ahmad fakhoury | Southern Illinois University (original) (raw)

Papers by ahmad fakhoury

Research paper thumbnail of Identification of genes differentially expressed during aflatoxin biosynthesis in Aspergillus flavus and Aspergillus parasiticus

Fungal Genetics and Biology, Jul 1, 2003

A complex regulatory network governs the biosynthesis of aflatoxin. While several genes involved ... more A complex regulatory network governs the biosynthesis of aflatoxin. While several genes involved in aflatoxin production are known, their action alone cannot account for its regulation. Arrays of clones from an Aspergillus flavus cDNA library and glass slide microarrays of ESTs were screened to identify additional genes. An initial screen of the cDNA clone arrays lead to the identification of

Research paper thumbnail of Transformation ofFusarium virguliforme,the Causal Agent of Sudden Death Syndrome of Soybean

Journal of Phytopathology, May 1, 2009

Fusarium virguliforme is a soil‐borne pathogen that causes sudden death syndrome (SDS) in soybean... more Fusarium virguliforme is a soil‐borne pathogen that causes sudden death syndrome (SDS) in soybean. SDS is an important disease that causes significant losses in soybean growing areas worldwide. Little is known about the interaction between F. virguliforme and soybean. We have developed a protoplast‐based fungal transformation system for F. virguliforme. One of the applications of the transformation system was the production of a green fluorescent protein (GFP)‐expressing fungal transformant. The GFP‐expressing fungus can be used to study fungal infection processes including fungal penetration, colonization, and spread, especially at the early stages of disease development. Furthermore, in an attempt to increase the genetic resources available to identify and characterize fungal virulence genes involved in the F. virguliforme‐soybean system, we generated random insertional mutations in F. virguliforme using restriction enzyme mediated integration.

Research paper thumbnail of <i>Amy1</i>, the α-Amylase Gene of <i>Aspergillus flavus</i>: Involvement in Aflatoxin Biosynthesis in Maize Kernels

Phytopathology, Oct 1, 1999

Aspergillus flavus is the causal agent of an ear and kernel rot in maize. In this study, we chara... more Aspergillus flavus is the causal agent of an ear and kernel rot in maize. In this study, we characterized an α-amylase-deficient mutant and assessed its ability to infect and produce aflatoxin in wounded maize kernels. The α-amylase gene Amy1 was isolated from A. flavus, and its DNA sequence was determined to be nearly identical to Amy3 of A. oryzae. When Amy1 was disrupted in an aflatoxigenic strain of A. flavus, the mutant failed to produce extracellular α-amylase and grew 45% the rate of the wild-type strain on starch medium. The mutant produced aflatoxin in medium containing glucose but not in a medium containing starch. The α-amylasedeficient mutant produced aflatoxin in maize kernels with wounded embryos and occasionally produced aflatoxin only in embryos of kernels with wounded endosperm. The mutant strain failed to produce aflatoxin when inoculated onto degermed kernels. In contrast, the wild-type strain produced aflatoxin in both the endosperm and embryo. These results suggest that α-amylase facilitates aflatoxin production and growth of A. flavus from a wound in the endosperm to the embryo. A 14-kDa trypsin inhibitor associated with resistance to A. flavus and aflatoxin in maize also inhibited the α-amylase from A. flavus, indicating that it is a bifunctional inhibitor. The inhibitor may have a role in resistance, limiting the growth of the fungus in the endosperm tissue by inhibiting the degradation of starch.

Research paper thumbnail of Inhibition of Growth of <i>Aspergillus flavus</i> and Fungal α-Amylases by a Lectin-Like Protein from <i>Lablab purpureus</i>

Molecular Plant-microbe Interactions, Aug 1, 2001

Aspergillus flavus is a fungal pathogen of maize causing an important ear rot disease when plants... more Aspergillus flavus is a fungal pathogen of maize causing an important ear rot disease when plants are exposed to drought and heat stress. Associated with the disease is the production of aflatoxins, which are a series of structurally related mycotoxins known to be carcinogenic. Previous research has suggested that the α-amylase of A. flavus promotes aflatoxin production in the endosperm of infected maize kernels. We report here the isolation and characterization of a 36-kDa α-amylase inhibitor from Lablab purpureus (AILP). AILP inhibited the α-amylases from several fungi but had little effect on those from animal and plant sources. The protein inhibited conidial germination and hyphal growth of A. flavus. The amino acid sequence indicated that AILP is similar to lectin members of a lectin-arcelin-α-amylase inhibitor family described in common bean and shown to be a component of plant resistance to insect pests. AILP also agglutinated papain-treated red blood cells from human and rabbit. These data indicate that AILP represents a novel variant in the lectin-arcelin-α-amylase inhibitor family of proteins having lectin-like and α-amylase inhibitory activity.

Research paper thumbnail of Genomics of Filamentous Fungi

Mycology [electronic resource], Dec 17, 2003

Research paper thumbnail of Fluopyram activates systemic resistance in soybean

Frontiers in Plant Science

The soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is a significant yield-limiting fa... more The soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is a significant yield-limiting factor in soybean production in the Midwestern US. Several management practices are implemented to mitigate yield losses caused by SCN, including using SDHI (succinate dehydrogenase inhibitors) fungicides delivered as seed treatments. A set of studies was conducted to evaluate the effect of two seed-applied succinate dehydrogenase inhibitors (SDHI) compounds, fluopyram and pydiflumetofen, on SCN population densities, plant injury, and plant growth. Cyst counts in untreated control and pydiflumetofen treated plants were 3.44 and 3.59 times higher than fluopyram, respectively, while egg counts were 8.25 and 7.06 times higher in control and pydiflumetofen. Next-generation sequencing was later employed to identify transcriptomic shifts in gene expression profiles in fluopyram and pydiflumetofen -treated seedlings. RNA expression patterns of seed treatments clustered by sampling time (5 DAP vs....

Research paper thumbnail of The Effect of Wheat on Soybean Cyst Nematode Populations in Double-Cropping Soybean Production

Research paper thumbnail of Characterization of Quinone Outside Inhibitor Fungicide Resistance in Cercospora sojina and Development of Diagnostic Tools for its Identification

Plant Disease, 2015

Frogeye leaf spot of soybean, caused by the fungus Cercospora sojina, reduces soybean yields in m... more Frogeye leaf spot of soybean, caused by the fungus Cercospora sojina, reduces soybean yields in most of the top-producing countries around the world. Control strategies for frogeye leaf spot can rely heavily on quinone outside inhibitor (QoI) fungicides. In 2010, QoI fungicide-resistant C. sojina isolates were identified in Tennessee for the first time. As the target of QoI fungicides, the cytochrome b gene present in fungal mitochondria has played a key role in the development of resistance to this fungicide class. The cytochrome b genes from three QoI-sensitive and three QoI-resistant C. sojina isolates were cloned and sequenced. The complete coding sequence of the cytochrome b gene was identified and found to encode 396 amino acids. The QoI-resistant C. sojina isolates contained the G143A mutation in the cytochrome b gene, a guanidine to cytosine transversion at the second position in codon 143 that causes an amino acid substitution of alanine for glycine. C. sojina-specific poly...

Research paper thumbnail of A panel of qPCR assays to detect and quantify soybean soil-borne pathogens

Letters in Applied Microbiology, Jan 23, 2023

Fusarium oxysporum,F. graminearum,F. acuminatum,F. equiseti,F. proliferatum,F. solani, and Rhizoc... more Fusarium oxysporum,F. graminearum,F. acuminatum,F. equiseti,F. proliferatum,F. solani, and Rhizoctonia solani are soil-borne fungal pathogens that cause substantial yield loss in a widespread list of crops worldwide. The objective of this study was to develop a panel of TaqMan assays for the detection and quantification of these six widespread soil-borne fungal species using real-time polymerase chain reaction (qPCR). The primers and probes were designed based on the intergenic spacer ribosomal RNA and translation elongation factor 1-alpha gene (tef1). These assays, although not multiplexed, can be performed simultaneously as they have similar reaction conditions, allowing more efficiency when targeting multiple pathogens in a sample. The assays presented high efficiency (94.3%-108.9%) and sensitivity, with a limit of detection of 0.05 picograms (50 femtograms) of target DNA. Results from an assay targeting 19 non-target and closely related species confirmed the specificity of the developed assays. The assays were also evaluated to detect the target species in different matrices, such as soil and plant material. This panel of qPCR assays is an additional tool that can be used by plant pathologists, microbiologists, plant breeders, diagnostic clinics, and other researchers interested in these fungal species.

Research paper thumbnail of Interactions and Aflatoxin Production

Environmental influences on maize-Aspergillus flavus

Research paper thumbnail of Interactions and Aflatoxin Production

Environmental influences on maize-Aspergillus flavus

Research paper thumbnail of Ecology and diversity of culturable fungal species associated with soybean seedling diseases in the Midwestern United States

Journal of Applied Microbiology, 2022

To isolate and characterize fungi associated with diseased soybean seedlings in Midwestern soybea... more To isolate and characterize fungi associated with diseased soybean seedlings in Midwestern soybean production fields and to determine the influence of environmental and edaphic factors on their incidence.

Research paper thumbnail of Reduction of Pythium Damping-off in Soybean by Biocontrol Seed Treatment

Plant Disease

Pythium spp. is one of the major groups of pathogens that cause seedling diseases on soybean, lea... more Pythium spp. is one of the major groups of pathogens that cause seedling diseases on soybean, leading to both pre- and post-emergence damping-off and root rot. More than 100 species have been identified within this genus, with P. irregulare, P. sylvaticum, P. ultimum var ultimum, and P. torulosum, being particularly important for soybean production given their aggressiveness, prevalence, and abundance in production fields. This study investigated the antagonistic activity of potential BCAs native to the US Midwest against Pythium spp. First, in vitro screening identified BCAs that inhibit P. ultimum var. ultimum growth. Scanning electron microscopy demonstrated evidence of mycoparasitism of all potential biocontrol isolates against the Pythium ultimum var. ultimum and P. torulosum, with the formation of appressorium-like structures, short-hyphal branches around host hyphae, hook-shaped structures, coiling, and parallel growth of the mycoparasite along the host hyphae. Based on these...

Research paper thumbnail of Rocha - SON2020 - Shifts in soil microbial communities in double-cropping soybeans - implications in the management of the soybean cyst nematode

In double cropping (DC) systems, fields have two or more crops growing in sequence in a single gr... more In double cropping (DC) systems, fields have two or more crops growing in sequence in a single growing season. For soybean, the crop is commonly planted following winter wheat. The soybean cyst nematode (SCN) (Heterodera glycines) is a major soybean pathogen. Several reports in the literature suggested potential suppressive effects of wheat residue on SCN population densities. A field trial was conducted in the 2017-2018 growing season to assess the effect of wheat on SCN populations in DC soybeans. In each field location (N=9), wheat (WT) was planted in strips alternating with strips maintained in fallow (FL) over winter. Soybean followed all strips after wheat harvest. Wheat strips had reduced SCN population densities compared to fallow strips, at soybean growth stage R1 and after soybean harvest. Three field locations with noted SCN suppression were selected for a metagenomics study. Ten subplots were selected (5 WT and 5 FL) from each location. A total of 90 soil samples were se...

Research paper thumbnail of Unraveling the microbial profile of the rhizosphere of SDS-suppressive soils in Soybean fields

Research paper thumbnail of Wheat Production Alters Soil Microbial Profiles and Enhances Beneficial Microbes in Double-Cropping Soybean

Frontiers in Agronomy, 2022

Plant-parasitic nematodes represent a substantial constraint on global food security by reducing ... more Plant-parasitic nematodes represent a substantial constraint on global food security by reducing the yield potential of all major crops. The soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is widely distributed across important soybean production areas of the U.S., being the major soybean yield-limiting factor, especially in the Midwestern U.S. Double cropped (DC) soybean is commonly planted following winter wheat. We previously reported double-cropping soybean fields with reduced SCN counts compared to fallow at both R1 growth stage (beginning of flowering) (−31.8%) and after soybean harvest (−32.7%). To test if higher counts of beneficial and SCN antagonistic microorganisms could be correlated with the suppression of SCN in fields previously planted with wheat, three field locations with noted SCN suppression were selected for a metagenomics study. Ten subplots were selected (5 wheat and 5 fallow pre-soybean) from each location. A total of 90 soil samples were selected:...

Research paper thumbnail of Weeds Hosting the Soybean Cyst Nematode (Heterodera glycines Ichinohe): Management Implications in Agroecological Systems

Agronomy, 2021

The soybean cyst nematode (SCN; Heterodera glycines Ichinohe) is a major soybean-yield-limiting s... more The soybean cyst nematode (SCN; Heterodera glycines Ichinohe) is a major soybean-yield-limiting soil-borne pathogen, especially in the Midwestern US. Weed management is recommended for SCN integrated management, since some weed species have been reported to be hosts for SCN. The increase in the occurrence of resistance to herbicides complicates weed management and may further direct ecological–evolutionary (eco–evo) feedbacks in plant–pathogen complexes, including interactions between host plants and SCN. In this review, we summarize weed species reported to be hosts of SCN in the US and outline potential weed–SCN management interactions. Plants from 23 families have been reported to host SCN, with Fabaceae including most host species. Out of 116 weeds hosts, 14 species have known herbicide-resistant biotypes to 8 herbicide sites of action. Factors influencing the ability of weeds to host SCN are environmental and edaphic conditions, SCN initial inoculum, weed population levels, and...

Research paper thumbnail of Hyperspectral Spectroscopy to Detect Different Responses of Two Soybean (Glycine Max) Cultivars to Charcoal Rot (Macrophomina Phaseolina) Toxin

Engenharia Agricola, 2021

Previous studies have confirmed the potential of hyperspectral spectroscopy (HS) to detect charco... more Previous studies have confirmed the potential of hyperspectral spectroscopy (HS) to detect charcoal rot toxin effect on soybean (Glycine max). However, there is no evidence that it can differentiate among difference responses to that effect. This research tested the potential of HS (900-2400 nm) to differentiate different responses to charcoal rot effect caused by the fungal pathogen Macrophomina phaseolina. Known susceptible 'Pharaoh' and resistant 'Spencer' cultivars of soybean seedlings at the V1 stage were tested, and HS reflectance was measured in each treatment at 0 h (just before toxin treatment), 24 h, and 36 h after toxin treatment. For both cultivars, reflectance generally decreased up to 10% over time for controls and increased for treated seedlings up to 22%. Pharaoh had significant increases in reflectance 24 and 36 h after treatment, while Spencer did not exhibit significant changes in reflectance over the 24 and 36-h periods. The ratio of Al-Ahmadi et al. (2018) was also evaluated. Spencer showed a higher ratio than Pharaoh. Based on our results, HS can be used to compare the relative resistance of soybean cultivars to charcoal rot and is a potential screening technique to classify soybean cultivars based on their resistance to charcoal rot toxin.

Research paper thumbnail of Impact of Wheat on Soybean Cyst Nematode Population Density in Double-Cropping Soybean Production

Frontiers in Plant Science, 2021

Double-cropping is defined as producing more than one crop on the same parcel of land in a single... more Double-cropping is defined as producing more than one crop on the same parcel of land in a single growing season. It is reported to have many benefits when incorporated in cropping systems, including improving soil health. In some double-cropping systems, soybean is planted following winter wheat. The soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is a major soybean pathogen, and several reports suggest suppressive effects of wheat on SCN populations. Field trials were conducted from 2017 to 2018 to investigate the effect of wheat on SCN populations in double-cropping soybean. Nine fields with three levels of initial SCN populations (low, moderate, and high) were selected in Illinois. Wheat was planted in strips alternating with strips-maintained weed-free and under fallow over winter and early spring. Soybean was planted in all strips after wheat harvest. SCN egg densities were acquired at four time points: wheat establishment, post-wheat/pre-soybean, mid-soybean (R1 gr...

Research paper thumbnail of Use of Dual RNA-seq for Systems Biology Analysis of Zea mays and Aspergillus flavus Interaction

Frontiers in Microbiology, 2020

The interaction between Aspergillus flavus and Zea mays is complex, and the identification of pla... more The interaction between Aspergillus flavus and Zea mays is complex, and the identification of plant genes and pathways conferring resistance to the fungus has been challenging. Therefore, the authors undertook a systems biology approach involving dual RNA-seq to determine the simultaneous response from the host and the pathogen. What was dramatically highlighted in the analysis is the uniformity in the development patterns of gene expression of the host and the pathogen during infection. This led to the development of a "stage of infection index" that was subsequently used to categorize the samples before downstream system biology analysis. Additionally, we were able to ascertain that key maize genes in pathways such as the jasmonate, ethylene and ROS pathways, were up-regulated in the study. The stage of infection index used for the transcriptomic analysis revealed that A. flavus produces a relatively limited number of transcripts during the early stages (0 to 12 h) of infection. At later stages, in A. flavus, transcripts and pathways involved in endosomal transport, aflatoxin production, and carbohydrate metabolism were up-regulated. Multiple WRKY genes targeting the activation of the resistance pathways (i.e., jasmonate, phenylpropanoid, and ethylene) were detected using causal inference analysis. This analysis also revealed, for the first time, the activation of Z. mays resistance genes influencing the expression of specific A. flavus genes. Our results show that A. flavus seems to be reacting to a hostile environment resulting from the activation of resistance pathways in Z. mays. This study revealed the dynamic nature of the interaction between the two organisms.

Research paper thumbnail of Identification of genes differentially expressed during aflatoxin biosynthesis in Aspergillus flavus and Aspergillus parasiticus

Fungal Genetics and Biology, Jul 1, 2003

A complex regulatory network governs the biosynthesis of aflatoxin. While several genes involved ... more A complex regulatory network governs the biosynthesis of aflatoxin. While several genes involved in aflatoxin production are known, their action alone cannot account for its regulation. Arrays of clones from an Aspergillus flavus cDNA library and glass slide microarrays of ESTs were screened to identify additional genes. An initial screen of the cDNA clone arrays lead to the identification of

Research paper thumbnail of Transformation ofFusarium virguliforme,the Causal Agent of Sudden Death Syndrome of Soybean

Journal of Phytopathology, May 1, 2009

Fusarium virguliforme is a soil‐borne pathogen that causes sudden death syndrome (SDS) in soybean... more Fusarium virguliforme is a soil‐borne pathogen that causes sudden death syndrome (SDS) in soybean. SDS is an important disease that causes significant losses in soybean growing areas worldwide. Little is known about the interaction between F. virguliforme and soybean. We have developed a protoplast‐based fungal transformation system for F. virguliforme. One of the applications of the transformation system was the production of a green fluorescent protein (GFP)‐expressing fungal transformant. The GFP‐expressing fungus can be used to study fungal infection processes including fungal penetration, colonization, and spread, especially at the early stages of disease development. Furthermore, in an attempt to increase the genetic resources available to identify and characterize fungal virulence genes involved in the F. virguliforme‐soybean system, we generated random insertional mutations in F. virguliforme using restriction enzyme mediated integration.

Research paper thumbnail of <i>Amy1</i>, the α-Amylase Gene of <i>Aspergillus flavus</i>: Involvement in Aflatoxin Biosynthesis in Maize Kernels

Phytopathology, Oct 1, 1999

Aspergillus flavus is the causal agent of an ear and kernel rot in maize. In this study, we chara... more Aspergillus flavus is the causal agent of an ear and kernel rot in maize. In this study, we characterized an α-amylase-deficient mutant and assessed its ability to infect and produce aflatoxin in wounded maize kernels. The α-amylase gene Amy1 was isolated from A. flavus, and its DNA sequence was determined to be nearly identical to Amy3 of A. oryzae. When Amy1 was disrupted in an aflatoxigenic strain of A. flavus, the mutant failed to produce extracellular α-amylase and grew 45% the rate of the wild-type strain on starch medium. The mutant produced aflatoxin in medium containing glucose but not in a medium containing starch. The α-amylasedeficient mutant produced aflatoxin in maize kernels with wounded embryos and occasionally produced aflatoxin only in embryos of kernels with wounded endosperm. The mutant strain failed to produce aflatoxin when inoculated onto degermed kernels. In contrast, the wild-type strain produced aflatoxin in both the endosperm and embryo. These results suggest that α-amylase facilitates aflatoxin production and growth of A. flavus from a wound in the endosperm to the embryo. A 14-kDa trypsin inhibitor associated with resistance to A. flavus and aflatoxin in maize also inhibited the α-amylase from A. flavus, indicating that it is a bifunctional inhibitor. The inhibitor may have a role in resistance, limiting the growth of the fungus in the endosperm tissue by inhibiting the degradation of starch.

Research paper thumbnail of Inhibition of Growth of <i>Aspergillus flavus</i> and Fungal α-Amylases by a Lectin-Like Protein from <i>Lablab purpureus</i>

Molecular Plant-microbe Interactions, Aug 1, 2001

Aspergillus flavus is a fungal pathogen of maize causing an important ear rot disease when plants... more Aspergillus flavus is a fungal pathogen of maize causing an important ear rot disease when plants are exposed to drought and heat stress. Associated with the disease is the production of aflatoxins, which are a series of structurally related mycotoxins known to be carcinogenic. Previous research has suggested that the α-amylase of A. flavus promotes aflatoxin production in the endosperm of infected maize kernels. We report here the isolation and characterization of a 36-kDa α-amylase inhibitor from Lablab purpureus (AILP). AILP inhibited the α-amylases from several fungi but had little effect on those from animal and plant sources. The protein inhibited conidial germination and hyphal growth of A. flavus. The amino acid sequence indicated that AILP is similar to lectin members of a lectin-arcelin-α-amylase inhibitor family described in common bean and shown to be a component of plant resistance to insect pests. AILP also agglutinated papain-treated red blood cells from human and rabbit. These data indicate that AILP represents a novel variant in the lectin-arcelin-α-amylase inhibitor family of proteins having lectin-like and α-amylase inhibitory activity.

Research paper thumbnail of Genomics of Filamentous Fungi

Mycology [electronic resource], Dec 17, 2003

Research paper thumbnail of Fluopyram activates systemic resistance in soybean

Frontiers in Plant Science

The soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is a significant yield-limiting fa... more The soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is a significant yield-limiting factor in soybean production in the Midwestern US. Several management practices are implemented to mitigate yield losses caused by SCN, including using SDHI (succinate dehydrogenase inhibitors) fungicides delivered as seed treatments. A set of studies was conducted to evaluate the effect of two seed-applied succinate dehydrogenase inhibitors (SDHI) compounds, fluopyram and pydiflumetofen, on SCN population densities, plant injury, and plant growth. Cyst counts in untreated control and pydiflumetofen treated plants were 3.44 and 3.59 times higher than fluopyram, respectively, while egg counts were 8.25 and 7.06 times higher in control and pydiflumetofen. Next-generation sequencing was later employed to identify transcriptomic shifts in gene expression profiles in fluopyram and pydiflumetofen -treated seedlings. RNA expression patterns of seed treatments clustered by sampling time (5 DAP vs....

Research paper thumbnail of The Effect of Wheat on Soybean Cyst Nematode Populations in Double-Cropping Soybean Production

Research paper thumbnail of Characterization of Quinone Outside Inhibitor Fungicide Resistance in Cercospora sojina and Development of Diagnostic Tools for its Identification

Plant Disease, 2015

Frogeye leaf spot of soybean, caused by the fungus Cercospora sojina, reduces soybean yields in m... more Frogeye leaf spot of soybean, caused by the fungus Cercospora sojina, reduces soybean yields in most of the top-producing countries around the world. Control strategies for frogeye leaf spot can rely heavily on quinone outside inhibitor (QoI) fungicides. In 2010, QoI fungicide-resistant C. sojina isolates were identified in Tennessee for the first time. As the target of QoI fungicides, the cytochrome b gene present in fungal mitochondria has played a key role in the development of resistance to this fungicide class. The cytochrome b genes from three QoI-sensitive and three QoI-resistant C. sojina isolates were cloned and sequenced. The complete coding sequence of the cytochrome b gene was identified and found to encode 396 amino acids. The QoI-resistant C. sojina isolates contained the G143A mutation in the cytochrome b gene, a guanidine to cytosine transversion at the second position in codon 143 that causes an amino acid substitution of alanine for glycine. C. sojina-specific poly...

Research paper thumbnail of A panel of qPCR assays to detect and quantify soybean soil-borne pathogens

Letters in Applied Microbiology, Jan 23, 2023

Fusarium oxysporum,F. graminearum,F. acuminatum,F. equiseti,F. proliferatum,F. solani, and Rhizoc... more Fusarium oxysporum,F. graminearum,F. acuminatum,F. equiseti,F. proliferatum,F. solani, and Rhizoctonia solani are soil-borne fungal pathogens that cause substantial yield loss in a widespread list of crops worldwide. The objective of this study was to develop a panel of TaqMan assays for the detection and quantification of these six widespread soil-borne fungal species using real-time polymerase chain reaction (qPCR). The primers and probes were designed based on the intergenic spacer ribosomal RNA and translation elongation factor 1-alpha gene (tef1). These assays, although not multiplexed, can be performed simultaneously as they have similar reaction conditions, allowing more efficiency when targeting multiple pathogens in a sample. The assays presented high efficiency (94.3%-108.9%) and sensitivity, with a limit of detection of 0.05 picograms (50 femtograms) of target DNA. Results from an assay targeting 19 non-target and closely related species confirmed the specificity of the developed assays. The assays were also evaluated to detect the target species in different matrices, such as soil and plant material. This panel of qPCR assays is an additional tool that can be used by plant pathologists, microbiologists, plant breeders, diagnostic clinics, and other researchers interested in these fungal species.

Research paper thumbnail of Interactions and Aflatoxin Production

Environmental influences on maize-Aspergillus flavus

Research paper thumbnail of Interactions and Aflatoxin Production

Environmental influences on maize-Aspergillus flavus

Research paper thumbnail of Ecology and diversity of culturable fungal species associated with soybean seedling diseases in the Midwestern United States

Journal of Applied Microbiology, 2022

To isolate and characterize fungi associated with diseased soybean seedlings in Midwestern soybea... more To isolate and characterize fungi associated with diseased soybean seedlings in Midwestern soybean production fields and to determine the influence of environmental and edaphic factors on their incidence.

Research paper thumbnail of Reduction of Pythium Damping-off in Soybean by Biocontrol Seed Treatment

Plant Disease

Pythium spp. is one of the major groups of pathogens that cause seedling diseases on soybean, lea... more Pythium spp. is one of the major groups of pathogens that cause seedling diseases on soybean, leading to both pre- and post-emergence damping-off and root rot. More than 100 species have been identified within this genus, with P. irregulare, P. sylvaticum, P. ultimum var ultimum, and P. torulosum, being particularly important for soybean production given their aggressiveness, prevalence, and abundance in production fields. This study investigated the antagonistic activity of potential BCAs native to the US Midwest against Pythium spp. First, in vitro screening identified BCAs that inhibit P. ultimum var. ultimum growth. Scanning electron microscopy demonstrated evidence of mycoparasitism of all potential biocontrol isolates against the Pythium ultimum var. ultimum and P. torulosum, with the formation of appressorium-like structures, short-hyphal branches around host hyphae, hook-shaped structures, coiling, and parallel growth of the mycoparasite along the host hyphae. Based on these...

Research paper thumbnail of Rocha - SON2020 - Shifts in soil microbial communities in double-cropping soybeans - implications in the management of the soybean cyst nematode

In double cropping (DC) systems, fields have two or more crops growing in sequence in a single gr... more In double cropping (DC) systems, fields have two or more crops growing in sequence in a single growing season. For soybean, the crop is commonly planted following winter wheat. The soybean cyst nematode (SCN) (Heterodera glycines) is a major soybean pathogen. Several reports in the literature suggested potential suppressive effects of wheat residue on SCN population densities. A field trial was conducted in the 2017-2018 growing season to assess the effect of wheat on SCN populations in DC soybeans. In each field location (N=9), wheat (WT) was planted in strips alternating with strips maintained in fallow (FL) over winter. Soybean followed all strips after wheat harvest. Wheat strips had reduced SCN population densities compared to fallow strips, at soybean growth stage R1 and after soybean harvest. Three field locations with noted SCN suppression were selected for a metagenomics study. Ten subplots were selected (5 WT and 5 FL) from each location. A total of 90 soil samples were se...

Research paper thumbnail of Unraveling the microbial profile of the rhizosphere of SDS-suppressive soils in Soybean fields

Research paper thumbnail of Wheat Production Alters Soil Microbial Profiles and Enhances Beneficial Microbes in Double-Cropping Soybean

Frontiers in Agronomy, 2022

Plant-parasitic nematodes represent a substantial constraint on global food security by reducing ... more Plant-parasitic nematodes represent a substantial constraint on global food security by reducing the yield potential of all major crops. The soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is widely distributed across important soybean production areas of the U.S., being the major soybean yield-limiting factor, especially in the Midwestern U.S. Double cropped (DC) soybean is commonly planted following winter wheat. We previously reported double-cropping soybean fields with reduced SCN counts compared to fallow at both R1 growth stage (beginning of flowering) (−31.8%) and after soybean harvest (−32.7%). To test if higher counts of beneficial and SCN antagonistic microorganisms could be correlated with the suppression of SCN in fields previously planted with wheat, three field locations with noted SCN suppression were selected for a metagenomics study. Ten subplots were selected (5 wheat and 5 fallow pre-soybean) from each location. A total of 90 soil samples were selected:...

Research paper thumbnail of Weeds Hosting the Soybean Cyst Nematode (Heterodera glycines Ichinohe): Management Implications in Agroecological Systems

Agronomy, 2021

The soybean cyst nematode (SCN; Heterodera glycines Ichinohe) is a major soybean-yield-limiting s... more The soybean cyst nematode (SCN; Heterodera glycines Ichinohe) is a major soybean-yield-limiting soil-borne pathogen, especially in the Midwestern US. Weed management is recommended for SCN integrated management, since some weed species have been reported to be hosts for SCN. The increase in the occurrence of resistance to herbicides complicates weed management and may further direct ecological–evolutionary (eco–evo) feedbacks in plant–pathogen complexes, including interactions between host plants and SCN. In this review, we summarize weed species reported to be hosts of SCN in the US and outline potential weed–SCN management interactions. Plants from 23 families have been reported to host SCN, with Fabaceae including most host species. Out of 116 weeds hosts, 14 species have known herbicide-resistant biotypes to 8 herbicide sites of action. Factors influencing the ability of weeds to host SCN are environmental and edaphic conditions, SCN initial inoculum, weed population levels, and...

Research paper thumbnail of Hyperspectral Spectroscopy to Detect Different Responses of Two Soybean (Glycine Max) Cultivars to Charcoal Rot (Macrophomina Phaseolina) Toxin

Engenharia Agricola, 2021

Previous studies have confirmed the potential of hyperspectral spectroscopy (HS) to detect charco... more Previous studies have confirmed the potential of hyperspectral spectroscopy (HS) to detect charcoal rot toxin effect on soybean (Glycine max). However, there is no evidence that it can differentiate among difference responses to that effect. This research tested the potential of HS (900-2400 nm) to differentiate different responses to charcoal rot effect caused by the fungal pathogen Macrophomina phaseolina. Known susceptible 'Pharaoh' and resistant 'Spencer' cultivars of soybean seedlings at the V1 stage were tested, and HS reflectance was measured in each treatment at 0 h (just before toxin treatment), 24 h, and 36 h after toxin treatment. For both cultivars, reflectance generally decreased up to 10% over time for controls and increased for treated seedlings up to 22%. Pharaoh had significant increases in reflectance 24 and 36 h after treatment, while Spencer did not exhibit significant changes in reflectance over the 24 and 36-h periods. The ratio of Al-Ahmadi et al. (2018) was also evaluated. Spencer showed a higher ratio than Pharaoh. Based on our results, HS can be used to compare the relative resistance of soybean cultivars to charcoal rot and is a potential screening technique to classify soybean cultivars based on their resistance to charcoal rot toxin.

Research paper thumbnail of Impact of Wheat on Soybean Cyst Nematode Population Density in Double-Cropping Soybean Production

Frontiers in Plant Science, 2021

Double-cropping is defined as producing more than one crop on the same parcel of land in a single... more Double-cropping is defined as producing more than one crop on the same parcel of land in a single growing season. It is reported to have many benefits when incorporated in cropping systems, including improving soil health. In some double-cropping systems, soybean is planted following winter wheat. The soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is a major soybean pathogen, and several reports suggest suppressive effects of wheat on SCN populations. Field trials were conducted from 2017 to 2018 to investigate the effect of wheat on SCN populations in double-cropping soybean. Nine fields with three levels of initial SCN populations (low, moderate, and high) were selected in Illinois. Wheat was planted in strips alternating with strips-maintained weed-free and under fallow over winter and early spring. Soybean was planted in all strips after wheat harvest. SCN egg densities were acquired at four time points: wheat establishment, post-wheat/pre-soybean, mid-soybean (R1 gr...

Research paper thumbnail of Use of Dual RNA-seq for Systems Biology Analysis of Zea mays and Aspergillus flavus Interaction

Frontiers in Microbiology, 2020

The interaction between Aspergillus flavus and Zea mays is complex, and the identification of pla... more The interaction between Aspergillus flavus and Zea mays is complex, and the identification of plant genes and pathways conferring resistance to the fungus has been challenging. Therefore, the authors undertook a systems biology approach involving dual RNA-seq to determine the simultaneous response from the host and the pathogen. What was dramatically highlighted in the analysis is the uniformity in the development patterns of gene expression of the host and the pathogen during infection. This led to the development of a "stage of infection index" that was subsequently used to categorize the samples before downstream system biology analysis. Additionally, we were able to ascertain that key maize genes in pathways such as the jasmonate, ethylene and ROS pathways, were up-regulated in the study. The stage of infection index used for the transcriptomic analysis revealed that A. flavus produces a relatively limited number of transcripts during the early stages (0 to 12 h) of infection. At later stages, in A. flavus, transcripts and pathways involved in endosomal transport, aflatoxin production, and carbohydrate metabolism were up-regulated. Multiple WRKY genes targeting the activation of the resistance pathways (i.e., jasmonate, phenylpropanoid, and ethylene) were detected using causal inference analysis. This analysis also revealed, for the first time, the activation of Z. mays resistance genes influencing the expression of specific A. flavus genes. Our results show that A. flavus seems to be reacting to a hostile environment resulting from the activation of resistance pathways in Z. mays. This study revealed the dynamic nature of the interaction between the two organisms.