Browsing by Subject "Fusarium"

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Biological control of Striga hermonthica (Del.) Benth. using formulated mycoherbicides under Sudan field conditions
(2008) Zahran, Eldur; Sauerborn, Joachim
Striga hermonthica is a parasitic flowering plant belonging to the family Orobanchaceae. It is a root parasite that attacks sorghum, maize, millet and several grass weeds in the semi-arid Tropics. In Sudan, Striga is widespread in irrigated and rainfed areas and considered the main biotic constraint in production of sorghum, the main staple food for the majority of Sudanese people. More than 500,000 hectares under rainfed cultivation are heavily infested with Striga, which commonly results in significant yield losses of 70 ? 100%. It has become obvious that there is no simple, fast and inexpensive solution to the Striga problem in Africa. Biological control is considered a potential cost-effective and environmentally safe means for reducing weed populations in crops, forests, or rangelands where low profit margins prevent large herbicide expenditure. Biological control using microorganisms (especially phytopathogenic fungi) showed a high efficacy in controlling S. hermonthica under controlled and field conditions. However, so far it did not come to practical field application. This could be attributed to environmental obstacles or due to the lack of appropriate delivery systems. The pathogenicity of two fungal isolates indigenous to Sudan (Fusarium nygamai [FN] and F. ?Abuharaz? [FA] isolate) against Striga has been studied using infected sorghum grains or a spore suspension as inoculum. These formulations were very effective in controlling Striga under controlled and natural conditions; however, a high level of fungal inoculum (approximately 800 kg ha-1 for the grain inoculum) would be required for effective control, which arises a lot of problems e.g. concerning sterilization and transportation. Such problems can be overcome by adopting an appropriate formulation technology. Granular formulations such as ?Pesta? and alginate pellets were found to be suitable delivery systems for controlling weeds. ?Pesta? granules are made by encapsulating bioagents in a gluten matrix. Alginate formulations are prepared by incorporating the biocontrol agent?s propagules in a sodium alginate solution, which is dripped to a calcium chloride or calcium gluconate solution. Alginate pellets are then formed by ionotrophic gelation. The main objectives of this study were: (a) to study the efficacy of the two Fusarium species in controlling Striga under field conditions using ?Pesta? and alginate formulations, (b) evaluate the effect on sorghum yield, (c) determine the optimum dose of the formulated material, (d) investigate the persistence of the formulated fungal isolates in the soil, and (e) study the efficacy of seed treatments as an alternative delivery system. Furthermore, for environmental safety reasons the newly isolated F. ?Abuharaz? isolate was tested for its ability to produce some of the most important mycotoxins. Harvested sorghum seeds out of the fungus-treated plots were also investigated for their mycotoxins content. A prerequisite to be able to formulate biocontrol fungi is the development of an inexpensive method of inoculum production that yields sufficient biomass containing viable, highly virulent propagules. Chlamydospores are the soil-persisting propagules of many Fusarium species and considered as ideal propagules to be used in granular formulations. For this reason, finding a medium suitable for the production of chlamydospores by the two Fusarium isolates was one of the specific objectives of this study. Different media were tested among them Special Nutrient-poor Broth (SNB) + yeast gave the highest number of chlamydospores (105 ml-1) in both isolates throughout the incubation period. However, both isolates generally did not form sufficient chlamydospores to be used within a bioherbicide formulation. Richard?s solution gave the highest number of microconidia (108 ml-1) after five days of incubation and hence it was selected as growth medium for formulation purposes throughout this study. FN and FA were successfully formulated in ?Pesta? and alginate granules amended either with 10% wheat flour or 6% sorghum flour or yeast extract. Alginate granules generally gave higher numbers of colony forming units (cfu) per g of formulated material compared to ?Pesta?. Alginate preparations amended with 6% sorghum flour or yeast extract had significantly higher cfu compared to the alginate formulation using 10% wheat flour. Yeast extract amendment further increased the number of cfu by about 38 and 32% for FN and FA, respectively, compared to sorghum amendment. In the first field experiment (2003/04), a screening for the suitable dose of ?Pesta? granules per planting hole to control Striga was conducted together with the investigation of a seed coating treatment as an alternative delivery system of the biocontrol agents. The ?Pesta? technology showed a potential to be used as a delivery system to control S. hermonthica under field conditions. Both ?Pesta?-formulated Fusarium isolates were able to delay Striga emergence, reduce the total number of Striga shoots and induce disease symptoms on all growth stages of Striga plants, irrespective to the dose and method of application used. The highest control efficacy was achieved by applying FA at 1.5g, which reduced the total number of parasite shoots by 82 % and the number of healthy Striga shoots by 88% compared to the untreated control. As a consequence, sorghum biomass and sorghum 100-seed weight were increased by 86 and 110%, respectively. FN and the combination of the fungal isolates were slightly less efficient in controlling the parasites. 1.5 g ?Pesta? granules per planting hole was found to be the optimum dosage for Striga management since increasing the dosage did not result in a significant improvement of control. In the second season (2004/05), the efficacy of alginate formulations amended with 10% wheat flour applied at 1.5g/planting hole was evaluated in addition to the ?Pesta? formulation for controlling Striga under field conditions. Alginate granules were able to delay Striga incidence significantly and reduce the total number of Striga shoots by 64 ? 78 % compared to the control early in the season. In contrast to the first season, fungal isolates formulated in ?Pesta? granules had no pronounced effect on delaying Striga emergence, however, ?Pesta?-granulated Fusarium species were able to reduce the total number of Striga shoots by 42 ? 55 % compared to the control early in the season. By the end of the season, the effect of both formulations on the total number of Striga shoots became negligible, but they significantly increased disease incidence on Striga shoots compared to the untreated control. FA formulated in ?Pesta? or alginate pellets was especially effective in this regard, causing disease in 74 and 80% of the Striga plants and reducing the total number of healthy Striga shoots by 55 and 60% compared to the control, respectively. FA applied as ?Pesta? granules was the most effective treatment in reducing Striga biomass by 58 % compared to the control which was positively reflected in an increased sorghum grain yield (63%) and sorghum straw yield (73%) compared to the control. The reduction of the efficacy of the ?Pesta? formulation in controlling Striga in the second season compared to the first season can probably be attributed to three reasons. These include a) climatic conditions, which differed from the first season in higher temperatures coupled with lower rainfall and low relative humidity, b) sodicity problems in the fields which might have affected the proliferation of the fungi in the soil and c) an inhibitory effect of the metabolites of the applied insecticide Sevin (active ingredient Carbaryl (1-naphthyl N-methylcarbamate)) on the virulence of soil fungi. Furthermore, an outdoor pot experiment was conducted to study the efficacy of alginate formulations with different amendments (wheat flour, sorghum flour and yeast extract) in comparison to the ?Pesta? formulation and seed treatments on controlling Striga. In this experiment, FA formulated as ?Pesta? granules was the most effective treatment and successfully inhibited Striga emergence until the end of the season. This was reflected in a significantly increased sorghum plant height (by 80%) and sorghum shoot dry weight (400%) compared to the negative control. Fusarium species in alginate granules also delayed Striga emergence and reduced the total number of Striga throughout the growing season. The best efficacy was obtained by FA, which reduced the total number of Striga shoots by 71% (using 10% wheat flour) and 84% (6% sorghum flour or yeast extract) compared to the control. Likewise, it reduced the proportion of healthy Striga shoots by 71%, 88% and 84%, respectively, and Striga biomass by 50%, 81%, and 89%, respectively. Alginate formulations generally also significantly increased sorghum plant height by up to 80% and sorghum shoot dry weight by 200 to 400% compared to the control. It can therefore be summarized that of the investigated fungal isolates and granular formulations FA formulated in ?Pesta? granules showed the best efficacy in controlling Striga under field and controlled conditions. The ability of FA to produce trichothecene mycotoxins that could be a hazard to humans or animals was assessed from samples of the fungus growing on autoclaved wheat grains. Additionally, samples of harvested sorghum seeds from various plots inoculated with the biocontrol agents were investigated for the content of trichothecene mycotoxins. None of the following toxins were either produced by FA or translocated to harvested sorghum seeds under field conditions: nivalenol, fusarenon X, deoxynivalenol, 15-actetyldeoxynivalenol, scirpentriol, monoacetoxyscirpenol, diacetoxyscirpenol, T-2 triol, HT-2 toxin, T-2 toxin and neosolaniol. A simple seed coating treatment using fine ?Pesta? granules and gum Arabic as adhesive material also showed a potential to control Striga in the first season experiment. It was able to reduce the total number of Striga shoots by more than 55% compared to the control. Consequently, sorghum biomass was increased by 54 ? 67% and sorghum100-seed weight by 70%. The effects were comparable to that caused by chemical control using 2,4-D. To improve the efficacy of seed coating under field conditions for the second season experiments, the effect of five types of adhesive materials (2 types of cellulose, 2 types of organic polymers and a clay) on growth and sporulation of FA and FN were tested in solid and liquid media. Cellulose 1 and 2 were found to enhance radial growth of both isolates but did not increase sporulation in liquid media. Organic polymer 2 was found to retard both radial growth and sporulation of the two isolates. Organic polymer 1 and clay significantly enhanced the production of chlamydospores, especially by FA, compared to Richard?s solution alone or amended with the other tested materials. For this reason one of them was suggested to be used for seed coating in addition to Arabic gum. Sorghum seeds were coated by a private company (SUET Saat- und Erntetechnik GmbH, Eschwege, Germany), using air-dried fungal propagules fermented on 1.5 % (w/v) sorghum straw. Arabic gum was observed to give a better coverage of sorghum seeds and higher cfu per seed (4 x 104 [FA] and 19 x104 cfu [FN]) compared to the second adhesive material. Coating sorghum seeds with the biocontrol agents did generally not result in a significant reduction of Striga shoots in the field, but some of the seed-coating treatments were very efficient in inducing disease symptoms on Striga shoots. Especially FN coated with Arabic gum and FA applied to the seeds in fine ?Pesta? granules increased the proportion of diseased Striga shoots significantly compared to the control by 79%. In the pot experiment, the fungal isolates applied as a seed coating neither resulted in a significant reduction of Striga shoots. Nevertheless, FA applied to sorghum seeds using the adhesive material provided by SUET reduced the total number of Striga plants by 52% compared to the control at the end of the season. FN in the same treatment significantly increased the proportion of diseased Striga by 77% at the end of the season. The effect of seed coating on increasing sorghum plant height and dry matter was lower than that caused by the granular formulations and not statistically significant compared to the control. From the obtained results it can be concluded that both granular formulations applied to the planting holes and seed coating can be used as effective delivery systems for biocontrol fungi and can be adopted under field conditions to reduce Striga infestation. However, the granular formulations showed a higher efficacy in controlling Striga. Inoculum type and concentration as well as nutritional amendments to the formulations should be further optimized in future investigations.
Breeding for resistance to Fusarium ear diseases in maize and small-grain cereals using genomic tools
(2021) Gaikpa, David Sewordor; Miedaner, Thomas
The world’s human and livestock population is increasing and there is the need to increase quality food production to achieve the global sustainable development goal 3, zero hunger by year 2030 (United Nations, 2015). However, biotic stresses such as Fusarium ear infections pose serious threat to cereal crop production. Breeding for host plant resistance against toxigenic Fusarium spp. is a sustainable way to produce more and safer cereal crops such as maize and small-grain winter cereals. Many efforts have been made to improve maize and small-grain cereals for ear rot (ER) and Fusarium head blight (FHB) resistances, using conventional and genomic techniques. Among small-grain cereals, rye had the shortest maturity period followed by the descendant, hexaploid triticale while both wheat species had the longest maturity period. In addition, rye and triticale were more robust to Fusarium infection and deoxynivalenol accumulation, making them safer grain sources for human and animal consumption. However, a few resistant cultivars have been produced by prolonged conventional breeding efforts in durum wheat and bread wheat. High genetic variation was present within each crop species and can be exploited for resistance breeding. In this thesis, the genetic architecture of FHB resistance in rye was investigated for the first time, by means of genome-wide association study (GWAS) and genomic prediction (GP). GWAS detected 15 QTLs for Fusarium culmorum head blight severity, of which two had major effects. Both weighted and unweighted GP approaches yielded higher prediction abilities than marker-assisted selection (MAS) for FHB severity, heading stage and plant height. Genomics-assisted breeding can shorten the duration of breeding rye for FHB resistance. In the past decade, genetic mapping and omics were used to identify a multitude of QTLs and candidate genes for ear rot resistances and mycotoxin accumulation in maize. The polygenic nature of resistance traits, high genotype x environment interaction, and large-scale phenotyping remain major bottlenecks to increasing genetic gains for ear rots resistance in maize. Phenotypic and molecular analyses of DH lines originating from two European flint landraces (“Kemater Landmais Gelb”, KE, and “Petkuser Ferdinand Rot”, PE) revealed high variation for Gibberella ear rot (GER) severity and three agronomic traits viz. number of days to female flowering, plant height and proportion of kernels per cob. By employing multi-SNP GWAS method, we found four medium-effect QTLs and many small-effect (10) QTLs for GER severity in combined DH libraries (when PCs used as fixed effects), none co-localized with the QTLs detected for the three agronomic traits analyzed. However, one major QTL was detected within KE DH library for GER severity. Two prioritized SNPs detected for GER resistance were associated with 25 protein-coding genes placed in various functional categories, which further enhances scientific knowledge on the molecular mechanisms of GER resistance. Genome-based approaches seems promising for tapping GER resistance alleles from European maize landraces for applied breeding. After several cycles of backcrossing and sufficient selection for agronomic adaptation traits, the resistant lines identified in this thesis can be incorporated into existing maize breeding programs to improve immunity against F. graminearum ear infection. Breeding progress can be faster using KE landrace than PE. A successful validation of QTLs identified in this thesis can pave way for MAS in rye and marker-assisted backcrossing in maize. Effective implementation of genomic selection requires proper design of the training and validation sets, which should include part of the current breeding population.
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.
Genomics-assisted breeding strategies for quantitative resistances to Northern corn leaf blight in maize (Zea mays L.) and Fusarium diseases in maize and in triticale (× Triticosecale Wittm.)
(2021) Galiano Carneiro, Ana Luísa; Miedaner, Thomas
Fusarium head blight (FHB) in triticale (× Triticosecale Wittm.), Gibberella ear rot (GER) and Northern corn leaf blight (NCLB) in maize (Zea mays L.) are devastating crop diseases causing yield losses and/or reducing grain quality worldwide. Resistance breeding is the most efficient and sustainable approach to reduce the damages caused by these diseases. For all three pathosystems, a quantitative inheritance based on many genes with small effects has been described in previous studies. Hence, this thesis aimed to assess the potential of genomics-assisted breeding strategies to reduce FHB, GER and NCLB in applied breeding programs. In particular, the objectives were to: (i) Dissect the genetic architecture underlying quantitative variation for FHB, GER and NCLB through different quantitative trait loci (QTL) and association mapping approaches; (ii) assess the potential of genomics-assisted selection to select superior triticale genotypes harboring FHB resistance; (iii) phenotype and characterize Brazilian resistance donors conferring resistance to GER and NCLB in multi-environment trials in Brazil and in Europe; and (iv) evaluate approaches for the introgression and integration of NCLB and GER resistances from tropical to adapted germplasm. The genome-wide association study (GWAS) conducted for FHB resistance in triticale revealed six QTL that reduced damages by 5 to 8%. The most prominent QTL identified in our study was mapped on chromosome 5B and explained 30% of the genotypic variance. To evaluate the potential of genomic selection (GS), we performed a five-fold cross-validation study. Here, weighted genomic selection increased the prediction accuracy from 0.55 to 0.78 compared to the non-weighted GS model, indicating the high potential of the weighted genomic selection approach. The successful application of GS requires large training sets to develop robust models. However, large training sets based on the target trait deoxynivalenol (DON) are usually not available. Due to the rather moderate correlation between FHB and DON, we recommend a negative selection based on genomic estimated breeding values (GEBVs) for FHB severity in early breeding stages. In the long-run, however, we encourage breeders to build and test GS calibrations for DON content in triticale. The genetic architecture of GER caused by Fusarium graminearum in maize was investigated in Brazilian tropical germplasm in multi-environment trials. We observed high genotype-by-environment interactions which requires trials in many environments for the identification of stable QTL. We identified four QTL that explained between 5 to 22% of the genotypic variance. Most of the resistance alleles identified in our study originated from the Brazilian tropical parents indicating the potential of this exotic germplasm as resistance source. The QTL located on chromosome bin 1.02 was identified both in Brazilian and in European trials, and across all six biparental populations. This QTL is likely stable, an important feature for its successful employment across different genetic backgrounds and environments. This stable QTL is a great candidate for validation and fine mapping, and subsequent introgression in European germplasm but possible negative linkage drag should be tackled. NCLB is another economically important disease in maize and the most devastating leaf disease in maize grown in Europe. Virulent races have already overcome the majority of known qualitative resistances. Therefore, a constant monitoring of S. turcica races is necessary to assist breeders on the choice of effective resistances in each target environment. We investigated the genetic architecture of NCLB in Brazilian tropical germplasm and identified 17 QTL distributed along the ten chromosomes of maize explaining 4 to 31% of the trait genotypic variance each. Most of the alleles reducing the infections originated from Brazilian germplasm and reduced NCLB between 0.3 to 2.5 scores in the 1-9 severity scale, showing the potential of Brazilian germplasm to reduce not only GER but also NCLB severity in maize. These QTL were identified across a wide range of environments comprising different S. turcica race compositions indicating race non-specific resistance and most likely stability. Indeed, QTL 7.03 and 9.03/9.04 were identified both in Brazil and in Europe being promising candidates for trait introgression. These major and stable QTL identified for GER and NCLB can be introgressed into elite germplasm by marker-assisted selection. Subsequently, an integration step is necessary to account for possible negative linkage drag. A rapid genomics-assisted breeding approach for the introgression and integration of exotic into adapted germplasm has been proposed in this thesis. Jointly, our results demonstrate the high potential of genomics-assisted breeding strategies to efficiently increase the quantitative resistance levels of NCLB in maize and Fusarium diseases in maize and in triticale. We identified favorable QTL to increase resistance levels in both crops. In addition, we successfully characterized Brazilian germplasm for GER and NCLB resistances. After validation and fine mapping, the introgression and integration of the QTL identified in this study might contribute to the release of resistant cultivars, an important pillar to cope with global food security.
Improving host resistance to Fusarium head blight in wheat (Triticum aestivum L.) and Gibberella ear rot in maize (Zea mays L.)
(2023) Akohoue, Félicien; Miedaner, Thomas
Fusarium head blight (FHB) in wheat and Fusarium (FER) and Gibberella ear rot (GER) in maize are major cereal diseases which reduce yield and contaminate kernels with several mycotoxins. In Europe, these diseases contribute to significant yield gaps and high mycotoxin risks across countries. However, existing management strategies related to agronomic practices are not fully effective, with some of them being cost-prohibitive. Enhancing host plant resistance is additionally required for managing the diseases more effectively and sustainably. Unfortunately, breeding for FHB resistance is challenged by complex interactions with morphological traits and the quantitative nature of the trait. In maize, available genetic resources have not been fully exploited to improve GER resistance in elite materials. In this work, we elucidated the complex interactions between FHB resistance and morphological traits, like plant height (PH) and anther retention (AR) in wheat. The effect of reduced height (Rht) gene Rht24 on AR and the contribution of genomic background (GB) to FHB resistance in semi-dwarf genotypes were also assessed. GB refers to all genomic loci, except major Rht genes, that affect the traits. To achieve this, 401 winter wheat cultivars were evaluated across five environments (location × year combination). All cultivars were genotyped using Illumina 25 K Infinium single-nucleotide polymorphism array. We performed correlation and path coefficient analysis, and combined single and multi-trait genome-wide association studies (GWAS). Our findings revealed significant genotypic correlations and path effects between FHB severity with PH and AR, which were controlled by several pleiotropic loci. FHB severity and PH shared both negatively and positively acting pleiotropic loci, while only positively acting pleiotropic loci were detected between FHB severity and AR. Rht-D1 is a major pleiotropic gene which exerted a negative effect on FHB resistance. These pleiotropic loci contribute to our understanding of the complex genetic basis of FHB resistance, and their exploitation can help to simultaneously select for FHB resistance with PH and AR. Contrary to Rht-D1b, Rht24b had no negative effect on FHB resistance and AR. This exhibits Rht24 as an important FHB-neutral Rht gene which can be integrated into breeding programs. Genomic estimated breeding values (GEBV) were calculated for each cultivar to assess GB. We observed highly negative GEBV for FHB severity within resistant wheat cultivars. Susceptible cultivars exhibited positive GEBV. Genomic prediction has a great potential and can be exploited by selecting for semi-dwarf winter wheat genotypes with higher FHB resistance due to their genomic background resistance. To tackle maize ear rot diseases, refined and stable quantitative trait loci (QTL) harboring candidate genes conferring resistances to FER and GER were identified. The effectiveness of introgression of two European flint landraces, namely “Kemater Gelb Landmais” (KE) and “Petkuser Ferdinand Rot” (PE) was evaluated. The prediction accuracy of using line performance as a predictor of hybrid performance for GER resistance was also evaluated within the two landraces. We applied a meta-QTL (MQTL) analysis based on 15 diverse SNP-based QTL mapping studies and performed gene expression analysis using published RNA-seq data on GER resistance. In total, 40 MQTL were identified, of which 14 most refined MQTL harbored promising candidate genes for use in breeding programs for improving FER and GER resistances. 28 MQTL were common to both FER and GER, with most of them being shared between silk (channel) and kernel resistances. This highlights the co-inheritance of FER and GER resistances as well as types of active resistance. Resistance genes can be transferred into elite cultivars by integrating refined MQTL into genomics-assisted breeding strategies. Afterwards, four GER resistant doubled haploid (DH) lines from both KE and PE landraces were crossed with two susceptible elite lines to generate six bi-parental populations with a total of 534 DH lines which were evaluated for GER resistance. GER severity within the six landrace-derived populations were reduced by 39−61% compared to the susceptible elite lines. Moderate to high genetic advance was observed within each population, and the use of KE landrace as a donor was generally more effective than PE landrace. This shows promise in enhancing resistance to GER in elite materials using the European flint landraces as donors. Furthermore, per se performance of 76 DH lines from both landraces was used to predict GER resistance of their corresponding testcrosses (TC). Moderate phenotypic and genomic prediction accuracy between TC and line per se performance was found for GER resistance. This implies that pre-selecting lines for GER resistance is feasible; however, TC should be additionally tested on a later selection stage to aim for GER-resistant hybrid cultivars.
Management of Fusarium graminearum-inoculated crop residues : effects on head blight, grain yield and grain quality of subsequent winter wheat crops
(2001) Yi, Cuilin; Aufhammer, Walter
On the experimental station Ihinger Hof of Hohenheim University field experiments with artificial inoculation were conducted. An isolation-strip experiment included strips of winter rape crops, separating non-inoculated test plots of wheat from inoculated wheat plots. For the main field experiment, maize or spring wheat were planted as pre-crops in rotations with winter wheat and different crop residue treatments were applied. Additional residue management greenhouse tests were conducted and treated equivalent to the field experiment. Results of isolation-strips field experiments with wheat showed that. Isolation strips of 2 m width reduced disease incidence on neighbour plots by more than 50%. A further increase in isolation strip width did improve the isolation effect, but the differences between isolation strip widths were comparatively small. The infection in test plots was not completely eliminated even with 8 m wide strips. Greenhouse tests of residue management showed that deeper residue incorporation effectively reduced the F. g. populations on residues. The application of nitrolime reduced the population level of F. g. On the contrary, fertilization with calcium ammonium nitrate promoted F. g. populations. Soaking the residues in a fungicide preparation eliminated F. g. on the residues completely. Residue management field experiments with artificial inoculation of pre-crops, there were no significant differences in infection level after either maize or wheat, and the infection level of winter wheat was especially high after maize for silage use in one year. The reductions of FHB incidence due to ploughing or nitrolime application were 27-32% or 31-59% compared with residues remaining on the surface or calcium ammonium nitrate fertilization, respectively. But at that moderate FHB infection level, the residue management hardly influenced wheat grain yield and technological grain quality.
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.
Phänotypische und molekulare Analyse von Kreuzungsnachkommen auf Resistenz gegen Ährenfusariosen bei Triticale (x Triticosecale Wittmack)
(2011) Großmann, Maren; Großmann, Maren
Fusarium head blight (FHB) is an epidemic disease of cereals but disease control still is insufficiently possible. Resistance breeding can be addressed as one approach to reduce the mycotoxin contents. Since 2006 the European Union had strict rules governing the values for Desoxinivalenol (DON) in food products. The extraordinary loads through mycotoxins in triticale, which are caused by FHB, lead to large reductions in grain yield and quality. Beyond the fact that it can cause health issues in animals, especially in pig feed, it may induce financial burdens for farming industries. Resistance against FHB is inherited quantitatively with mainly additive effects and therefore is only recorded by complex field experiments. Molecular markers accelerate resistance breeding and enable specific introgressions of favorable QTL. The main topic of this thesis is to show the application possibilities of molecular markers for the investigation of FHB resistance in triticale. In detail this study aims to analyze factors leading to a minimized mycotoxin accumulation in perennial field trials at several locations. Furthermore genetic maps of two different crossing populations were compiled and QTL mapping for ear appearance, plant height, DON content and FHB resistance was implemented. For all populations significant varieties had been displayed for all characteristics. The average values for FHB rate were between 8 and 43 % depending on year and location. The heritability?s showed high values for each population (h2 = 0.7 ? 0.82). The substantial genotype-environment-interaction pointed out the importance of field experiments. Furthermore no significant correlation was obtained between ear appearance and plant height. Due to the fact that the correlation factor between DON-content and FHB rate was very low (r = 0.32) to moderate (r = 0,65). Therefore no assumption about the DON-content subjected to the FHB rate could be made. The mapping of both populations LASKO x ALAMO and LASKO x TRIMESTER has been accomplished with SSR- and DArT-markers. The rate of polymorphism could be increased from 7 - 12 % respectively to 9 - 18 %. Genetic maps were constructed with length of 1.815 and 1.407 cM. They indicated 5.19 and 4.54 cM in their average marker distances. Throughout the QTL mapping several QTL were detected (6 QTL for ear appearance, 4 QTL for plant height, 1 QTL for DON-content and 4 QTL for FHB rate). These QTL explained 8 to 66 % of the phenotypical variance. In addition crossing populations of LASKO x TRIMESTER showed 2 QTL for ear appearance, 1 for plant height and 2 QTL for FHB resistance, which could explain 3 to 41 % of the phenotypical variance. This was also observed in several other QTL-surveys for wheat. Prospectively using molecular markers and genetically engineered methods will increase the research and development of resistant varieties and lead to a reduced mycotoxin accumulation. So far neither genetic maps nor any QTL studies for FHB resistance or DON-content in triticale have been published. Due to the fact that resistance is transmitted by several genes numerous artificial inoculations have to be carried out during breeding. Additive gene effects are useful to combine several resistance genes from different parents. Established methods in plant breeding such as recurrent selection, phenotypical and marker-based selection are successfully in use to reduce FHB- symptoms and to reduce the mycotoxin value but these methods have to be improved. The results of this thesis are promising for a superior resistance breeding in the future.
Resistance of Maize (Zea mays L.) Against the European Corn Borer (Ostrinia nubilalis Hb.) and its Association with Mycotoxins Produced by Fusarium spp.
(2004) Magg, Thomas; Melchinger, Albrecht E.
The European corn borer (ECB, Ostrinia nubilalis Hübner) is a major pest of maize (Zea mays L.) in Europe and continues to spread to northern maize growing regions. The ECB severely affects commercial maize production by decreasing yield stability. In addition, damaged plants often show an increased susceptibility to secondary infections caused by Fusarium spp.. Information about the potential of Bt hybrids (event 176, MON810) to reduce yield losses and mycotoxin contamination under Central European growing conditions is still lacking. However, such monogenic resistances with a strong negative effect on the ECB will break down rapidly. Improving the natural host plant resistance of maize could provide an economical and ecological tool for an integrated pest management system. The overall goal of this study was to evaluate alternative breeding strategies for improving resistance of maize against ECB damage and Fusarium spp.. The objectives were to (1) initiate a selection experiment in the early maturing European flint pool and evaluate a breeding program for ECB resistance in the European dent pool, (2) compare the efficiency of host plant resistance vs. Bt resistance in maize, (3) determine Fusarium-caused mycotoxin contamination of maize genotypes with improved host plant resistance to ECB, and (4) study the association between important agronomic traits, ECB resistance traits, and mycotoxin concentration in early European maize germplasm. The goal of the Hohenheim ECB breeding program, initiated in 1992, was to select lines with improved per se and testcross performance for multiple agronomic traits and ECB resistance. In the standard breeding scheme, line development started from a segregating S1 population. Genotypes were evaluated for their line per se ECB resistance in generations S1, S3, and S5. Lines from the S2, S4, and S5 generations were testcrossed and evaluated for their agronomic performance. Selection was based on ECB resistance and TC performance for grain yield and maturity. In order to compare transgenic Bt maize hybrids carrying event 176 or MON810 with their isogenic counterparts and commercial hybrids or experimental hybrids, field trials in multiple environments were conducted in 1998 to 2000. Furthermore, a laboratory bioassay with neonate ECB larvae was performed to assess mortality and subsequently the level of Bt antibiosis present in the used hybrids of 1998. Resistance traits such as damage rating of stalks, number of damaged plants, and number of larvae per plant were assessed exclusively in manually ECB infested plots. Grain yield, grain dry matter content and plant height were determined in the insecticide protected and the ECB infested main plots. In addition, grain samples from each subplot were drawn at random and analyzed separately for Fusarium mycotoxins such as type B trichothecenes (DON, NIV), Zearalenon (ZEN), Fumonisins (FUM), and Moniliformin (MON). The inbred lines displayed a significant genotypic variance for all ECB resistance traits evaluated. However, in the further course of selection and topcross testing, most dent and flint lines, especially those displaying improved resistance to ECB larvae feeding, were discarded because of their poor agronomic performance. Negative correlations between grain yield, early maturity and the damage rating of stalks were identified. However, three dent lines (P028, P029, P030) with moderate resistance to ECB were developed. In all experiments, Bt hybrids were superior to other hybrids in the control of ECB larvae. Non-Bt hybrids displayed a significant genotypic variance for all evaluated resistance traits; grain yield reductions ranged from 8.6 to 21.8% under manual infestation of ECB. All evaluated resistance traits were highly significantly correlated with each other and showed significant negative correlations to grain yield reduction. Bt hybrids did not differ from their isogenic counterparts for most agronomic traits. Highly significant location and genotype × location interactions were identified for all mycotoxins evaluated, except MON. MON concentration doubled under manual infestation of ECB compared to insecticide protected conditions and a similar trend was found for FUM. Bt hybrids displayed significantly lower MON concentrations than non-Bt hybrids and significantly lower DON concentrations than their isogenic counterparts under ECB infestation. Highly significant correlations between ECB resistance traits and MON were found. However, a significant genotypic variance was observed for DON, 15-A-DON, FUM, and MON concentrations, suggesting variation for resistance against Fusarium spp. in current elite hybrids. By combining different sources of monogenic Bt resistance and quantitatively inherited resistances to ECB, it may be possible to develop hybrids with multiple resistance by pyramiding the underlying genes in one genotype. Therefore, further research is required to identify new sources of ECB resistance and new breeding strategies should be developed. Furthermore, there is indication that an improved resistance against Fusarium spp. possesses a greater potential for reducing mycotoxin contamination of maize kernels than a high level of ECB resistance. Since resistance to ECB and resistance to Fusarium spp. are inherited fairly independently, simultaneous improvement of both resistances seems to be necessary for improving the stability and quality of future maize hybrids.