Browsing by Subject "Mycotoxins"

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Analyzing resistance to ergot caused by Claviceps purpurea [Fr.] Tul. and alkaloid contamination in winter rye (Secale cereale L.)
(2022) Kodisch, Anna; Miedaner, Thomas
Ergot caused by Claviceps purpurea [Fr.] Tul. is one of the oldest well-known plant diseases leading already in medieval times to severe epidemic outbreaks. After the occurrence of honeydew, the characteristic ergot bodies called sclerotia are formed on the ear. These are containing toxic ergot alkaloids (EAs). Strict limits are set within the European Union. Rye (Secale cereale L.) as cross-pollinating crop is particularly vulnerable to ergot since the competitive situation of fungal spores and pollen during flowering. Nevertheless, even today the threat is real as agricultural practice is changing and screening studies revealed EAs in samples of the whole cereal value chain frequently. The aims were to establish a harmonized method to test ergot resistance and EA contamination in winter rye, to clarify major significant factors and their relevance and to reveal the suitability of one commercial ELISA test. Further, effort was paid to examine the covariation of ergot amount and EA content considering different factors because of prospective legislative changes. Genotypes showed significant variation for ergot severity and pollen-fertility restoration after natural infection as well as artificial inoculation whereas a high positive correlation could be found between both treatments. Additionally, variances of environment, general combining ability (GCA), specific combining ability (SCA), and interactions were significant. Although male pollen-fertility restoration was of utmost importance, the female component was also significant. This illustrates that apart from promising selection of high restoration ability the maternal restorability could be exploited in future breeding programs especially when a high pollen amount is already reached. A large-scale calibration study was performed to clarify the covariation of ergot severity, EA content (HPLC, ELISA) considering genotypes, locations, countries, years, and isolates. EA profile was rather stable across country-specific isolates although large differences regarding the EA content were detected. The moderate covariation between ergot severity and EA content (HPLC) indicates that a reliable prediction of the EA content based on ergot severity is not possible what can also not be explained by grouping effects of the factors. Further, EAs seem not to act as virulence factor in the infection process since EA content showed no relationship to disease severity. Additionally, the missing correlation of ELISA and HPLC leads to the conclusion that the ELISA is not an appropriate tool what can be used safely to screen samples regarding ergot in the daily life. The genetic variation of male-sterile CMS-single crosses was analysed in a special design without pollen in field and greenhouse to identify resistance mechanisms and to clarify whether ergot can be reduced in the female flower. At this, comparison of needle and spray inoculation revealed medium to high correlations illustrating that both methods were suitable for this research. Significant environment and genotype by environment interaction variances were detected. So, testing across several environments is necessary also without pollen. Further, small but significant genotypic variation and identification of one more ergot-resilient candidate revealed that selection of female lines could be promising to further reduce ergot. The EA content was lower for less susceptible genotypes. Thus, EA content can be considerably reduced by breeding. A strong positive correlation could be found for ergot severity and EA content when analysing 15 factorial single crosses. The male pollen-fertility restoration was also here the most relevant component but the female component contributed an obviously higher proportion for the EA content than for ergot severity. In conclusion, this thesis demonstrate that implementing of a high and environmental stable male fertility restoration ability via exotic Rf genes can effectively reduce ergot although also the female restorability enables great opportunities. The unpredictable covariation between ergot amount and EA content illustrates that both traits have to be assessed, in particular the EA content by a valid HPLC approach to guarantee food and feed safety.
Effects of non-adapted quantitative trait loci (QTL) for Fusarium head blight resistance on European winter wheat and Fusarium isolates
(2010) Ohe, Christiane von der; Miedaner, Thomas
Fusarium head blight (FHB), caused by Fusarium graminearum and F. culmorum, is a devastating disease responsible for tremendous damage in wheat fields and contamination of grain with mycotoxins deoxynivalenol (DON) and nivalenol (NIV), rendering the harvest unsafe for human and animal consumption. The variability of Fusarium populations is high and changes in aggressiveness, chemotypes or species within and among Fusarium populations are known. Stable FHB resistance combined with high yield is one main target in wheat breeding programs. Mapping studies detected several quantitative trait loci (QTL) for FHB resistance in non-adapted sources, such as Sumai3 from China. The two most important and commonly used major QTL are located on chromosome 3BS (Fhb1) und 5A (Qfhs.ifa-5A). However, negative side effects of non-adapted resistance sources introgressed in elite winter wheat material are feared in Europe. Furthermore, the stability of the QTL effect against changing Fusarium populations is unknown. The objectives of this research were to analyze whether (1) the QTL Fhb1 and Qfhs.ifa-5A introgressed from a non-adapted resistance source into two winter wheat varieties have possible side effects on agronomic and quality performance, (2) 3-ADON and 15-ADON chemotypes are significantly different in their aggressiveness and DON production, (3) competition among Fusarium isolates in mixtures exists, and if so, how the resistant host will influence this competition. In conclusion, both resistance QTL are effective and stable in elite spring and winter wheat backgrounds. For improvement of FHB resistance both QTL are valuable, but Qfhs.ifa-5A would suffice for European breeding programs. Due to chemotype shifts, 3-ADON isolates could pose a greater risk to food safety than 15-ADON but breeding and use of highly resistant lines can reduce the risks associated with DON in wheat. Accordingly, resistant spring wheat lines were less affected by the tested Fusarium isolates and mixtures and, therefore, confirmed a high stability of these QTL. Directed selection of highly aggressive isolates due to the resistance QTL seems to be unlikely in the short term.
Genetic variation in early maturing European maize germplasm for resistance to ear rots and mycotoxin contamination caused by Fusarium spp.
(2010) Bolduan, Christof; Melchinger, Albrecht E.
Ear rots of maize, caused by Fusarium spp., are of major concern because they lead to losses in grain yield and contamination with mycotoxins which harm animals and humans. In the absence of other strategies, breeding maize for genetic resistance is currently the most promising avenue to control these rots and mycotoxin accumulation. The predominant pathogens in Central Europe are F. graminearum, the causative agent of Gibberella ear rot (GER), and F. verticillioides, the causative agent of Fusarium ear rot (FER). GER causes contamination with deoxynivalenol (DON), nivalenol and zearalenone (ZEA), whereas FER causes contamination with fumonisins (FUM). Information on the resistance to GER and FER and mycotoxin contamination is lacking for maize adapted to the cooler climatic conditions of Central Europe. In this study we investigated (1) the resistance of early maturing European elite inbred lines against GER and FER and contamination of mycotoxins, (2) the genetic variances and heritabilities for ear rot ratings and mycotoxin concentrations, (3) the correlations of ear rot ratings with mycotoxin concentrations, (4) the correlations between line per se (LP) and testcross performance (TP) for GER rating and DON concentration, (5) the aggressiveness of and mycotoxins produced by different isolates of F. graminearum and F. verticillioides, and (6) the potential of near infrared spectroscopy (NIRS) to estimate concentrations of DON and FUM in maize grains under artificial inoculation. Significant genotypic variances and moderate to high heritabilities were found for GER, DON and ZEA among the inbred lines and for GER and DON among the testcrosses, as well as for FER and FUM among the inbred lines. Further, genotype x environment interaction variances were significant for all traits except FUM. Thus, the results underlined the presence of ample genotypic variation and the need to conduct multi-environment tests for reliable identification of resistant genotypes. Ear rot ratings and mycotoxin production of eight isolates each of F. graminearum and F. verticillioides differed significantly. Even though, isolate x inbred interactions were significant only in the case of F. graminearum, and no rank reversals occurred among the tested inbred lines. Most isolates differentiated the susceptible inbreds from the resistant ones for severity ratings. However, the differences between the two groups were smaller for the less aggressive isolates. Therefore, we recommend using a single, environmentally stable and sufficiently aggressive isolate for resistance screenings under artificial inoculation. Strong correlations between ear rot severity and mycotoxin concentrations indicated that selection for low ear rot severity under artificial inoculation will result in high correlated selection response for low mycotoxin concentration, particularly for GER and DON. Selection for ear rot severity is less resource-demanding and quicker than selection for mycotoxin concentration. Thus, it enables the breeder to maximize selection gain for a given budget. However, the selected elite material should be evaluated for mycotoxin concentrations in order to avoid ?false positives?. In this regard, NIRS showed high potential to predict DON concentrations in grain obtained from artificially inoculated maize. Compared to the commonly employed ELISA assay, NIRS assays are considerably cheaper, because no mycotoxin extractions and test kits are needed. We observed moderate positive correlations between GER and FER, and identified inbreds combining resistance to both ear rots. Therefore, selection for resistance to one pathogen is expected to result in indirect response to the other. Nevertheless, in advanced stages of each breeding cycle, lines preselected for other agronomically important traits should be evaluated for resistance to both pathogens. Genotypic variances for GER and DON were generally higher in LP than TP. Thus, assuming identical selection intensities for each scheme, the expected response to selection for LP should be higher than for TP. However, owing to moderate correlations between LP and TP for GER and DON, selection based on LP is not sufficient, because the ultimate goal is to develop resistant hybrids. Therefore, a multi-stage selection procedure is recommended with evaluation of agronomically promising lines for GER in only one environment in order to eliminate highly susceptible lines, followed by evaluation of TP of the selected lines for GER with one tester of moderate to high resistance level from the opposite heterotic pool in two to three environments.
Genetics of resistance to ear diseases and mycotoxin accumulation in the pathosystems maize/Fusarium and wheat/Fusarium
(2010) Messerschmidt, Martin; Miedaner, Thomas
Infection of ears of maize with Fusarium graminearum (FG) reduces yield and, more important, contaminate the harvest with mycotoxins. F. verticillioides (FV) is an economically important cause of ear rot. Among other mycotoxins, FV produces the fumonisins (FUM) and FG produces deoxynivalenol (DON) and zearalenone (ZEA). All three mycotoxins are harmful to humans and animals. Therefore, the European Union released legally enforceable limits. One alternative to reduce ear rot severity and mycotoxin concentrations is breeding and growing varieties resistant to Fusarium infections. However, few is known about breeding parameters for resistance to Fusarium infections and mycotoxin accumulation in European maize breeding material. The main objective of this thesis was to draw conclusions for breeding of resistance to ear rot and mycotoxin accumulation with special attention on three European maize maturity groups. We investigated methodical aspects like (1) the comparison of natural and artificial inoculation to evaluate ear rot resistance and (2) the necessity of separate testing of FV and FG. Furthermore, quantitative-genetic parameters like heritabilities and correlations were estimated to draw conclusions about (3a) genetic variation in line and testcross performance and the relationships (3b) between ear rot severity and mycotoxin concentrations in lines and testcrosses and (3c) between line and testcross performance. Three maturity groups (early, mid-late, late) each comprising about 150 maize inbred lines were evaluated for ear rot resistance to FV. The same genotypes of the early maturity group were additionally evaluated for resistance to FG in separate, but adjacent trials. Field evaluation was conducted in two to six environments with silk channel inoculation and natural infection, respectively. In the late maturity group kernel inoculation was conducted additionally. Out of the 150 lines, 50 to 60 lines per maturity group were crossed with two unrelated testers of the opposite heterotic group. The concentrations of toxins FUM, DON and ZEA of the chosen lines and their testcrosses were analyzed by immunotests. Despite significant genotypic differences among the inbred lines after inoculation or natural infections, inoculation was found to be superior due to easier visual differentiation and increased accuracy. Therefore, inoculation should be conducted. In the late maturity group silk channel inoculation (simulating infection over the silks) and kernel inoculation (simulating secondary infection after wounding) were appropriate since both caused similar ear rot severity. However, both inoculation methods should be tested separately due to only moderate correlations between them. In the early maturity group resistance to FG or FV should be tested separately due to moderate correlations. Significant genotypic variances in large sets and subsets of lines and also in testcrosses revealed that there is genetic variation in all maturity groups and also within heterotic groups. In the flint group less lines were resistant to FV and FG than in dents indicating that resistance needs improvement, i.e. by introgression of resistance alleles followed by recurrent selection. Significant genotype x environment interactions may complicate selection and, therefore, multi-environmental trials are required for an accurate selection. High genotypic correlations between ear rot rating and mycotoxin concentrations were found among lines and testcrosses. The cost efficient indirect selection for mycotoxin concentrations based on ear rot rating could increase response to selection by testing more genotypes and/or in more test environments assuming a fixed budget. This should increase selection intensity and/or heritability. Moderate genotypic correlations between line and testcross performance were. One moderately to highly susceptible tester is sufficient due to high genotypic correlations between testcrosses of different testers. Both indicates a mainly additive gene action, but also non-additive gene action may play a role in some crosses. Selection for testcross performance based on line performance was less effective when calculating relative efficiencies. Different scenarios have been identified: (1) In Central Europe mainly resistance to ear rot in lines needs to be tested to ensure high seed quality, whereas resistance in testcrosses is not important due to low natural infection. (2) In Southern Europe, where high natural infections occur regularly, parallel selection for resistance to ear rot in lines and testcrosses is important. One susceptible tester should be used for creation of testcrosses. For selection in lines all parental lines should be inoculated but only lines selected out of testcrosses for agronomic traits would be rated afterwards saving resources. This is feasible due to later harvest date of lines than of testcrosses.
Molecular mapping of resistance and aggressiveness in the cereal/Fusarium head blight pathosystem
(2016) Kalih, Rasha; Miedaner, Thomas
Fusarium head blight (FHB) is one of the most destructive fungal diseases in small-grain cereals worldwide causing significant yield losses and contamination of grain with mycotoxins e.g., deoxynivalenol (DON). This renders the grain unsuitable for human consumption and animal feeding. Exploring the genetic mechanism of FHB resistance is considered the key tool for modern cereal breeding activities. Triticale, the intergeneric hybrid between wheat and rye, is an important cereal crop in Poland and Germany. Resistance breeding using genetic mapping to identify quantitative-trait loci (QTL) associated with FHB resistance represents the best strategy for controlling the disease. In parallel, understanding the mechanism of aggressiveness and DON production of F. graminearum will be a significant contribution to improve FHB management. The objectives of the present work were (1) identification of QTL related to FHB resistance in triticale, together with the analysis of the correlation of FHB severity with other related traits such as plant height and heading stage, (2) correlation between DON production and FHB severity, (3) mapping of dwarfing gene Ddw1 in triticale and studying its effect on FHB resistance, plant height and heading stage, (4) detection of SNPs in candidate genes associated with aggressiveness and DON production of a large Fusarium graminearum population in bread wheat. To study the genetic architecture of FHB resistance in triticale, five doubled-haploid (DH) triticale populations with 120 to 200 progenies were successfully tested under field conditions by inoculation with Fusarium culmorum (FC46) in multiple environments. All genotypes were evaluated for FHB resistance, plant height and heading stage. DArT markers were used to genotype triticale populations. Significant genotypic variances (P<0.001) were observed for FHB severity in all populations combined with high heritability. Twenty-two QTLs for FHB resistance in triticale were reported with two to five QTL per population, thus confirming the quantitative inheritance of FHB resistance in triticale. The most prominent (R2 ≥ 35%) QTLs were located on chromosomes 6A, 3B, 4R, and 5R. QTLs for plant height and heading stage were also detected in our work, some of them were overlapping with QTLs for FHB resistance. Correlation between FHB severity, DON content and Fusarium damaged kernels (FDK) in triticale was studied in the population Lasko x Alamo. Significant genotypic variance was detected for all traits. However, low correlation between FHB severity and DON content (r=0.31) was found. Interestingly, correlation between FHB severity and FDK rating was considerably higher (r=0.57). For FHB severity, two QTLs were detected in this population. A QTL located on chromosome 2A with minor effect for FHB severity was also a common QTL for DON content and FDK rating and explained ≥34% of genotypic variance for these two traits. A second QTL on chromosome 5R was a major QTL but it has no effect on DON content or FDK rating. For analyzing the rye dwarfing gene Ddw1 derived from the father Pigmej, 199 (DH) progenies were genotyped with DArT markers and in addition with conserved ortholog set (COS) markers linked to the Ddw1 locus in rye. QTL analyses detected three, four, and six QTLs for FHB severity, plant height and heading stage, respectively. Two specific markers tightly linked with Ddw1 on rye chromosome 5R explained 48, 77, and 71 % of genotypic variation for FHB severity, plant height, and heading stage, respectively. This is strong evidence, that we indeed detected the rye gene Ddw1 in this triticale population. Another objective was to highlight the association between quantitative variation of aggressiveness and DON production of 152 F. graminearum isolates with single nucleotide polymorphism (SNP) markers in seven candidate genes. One to three significant SNPs (P < 0.01 using cross-validation) were associated to FHB severity in four genes (i.e., Gmpk1, Mgv1, TRI6, and Erf2). For DON content, just one significant SNP was detected in the gene Mgv1 explaining 6.5% of the total genotypic variance. In conclusion, wide genetic variation in FHB resistance in triticale has been observed in five populations. QTL mapping analyses revealed twenty-two QTLs for FHB resistance derived from wheat and rye genomes. QTLs located on the rye genome were reported here for the first time and they are a new source for FHB resistance in triticale. In parallel, analysis of the diversity of four pathogenicity genes in F. graminearum is an important first step in inferring the genetic network of pathogenicity in this fungal pathogen.
Occurrence of mycotoxins in pulses
(2022) Acuña‐Gutiérrez, Catalina; Jiménez, Víctor M.; Müller, Joachim
Pulses, dry grains of the Fabaceae family used for food and feed, are particularly important agricultural products with increasing commercial and nutritional relevance. Similar to other plant commodities, pulses can be affected by fungi in the field and during postharvest. Some of these fungi produce mycotoxins, which can seriously threaten human and animal health by causing acute poisoning and chronic effects. In this review, information referring to the analysis and occurrence of these compounds in pulses is summarized. An overview of the aims pursued, and of the methodologies employed for mycotoxin analysis in the different reports is presented, followed by a comprehensive review of relevant articles on mycotoxins in pulses, categorized according to the geographical region, among other considerations. Moreover, special attention was given to the effect of climatic conditions on microorganism infestation and mycotoxin accumulation. Furthermore, the limited literature available was considered to look for possible correlations between the degree of fungal infection and the mycotoxin incidence in pulses. In addition, the potential effect of certain phenolic compounds on reducing fungi infestation and mycotoxin accumulation was reviewed with examples on beans. Emphasis was also given to a specific group of mycotoxins, the phomopsins, that mainly impact lupin. Finally, the negative consequences of mycotoxin accumulation on the physiology and development of contaminated seeds and seedlings are presented, focusing on the few reports available on pulses. Given the agricultural and nutritional potential that pulses offer for human well-being, their promotion should be accompanied by attention to food safety issues, and mycotoxins might be among the most serious threats.