1,721,215 research outputs found
Biological detoxification of the mycotoxin deoxynivalenol and its use in genetically engineered crops and feed additives
Full text linkDeoxynivalenol (DON) is the major mycotoxin produced by Fusarium fungi in grains. Food and feed contaminated with DON pose a health risk to humans and livestock. The risk can be reduced by enzymatic detoxification. Complete mineralization of DON by microbial cultures has rarely been observed and the activities turned out to be unstable. The detoxification of DON by reactions targeting its epoxide group or hydroxyl on carbon 3 is more feasible. Microbial strains that de-epoxidize DON under anaerobic conditions have been isolated from animal digestive system. Feed additives claimed to de-epoxidize trichothecenes enzymatically are on the market but their efficacy has been disputed. A new detoxification pathway leading to 3-oxo-DON and 3-epi-DON was discovered in taxonomically unrelated soil bacteria from three continents; the enzymes involved remain to be identified. Arabidopsis, tobacco, wheat, barley, and rice were engineered to acetylate DON on carbon 3. In wheat expressing DON acetylation activity, the increase in resistance against Fusarium head blight was only moderate. The Tri101 gene from Fusarium sporotrichioides was used; Fusarium graminearum enzyme which possesses higher activity towards DON would presumably be a better choice. Glycosylation of trichothecenes occurs in plants, contributing to the resistance of wheat to F. graminearum infection. Marker-assisted selection based on the trichothecene-3-O-glucosyltransferase gene can be used in breeding for resistance. Fungal acetyltransferases and plant glucosyltransferases targeting carbon 3 of trichothecenes remain promising candidates for engineering resistance against Fusarium head blight. Bacterial enzymes catalyzing oxidation, epimerization, and less likely de-epoxidation of DON may extend this list in future
Diophantine Equations Relating Sums and Products of Positive Integers: Computation-Aided Study of Parametric Solutions, Bounds, and Distinct-Term Solutions
No full textDiophantine equations ∏i=1nxi=F∑i=1nxi with xi,F∈ℤ+ associate the products and sums of n natural numbers. Only special cases have been studied so far. Here, we provide new parametric solutions depending on F and the divisors of F or F2. One of these solutions shows that the equation of any degree with any F is solvable. For n = 2, exactly two solutions exist if and only if F is a prime. These solutions are (2F, 2F) and (F + 1, F(F + 1)). We generalize an upper bound for the sum of solution terms from n = 3 established by Crilly and Fletcher in 2015 to any n to be (F+1)(F+n−1) and determine a lower bound to be nnFn−1. Confining the solutions to n-tuples consisting of distinct terms, equations of the 4th degree with any F are solvable but equations of the 5th to 9th degree are not. An upper bound for the sum of terms of distinct-term solutions is conjectured to be (F+1)[F+(n−2)(n−1)!/2+1]/(n−2)!. The conjecture is supported by computation, which also indicates that the upper bound equals the largest sum of solution terms if and only if F=(n+k−2)(n−2)!−1, k∈ℤ+. Computation provides further insights into the relationships between F and the sum of terms of distinct-term solutions
Components of variance in transcriptomics based on electrophoretic separation of cDNA fragments (cDNA-AFLP)
No full textThe sources of variance and errors in transcriptomics based on the electrophoretic separation of amplified cDNA fragments were investigated using cDNA-amplified fragment length polymorphism (AFLP). Transcriptome profiles of the plant-pathogenic fungus Verticillium longisporum were generated by a standard cDNA-AFLP protocol followed by electrophoretic separation of amplified DNA fragments in flatbed polyacrylamide gels with fluorescence detection as well as by capillary electrophoresis (DNA sequencer). The total variance was partitioned into contributions of cDNA synthesis, adapter ligation, preamplification, amplification, and electrophoresis. Parameters of computer-aided peak recognition and matching were investigated and strategies improving matching success based on double passage with different signal intensity thresholds were developed. The overall quality of data was similar for cDNA-AFLP and microarray hybridization. Variance of cDNA-AFLP was independent of signal intensity, whereas microarray data showed higher variance for low-intensity signals. Capillary electrophoresis significantly reduced the number of wrongly matched and unmatched signals as compared with flatbed gels. These results are also likely to apply to related electrophoresis-based transcriptome analysis techniques such as mRNA differential display.German Research Foundation [DFG 546
AFLP fingerprinting of sesame (Sesamum indicum L.) cultivars: identification, genetic relationship and comparison of AFLP informativeness parameters
No full textAmplified fragments length polymorphism (AFLP) was used to distinguish 20 cultivars of sesame (Sesamum indicum L.) and to elucidate the genetic relationship among these genotypes. The data were also used to estimate the usefulness of parameters currently used to assess the informativeness of molecular markers. A total of 339 markers were obtained using 8 primer combinations. Of the bands, 91% were polymorphic. Five primer combinations were able to distinguish all 20 cultivars used. None of the remaining three primer combinations could distinguish all accessions if used alone, but using all three combinations reduced the probability of a random match to 5 x 10(-5). Polymorphic information content (PIC), resolving power (Rp) and marker index (MI) of each primer combination failed to correlate significantly with the number of genotypes resolved. Jaccard's similarity coefficients ranged from 0.31 to 0.78. Fifteen cultivars were grouped by four UPGMA-clusters supported by bootstrapping values larger than 0.70. The grouping pattern was similar to the grouping generated by principal coordinate analysis. The results demonstrated that AFLP-based fingerprints can be used to identify unequivocally sesame genotypes, which is needed for cultivar identification and for the assessment of the genetic variability of breeding stocks. We recommend to use the number of cultivars identified by a primer combination instead of PIC, Rp and MI; and to calculate the maximal, instead of average probability of identical match by chance in the assessment of the informativeness of a marker for cultivar identification
Role of zearalenone lactonase in protection of Gliocladium roseum from fungitoxic effects of the mycotoxin zearalenone
No full textZearalenone is a mycotoxin with estrogenic effects on mammals that is produced by several species of Fusarium. We found that zearalenone and its derivatives inhibit the growth of filamentous fungi on solid media at concentrations of alpha-zearalenol > beta-zearalenol. The mycoparasitic fungus Gliocladium roseum produces a zearalenone-specific lactonase which catalyzes the hydrolysis of zearalenone, followed by a spontaneous decarboxylation. The growth of G. roseum was not inhibited by zearalenone, and the lactonase may protect G. roseum from the toxic effects of this mycotoxin. We inactivated zes2, the gene encoding zearalenone lactonase in G. roseum, by inserting a hygromycin resistance cassette into the coding sequence of the gene by means of Agrobacterium tumefaciens-mediated genetic transformation. The zes2 disruption mutants could not hydrolyze the lactone bond of zearalenone and were more sensitive to zearalenone. These data are consistent with a hypothesis that resorcylic acid lactones exemplified by zearalenone act to reduce growth competition by preventing competing fungi from colonizing substrates occupied by zearalenone producers and suggest that they may play a role in fungal defense against mycoparasites
Simultaneous detection of <it>Fusarium culmorum </it>and <it>F. graminearum </it>in plant material by duplex PCR with melting curve analysis
No full textAbstract Background Fusarium head blight (FHB) is a disease of cereal crops, which has a severe impact on wheat and barley production worldwide. Apart from reducing the yield and impairing grain quality, FHB leads to contamination of grain with toxic secondary metabolites (mycotoxins), which pose a health risk to humans and livestock. The Fusarium species primarily involved in FHB are F. graminearum and F. culmorum. A key prerequisite for a reduction in the incidence of FHB is an understanding of its epidemiology. Results We describe a duplex-PCR-based method for the simultaneous detection of F. culmorum and F. graminearum in plant material. Species-specific PCR products are identified by melting curve analysis performed in a real-time thermocycler in the presence of the fluorescent dye SYBR Green I. In contrast to multiplex real-time PCR assays, the method does not use doubly labeled hybridization probes. Conclusion PCR with product differentiation by melting curve analysis offers a cost-effective means of qualitative analysis for the presence of F. culmorum and F. graminearum in plant material. This method is particularly suitable for epidemiological studies involving a large number of samples.</p
A Comprehensive Review of Hypotheses About the Biological Function of Zearalenone, and a New Hypothesis for the Function of Resorcylic and Dihydroxyphenylacetic Macrolactones in Fungi
No full textThe special metabolite of Fusarium spp. zearalenone (ZEN) exerts estrogenic effects on mammals, stimulates plant growth, stimulates sexual development in fungi, and inhibits fungal growth. These activities inspired hypotheses about the biological function of ZEN. We briefly review the discovery of ZEN and its implications. The main subject of this review is a critical assessment of the hypotheses that ZEN is a fungal hormone, a plant hormone, a virulence factor, or a fungal defense metabolite. Conceptual and technical issues related to testing these hypotheses, such as inadequate analytical methods, confusion of incidental effects with biological functions, and lack of normalization, are illuminated. Based on these considerations, gene knockout experiments, and on the effects of biotic interactions on ZEN synthesis, we argue that ZEN is a defense metabolite protecting Fusarium spp. against mycoparasites and competitors. Similar reasoning and published data suggest that the Fusarium metabolite fusaristatin A fulfils the same function. Fungi produce many macrolactones of resorcylic acid (RALs) and dihydroxyphenylacetic acid (DHPLs) with properties similar to ZEN. Their widespread occurrence, antifungal activity, and further considerations prompt us to hypothesize that the fundamental function of fungal RALs and DHPLs lies in defense and interference competition
Genetic relationship and diversity in a sesame (<it>Sesamum indicum L</it>.) germplasm collection using amplified fragment length polymorphism (AFLP)
No full textAbstract Background Sesame is an important oil crop in tropical and subtropical areas. Despite its nutritional value and historic and cultural importance, the research on sesame has been scarce, particularly as far as its genetic diversity is concerned. The aims of the present study were to clarify genetic relationships among 32 sesame accessions from the Venezuelan Germplasm Collection, which represents genotypes from five diversity centres (India, Africa, China-Korea-Japan, Central Asia and Western Asia), and to determine the association between geographical origin and genetic diversity using amplified fragment length polymorphism (AFLP). Results Large genetic variability was found within the germplasm collection. A total of 457 AFLP markers were recorded, 93 % of them being polymorphic. The Jaccard similarity coefficient ranged from 0.38 to 0.85 between pairs of accessions. The UPGMA dendrogram grouped 25 of 32 accessions in two robust clusters, but it has not revealed any association between genotype and geographical origin. Indian, African and Chinese-Korean-Japanese accessions were distributed throughout the dendrogram. A similar pattern was obtained using principal coordinates analysis. Genetic diversity studies considering five groups of accessions according to the geographic origin detected that only 20 % of the total diversity was due to diversity among groups using Nei's coefficient of population differentiation. Similarly, only 5% of the total diversity was attributed to differences among groups by the analysis of molecular variance (AMOVA). This small but significant difference was explained by the fact that the Central Asia group had a lower genetic variation than the other diversity centres studied. Conclusion We found that our sesame collection was genetically very variable and did not show an association between geographical origin and AFLP patterns. This result suggests that there was considerable gene flow among diversity centres. Future germplasm collection strategies should focus on sampling a large number of plants. Covering many diversity centres is less important because each centre represents a major part of the total diversity in sesame, Central Asia centre being the only exception. The same recommendation holds for the choice of parents for segregant populations used in breeding projects. The traditional assumption that selecting genotypes of different geographical origin will maximize the diversity available to a breeding project does not hold in sesame.</p
Research article
No full textBackground: Diversity estimates in cultivated plants provide a rationale for conservation strategies and support the selection of starting material for breeding programs. Diversity measures applied to crops usually have been limited to the assessment of genome polymorphism at the DNA level. Occasionally, selected morphological features are recorded and the content of key chemical constituents determined, but unbiased and comprehensive chemical phenotypes have not been included systematically in diversity surveys. Our objective in this study was to assess metabolic diversity in sesame by nontargeted metabolic profiling and elucidate the relationship between metabolic and genome diversity in this crop.Results: Ten sesame accessions were selected that represent most of the genome diversity of sesame grown in India, Western Asia, Sudan and Venezuela based on previous AFLP studies. Ethanolic seed extracts were separated by HPLC, metabolites were ionized by positive and negative electrospray and ions were detected with an ion trap mass spectrometer in full-scan mode for m/z from 50 to 1000. Genome diversity was determined by Amplified Fragment Length Polymorphism (AFLP) using eight primer pair combinations. The relationship between biodiversity at the genome and at the metabolome levels was assessed by correlation analysis and multivariate statistics. Conclusion: Patterns of diversity at the genomic and metabolic levels differed, indicating that selection played a significant role in the evolution of metabolic diversity in sesame. This result implies that when used for the selection of genotypes in breeding and conservation, diversity assessment based on neutral DNA markers should be complemented with metabolic profiles. We hypothesize that this applies to all crops with a long history of domestication that possess commercially relevant traits affected by chemical phenotypes
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