morphology of Conoideocrella tenuis. (A,B) Telemorphic stroma containing perithecia; (C) Perithecium; (D–H) Mature asci with developing asci; (I) Obverse of colonies on PDA at 25 °C after 21 days; (J) Reverse of colonies on PDA at 25 °C after 21 days; (K–O) Conidia of hirsutella-like asexual stage on PDA. Scale bars: 1 mm (A,B); 200 µm (C); 50 µm (D,E); 25 µm (F); 50 µm (G); 20 µm (H); 1 cm (I,J); 10 µm (K–M); and 5 µm (N,O). Full article ">Figure 3
Morphology of Conoideocrella fenshuilingensis. (A,B) Telemorphic stroma containing perithecia; (C,D) Perithecium; (E–L) Mature asci with developing asci. Scale bars: 1 mm (A,B); 200 µm (C–G); 100 µm (H); 150 µm (I); 100 µm (J); 30 µm (K); and 20 µm (L). Full article ">Figure 4
Morphology of Moelleriella jinuoana. (A–C) Anamorphic stromata containing conidiomata; (D,E) Section of stromata showing conidiomata; (F,G) Phialides with conidia at the tips; (H) Conidia; (I) Obverse of colonies on PDA at 25 °C after 21 days; (J) Reverse of colonies on PDA at 25 °C after 21 days. Scale bars: 1 mm (A); 0.5 mm (B,C); 100 µm (D,E); 20 µm (F,G); 10 µm (H); and 2 mm (I,J). Full article ">Figure 5
Morphology of Moelleriella longzhuensis. (A–C) Anamorphic stromata containing conidiomata; (D,E) Section of stromata showing conidiomata; (F) Phialides with conidia at the tips; (G) Conidia; (H) Obverse of colonies on PDA at 25 °C after 21 days; (I) Reverse of colonies on PDA at 25 °C after 21 days. Scale bars: 1 mm (A); 0.5 mm (B,C); 50 µm (D); 100 µm (E); 20 µm (F,G); and 2 mm (H,I). Full article ">

17 pages, 4219 KiB

Open AccessArticle

Neutrophil Oxidative Burst Profile Is Related to a Satisfactory Response to Itraconazole and Clinical Cure in Feline Sporotrichosis

by Luisa Helena Monteiro de Miranda, Marta de Almeida Santiago, Julia Frankenfeld, Erica Guerino dos Reis, Rodrigo Caldas Menezes, Sandro Antonio Pereira, Isabella Dib Ferreira Gremião, Regina Hofmann-Lehmann and Fátima Conceição-Silva

J. Fungi 2024, 10(6), 422; https://doi.org/10.3390/jof10060422 - 14 Jun 2024

Viewed by 773

Abstract

Despite the central role of cats in the transmission and amplification of Sporothrix, studies regarding immune response in feline sporotrichosis are scarce. In cats with sporotrichosis, neutrophil-rich lesions are usually associated to good general condition and lower fungal burden. However, the role [...] Read more.

Despite the central role of cats in the transmission and amplification of Sporothrix, studies regarding immune response in feline sporotrichosis are scarce. In cats with sporotrichosis, neutrophil-rich lesions are usually associated to good general condition and lower fungal burden. However, the role of neutrophils in anti-Sporothrix immunity has been little explored in cats. Thus, the aim of this study was to evaluate the neutrophil oxidative burst in the blood of cats with sporotrichosis. Cats with sporotrichosis included in the study were treated with itraconazole (ITZ) alone or combined with potassium iodide (KI). The neutrophil oxidative burst was evaluated through a flow-cytometry-based assay using dihydrorhodamine 123 (background) and stimulation with Zymosan and heat-killed Sporothrix yeasts. The cure rate was 50.0% in cats under treatment with ITZ monotherapy and 90.9% in cats treated with ITZ + KI (p = 0.014), endorsing the combination therapy as an excellent alternative for the treatment of feline sporotrichosis. Higher percentages of Sporothrix-stimulated neutrophils were associated with good general condition (p = 0.003). Higher percentages of Sporothrix- (p = 0.05) and Zymosan-activated (p = 0.014) neutrophils before and early in the treatment were related to clinical cure in ITZ-treated cats. The correlation between oxidative burst and successful use of KI could not be properly assessed given the low number of failures (n = 2) in this treatment group. Nasal mucosa involvement, typically linked to treatment failure, was related to lower percentages of activated neutrophils in the background at the treatment outcome (p = 0.02). Our results suggest a beneficial role of neutrophils in feline sporotrichosis and a positive correlation between neutrophil activation and the cure process in ITZ-treated cats. Full article

(This article belongs to the Special Issue New Perspectives on Sporothrix and Sporotrichosis)

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Figure 1
Feline sporotrichosis. Skin lesions in cats presented at Lapclin-Dermzoo/INI/Fiocruz, Rio de Janeiro, Brazil. Disseminated skin lesions (A); Multiple skin ulcers on the face (B); Skin ulcers on the forelimb (C); Swelling over the bridge of the nose (D). Full article ">Figure 2
Feline sporotrichosis. Response to treatment. Clinical cure. Cat presenting multiple skin ulcers on the face before the beginning of the treatment (A) and complete healing after eighteen weeks of treatment with the combination of itraconazole and KI (B); Cat presenting swelling over the bridge of the nose and a skin ulcer at the palmar side of the left forelimb before the beginning of the treatment (C) and complete healing and the remission of clinical signs after twenty-nine weeks of treatment with itraconazole (D); Cat presenting ulcerated lesion on the right forelimb pad before the beginning of the treatment (E) and complete healing after sixteen weeks of treatment with itraconazole (F). Full article ">Figure 3
Feline sporotrichosis. Response to treatment. Treatment failure. Cat presenting swelling and ulceration on the bridge of the nose and periocular skin lesions before the beginning of the treatment (A); and the persistence of the skin lesion on the bridge of the nose after eight weeks of treatment with itraconazole (B); Cat presenting skin ulcer lateral to the nose and nasal planum before the beginning of the treatment (C); and persistence of the lesion with the swelling of the nose and the appearance of multiple nodules on the left ear after sixteen weeks of treatment with the combination of itraconazole and KI (D). Full article ">Figure 4
The neutrophil oxidative burst test performed on the peripheral blood of cats with sporotrichosis demonstrates a significant variation in some neutrophil oxidative burst parameters, depending on clinical and therapeutic features. Good general condition is associated (pMW = 0.003) with a higher percentage of Sporothrix-stimulated neutrophils at T1 (A); Clinical cure with combination therapy is associated (pMW = 0.035) with a higher stimulation index in Sporothrix-stimulated neutrophils at T2 in comparison to monotherapy (B); Clinical cure with monotherapy is associated (pMW = 0.05) with a higher percentage of Sporothrix-stimulated neutrophils at T1 in comparison to treatment failure (C); The percentage of Zymosan-stimulated neutrophils is significantly higher (pF = 0.014) in T2 in comparison to T1 in clinical cure in comparison to treatment failure (D); The percentage of positive neutrophils in the background control at T2 is significantly higher (pMW = 0.03) in clinical cure with monotherapy in comparison to treatment failure (E); Nasal mucosa involvement is associated (pMW = 0.02) with a lower percentage of positive neutrophils in the background control at T3 in cats submitted to the monotherapy (F). Full article ">

12 pages, 1889 KiB

Open AccessCommunication

Comprehensive Characterization of Tuber maculatum, New in Uruguay: Morphological, Molecular, and Aromatic Analyses

by Francisco Kuhar, Eva Tejedor-Calvo, Alejandro Sequeira, David Pelissero, Mariana Cosse, Domizia Donnini and Eduardo Nouhra

J. Fungi 2024, 10(6), 421; https://doi.org/10.3390/jof10060421 - 14 Jun 2024

Viewed by 997

Abstract

Although only a few species of Tuber account for the major truffle sales volume, many species that are not considered delicacies are finding their way to the market, especially in regions where the traditionally appreciated ones do not occur. This is the case [...] Read more.

Although only a few species of Tuber account for the major truffle sales volume, many species that are not considered delicacies are finding their way to the market, especially in regions where the traditionally appreciated ones do not occur. This is the case for whitish truffles. Specimens of whitish truffles were collected in pecan (Carya illinoinensis) orchards in Uruguay in October 2021. Morphological and molecular methods were used to characterize and assess their identity as Tuber maculatum Vittad. An SPME extraction of volatile compounds and GC–MS analyses were performed to characterize the aromatic profile of these specimens and evaluate their potential applications. Among the 60 VOCs detected, 3-octenone (mushroom odor), 3-octanol (moss, nut, mushroom odor), and 2H-pyran-2-one (no odor), followed by octen-1-ol-acetate (no odor) and 2-undecanone (orange, fresh, green odor) were the major compounds in T. maculatum fruiting bodies. The attributes of exotic edible mushrooms of commercial value in the region are highlighted. In particular, this work emphasizes the characteristics of truffles as a byproduct of pecan cultivation. Full article

(This article belongs to the Special Issue New Perspectives on Tuber Fungi)

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17 pages, 6302 KiB

Open AccessArticle

Photoreactivation Activities of Rad5, Rad16A and Rad16B Help Beauveria bassiana to Recover from Solar Ultraviolet Damage

by Xin-Cheng Luo, Lei Yu, Si-Yuan Xu, Sheng-Hua Ying and Ming-Guang Feng

J. Fungi 2024, 10(6), 420; https://doi.org/10.3390/jof10060420 - 13 Jun 2024

Viewed by 951

Abstract

In budding yeast, Rad5 and Rad7-Rad16 play respective roles in the error-free post-replication repair and nucleotide excision repair of ultraviolet-induced DNA damage; however, their homologs have not yet been studied in non-yeast fungi. In the fungus Beauveria bassiana, a deficiency in the [...] Read more.

In budding yeast, Rad5 and Rad7-Rad16 play respective roles in the error-free post-replication repair and nucleotide excision repair of ultraviolet-induced DNA damage; however, their homologs have not yet been studied in non-yeast fungi. In the fungus Beauveria bassiana, a deficiency in the Rad7 homolog, Rad5 ortholog and two Rad16 paralogs (Rad16A/B) instituted an ability to help the insect-pathogenic fungus to recover from solar UVB damage through photoreactivation. The fungal lifecycle-related phenotypes were not altered in the absence of rad5, rad16A or rad16B, while severe defects in growth and conidiation were caused by the double deletion of rad16A and rad16B. Compared with the wild-type and complemented strains, the mutants showed differentially reduced activities regarding the resilience of UVB-impaired conidia at 25 °C through a 12-h incubation in a regime of visible light plus dark (L/D 3:9 h or 5:7 h for photoreactivation) or of full darkness (dark reactivation) mimicking a natural nighttime. The estimates of the median lethal UVB dose LD₅₀ from the dark and L/D treatments revealed greater activities of Rad5 and Rad16B than of Rad16A and additive activities of Rad16A and Rad16B in either NER-dependent dark reactivation or photorepair-dependent photoreactivation. However, their dark reactivation activities were limited to recovering low UVB dose-impaired conidia but were unable to recover conidia impaired by sublethal and lethal UVB doses as did their photoreactivation activities at L/D 3:9 or 5:7, unless the night/dark time was doubled or further prolonged. Therefore, the anti-UV effects of Rad5, Rad16A and Rad16B in B. bassiana depend primarily on photoreactivation and are mechanistically distinct from those for their yeast homologs. Full article

(This article belongs to the Special Issue Interactions between Filamentous Fungal Pathogens and Hosts: 2nd Edition)

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Recognition of Rad5, Rad16A and Rad16B in B. bassiana. (A) Comparison of domain architectures for Rad5 and Rad16 homologs of B. bassiana (Bb) and S. cerevisiae (Sc). Associated with each NLS motif is the maximal probability (decimal value) predicted. (B) Relative transcript (RT) levels for rad5, rad16A and rad16B expressed in the WT strain during 7-day incubation (versus day 2) on SDAY at 25 °C and 12:12 (L/D) photoperiod. (C) Laser scanning confocal microscopic images (scale: 5 μm) for subcellular localization of Rad5-GFP, Rad16A-GFP and Rad16B-GFP, which were expressed in the WT hyphae stained with DAPI. The bright, expressed (green), stained (red) and merged views of each microscopic field are shown in images 1, 2, 3 and 4, respectively. (D) N/C-GFI ratios of Rad5-GFP, Rad16A-GFP and Rad16B-GFP fusion proteins. Different uppercase letters denote a significant difference of p < 0.01 (Tukey’s test). Error bars: standard deviations (SDs) from three cDNA samples (C) or 15 hyphal cells (D) analyzed. Full article ">Figure 2
Y2H assays for protein–protein interactions. (A–D) Interactions of Rad5 (E3 ubiquitin ligase) with Mms2 (E2 ubiquitin-conjugating enzyme), Rad16B with Rad23, and Rad16A and Rad16B with the Elc1-like E3 ubiquitin ligase. The constructed diploids during 3 days of incubation at 30 °C grew as well as positive control on quadruple-dropout plates (right in each image) after inoculation with 5 × 102 (C3), 5 × 103 (C2) and 5 × 104 (C1) cells. Full article ">Figure 3
Roles of Rad5, Rad16A and Rad16B in B. bassiana lifecycle. (A) Diameters of fungal colonies (DM, deletion mutant; DDM, double-deletion mutant; CM, complementation mutant) incubated at the optimal regime of 25 °C and 12:12 (L/D) for 7 days on the plates of 1/4 SDAY, CDA and CDAs amended with different carbon or nitrogen sources. (B) Relative growth inhibition (RGI) of fungal colonies after 7-day incubation at 25 °C on CDA plates containing indicated concentrations of different chemical stressors. The colony growth was initiated with ~103 conidia. (C) Conidial yields in the SDAY cultures incubated for 5, 7 and 9 days at the optimal regime after each culture was initiated by spreading 100 μL of conidial suspension (107 conidia/mL). (D) Estimates of GT50 as an index of conidial viability. (E) Estimates of LT50 as a virulence index of fungal strains against G. mellonella larvae through normal cuticle infection or cuticle-bypassing infection (intrahemocoel injection). p < 0.01 ** or 0.001 *** in Tukey’s test. Error bars: SDs from three independent replicates. Full article ">Figure 4
Comparative activities of Rad5, Rad16A and Rad16B in dark reactivation of B. bassiana conidia impaired by UVB to different degrees. (A) Microscopic images (scale: 20 μm) for germination status of UVB-impaired conidia after 12, 24 and 40 h of dark incubation at 25 °C. (B) Trends of conidial survival indices in the 12, 24 and 40 h dark treatments after exposure to gradient UVB doses. (C) LD50 values estimated as an index of NER activity from the survival trends fitted in different dark treatments. Different uppercase letters denote significant differences of p < 0.01 (Tukey’s HSD). Error bars: SDs from three independent replicates. Full article ">Figure 5
Comparative activities of Rad5, Rad16A and Rad16B in photoreactivation of B. bassiana conidia irradiated at gradient UVB doses. (A) Microscopic images (scale: 20 μm) for germination status of UVB-impaired conidia incubated at 25 °C for a total of 12 or 24 h in L/D 3:9, 5:7, 3:21 and 5:19 post-irradiation at 0.2 or 0.4 J/cm2, respectively. (B) Trends of conidial survival indices photoreactivated in the L/D treatments after conidia were irradiated at gradient UVB doses. (C) LD50 values estimated as an index of photorepair activity from the survival trends fitted in different L/D treatments. Different uppercase letters denote significant differences of p < 0.01 (Tukey’s HSD). Error bars: SDs from three independent replicates. Full article ">

25 pages, 4702 KiB

Open AccessArticle

Microbial-Based Biofungicides Mitigate the Damage Caused by Fusarium oxysporum f. sp. cubense Race 1 and Improve the Physiological Performance in Banana

by Luisa Fernanda Izquierdo-García, Sandra Lorena Carmona-Gutiérrez, Carlos Andrés Moreno-Velandia, Andrea del Pilar Villarreal-Navarrete, Diana Marcela Burbano-David, Ruth Yesenia Quiroga-Mateus, Magda Rocío Gómez-Marroquín, Gustavo Adolfo Rodríguez-Yzquierdo and Mónica Betancourt-Vásquez

J. Fungi 2024, 10(6), 419; https://doi.org/10.3390/jof10060419 - 12 Jun 2024

Viewed by 1159

Abstract

Fusarium wilt of banana (FWB) is the most limiting disease in this crop. The phytosanitary emergency caused by FWB since 2019 in Colombia has required the development of ecofriendly control methods. The aim of this study was to test the effectiveness of microbial-based [...] Read more.

Fusarium wilt of banana (FWB) is the most limiting disease in this crop. The phytosanitary emergency caused by FWB since 2019 in Colombia has required the development of ecofriendly control methods. The aim of this study was to test the effectiveness of microbial-based biofungicides against FWB caused by Fusarium oxysporum f. sp. cubense race 1 (Foc R1) and correlate such effect with plant physiological parameters. Five Trichoderma (T1 to T4 and T9) and four Bacillus (T5 to T8)-based biofungicides were evaluated in pot experiments. In vitro, dual confrontation tests were also carried out to test whether the in vitro effects on Foc growth were consistent with the in vivo effects. While Trichoderma-based T3, T4, and T9, and Bacillus-based T8, significantly reduced the growth of Foc R1 in vitro, Trichoderma-based T1, T3, T4, and T9 temporarily reduced the Foc population in the soil. However, the incidence progress of FWB was significantly reduced by Bacterial-based T7 (74% efficacy) and Trichoderma-based T2 (50% efficacy). The molecular analysis showed that T7 prevented the inner tissue colonization by Foc R1 in 80% of inoculated plants. The T2, T4, T7, and T9 treatments mitigated the negative effects caused by Foc R1 on plant physiology and growth. Our data allowed us to identify three promising treatments to control FWB, reducing the progress of the disease, delaying the colonization of inner tissue, and mitigating physiological damages. Further studies should be addressed to determine the modes of action of the biocontrol agents against Foc and validate the utilization in the field. Full article

(This article belongs to the Special Issue Plant Fungal Diseases and Crop Protection)

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Effect of microbial biofungicides (T1–T9) on Fusarium wilt incidence progress in banana plants. Blue marks represent the original data. Red marks represent the average incidence (n = 9). The black line shows the predicted incidence by the logistic growth model as fitted for each treatment. The parameters of the model are shown in <a href="#jof-10-00419-t002" class="html-table">Table 2</a>. T10: negative control. Full article ">Figure 2
Effect of microbial biofungicides on Fusarium wilt severity in banana plants. The graphics in the top panel show the effects of biofungicides on the area under the disease intensity index progress curve (AUDPC DII) at the end of each replicate (R1–R3) of the experiment (77, 72, and 68 days after transplant for R1, R2, and R3, respectively). Bars on the columns represent the standard deviation of the mean (n = 3). Columns with the same letter are not significantly different according to Tukey’s test (α = 0.05). Biofungicides: T1–T9. Negative control: T10. The images show the representative state of the plants at the end of the experiment in replicates 2 (R2) and 3 (R3) of the entire experiment. Full article ">Figure 3
Electrophoresis in agarose gel (1.4%) used for verification of PCR amplification of the marker SIX6b (specific for Foc R1) on DNA extracted from banana plant samples in the first replicate of the experiment. Full article ">Figure 4
Chlorophyll index reduction in banana plants at 70 days after transplantation in soil artificially inoculated with Foc R1 and treated with biofungicides. Bars on the columns represent the standard deviation of the mean (n = 3). Full article ">Figure 5
Plant height reduction of banana plants grown in soil artificially inoculated with Foc R1 at 70 days after transplant. Bars on the columns represent the standard deviation of the mean (n = 3). Full article ">Figure 6
Pseudostem diameter reduction of banana plants at 70 days after transplantation in soil artificially inoculated with Foc R1. Bars on the columns represent the standard deviation of the mean (n = 3). Full article ">Figure 7
In vitro antagonism test of the microbial fungicides against Foc R1. The inhibition of Foc growth was calculated on the basis of the diameter of the colony measured one week after incubation at 25 °C. Full article ">

22 pages, 2797 KiB

Open AccessReview

Bioprocess of Gibberellic Acid by Fusarium fujikuroi: The Challenge of Regulation, Raw Materials, and Product Yields

by Aranza Hernández Rodríguez, Adrián Díaz Pacheco, Shirlley Elizabeth Martínez Tolibia, Yazmin Melendez Xicohtencatl, Sulem Yali Granados Balbuena and Víctor Eric López y López

J. Fungi 2024, 10(6), 418; https://doi.org/10.3390/jof10060418 - 12 Jun 2024

Cited by 1 | Viewed by 1285

Abstract

Gibberellic acid (GA₃) is a tetracyclic diterpenoid carboxylic acid synthesized by the secondary metabolism of Fusarium fujikuroi. This phytohormone is widely studied due to the advantages it offers as a plant growth regulator, such as growth stimulation, senescence delay, flowering [...] Read more.

Gibberellic acid (GA₃) is a tetracyclic diterpenoid carboxylic acid synthesized by the secondary metabolism of Fusarium fujikuroi. This phytohormone is widely studied due to the advantages it offers as a plant growth regulator, such as growth stimulation, senescence delay, flowering induction, increased fruit size, and defense against abiotic or biotic stress, which improve the quality and yield of crops. Therefore, GA₃ has been considered as an innovative strategy to improve agricultural production. However, the yields obtained at large scale are insufficient for the current market demand. This low productivity is attributed to the lack of adequate parameters to optimize the fermentation process, as well as the complexity of its regulation. Therefore, this article describes the latest advances for potentializing the GA₃ production process, including an analysis of its origins from crops, the benefits of its application, the related biosynthetic metabolism, the maximum yields achieved from production processes, and their association with genetic engineering techniques for GA₃ producers. This work provides a new perspective on the critical points of the production process, in order to overcome the limits surrounding this modern line of bioengineering. Full article

(This article belongs to the Special Issue Recent Advances in Fungal Secondary Metabolism, 2nd Edition)

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18 pages, 6191 KiB

Open AccessArticle

Plant Growth Promotion and Biological Control against Rhizoctonia solani in Thai Local Rice Variety “Chor Khing” Using Trichoderma breve Z2-03

by Warin Intana, Nakarin Suwannarach, Jaturong Kumla, Prisana Wonglom and Anurag Sunpapao

J. Fungi 2024, 10(6), 417; https://doi.org/10.3390/jof10060417 - 11 Jun 2024

Cited by 1 | Viewed by 958

Abstract

Several strains of Trichoderma are applied in the field to control plant diseases due to their capacity to suppress fungal pathogens and control plant diseases. Some Trichoderma strains also are able to promote plant growth through the production of indole-3-acetic acid (IAA). In [...] Read more.

Several strains of Trichoderma are applied in the field to control plant diseases due to their capacity to suppress fungal pathogens and control plant diseases. Some Trichoderma strains also are able to promote plant growth through the production of indole-3-acetic acid (IAA). In southern Thailand, the local rice variety “Chor Khing” is mainly cultivated in the Songkhla province; it is characterized by slow growth and is susceptible to sheath blight caused by Rhizoctonia solani. Therefore, this research aimed to screen Trichoderma species with the ability to promote plant growth in this rice variety and enact biological control against R. solani. A total of 21 Trichoderma isolates were screened for indole compound production using the Salkowski reagent. The Z2-03 isolate reacted positively to the Salkowski reagent, indicating the production of the indole compound. High-performance liquid chromatography (HPCL) confirmed that Z2-03 produced IAA at 35.58 ± 7.60 μg/mL. The cell-free culture filtrate of the potato dextrose broth (CF) of Z2-03 induced rice germination in rice seeds, yielding root and shoot lengths in cell-free CF-treated rice that were significantly higher than those of the control (distilled water and culture broth alone). Furthermore, inoculation with Trichoderma conidia promoted rice growth and induced a defense response against R. solani during the seedling stage. Trichoderma Z2-03 displayed an antifungal capacity against R. solani, achieving 74.17% inhibition (as measured through dual culture assay) and the production of siderophores on the CAS medium. The pot experiment revealed that inoculation with the Trichoderma sp. Z2-03 conidial suspension increased the number of tillers and the plant height in the “Chor Khing” rice variety, and suppressed the percentage of disease incidence (PDI). The Trichoderma isolate Z2-03 was identified, based on the morphology and molecular properties of ITS, translation elongation factor 1-alpha (tef1-α), and RNA polymerase 2 (rpb2), as Trichoderma breve Z2-03. Our results reveal the ability of T. breve Z2-03 to act as a plant growth promoter, enhancing growth and development in the “Chor Khing” rice variety, as well as a biological control agent through its competition and defense induction mechanism in this rice variety. Full article

(This article belongs to the Special Issue Soil Fungi and Their Role in Plant Growth)

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15 pages, 3451 KiB

Open AccessArticle

Natural Prevalence, Molecular Characteristics, and Biological Activity of Metarhizium rileyi (Farlow) Isolated from Spodoptera frugiperda (J. E. Smith) Larvae in Mexico

by Yordanys Ramos, Samuel Pineda-Guillermo, Patricia Tamez-Guerra, Alonso Alberto Orozco-Flores, José Isaac Figueroa de la Rosa, Selene Ramos-Ortiz, Juan Manuel Chavarrieta-Yáñez and Ana Mabel Martínez-Castillo

J. Fungi 2024, 10(6), 416; https://doi.org/10.3390/jof10060416 - 8 Jun 2024

Viewed by 903

Abstract

Entomopathogenic fungi have been considered potential biological control agents against the fall armyworm Spodoptera frugiperda (J. E. Smith), the world’s most important pest of maize. In this study, we evaluated the natural infection, molecular characteristics, and biological activity of Metarhizium rileyi (Farlow) isolated [...] Read more.

Entomopathogenic fungi have been considered potential biological control agents against the fall armyworm Spodoptera frugiperda (J. E. Smith), the world’s most important pest of maize. In this study, we evaluated the natural infection, molecular characteristics, and biological activity of Metarhizium rileyi (Farlow) isolated from S. frugiperda larvae of this insect, collected from maize crops in five Mexican locations. Natural infection ranged from 23% to 90% across all locations analyzed. Twenty-four isolates were evaluated on S. frugiperda second instars at a concentration of 1.0 × 10⁸ conidia/mL, causing 70% to 98.7% mortality and 60.5% to 98.7% sporulation. Isolates T9-21, Z30-21, PP48-21, and L8-22 were selected to determine their phylogenetic relationships by β-tubulin gene analysis and to compare median lethal concentration (CL₅₀), median lethal time (LT₅₀), and larval survival. These isolates were grouped into three clades. The T9-21, PP48-21, and J10-22 isolates were closely related (clade A), but phylogenetically distant from Z30-21 (clade B) and L8-22 (clade C) isolates. These genetic differences were not always reflected in their pathogenicity characteristics since no differences were observed among the LC₅₀ values. Furthermore, isolates T9-21, J10-22, and L8-22 were the fastest to kill S. frugiperda larvae, causing lower survival rates. We conclude that native M. rileyi isolates represent an important alternative for the biocontrol of S. frugiperda. Full article

(This article belongs to the Section Fungal Pathogenesis and Disease Control)

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Morphological characteristics of M. rileyi. (A–E) Sporulation produced by isolates T9-21, Z30-21, PP48-21, J10-22, and L8-22 on S. frugiperda larvae, respectively, 8 days after inoculation. Whitish mycelium followed by light green spores can be observed. (F) M. rileyi colony on MPYA culture medium of 14 days after inoculation. A dense whitish cover is observed initially, which later turns into a light green colour. (G) Phialides in whorls and conidiophores. (H) Ovoid conidia dispersed and some of them form short chains. Full article ">Figure 2
Phylogenetic tree inferred using the neighbor-joining method of 13 β tubulin sequences from entomopathogenic fungi isolates. Five sequences of M. rileyi (T9-21, Z30-21, PP48-21, J10-22, and L8-22 isolates) obtained from S. frugiperda larvae in five locations of the Michoacán state, Mexico, were clustered using the Tamura 3 model. Branch length is presented as the number of base substitutions per site. Three clades (A, B, and C) were grouped with M. rileyi sequences. Full article ">Figure 3
Gehan–Breslow and Kaplan–Meier survival curves of second-instar larvae of S. frugiperda after inoculation with a concentration of 1.0 × 108 conidia/mL of five isolates of M. rileyi. Different letters indicate significant differences among isolates (log-rank test, p < 0.05). Full article ">

22 pages, 14933 KiB

Open AccessArticle

The Performance and Evolutionary Mechanism of Ganoderma lucidum in Enhancing Selenite Tolerance and Bioaccumulation

by Mengmeng Xu, Qi Meng, Song Zhu, Ruipeng Yu, Lei Chen, Guiyang Shi, Ka-Hing Wong, Daming Fan and Zhongyang Ding

J. Fungi 2024, 10(6), 415; https://doi.org/10.3390/jof10060415 - 8 Jun 2024

Viewed by 1034

Abstract

Background: Selenium (Se) pollution poses serious threats to terrestrial ecosystems. Mushrooms are important sources of Se with the potential for bioremediation. Pre-eminent Se resources must possess the ability to tolerate high levels of Se. To obtain Se-accumulating fungi, we isolated selenite-tolerance-enhanced Ganoderma lucidum [...] Read more.

Background: Selenium (Se) pollution poses serious threats to terrestrial ecosystems. Mushrooms are important sources of Se with the potential for bioremediation. Pre-eminent Se resources must possess the ability to tolerate high levels of Se. To obtain Se-accumulating fungi, we isolated selenite-tolerance-enhanced Ganoderma lucidum JNUSE-200 through adaptive evolution. Methods: The molecular mechanism responsible for selenite tolerance and accumulation was explored in G. lucidum JNUSE-200 by comparing it with the original strain, G. lucidum CGMCC 5.26, using a combination of physiological and transcriptomic approaches. Results: G. lucidum JNUSE-200 demonstrated tolerance to 200 mg/kg selenite in liquid culture and exhibited normal growth, whereas G. lucidum CGMCC 5.26 experienced reduced growth, red coloration, and an unpleasant odor as a result of exposure to selenite at the same concentration. In this study, G. lucidum JNUSE-200 developed a triple defense mechanism against high-level selenite toxicity, and the key genes responsible for improved selenite tolerance were identified. Conclusions: The present study offers novel insights into the molecular responses of fungi towards selenite, providing theoretical guidance for the breeding and cultivation of Se-accumulating varieties. Moreover, it significantly enhances the capacity of the bio-manufacturing industry and contributes to the development of beneficial applications in environmental biotechnology through fungal selenite transformation bioprocesses. Full article

(This article belongs to the Special Issue Edible and Medicinal Macrofungi, 3rd Edition)

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74 pages, 2577 KiB

Open AccessReview

An In-Depth Study of Phytopathogenic Ganoderma: Pathogenicity, Advanced Detection Techniques, Control Strategies, and Sustainable Management

by Samantha C. Karunarathna, Nimesha M. Patabendige, Wenhua Lu, Suhail Asad and Kalani K. Hapuarachchi

J. Fungi 2024, 10(6), 414; https://doi.org/10.3390/jof10060414 - 7 Jun 2024

Cited by 1 | Viewed by 1366

Abstract

Phytopathogenic Ganoderma species pose a significant threat to global plant health, resulting in estimated annual economic losses exceeding USD (US Dollars) 68 billion in the agriculture and forestry sectors worldwide. To combat this pervasive menace effectively, a comprehensive understanding of the biology, ecology, [...] Read more.

Phytopathogenic Ganoderma species pose a significant threat to global plant health, resulting in estimated annual economic losses exceeding USD (US Dollars) 68 billion in the agriculture and forestry sectors worldwide. To combat this pervasive menace effectively, a comprehensive understanding of the biology, ecology, and plant infection mechanisms of these pathogens is imperative. This comprehensive review critically examines various aspects of Ganoderma spp., including their intricate life cycle, their disease mechanisms, and the multifaceted environmental factors influencing their spread. Recent studies have quantified the economic impact of Ganoderma infections, revealing staggering yield losses ranging from 20% to 80% across various crops. In particular, oil palm plantations suffer devastating losses, with an estimated annual reduction in yield exceeding 50 million metric tons. Moreover, this review elucidates the dynamic interactions between Ganoderma and host plants, delineating the pathogen’s colonization strategies and its elicitation of intricate plant defense responses. This comprehensive analysis underscores the imperative for adopting an integrated approach to Ganoderma disease management. By synergistically harnessing cultural practices, biological control, and chemical treatments and by deploying resistant plant varieties, substantial strides can be made in mitigating Ganoderma infestations. Furthermore, a collaborative effort involving scientists, breeders, and growers is paramount in the development and implementation of sustainable strategies against this pernicious plant pathogen. Through rigorous scientific inquiry and evidence-based practices, we can strive towards safeguarding global plant health and mitigating the dire economic consequences inflicted by Ganoderma infections. Full article

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15 pages, 4715 KiB

Open AccessArticle

Active Prevalence of Fusarium falciforme and F. acutatum Causing Basal Rot of Onion in Maharashtra, India

by Ram Dutta, Krishnappa Jayalakshmi, Auji Radhakrishna, Satish Kumar and Vijay Mahajan

J. Fungi 2024, 10(6), 413; https://doi.org/10.3390/jof10060413 - 7 Jun 2024

Viewed by 1209

Abstract

Over the past decade, there have been accumulating reports from researchers, farmers, and field extension personnel on the increasing incidence and spread of onion basal rot in India. Onion basal rot disease is mainly caused by Fusarium spp. This study aimed to validate [...] Read more.

Over the past decade, there have been accumulating reports from researchers, farmers, and field extension personnel on the increasing incidence and spread of onion basal rot in India. Onion basal rot disease is mainly caused by Fusarium spp. This study aimed to validate the information on the active prevalence of F. falciforme and F. acutatum causing Fusarium basal rot (FBR) in Maharashtra. A survey was conducted, and the infected plants/bulbs were collected from fields of 38 locations comprising five districts of Maharashtra, namely, Nashik, Aurangabad, Solapur, Ahmednagar, and Pune, in 2023. This disease was prevalent in high-moisture and high-oil-temperature conditions and the symptoms were observed in most of the fields, with the FBR incidence ranging from 17 to 41%. The available data of basal rot incidence from 1998 to 2022 were analyzed, based on which the prevalence of FBR was 11–50%. Tissue from the infected samples of onion bulbs was used for the isolation. The identification was performed based on colony morphology and microscopic features and confirmed through molecular markers using ITS and Tef-1α gene primers. Of the ten Fusarium isolates collected from selected locations, six species were confirmed as F. acutatum and four as F. falciforme. The pathogenicity tests performed with onion seedlings and bulbs under moist conditions proved that both F. acutatum and F. falciforme independently could cause basal rot disease symptoms but with different degrees of virulence. Koch’s postulates were confirmed by reisolating the same pathogens from the infected plants. Thus, the active prevalence of FBR was confirmed in Maharashtra and also, to the best of our knowledge, this is the first report of F. falciforme and F. acutatum causing basal rot of onion independently in Maharashtra, India. Full article

(This article belongs to the Special Issue Management of Postharvest Fungal Diseases of Fruits and Vegetables)

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Disease incidence of Fusarium basal rot disease in the Maharashtra state of India from 1998 to 2022. Full article ">Figure 2
Disease incidence of Fusarium basal rot disease in the Maharashtra state of India during 2023. Full article ">Figure 3
Map showing the onion basal rot incidence in Maharashtra. Full article ">Figure 4
Varying symptoms of Fusarium Basal rot of onion: (a) FBR-infected field. (b) Single plant from the field with leaf dieback, rotting of the basal plate, and loss of roots. (c) Intact infected plant and split open bulb showing soft watery decay. Full article ">Figure 5
Colony morphology and microscopic characterization of the Fusarium spp. isolates: (A) top view of mycelia, (B) bottom view, (C) microscopic field (1000×), (a) macroconidia, (b) microconidia, and (c) spore-bearing structure (sporophyte). Full article ">Figure 6
Phylogenetic tree showing the relationship of 10 Fusarium isolates based on tef1-α gene sequence using the Maximum Likelihood method. The percentage of replicate trees in which the linked taxa clustered together in the bootstrap test (1000 replicates). Colletotrichum fructicola and Colletotrichum gloeosporioides were used as an out-group (* isolates included in this study). Full article ">

15 pages, 2780 KiB

Open AccessArticle

Investigation of the Microbial Diversity in the Oryza sativa Cultivation Environment and Artificial Transplantation of Microorganisms to Improve Sustainable Mycobiota

by Yeu-Ching Shi, Yu-Juan Zheng, Yi-Ching Lin, Cheng-Hao Huang, Tang-Long Shen, Yu-Chia Hsu and Bao-Hong Lee

J. Fungi 2024, 10(6), 412; https://doi.org/10.3390/jof10060412 - 6 Jun 2024

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Abstract

Rice straw is not easy to decompose, it takes a long time to compost, and the anaerobic bacteria involved in the decomposition process produce a large amount of carbon dioxide (CO₂), indicating that applications for rice straw need to be developed. [...] Read more.

Rice straw is not easy to decompose, it takes a long time to compost, and the anaerobic bacteria involved in the decomposition process produce a large amount of carbon dioxide (CO₂), indicating that applications for rice straw need to be developed. Recycling rice straw in agricultural crops is an opportunity to increase the sustainability of grain production. Several studies have shown that the probiotic population gradually decreases in the soil, leading to an increased risk of plant diseases and decreased biomass yield. Because the microorganisms in the soil are related to the growth of plants, when the soil microbial community is imbalanced it seriously affects plant growth. We investigated the feasibility of using composted rice stalks to artificially cultivate microorganisms obtained from the Oryza sativa-planted environment for analyzing the mycobiota and evaluating applications for sustainable agriculture. Microbes obtained from the water-submerged part (group-A) and soil part (group-B) of O. sativa were cultured in an artificial medium, and the microbial diversity was analyzed with internal transcribed spacer sequencing. Paddy field soil was mixed with fermented paddy straw compost, and the microbes obtained from the soil used for O. sativa planting were designated as group-C. The paddy fields transplanted with artificially cultured microbes from group-A were designated as group-D and those from group-B were designated as group-E. We found that fungi and yeasts can be cultured in groups-A and -B. These microbes altered the soil mycobiota in the paddy fields after transplantation in groups-D and -E compared to groups-A and -B. Development in O. sativa post treatment with microbial transplantation was observed in the groups-D and -E compared to group-C. These results showed that artificially cultured microorganisms could be efficiently transplanted into the soil and improve the mycobiota. Phytohormones were involved in improving O. sativa growth and rice yield via the submerged part-derived microbial medium (group-D) or the soil part-derived microbial medium (group-E) treatments. Collectively, these fungi and yeasts may be applied in microbial transplantation via rice straw fermentation to repair soil mycobiota imbalances, facilitating plant growth and sustainable agriculture. These fungi and yeasts may be applied in microbial transplantation to repair soil mycobiota imbalances and sustainable agriculture. Full article

(This article belongs to the Section Fungal Evolution, Biodiversity and Systematics)

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18 pages, 2344 KiB

Open AccessReview

Applications of the Methylotrophic Yeast Komagataella phaffii in the Context of Modern Biotechnology

by Lidia Maria Pepe de Moraes, Henrique Fetzner Marques, Viviane Castelo Branco Reis, Cintia Marques Coelho, Matheus de Castro Leitão, Alexsandro Sobreira Galdino, Thais Paiva Porto de Souza, Luiza Cesca Piva, Ana Laura Alfonso Perez, Débora Trichez, João Ricardo Moreira de Almeida, Janice Lisboa De Marco and Fernando Araripe Gonçalves Torres

J. Fungi 2024, 10(6), 411; https://doi.org/10.3390/jof10060411 - 6 Jun 2024

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Abstract

Komagataella phaffii (formerly Pichia pastoris) is a methylotrophic yeast widely used in laboratories around the world to produce recombinant proteins. Given its advantageous features, it has also gained much interest in the context of modern biotechnology. In this review, we present the [...] Read more.

Komagataella phaffii (formerly Pichia pastoris) is a methylotrophic yeast widely used in laboratories around the world to produce recombinant proteins. Given its advantageous features, it has also gained much interest in the context of modern biotechnology. In this review, we present the utilization of K. phaffii as a platform to produce several products of economic interest such as biopharmaceuticals, renewable chemicals, fuels, biomaterials, and food/feed products. Finally, we present synthetic biology approaches currently used for strain engineering, aiming at the production of new bioproducts. Full article

(This article belongs to the Special Issue New Perspectives on Industrial Yeasts)

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19 pages, 5433 KiB

Open AccessArticle

Biological Control of Root Rot of Strawberry by Bacillus amyloliquefaciens Strains CMS5 and CMR12

by Ruixian Yang, Ping Liu, Wenyu Ye, Yuquan Chen, Daowei Wei, Cuicui Qiao, Bingyi Zhou and Jingyao Xiao

J. Fungi 2024, 10(6), 410; https://doi.org/10.3390/jof10060410 - 6 Jun 2024

Cited by 1 | Viewed by 1107

Abstract

Strawberry root rot caused by Fusarium solani is one of the main diseases of strawberries and significantly impacts the yield and quality of strawberry fruit. Biological control is becoming an alternative method for the control of plant diseases to replace or decrease the [...] Read more.

Strawberry root rot caused by Fusarium solani is one of the main diseases of strawberries and significantly impacts the yield and quality of strawberry fruit. Biological control is becoming an alternative method for the control of plant diseases to replace or decrease the application of traditional chemical fungicides. To obtain antagonistic bacteria with a high biocontrol effect on strawberry root rot, over 72 rhizosphere bacteria were isolated from the strawberry rhizosphere soil and screened for their antifungal activity against F. solani by dual culture assay. Among them, strains CMS5 and CMR12 showed the strongest inhibitory activity against F. solani (inhibition rate 57.78% and 65.93%, respectively) and exhibited broad-spectrum antifungal activity. According to the phylogenetic tree based on 16S rDNA and gyrB genes, CMS5 and CMR12 were identified as Bacillus amyloliquefaciens. Lipopeptide genes involved in surfactin, iturin, and fengycin biosynthesis were detected in the DNA genomes of CMS5 and CMR12 by PCR amplification. The genes related to the three major lipopeptide metabolites existed in the DNA genome of strains CMS5 and CMR12, and the lipopeptides could inhibit the mycelial growth of F. solani and resulted in distorted hyphae. The inhibitory rates of lipopeptides of CMS5 and CMR12 on the spore germination of F. solani were 61.00% and 42.67%, respectively. The plant-growth-promoting (PGP) traits in vitro screening showed that CMS5 and CMR12 have the ability to fix nitrogen and secreted indoleacetic acid (IAA). In the potting test, the control efficiency of CMS5, CMR12 and CMS5+CMR12 against strawberry root rot were 65.3%, 67.94% and 88.00%, respectively. Furthermore, CMS5 and CMR12 enhanced the resistance of strawberry to F. solani by increasing the activities of defense enzymes MDA, CAT and SOD. Moreover, CMS5 and CMR12 significantly promoted the growth of strawberry seedlings such as root length, seedling length and seedling fresh weight. This study revealed that B. amyloliquefaciens CMS5 and CMR12 have high potential to be used as biocontrol agents to control strawberry root rot. Full article

(This article belongs to the Special Issue Plant Fungal Diseases and Crop Protection)

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