scholarly journals Structural and Functional Characterization of the TRI101 Trichothecene 3-O-Acetyltransferase from Fusarium sporotrichioides and Fusarium graminearum

2007 ◽  
Vol 283 (3) ◽  
pp. 1660-1669 ◽  
Author(s):  
Graeme S. Garvey ◽  
Susan P. McCormick ◽  
Ivan Rayment
Keyword(s):  
Fusarium Graminearum ◽  
Coenzyme A ◽  
Functional Characterization ◽  
Crop Protection ◽  
Three Dimensional ◽  
Transgenic Crop ◽  
Kinetic Properties ◽  
Fusarium Sporotrichioides ◽  
Head Blight

Fusarium head blight (FHB) is a plant disease with serious economic and health impacts. It is caused by fungal species belonging to the genus Fusarium and the mycotoxins they produce. Although it has proved difficult to combat this disease, one strategy that has been examined is the introduction of an indigenous fungal protective gene into cereals such as wheat barley and rice. Thus far the gene of choice has been tri101 whose gene product catalyzes the transfer of an acetyl group from acetyl coenzyme A to the C3 hydroxyl moiety of several trichothecene mycotoxins. In vitro this has been shown to reduce the toxicity of the toxins by ∼100-fold but has demonstrated limited resistance to FHB in transgenic cereal. To understand the molecular basis for the differences between in vitro and in vivo resistance the three-dimensional structures and kinetic properties of two TRI101 orthologs isolated from Fusarium sporotrichioides and Fusarium graminearum have been determined. The kinetic results reveal important differences in activity of these enzymes toward B-type trichothecenes such as deoxynivalenol. These differences in activity can be explained in part by the three-dimensional structures for the ternary complexes for both of these enzymes with coenzyme A and trichothecene mycotoxins. The structural and kinetic results together emphasize that the choice of an enzymatic resistance gene in transgenic crop protection strategies must take into account the kinetic profile of the selected protein.

Impact of the biofungicide tetramycin on the development of Fusarium head blight, grain yield and deoxynivalenol accumulation in wheat

2020 ◽  
Vol 13 (2) ◽  
pp. 235-246
Author(s):  
W.Q. Shi ◽  
L.B. Xiang ◽  
D.Z. Yu ◽  
S.J. Gong ◽  
L.J. Yang
Keyword(s):  
Fusarium Head Blight ◽  
Fusarium Graminearum ◽  
Spore Germination ◽  
Field Experiments ◽  
Synergistic Effects ◽  
Field Trials ◽  
Head Blight ◽  
Mycelium Growth ◽  
Key Aspects

Fusarium graminearum causes Fusarium head blight (FHB), a devastating disease that leads to extensive yield and quality loss in wheat and barley production. Integrated pest management (IPM) is required to control this disease and biofungicides, such as tetramycin, could be a novel addition to IPM strategies. The current study investigated in vitro tetramycin toxicity in Fusarium graminearum and evaluated its effectiveness for the control of Fusarium head blight FHB. Tetramycin was shown to affect three key aspects of Fusarium pathogenicity: spore germination, mycelium growth and deoxynivalenol (DON) production. The in vitro results indicated that tetramycin had strong inhibitory activity on the mycelial growth and spore germination. Field trials indicated that tetramycin treatment resulted in a significant reduction in both the FHB disease index and the level of DON accumulation. The reduced DON content in harvested grain was correlated with the amount of Tri5 mRNA determined by qRT-PCR. Synergistic effects between tetramycin and metconazole, in both the in vitro and field experiments were found. Tetramycin could provide an alternative option to control FHB.

PCR-RFLP for detection of Fusarium graminearum genotypes with resistance to phenamacril

Plant Disease ◽  
2020 ◽  
Author(s):  
Jiao-Sheng Li ◽  
Luo-Yu Wu ◽  
Hui Zhang ◽  
Xiu-Shi Song ◽  
Jian-Xin Wang ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Conventional Method ◽  
Resistance Mutations ◽  
Head Blight ◽  
Fusarium Asiaticum ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Resistant Strains ◽  
Codon Mutation

Phenamacril is a cyanoacrylate fungicide that provides excellent control of Fusarium head blight (FHB) or wheat scab, which is caused predominantly by Fusarium graminearum and Fusarium asiaticum. Previous studies revealed that codon mutations of the myosin-5 gene of Fusarium spp. conferred resistance to phenamacril in vitro lab experiments. In this study, PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) was developed to detect three common mutations (A135T, GCC to ACC at codon 135; S217L, TCA to TTA at codon 217, and E420K, GAA to AAA at codon 420) in F. graminearum induced by fungicide domestication in vitro. PCR products of 841 bp (for mutation of A135T), 802 bp (for mutation of S217L) or 1649 bp (for mutation of E420K) in myosin-5 gene were amplified respectively by appropriate primer pairs. Restriction enzyme KpnⅠ, TasⅠ or DraⅠ was used to distinguish phenamacril-sensitive and -resistant strains with mutation genotypes of A135T, S217L and E420K, respectively. KpnⅠ digested the 841 bp PCR products of phenamacri-resistant strains with codon mutation A135T into two fragments of 256 bp and 585 bp. In contrast, KpnⅠ did not digest the PCR products of sensitive strains. TasⅠ digested the 802 bp PCR products of phenamacril-strains with codon mutation S217L into three fragments of 461 bp, 287bp and 54 bp. In contrast, TasⅠ digestion of the 802 bp PCR products of phenamacril-sensitive strains resulted in only two fragments of 515bp and 287bp. DraⅠ digested the 1649 bp PCR products of phenamacril-resistant strains with codon mutation E420K into two fragments of 932 bp and 717 bp, while the PCR products of phenamacril-sensitive strains was not digested. The three genotypes of resistance mutations were determined by analyzing electrophoresis patterns of the digestion fragments of PCR products. The PCR-RFLP method was evaluated on 48 phenamacril-resistant strains induced by fungicide domestication in vitro and compared with the conventional method (mycelial growth on fungicide-amended agar). The accuracy of the PCR-RFLP method for detecting the three resistant mutation genotypes of F. graminearum to phenamacril was 95.12% compared with conventional method. Bioinformatics analysis revealed that the PCR-RFLP method could also be used to detect the codon mutations of A135T and E420K in F. asiaticum.

Chemotype and aggressiveness evaluation of Fusarium graminearum and Fusarium culmorum isolates from wheat fields in Wisconsin

Plant Disease ◽  
2021 ◽  
Author(s):  
Brian Mueller ◽  
Carol Groves ◽  
Damon L. Smith
Keyword(s):  
Growth Rate ◽  
Fusarium Head Blight ◽  
Fusarium Graminearum ◽  
Fusarium Culmorum ◽  
Head Blight ◽  
Growing Seasons ◽  
Progress Curve ◽  
Sporulation Capacity ◽  
Derivatives Of

Fusarium graminearum commonly causes Fusarium head blight (FHB) on wheat, barley, rice, and oats. Fusarium graminearum produces nivalenol and deoxynivalenol (DON) and forms derivatives of DON based on its acetylation sites. The fungus is profiled into chemotypes based on DON derivative chemotypes (3 acetyldeoxynivalenol (3ADON) chemotype; 15 acetyldeoxynivalenol (15ADON) chemotype) and/or the nivalenol (NIV) chemotype. The current study assessed the Fusarium population found on wheat and the chemotype profile of the isolates collected from 2016 and 2017 in Wisconsin. Fusarium graminearum was isolated from all locations sampled in both 2016 and 2017. Fusarium culmorum was isolated only from Door County in 2016. Over both growing seasons, 91% of isolates were identified as the 15ADON chemotype while 9% of isolates were identified as the 3ADON chemotype. Aggressiveness was quantified by area under disease progress curve (AUDPC). The isolates with the highest AUDPC values were from the highest wheat producing cropping districts in the state. Deoxynivalenol production in grain and sporulation and growth rate in vitro were compared to aggressiveness in the greenhouse. Our results showed that 3ADON isolates in Wisconsin were among the highest in sporulation capacity, growth rate, and DON production in grain. However, there were no significant differences in aggressiveness between the 3ADON and 15ADON isolates. The results of this research detail the baseline frequency and distribution of 3ADON and 15ADON chemotypes observed in Wisconsin. Chemotype distributions within populations of F. graminearum in Wisconsin should continue to be monitored in the future.

scholarly journals One Small RNA of Fusarium graminearum Targets and Silences CEBiP Gene in Common Wheat

2019 ◽  
Vol 7 (10) ◽  
pp. 425 ◽  
Author(s):  
Jiao Jian ◽  
Xu Liang
Keyword(s):  
Fusarium Graminearum ◽  
Small Rna ◽  
Pathogenic Fungus ◽  
Effective Means ◽  
Transcriptional Level ◽  
Head Blight ◽  
Protein Coding ◽  
Pathogen Invasion

The pathogenic fungus Fusarium graminearum (F. graminearum), causing Fusarium head blight (FHB) or scab, is one of the most important cereal killers worldwide, exerting great economic and agronomic losses on global grain production. To repress pathogen invasion, plants have evolved a sophisticated innate immunity system for pathogen recognition and defense activation. Simultaneously, pathogens continue to evolve more effective means of invasion to conquer plant resistance systems. In the process of co-evolution of plants and pathogens, several small RNAs (sRNAs) have been proved in regulating plant immune response and plant-microbial interaction. In this study, we report that a F. graminearum sRNA (Fg-sRNA1) can suppress wheat defense response by targeting and silencing a resistance-related gene, which codes a Chitin Elicitor Binding Protein (TaCEBiP). Transcriptional level evidence indicates that Fg-sRNA1 can target TaCEBiP mRNA and trigger silencing of TaCEBiP in vivo, and in Nicotiana benthamiana (N. benthamiana) plants, Western blotting experiments and YFP Fluorescence observation proofs show that Fg-sRNA1 can suppress the accumulation of protein coding by TaCEBiP gene in vitro. F. graminearum PH-1 strain displays a weakening ability to invasion when Barley stripe mosaic virus (BSMV) vector induces effective silencing Fg-sRNA1 in PH-1 infected wheat plants. Taken together, our results suggest that a small RNA from F. graminearum can target and silence the wheat TaCEBiP gene to enhance invasion of F. graminearum.

scholarly journals Congenital Lipoid Adrenal Hyperplasia: Functional Characterization of Three Novel Mutations in the STAR Gene

2010 ◽  
Vol 95 (3) ◽  
pp. 1301-1308 ◽  
Author(s):  
Susanne Bens ◽  
Angelika Mohn ◽  
Bilgin Yüksel ◽  
Alexandra E. Kulle ◽  
Matthias Michalek ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Regulatory Protein ◽  
Three Dimensional ◽  
Binding Pocket ◽  
Adrenal Hyperplasia ◽  
Turkish Patient ◽  
Star Protein ◽  
Sex Development ◽  
Congenital Lipoid Adrenal Hyperplasia

Abstract Context: The steroidogenic acute regulatory protein (StAR) has been shown to be essential for steroidogenesis by mediating cholesterol transfer into mitochondria. Inactivating StAR mutations cause the typical clinical picture of congenital lipoid adrenal hyperplasia. Objective: The objective of the investigation was to study the functional and structural consequences of three novel StAR mutations (p.N148K in an Italian patient; p.P129fs and p.Q128R in a Turkish patient). Methods and Results: Transient in vitro expression of the mutant proteins together with P450 side-chain cleavage enzyme, adrenodoxin, and adrenodoxin reductase yielded severely diminished cholesterol conversion of the p.N148K mutant, the combined p.P129fs and p.Q128R mutant, and the p.P129fs mutant by itself. The p.Q128R mutant led to a higher cholesterol conversion than the wild-type StAR protein. As derived from three-dimensional protein modeling, the residue N148 is lining the ligand cavity of StAR. A positively charged lysine residue at position 148 disturbs the hydrophobic cluster formed by the α4-helix and the sterol binding pocket. The frame shift mutation p.P129fs truncates the StAR protein. Residue p.Q128 is situated at the surface of the molecule and is not part of any functionally characterized region of the protein. Conclusion: The mutations p.N148K and p.P129fs cause adrenal insufficiency in both cases and lead to a disorder of sex development with complete sex reversal in the 46, XY case. The mutation p.Q128R, which is not relevant for the patient’s phenotype, is the first reported variant showing a gain of function. We speculate that the substitution of hydrophilic glutamine with basic arginine at the surface of the molecule may accelerate cholesterol transfer.

scholarly journals Investigating Useful Properties of Four Streptomyces Strains Active against Fusarium graminearum Growth and Deoxynivalenol Production on Wheat Grains by qPCR

Toxins ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 560
Author(s):  
Elena Maria Colombo ◽  
Andrea Kunova ◽  
Claudio Gardana ◽  
Cristina Pizzatti ◽  
Paolo Simonetti ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Plant Pathogens ◽  
Fungal Growth ◽  
Toxin Production ◽  
Fungal Mycelium ◽  
Head Blight ◽  
Wheat Grains ◽  
Streptomyces Spp ◽  
Dual Plate

Streptomyces spp. can be exploited as biocontrol agents (BCAs) against plant pathogens such as Fusarium graminearum, the main causal agent of Fusarium head blight (FHB) and against the contamination of grains with deoxynivalenol (DON). In the present research, four Streptomyces strains active against F. graminearum in dual plate assays were characterized for their ability to colonize detached wheat grains in the presence of F. graminearum and to limit DON production. The pathogen and BCA abundance were assessed by a quantitative real-time PCR, while DON production was assessed by HPLC quantification and compared to ergosterol to correlate the toxin production to the amount of fungal mycelium. Fungal growth and mycotoxin production were assessed with both co-inoculation and late inoculation of the BCAs in vitro (three days post-Fusarium inoculation) to test the interaction between the fungus and the bacteria. The level of inhibition of the pathogen and the toxin production were strain-specific. Overall, a higher level of DON inhibition (up to 99%) and a strong reduction in fungal biomass (up to 71%) were achieved when streptomycetes were co-inoculated with the fungus. This research enabled studying the antifungal efficacy of the four Streptomyces strains and monitoring their development in DON-inducing conditions.

scholarly journals Phenotypic and functional characterization of two bovine mammary epithelial cell lines in 2D and 3D models

2016 ◽  
Vol 310 (5) ◽  
pp. C348-C356 ◽  
Author(s):  
Magdalena Arévalo Turrubiarte ◽  
Marie-Hélène Perruchot ◽  
Laurence Finot ◽  
Frédérique Mayeur ◽  
Frédéric Dessauge
Keyword(s):  
Epithelial Cell ◽  
Cell Lines ◽  
Functional Characterization ◽  
Three Dimensional ◽  
Mammary Epithelial ◽  
Specific Cell ◽  
Alveolar Cells ◽  
Bovine Mammary Epithelial Cells ◽  
Precise Knowledge

Immortalized bovine mammary epithelial cells (BME-UV1) and immortalized bovine mammary alveolar cells (MAC-T) have been extensively used as in vitro cell models to understand milk production in dairy cows. Precise knowledge about their phenotype and performance remains, however, unknown. This study aims to characterize MAC-T and BME-UV1 profiles when cultured in two-dimensional adherent, three-dimensional adherent (Matrigel), and three-dimensional no adherent [ultralow attachment (ULA)] supports. MAC-T and BME-UV1 were compared according to their proliferation capacities and to specific cell surface markers CD24, CD326 [epithelial cell adhesion molecule (EpCAM)], CD10, and integrin CD49f (α-6). Cytokeratin (CK14 and CK19), signal transducer and activator of transcription 5, and other proteins (occludin and cadherin-1) were analyzed. BME-UV1 in ULA support expressed higher CD49f marker. A different intensity of CD49 staining allowed the discrimination between the two cell lines in adherent condition. CD10, EpCAM, and CK19 expressions show that BME-UV1 cells have luminal capacity, while MAC-T has a myoepithelial profile with a high expression of CK14. BME-UV1 cells possess a closer committed progenitor profile due to their higher expression in aldehyde dehydrogenase and EpCAM. We observed that BME-UV1 cells have a better capacity to form spherical structures, mammospheres, in Matrigel than MAC-T, which was confirmed by the higher mammosphere area. In the ULA condition, BME-UV1 proliferated over the 6 days of culture. Taken together, our results clearly confirm the BME-UV1 luminal profile and MAC-T ductal/myoepithelial-like phenotype.

scholarly journals Functional Characterization of Three-Dimensional Cortical Cultures for In Vitro Modeling of Brain Networks

iScience ◽  
2020 ◽  
Vol 23 (8) ◽  
pp. 101434
Author(s):  
Yu-Ting L. Dingle ◽  
Volha Liaudanskaya ◽  
Liam T. Finnegan ◽  
Kyler C. Berlind ◽  
Craig Mizzoni ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Brain Networks ◽  
Three Dimensional ◽  
In Vitro Modeling ◽  
Cortical Cultures

scholarly journals Variation Among Isolates of Fusarium graminearum Associated with Fusarium Head Blight in North Carolina

Plant Disease ◽  
2001 ◽  
Vol 85 (4) ◽  
pp. 404-410 ◽  
Author(s):  
Scott L. Walker ◽  
Steven Leath ◽  
Winston M. Hagler ◽  
J. Paul Murphy
Keyword(s):  
North Carolina ◽  
Fusarium Head Blight ◽  
Fusarium Graminearum ◽  
Phenotypic Diversity ◽  
In Vitro Growth ◽  
Head Blight ◽  
Potential Threat ◽  
Previous Crop

Fusarium head blight (FHB) can reduce yield of wheat and decrease the value of harvested grain by accumulation of detrimental toxins. Understanding the variability of the fungal population associated with infection could improve disease control strategies. Sixty-six isolates of Fusarium graminearum associated with FHB were collected in North Carolina and tested for in vitro growth rate, in vitro production of deoxynivalenol (DON) and zearalenone, and pathogenicity on three cultivars of soft red winter wheat. Significant differences among isolates were found for all three traits. Randomly Amplified Polymorphic DNA (RAPD) analysis revealed high levels of genotypic diversity among isolates. Isolates of F. graminearum, F. culmorum, and F. avenaceum acquired from the Pennsylvania State University Fusarium Center were included for comparison in all tests. In vivo levels of DON were measured for the five isolates associated with the highest levels of disease and the five isolates associated with the lowest levels of disease, and no significant differences were found. However, all ten isolates produced detectable levels of DON in vivo. Mean disease ratings ranged from 3.4 to 96.4%, in vitro (DON) levels ranged from 0 to 7176.2 ppm, and zearalenone ranged from 0 to 354.7 ppm, among isolates. A multiple regression model using in vitro growth, in vitro DON, and zearalenone production, collection location, wheat cultivar of isolate origin, plot, tillage conditions, and previous crop as independent variables and percent blighted tissue as the dependent variable was developed. The cumulative R2 value for the model equaled 0.27 with in vitro rate of growth making the largest contribution. Analysis of phenotype and genotype among isolates demonstrated diversity in a single plot, in a single location, and in North Carolina. Genotypic and phenotypic diversity were significant under both conventional and reduced tillage conditions, and diversity was high regardless of whether the previous crop had been a host or non-host for F. graminearum. These data indicate a variable pathogen population of F. graminearum exists in North Carolina, and members of this population can be both highly pathogenic on wheat and produce high levels of detrimental toxins, indicating a potential threat for problems with FHB within the state.

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