Research

15 pages, 3109 KiB

Open AccessArticle

MinION Sequencing of Fungi in Sub-Saharan African Air and a Novel LAMP Assay for Rapid Detection of the Tropical Phytopathogenic Genus Lasiodiplodia

by Kevin M. King, Gail G. M. Canning and Jonathan S. West

Pathogens 2024, 13(4), 330; https://doi.org/10.3390/pathogens13040330 - 17 Apr 2024

Viewed by 441

Abstract

To date, there have been no DNA-based metabarcoding studies into airborne fungi in tropical Sub-Saharan Africa. In this initial study, 10 air samples were collected onto Vaseline-coated acrylic rods mounted on drones flown at heights of 15–50 meters above ground for 10–15 min [...] Read more.

To date, there have been no DNA-based metabarcoding studies into airborne fungi in tropical Sub-Saharan Africa. In this initial study, 10 air samples were collected onto Vaseline-coated acrylic rods mounted on drones flown at heights of 15–50 meters above ground for 10–15 min at three sites in Ghana. Purified DNA was extracted from air samples, the internal transcribed spacer (ITS) region was amplified using fungal-specific primers, and MinION third-generation amplicon sequencing was undertaken with downstream bioinformatics analyses utilizing GAIA cloud-based software (at genus taxonomic level). Principal coordinate analyses based on Bray–Curtis beta diversity dissimilarity values found no clear evidence for the structuring of fungal air communities, nor were there significant differences in alpha diversity, based on geographic location (east vs. central Ghana), underlying vegetation type (cocoa vs. non-cocoa), or height above ground level (15–23 m vs. 25–50 m), and despite the short flight times (10–15 min), ~90 operational taxonomic units (OTUs) were identified in each sample. In Ghanaian air, fungal assemblages were skewed at the phylum taxonomic level towards the ascomycetes (53.7%) as opposed to basidiomycetes (24.6%); at the class level, the Dothideomycetes were predominant (29.8%) followed by the Agaricomycetes (21.8%). The most common fungal genus in Ghanaian air was cosmopolitan and globally ubiquitous Cladosporium (9.9% of reads). Interestingly, many fungal genera containing economically important phytopathogens of tropical crops were also identified in Ghanaian air, including Corynespora, Fusarium, and Lasiodiplodia. Consequently, a novel loop-mediated isothermal amplification (LAMP) assay, based on translation elongation factor-1α sequences, was developed and tested for rapid, sensitive, and specific detection of the fungal phytopathogenic genus Lasiodiplodia. Potential applications for improved tropical disease management are considered. Full article

(This article belongs to the Special Issue Fungal Pathogens of Crops)

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14 pages, 2107 KiB

Open AccessArticle

Searching for the Virulence-contributing Genes of the Magnaporthe oryzae by Transcriptome Analysis

by Jitao Hu, Linying Li, Yuqing He, Gaojie Hong and Chi Zhang

Pathogens 2024, 13(2), 105; https://doi.org/10.3390/pathogens13020105 - 25 Jan 2024

Viewed by 872

Abstract

Magnaporthe oryzae is a fungal pathogen that causes rice blast. Plant metabolites such as plant hormones and phytoalexin can promote or inhibit the rice blast infection. To study the effect of plant metabolites on M. oryzae, we selected salicylic acid (SA), abscisic [...] Read more.

Magnaporthe oryzae is a fungal pathogen that causes rice blast. Plant metabolites such as plant hormones and phytoalexin can promote or inhibit the rice blast infection. To study the effect of plant metabolites on M. oryzae, we selected salicylic acid (SA), abscisic acid (ABA), and a phytoalexin sakuranetin to treat M. oryzae grown on the medium. Through the analysis of transcriptome data, 185 and 38 genes, 803 and 156 genes, and 1525 and 428 genes were up- or down-regulated after SA, ABA, or sakuranetin treatment. Among these differentially expressed genes (DEGs), most of them were annotated to the cellular process and metabolic process in the biological process category and binding and catalytic activity in the molecular function category by GO analysis. According to KEGG pathway analysis, metabolism is the pathway with the highest number of DEGs, and the main enriched pathway is carbohydrate, lipid, and amino acid metabolism. In addition, we also found two ABA-induced up-regulated genes that may contribute to M. oryzae infection from the transcriptome data. We verified their expressions in M. oryzae that infected rice. Full article

(This article belongs to the Special Issue Fungal Pathogens of Crops)

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15 pages, 2264 KiB

Open AccessArticle

Expression of Genes Involved in Anthracnose Resistance in Chili (Capsicum baccatum) ‘PBC80’-Derived Recombinant Inbred Lines

by Wassana Kethom, Paul W. J. Taylor and Orarat Mongkolporn

Pathogens 2023, 12(11), 1306; https://doi.org/10.3390/pathogens12111306 - 01 Nov 2023

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Abstract

Chili anthracnose has long been a threat to chili production worldwide. Capsicum baccatum ‘PBC80’ has been identified as a source of resistance to anthracnose. Recently, a QTL for ripe fruit resistance from ‘PBC80’-derived RILs was located on chromosome 4 (123 Mb) and contained [...] Read more.

Chili anthracnose has long been a threat to chili production worldwide. Capsicum baccatum ‘PBC80’ has been identified as a source of resistance to anthracnose. Recently, a QTL for ripe fruit resistance from ‘PBC80’-derived RILs was located on chromosome 4 (123 Mb) and contained over 80 defense-related genes. To identify the genes most related to anthracnose resistance, a fine map of the QTL region was developed using single-marker analysis. Nine genes were selected from the new QTL (1.12 Mb) to study their expression after being challenged with Colletotrichum scovillei ‘MJ5’ in two different RIL genotypes (Resistance/Resistance or R/R and Susceptible/Susceptible or S/S) at 0, 6 and 12 h. Of the nine genes, LYM2, CQW23_09597, CLF, NFXL1, and PR-14 were significantly up-regulated, compared to the control, in the R/R genotype. ERF was up-regulated in both chili genotypes. However, the expression was relatively and constantly low in the S/S genotype. Most up-regulated genes reached the highest peak (2.3–4.5 fold) at 6 h, except for ERF, which had the highest peak at 12 h (6.4 fold). The earliest and highest expressed gene was a pathogen receptor, LYM2. Full article

(This article belongs to the Special Issue Fungal Pathogens of Crops)

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18 pages, 4130 KiB

Open AccessArticle

The Green Synthesis of Silver Nanoparticles from Avena fatua Extract: Antifungal Activity against Fusarium oxysporum f.sp. lycopersici

by Ahmad Kaleem Qureshi, Umar Farooq, Qaiser Shakeel, Sajjad Ali, Sarfraz Ashiq, Sohail Shahzad, Muhammad Tariq, Mahmoud F. Seleiman, Aftab Jamal, Muhammad Farhan Saeed and Barbara Manachini

Pathogens 2023, 12(10), 1247; https://doi.org/10.3390/pathogens12101247 - 16 Oct 2023

Cited by 2 | Viewed by 1330

Abstract

Using plant extracts as eco-friendly reducing and stabilizing agents for the synthesis of nanoparticles has gained significant attention in recent years. The current study explores the green synthesis of silver nanoparticles (AgNPs) using the Avena fatua extract and evaluates their antifungal activity against [...] Read more.

Using plant extracts as eco-friendly reducing and stabilizing agents for the synthesis of nanoparticles has gained significant attention in recent years. The current study explores the green synthesis of silver nanoparticles (AgNPs) using the Avena fatua extract and evaluates their antifungal activity against Fusarium oxysporum f.sp. lycopersici (Fol), a fungal plant pathogen. A green and sustainable approach was adopted to synthesize silver nanoparticles before these nanoparticles were employed for anti-fungal activity. The primary indication that AgNPs had formed was performed using UV-vis spectroscopy, where a strong peak at 425 nm indicated the effective formation of these nanoparticles. The indication of important functional groups acting as reducing and stabilizing agents was conducted using the FTIR study. Additionally, morphological studies were executed via SEM and AFM, which assisted with more effectively analyzing AgNPs. Crystalline behavior and size were estimated using powder XRD, and it was found that AgNPs were highly crystalline, and their size ranged from 5 to 25 nm. Synthesized AgNPs exhibited significant antifungal activity against Fol at a concentration of 40 ppm. Furthermore, the inhibitory index confirmed a positive correlation between increasing AgNPs concentration and exposure duration. This study suggests that the combined phytochemical mycotoxic effect of the plant extract and the smaller size of synthesized AgNPs were responsible for the highest penetrating power to inhibit Fol growth. Moreover, this study highlights the potential of using plant extracts as reducing and capping agents for the green synthesis of AgNPs with antifungal properties. The study concludes that A. fatua extract can synthesize antifungal AgNPs as a sustainable approach with robust antifungal efficacy against Fol, underscoring their promising potential for integration into plant protection strategies. Full article

(This article belongs to the Special Issue Fungal Pathogens of Crops)

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14 pages, 876 KiB

Open AccessCommunication

Identification and Pathogenicity of Fusarium Isolated from Soybean in Poland

by Hanna Olszak-Przybyś, Grażyna Korbecka-Glinka and Elżbieta Patkowska

Pathogens 2023, 12(9), 1162; https://doi.org/10.3390/pathogens12091162 - 14 Sep 2023

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Abstract

Fungi belonging to the Fusarium genus are commonly isolated from soybean plants and seeds but not all of them are pathogenic. The aim of this study was to compare the pathogenicity among different Fusarium isolates obtained from soybean plants with disease symptoms originating [...] Read more.

Fungi belonging to the Fusarium genus are commonly isolated from soybean plants and seeds but not all of them are pathogenic. The aim of this study was to compare the pathogenicity among different Fusarium isolates obtained from soybean plants with disease symptoms originating from an experimental field located in the southeast of Poland. Nineteen fungal isolates were selected for the pathogenicity assay, including eight isolates of F. oxysporum, six isolates of F. graminearum, four isolates of F. culmorum and one isolate of F. redolens. Species identification of these isolates was carried out using microscopic methods and sequencing of two genes: translation elongation factor 1-alpha (TEF1) and RNA polymerase second largest subunit (RPB2). To our knowledge, this is the first report of F. redolens being isolated from soybean in Europe. The pathogenicity test was set up by fungal inoculation of healthy soybean seeds of three cultivars: Abelina, Atlanta and Mavka. Symptoms were assessed seven days after inoculation. Disease area percentage of Fusarium inoculated seeds was significantly higher compared to uninoculated control. Nineteen isolates differed in their aggressiveness as the median disease area percentage ranged between 5.0 and 88.0% depending on isolate. The obtained isolates of four Fusarium species may be used in the future screening of soybean cultivars for resistance to these pathogens. Full article

(This article belongs to the Special Issue Fungal Pathogens of Crops)

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11 pages, 286 KiB

Open AccessArticle

Influence of Integrated Management Strategies on Soybean Sudden Death Syndrome (SDS) Root Infection, Foliar Symptoms, Yield and Net Returns

by Mariama T. Brown, Daren S. Mueller, Yuba R. Kandel and Darcy E. P. Telenko

Pathogens 2023, 12(7), 913; https://doi.org/10.3390/pathogens12070913 - 06 Jul 2023

Viewed by 1347

Abstract

Three soybean field trials were conducted in Indiana to evaluate the integration of seed treatment, cultivar selection, and seeding rate on sudden death syndrome (SDS) root rot, pathogen load in the root, foliar symptoms, yield, and net return. Two soybean cultivars, one moderately [...] Read more.

Three soybean field trials were conducted in Indiana to evaluate the integration of seed treatment, cultivar selection, and seeding rate on sudden death syndrome (SDS) root rot, pathogen load in the root, foliar symptoms, yield, and net return. Two soybean cultivars, one moderately resistant and one susceptible to SDS, were planted at three seeding rates (272,277 seeds/ha, 346,535 seeds/ha, and 420,792 seeds/ha). Fluopyram and pydiflumetofen seed treatments were applied to both cultivars, and the cultivars were then compared with a control. Low foliar SDS disease pressure was observed in our study. Seed treatment with either fluopyram or pydiflumetofen and the use of a moderately resistant cultivar decreased Fusarium virguliforme DNA concentration in the root relative to the control and the use of a susceptible cultivar. Fluopyram significantly reduced visual root rot severity by 8.8% and increased yield by 105 kg/ha relative to the control but was not different from pydiflumetofen. However, pydiflumetofen performed the same as the control with respect to root rot severity and yield. Findings from this study support the use of a seed treatment to protect roots from infection and the use of a moderately resistant cultivar planted at a seeding rate of 346,535 seeds/ha to protect yield and maximize net returns when a field has low foliar SDS pressure. Full article

(This article belongs to the Special Issue Fungal Pathogens of Crops)

13 pages, 3143 KiB

Open AccessCommunication

Multi-Trait Genome-Wide Association Studies of Sorghum bicolor Regarding Resistance to Anthracnose, Downy Mildew, Grain Mold and Head Smut

by Ezekiel Ahn, Louis K. Prom and Clint Magill

Pathogens 2023, 12(6), 779; https://doi.org/10.3390/pathogens12060779 - 30 May 2023

Cited by 1 | Viewed by 1337

Abstract

Multivariate linear mixed models (mvLMMs) are widely applied for genome-wide association studies (GWAS) to detect genetic variants affecting multiple traits with correlations and/or different plant growth stages. Subsets of multiple sorghum populations, including the Sorghum Association Panel (SAP), the Sorghum Mini Core Collection [...] Read more.

Multivariate linear mixed models (mvLMMs) are widely applied for genome-wide association studies (GWAS) to detect genetic variants affecting multiple traits with correlations and/or different plant growth stages. Subsets of multiple sorghum populations, including the Sorghum Association Panel (SAP), the Sorghum Mini Core Collection and the Senegalese sorghum population, have been screened against various sorghum diseases such as anthracnose, downy mildew, grain mold and head smut. Still, these studies were generally performed in a univariate framework. In this study, we performed GWAS based on the principal components of defense-related multi-traits against the fungal diseases, identifying new potential SNPs (S04_51771351, S02_66200847, S09_47938177, S08_7370058, S03_72625166, S07_17951013, S04_66666642 and S08_51886715) associated with sorghum’s defense against these diseases. Full article

(This article belongs to the Special Issue Fungal Pathogens of Crops)

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20 pages, 39086 KiB

Open AccessArticle

Whole Genome Sequencing and Comparative Genomics of Indian Isolates of Wheat Spot Blotch Pathogen Bipolaris sorokiniana Reveals Expansion of Pathogenicity Gene Clusters

by Sagar Yadav, Zarrine Raazi, Sheelavanta Matha Shivaraj, Deepika Somani, Ramya Prashant, Abhijeet Kulkarni, Rajeev Kumar, Suma Biradar, Shreenivas Desai and Narendra Kadoo

Pathogens 2023, 12(1), 1; https://doi.org/10.3390/pathogens12010001 - 20 Dec 2022

Viewed by 1942

Abstract

Spot blotch is a highly destructive disease in wheat caused by the fungal pathogen Bipolaris sorokiniana (teleomorph, Cochliobolus sativus). It is prevalent in warm and humid areas, including Africa, Asia, Latin America, and the USA. In the present study, twelve isolates of [...] Read more.

Spot blotch is a highly destructive disease in wheat caused by the fungal pathogen Bipolaris sorokiniana (teleomorph, Cochliobolus sativus). It is prevalent in warm and humid areas, including Africa, Asia, Latin America, and the USA. In the present study, twelve isolates of B. sorokiniana were collected from wheat fields in three different geographical locations in India. The pathogenicity of seven sporulating isolates was assessed on ‘DDK 1025’, a spot blotch-susceptible wheat variety under greenhouse conditions. The isolate ‘D2’ illustrated the highest virulence, followed by ‘SI’ and ‘BS52’. These three isolates were sequenced using the Illumina HiSeq1000 platform. The estimated genome sizes of the isolates BS52, D2, and SI were 35.19 MB, 39.32 MB, and 32.76 MB, with GC contents of 48.48%, 50.43%, and 49.42%, respectively. The numbers of pathogenicity genes identified in BS52, D2, and SI isolates were 2015, 2476, and 2018, respectively. Notably, the isolate D2 exhibited a relatively larger genome with expanded arsenals of Biosynthetic Gene Clusters (BGCs), CAZymes, secretome, and pathogenicity genes, which could have contributed to its higher virulence among the tested isolates. This study provides the first comparative genome analysis of the Indian isolates of B. sorokiniana using whole genome sequencing. Full article

(This article belongs to the Special Issue Fungal Pathogens of Crops)

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