Expression of 3-OH trichothecene acetyltransferase in barley (Hordeum vulgare L.) and effects on deoxynivalenol

Manoharan, Muthusamy; Dahleen, Lynn S.; Höhn, Thomas; Neate, Stephen M.; Yu, Xiaohong; Alexander, Nancy J.; McCormick, Susan P.; Bregitzer, Phil; Schwarz, Paul; Horsley, Richard D.

doi:10.1016/j.plantsci.2006.07.004

Cited by 44 publications

(34 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…7). Kinetic Analysis of FsTRI101 and FgTRI101-Previous studies have demonstrated that TRI101 isolated from different trichothecene producing organisms are able to acetylate both A and B type trichothecene mycotoxins (13,18,20,21,38). However, these earlier studies were limited to end point assays that did not evaluate the catalytic rate of each ortholog.…”

Section: Resultsmentioning

confidence: 99%

“…The tri101 gene from F. sporotrichioides has been isolated and transgenically inserted into yeast (18), tobacco (19), wheat (20), and barley (21). Likewise, the tri101 gene from F. graminearum has been transgenically inserted into Saccharomyces pombe (13) and rice (22).…”

mentioning

confidence: 99%

“…Unfortunately, field trials of transgenic wheat and barley expressing the tri101 gene from F. sporotrichioides did not show resistance to F. graminearum. It was concluded that the intense disease pressure in the field trials overwhelmed the resistance provided by FsTri101 and perhaps was compounded by the use of a tri101 gene from a T-2 producing organism rather than one from a DON producing Fusarium strain (21).…”

mentioning

confidence: 99%

See 2 more Smart Citations

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

Garvey

McCormick

Rayment

2008

Journal of Biological Chemistry

Self Cite

101

View full text Add to dashboard Cite

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 ϳ100-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.

show abstract

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

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

Garvey

McCormick

Rayment

2008

Journal of Biological Chemistry

Self Cite

101

View full text Add to dashboard Cite

show abstract

“…Rice roots subjected to DON showed reduced growth, while roots expressing TRI101 were not inhibited and grew similarly to unexposed wild-type roots. Barley transformed with FsTRI101 had reduced FHB in greenhouse assays, although field tests showed that transgenic plants were as susceptible to FHB as the controls (18). These noted works all illustrate the great potential of utilizing trichothecene acetyltransferase genes, such as TRI101, to detoxify DON.…”

mentioning

confidence: 90%

Bioprospecting for Trichothecene 3- O -Acetyltransferases in the Fungal Genus Fusarium Yields Functional Enzymes with Different Abilities To Modify the Mycotoxin Deoxynivalenol

Khatibi

Newmister

Rayment

et al. 2011

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

The trichothecene mycotoxin deoxynivalenol (DON) is a common contaminant of small grains, such as wheat and barley, in the United States. New strategies to mitigate the threat of DON need to be developed and implemented. TRI101 and TRI201 are trichothecene 3-O-acetyltransferases that are able to modify DON and reduce its toxicity. Recent work has highlighted differences in the activities of TRI101 from two different species of Fusarium (F. graminearum and F. sporotrichioides), but little is known about the relative activities of TRI101/TRI201 enzymes produced by other species of Fusarium. We cloned TRI101 or TRI201 genes from seven different species of Fusarium and found genetic identity between sequences ranging from 66% to 98%. In vitro feeding studies using transformed yeast showed that all of the TRI101/TRI201 enzymes tested were able to acetylate DON; conversion of DON to 3-acetyl-deoxynivalenol (3ADON) ranged from 50.5% to 100.0%, depending on the Fusarium species from which the gene originated. A time course assay showed that the rate of acetylation varied from species to species, with the gene from F. sporotrichioides having the lowest rate. Steady-state kinetic assays using seven purified enzymes produced catalytic efficiencies for DON acetylation ranging from 6.8 ؋ 10 4 M ؊1 ⅐ s ؊1 to 4.7 ؋ 10 6 M ؊1 ⅐ s ؊1. Thermostability measurements for the seven orthologs ranged from 37.1°C to 43.2°C. Extended sequence analysis of portions of TRI101/TRI201 from 31 species of Fusarium (including known trichothecene producers and nonproducers) suggested that other members of the genus may contain functional TRI101/TRI201 genes, some with the potential to outperform those evaluated in the present study.

show abstract

“…Therefore, DON detoxification should also be considered together with the transgenic strategies of ZEN decontamination. Although genes for irreversible detoxification of trichothecenes are not currently available, numerous efforts are being made to solve the problems of DON contamination (6,14,15,19,23,24). In combination with such progress in biotechnological detoxification of DON, the transgenic system of detoxification of ZEN demonstrated in this paper will contribute significantly to solving the mycotoxin problems in maize and other cereals.…”

Section: Discussionmentioning

confidence: 98%

Reduced Contamination by the Fusarium Mycotoxin Zearalenone in Maize Kernels through Genetic Modification with a Detoxification Gene

Igawa

Takahashi-Ando

Ochiai

et al. 2007

Appl Environ Microbiol

View full text Add to dashboard Cite

Maize is subject to ear rot caused by toxigenic Aspergillus and Fusarium species, resulting in contamination with aflatoxins, fumonisins, trichothecenes, and zearalenone (ZEN). The trichothecene group and ZEN mycotoxins are produced by the cereal pathogen Fusarium graminearum. A transgenic detoxification system for the elimination of ZEN was previously developed using an egfp::zhd101 gene (gfzhd101), encoding an enhanced green fluorescent protein fused to a ZEN-degrading enzyme. In this study, we produced a transgenic maize line expressing an intact copy of gfzhd101 and examined the feasibility of transgene-mediated detoxification in the kernels. ZEN-degrading activity has been detected in transgenic kernels during seed maturation (for a period of 6 weeks after pollination). The level of detoxification activity was unaltered after an additional storage period of 16 weeks at 6°C. When the seeds were artificially contaminated by immersion in a ZEN solution for 48 h at 28°C, the total amount of the mycotoxin in the transgenic seeds was uniformly reduced to less than 1/10 of that in the wild type. The ZEN in the transgenic maize kernels was also efficiently decontaminated under conditions of lower water activity (a w ) and temperature; e.g., 16.9 g of ZEN was removed per gram of seed within 48 h at an a w of 0.90 at 20°C. F. graminearum infection assays demonstrated an absence of ZEN in the transgenic maize seeds, while the mycotoxin accumulated in wild-type kernels under the same conditions. Transgenemediated detoxification may offer simple solutions to the problems of mycotoxin contamination in maize.

show abstract

Expression of 3-OH trichothecene acetyltransferase in barley (Hordeum vulgare L.) and effects on deoxynivalenol

Cited by 44 publications

References 29 publications

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

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

Bioprospecting for Trichothecene 3- O -Acetyltransferases in the Fungal Genus Fusarium Yields Functional Enzymes with Different Abilities To Modify the Mycotoxin Deoxynivalenol

Reduced Contamination by the Fusarium Mycotoxin Zearalenone in Maize Kernels through Genetic Modification with a Detoxification Gene

Contact Info

Product

Resources

About