scholarly journals Selection of Fusarium Trichothecene Toxin Genes for Molecular Detection Depends on TRI Gene Cluster Organization and Gene Function

Toxins ◽  
2019 ◽  
Vol 11 (1) ◽  
pp. 36 ◽  
Author(s):  
Ria Villafana ◽  
Amanda Ramdass ◽  
Sephra Rampersad
Keyword(s):  
Gene Cluster ◽  
Fungal Pathogens ◽  
Fusarium Species ◽  
Pcr Amplification ◽  
Trichothecene Mycotoxin ◽  
Cluster Organization ◽  
Food And Feed ◽  
Multiple Species ◽  
Selection Of

Food security is a global concern. Fusarium are among the most economically important fungal pathogens because they are ubiquitous, disease management remains a challenge, they produce mycotoxins that affect food and feed safety, and trichothecene mycotoxin production can increase the pathogenicity of some Fusarium species depending on the host species. Although trichothecenes may differ in structure by their patterns of hydroxylation or acetylation, these small changes have a significant impact on toxicity and the biological activity of these compounds. Therefore, detecting and identifying which chemotype is present in a given population are important to predicting the specific toxins that may be produced and, therefore, to evaluating the risk of exposure. Due to the challenges of inducing trichothecene production by Fusarium isolates in vitro for subsequent chemical analysis, PCR assays using gene-specific primers, either singly or in combination, designed against specific genes of the trichothecene gene cluster of multiple species of Fusarium have been developed. The establishment of TRI genotypes that potentially correspond to a specific chemotype requires examination of an information and knowledge pipeline whose critical aspects in sequential order are: (i) understanding the TRI gene cluster organization which differs according to Fusarium species under study; (ii) knowledge of the re-arrangements to the core TRI gene cluster over evolutionary time, which also differs according to Fusarium species; (iii) the functions of the TRI genes in the biosynthesis of trichothecene analogs; and (iv) based on (i)–(iii), selection of appropriate target TRI gene(s) for primer design in PCR amplification for the Fusarium species under study. This review, therefore, explains this pipeline and its connection to utilizing TRI genotypes as a possible proxy to chemotype designation.

scholarly journals In Vitro Analysis of Deoxynivalenol Influence on Steroidogenesis in Prostate

Toxins ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 685
Author(s):  
Kinga Anna Urbanek ◽  
Karolina Kowalska ◽  
Dominika Ewa Habrowska-Górczyńska ◽  
Kamila Domińska ◽  
Agata Sakowicz ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Fusarium Species ◽  
Male Rats ◽  
Trichothecene Mycotoxin ◽  
Normal Prostate ◽  
Prostate Cells ◽  
Expression Of Genes ◽  
Food And Feed ◽  
Vitro Analysis

Deoxynivalenol (DON) is a type-B trichothecene mycotoxin produced by Fusarium species, reported to be the most common mycotoxin present in food and feed products. DON is known to affect the production of testosterone, follicle stimulating hormone (FSH) and luteinizing hormone (LH) in male rats, consequently affecting reproductive endpoints. Our previous study showed that DON induces oxidative stress in prostate cancer (PCa) cells, however the effect of DON on the intratumor steroidogenesis in PCa and normal prostate cells was not investigated. In this study human normal (PNT1A) and prostate cancer cell lines with different hormonal sensitivity (PC-3, DU-145, LNCaP) were exposed to DON treatment alone or in combination with dehydroepiandrosterone (DHEA) for 48 h. The results of the study demonstrated that exposure to DON alone or in combination with DHEA had a stimulatory effect on the release of estradiol and testosterone and also affected progesterone secretion. Moreover, significant changes were observed in the expression of genes related to steroidogenesis. Taken together, these results indicate that DON might affect the process of steroidogenesis in the prostate, demonstrating potential reproductive effects in humans.

scholarly journals The Effect of HT-2 Toxin on Ovarian Steroidogenesis and Its Response to IGF-I, Leptin and Ghrelin in Rabbits

2017 ◽  
pp. 705-708 ◽  
Author(s):  
A. KOLESAROVA ◽  
N. MARUNIAKOVA ◽  
A. KADASI ◽  
M. HALENAR ◽  
M. MARAK ◽  
...  
Keyword(s):  
Growth Factor ◽  
Fusarium Species ◽  
Endocrine System ◽  
Control Group ◽  
Trichothecene Mycotoxin ◽  
Ovarian Steroidogenesis ◽  
Estradiol Secretion ◽  
Igf I ◽  
17Β Estradiol

T-2 toxin and its metabolite HT-2 toxin are one of the most toxic mycotoxins of type A-trichothecenes, which are produced mainly by Fusarium species. Therefore, study of Fusarium toxins T-2 toxin and HT-2 toxin is an essential issue because they could also play role in failures of reproductive functions as well as endocrine system of domestic animals. Assessment of the effect of A-trichothecene mycotoxin HT-2 toxin alone or combined with insulin-like growth factor (IGF-I), leptin and ghrelin on estradiol secretion by rabbit ovarian fragments in vitro was done. Rabbit ovarian fragments were incubated without (control group) or with HT-2 toxin, or its combinations with IGF-I, leptin and ghrelin at various concentrations for 24 h. Secretion of 17β-estradiol was determined by ELISA. Firstly, HT-2 toxin at the doses 10 and 100 ng.ml-1, but not at 1 ng.ml-1 decreased 17β-estradiol secretion by ovarian fragments. Secondly, 17β-estradiol secretion was not affected by HT-2 toxin exposure combined with growth factor IGF-I, metabolic hormones leptin and ghrelin. In conclusion, HT-2 toxin has potent direct dose-dependent effects on ovarian steroidogenesis in rabbits. These direct effects of HT-2 mycotoxin on ovarian steroidogenesis could impact negatively on the reproductive performance of rabbits.

scholarly journals Rhizosphere-enriched microbes as a pool to design synthetic communities for reproducible beneficial outputs

2018 ◽  
Author(s):  
Maria-Dimitra Tsolakidou ◽  
Ioannis A. Stringlis ◽  
Natalia Fanega-Sleziak ◽  
Stella Papageorgiou ◽  
Antria Tsalakou ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Fungal Pathogens ◽  
Growth Parameters ◽  
Tomato Plants ◽  
Beneficial Effects ◽  
Negative Effect ◽  
Tomato Growth ◽  
Selection Of

AbstractComposts represent a sustainable way to suppress diseases and improve plant growth. Identification of compost-derived microbial communities enriched in the rhizosphere of plants and characterization of their traits, could facilitate the design of microbial synthetic communities (SynComs) that upon soil inoculation could yield consistent beneficial effects towards plants. Here, we characterized a collection of compost-derived bacteria, previously isolated from tomato rhizosphere, forin vitroantifungal activity against soil-borne fungal pathogens and for their potential to change growth parameters inArabidopsis. We further assessed root-competitive traits in the dominant rhizospheric genusBacillus. Certain isolated rhizobacteria displayed antifungal activity against the tested pathogens and affected growth ofArabidopsis, and Bacilli members possessed several enzymatic activities. Subsequently, we designed two SynComs with different composition and tested their effect onArabidopsisand tomato growth and health. SynCom1, consisting of different bacterial genera, displayed negative effect onArabidopsis in vitro, but promoted tomato growth in pots. SynCom2, consisting of Bacilli, didn’t affectArabidopsisgrowth, enhanced tomato growth and suppressed Fusarium wilt symptoms. Overall, we found selection of compost-derived microbes with beneficial properties in the rhizosphere of tomato plants, and observed that application of SynComs on poor substrates can yield reproducible plant phenotypes.

scholarly journals Testing Methods Affecting the Antagonistic Ability of  Pseudomonas Biocontrol Strains

2002 ◽  
Vol 51 (1-2) ◽  
pp. 107-114 ◽  
Author(s):  
I. Jevcsák ◽  
Bálint Oldal ◽  
L. Ködöböcz ◽  
Keyword(s):  
Rhizoctonia Solani ◽  
Fusarium Solani ◽  
Fungal Pathogens ◽  
Culture Media ◽  
Colony Growth ◽  
Antagonistic Effect ◽  
Bacterial Strains ◽  
Testing Methods ◽  
Selection Of

The antagonistic effect of thirteen Pseudomonas aeruginosa and thirteen strains of other Pseudomonas species was studied on the soil-borne phytopathogenic Rhizoctonia solani and Fusarium solani fungi.  The inhibition of pathogen colony growth was tested with two different in vitro techniques using the same type of culture media. In case of the spread slant technique the antagonists induced a significantly stronger inhibition on the growth of pathogens than in case of spot transfer. Among the 26 investigated Pseudomonas strains, P. aeruginosa strains were generally more effective against the fungal pathogens. Rhizoctonia solani proved to be affected to a greater extent by the bacterial strains studied than the Fusarium solani representative. The possibility of in vitro strain selection of biocontrol microbes is being further discussed .

scholarly journals Identification of a Chitinase-modifying Protein from Fusarium verticillioides

2011 ◽  
Vol 286 (41) ◽  
pp. 35358-35366 ◽  
Author(s):  
Todd A. Naumann ◽  
Donald T. Wicklow ◽  
Neil P. J. Price
Keyword(s):  
Fungal Pathogens ◽  
Fusarium Verticillioides ◽  
Peptide Bond ◽  
Host Protein ◽  
Ear Rot ◽  
Reaction Products ◽  
Lectin Domain ◽  
Multiple Species ◽  
Class Iv

Chitinase-modifying proteins (cmps) are proteases secreted by fungal pathogens that truncate the plant class IV chitinases ChitA and ChitB during maize ear rot. cmp activity has been characterized for Bipolaris zeicola and Stenocarpella maydis, but the identities of the proteases are not known. Here, we report that cmps are secreted by multiple species from the genus Fusarium, that cmp from Fusarium verticillioides (Fv-cmp) is a fungalysin metalloprotease, and that it cleaves within a sequence that is conserved in class IV chitinases. Protein extracts from Fusarium cultures were found to truncate ChitA and ChitB in vitro. Based on this activity, Fv-cmp was purified from F. verticillioides. N-terminal sequencing of truncated ChitA and MALDI-TOF-MS analysis of reaction products showed that Fv-cmp is an endoprotease that cleaves a peptide bond on the C-terminal side of the lectin domain. The N-terminal sequence of purified Fv-cmp was determined and compared with a set of predicted proteins, resulting in its identification as a zinc metalloprotease of the fungalysin family. Recombinant Fv-cmp also truncated ChitA, confirming its identity, but had reduced activity, suggesting that the recombinant protease did not mature efficiently from its propeptide-containing precursor. This is the first report of a fungalysin that targets a nonstructural host protein and the first to implicate this class of virulence-related proteases in plant disease.

Quantification and assessment of viability of Cryptococcus neoformans by LightCycler amplification of capsule gene mRNA

2004 ◽  
Vol 53 (12) ◽  
pp. 1201-1206 ◽  
Author(s):  
Muhammad Amjad ◽  
Najla Kfoury ◽  
Raymond Cha ◽  
Reem Mobarak
Keyword(s):  
Cryptococcus Neoformans ◽  
Internal Control ◽  
Fungal Pathogens ◽  
Pcr Amplification ◽  
Plasmid Vector ◽  
Antifungal Drugs ◽  
Yeast Cells ◽  
Rt Pcr ◽  
Gene Mrna

Cryptococcus neoformans is an opportunistic fungal pathogen. It infects the central nervous system causing meningitis, which is fatal if untreated, especially in AIDS and immunosuppressed patients. In this study a method of quantification and assessment of viability of C. neoformans by LightCycler RT-PCR amplification of the capsule gene mRNA is established. The sequence of primers and probes were derived from C. neoformans capsular CAP10 gene mRNA (GenBank accession number AF144574), and were species specific. Agarose gel electrophoresis analysis of LightCycler RT-PCR product showed a single band of 223 bp in length. In order to develop an internal control a 223 bp exon fragment of capsule mRNA was cloned in the pCR2.1 plasmid vector and RNA was generated by in vitro transcription. To determine the sensitivity of the assay, serial dilutions of in vitro-transcribed RNA with known concentrations and copy numbers, and serially diluted cultures of viable and nonviable C. neoformans were used. Under optimal conditions as little as 0.472 fg of capsule mRNA could be detected, corresponding to 1–10 c.f.u. ml−1 of the sample. No amplification was observed from up to105 heat/UV radiation-killed yeast cells and RNA of other bacterial and fungal pathogens and human genomic DNA or RNA. The amplification of capsule mRNA represents a sensitive, specific and quantitative means of detection of viable C. neoformans in clinical specimens and can be useful in the evaluation of the therapeutic efficacy of antifungal drugs in the treatment of C. neoformans meningitis.

scholarly journals Arabidopsis MAPKKK δ-1 is required for full immunity against bacterial and fungal infection

2019 ◽  
Vol 71 (6) ◽  
pp. 2085-2097 ◽  
Author(s):  
Tomoya Asano ◽  
Thi Hang-Ni Nguyen ◽  
Michiko Yasuda ◽  
Yasir Sidiq ◽  
Kohji Nishimura ◽  
...  
Keyword(s):  
Disease Resistance ◽  
Fungal Infection ◽  
Pseudomonas Syringae ◽  
Fusarium Species ◽  
Bacterial Pathogen ◽  
Mitogen Activated Protein Kinase ◽  
Trichothecene Mycotoxin ◽  
Mitogen Activated Protein

Abstract The genome of Arabidopsis encodes more than 60 mitogen-activated protein kinase kinase (MAPKK) kinases (MAPKKKs); however, the functions of most MAPKKKs and their downstream MAPKKs are largely unknown. Here, MAPKKK δ-1 (MKD1), a novel Raf-like MAPKKK, was isolated from Arabidopsis as a subunit of a complex including the transcription factor AtNFXL1, which is involved in the trichothecene phytotoxin response and in disease resistance against the bacterial pathogen Pseudomonas syringae pv. tomato DC3000 (PstDC3000). A MKD1-dependent cascade positively regulates disease resistance against PstDC3000 and the trichothecene mycotoxin-producing fungal pathogen Fusarium sporotrichioides. MKD1 expression was induced by trichothecenes derived from Fusarium species. MKD1 directly interacted with MKK1 and MKK5 in vivo, and phosphorylated MKK1 and MKK5 in vitro. Correspondingly, mkk1 mutants and MKK5RNAi transgenic plants showed enhanced susceptibility to F. sporotrichioides. MKD1 was required for full activation of two MAPKs (MPK3 and MPK6) by the T-2 toxin and flg22. Finally, quantitative phosphoproteomics suggested that an MKD1-dependent cascade controlled phosphorylation of a disease resistance protein, SUMO, and a mycotoxin-detoxifying enzyme. Our findings suggest that the MKD1–MKK1/MKK5–MPK3/MPK6-dependent signaling cascade is involved in the full immune responses against both bacterial and fungal infection.

scholarly journals Trichothecene-Producing Fusarium Species Isolated from Soybean Roots in Ethiopia and Ghana and their Pathogenicity on Soybean

Plant Disease ◽  
2019 ◽  
Vol 103 (8) ◽  
pp. 2070-2075 ◽  
Author(s):  
Glen L. Hartman ◽  
Susan P. McCormick ◽  
Kerry O’Donnell
Keyword(s):  
Sequence Data ◽  
Fusarium Species ◽  
Phylogenetic Analyses ◽  
Negative Control ◽  
Sub Saharan Africa ◽  
Trichothecene Mycotoxin ◽  
Fusarium Spp ◽  
Fusarium Sp ◽  
Soybean Roots

Numerous pathogen surveys have reported that diverse Fusarium spp. threaten soybean production in North and South America. However, little research has been conducted to characterize Fusarium pathogens of soybean in sub-Saharan Africa. Our objectives were to (i) identify Fusarium spp. isolated from discolored root segments of soybean grown in Ethiopia and Ghana using DNA sequence data, (ii) determine whether isolates nested in the Fusarium incarnatum-equiseti and F. sambucinum species complexes (FIESC and FSAMSC, respectively) produced trichothecene mycotoxins in vitro, and (iii) test these isolates for pathogenicity on soybean. Molecular phylogenetic analyses revealed that the trichothecene mycotoxin-producing isolates comprised three undescribed species within the FIESC and FSAMSC. Mycotoxin type B trichothecene 4,15-diacetylnivalenol or T-2 toxin and related type A neosolaniol trichothecenes were produced by 18 of the 21 isolates. Of the 12 isolates from Ethiopia and Ghana tested for their impact on seed germination, 5, comprising two undescribed phylospecies (i.e., Fusarium sp. number 3 and Fusarium sp. FIESC 2,) completely inhibited germination, whereas 4 caused no reduction in germination. Root lesions induced by all 12 isolates were greater than the uninoculated negative control. Additional variation among the isolates was reflected in differences (α = 0.05) in lesion lengths, which ranged from 34 to 67% of total root length. This is the first report characterizing FIESC and FSAMSC isolates from soybean roots in Ethiopia and Ghana.

scholarly journals Burkholderia gladioli CGB10: A Novel Strain Biocontrolling the Sugarcane Smut Disease

2020 ◽  
Vol 8 (12) ◽  
pp. 1943
Author(s):  
Guobing Cui ◽  
Kai Yin ◽  
Nuoqiao Lin ◽  
Meiling Liang ◽  
Chengwei Huang ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Gene Cluster ◽  
Inhibitory Activity ◽  
Fungal Pathogens ◽  
Molecular Mechanisms ◽  
Biocontrol Agent ◽  
Burkholderia Species ◽  
Sporisorium Scitamineum ◽  
Burkholderia Gladioli

In this study, we isolated an endophytic Burkholderia gladioli strain, named CGB10, from sugarcane leaves. B. gladioli CGB10 displayed strong inhibitory activity against filamentous growth of fungal pathogens, one of which is Sporisorium scitamineum that causes sugarcane smut, a major disease affecting the quality and production of sugarcane in tropical and subtropical regions. CGB10 could effectively suppress sugarcane smut under field conditions, without itself causing any obvious damage or disease, thus underscoring a great potential as a biocontrol agent (BCA) for the management of sugarcane smut. A toxoflavin biosynthesis and transport gene cluster potentially responsible for such antifungal activity was identified in the CGB10 genome. Additionally, a quorum-sensing gene cluster was identified too and compared with two close Burkholderia species, thus supporting an overall connection to the regulation of toxoflavin synthesis therein. Overall, this work describes the in vitro and field Sporisorium scitamineum biocontrol by a new B. gladioli strain, and reports genes and molecular mechanisms potentially involved.

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