Biological control of seedling blight of wheat caused by Fusarium graminearum with beneficial rhizosphere microorganisms

Fusarium graminearum is associated with the cereal damping-off complex which reduces germination, seedling stand and yield. Fifty-two bacterial strains and six Trichoderma spp. isolated from the wheat rhizosphere were evaluated for biocontrol of seedling blight of wheat caused by F. graminearum. Their potential as biocontrol agents was tested in vitro and in the greenhouse. Isolates varied in their ability to inhibit the mycelial growth of F. graminearum in agar plate bioassays by 0–79%. This parameter was not related with biocontrol efficacy of in vivo assays. In greenhouse trials, all isolates were initially evaluated for reducing disease on wheat cultivars Klein Centauro (moderately resistant to F. graminearum) and Pro INTA Oasis (susceptible) planted in sterilized soil artificially infested with the pathogen. Among the 25 bacteria and six fungal isolates that exhibited a pronounced suppressive effect, the most efficient 10 for both cultivars were further assayed on eight cultivars (Buck Candil, Buck Catriel, Buck Chambergo, Buck Poncho, Buck Topacio, Klein Cacique, Klein Centauro and Pro INTA Oasis) potted in cultivated–inoculated soil. Three weeks after sowing, plant stand, percentage of diseased emerging seedlings, plant height and dry weight were evaluated. Among the antagonists only Stenotrophomonas maltophilia was significantly better than the control for the average of the eight cultivars for plant stand, height and dry weight. Stenotrophomonas maltophilia also caused a non-significant decrease in the percentage of diseased plants. Three strains of Bacillus cereus and one isolate of Trichoderma harzianum gave also a good control in some cultivars. The ability of these isolates to affect the infection of wheat seedlings by F. graminearum may be of potential value in field trials.     

Potential for control of seedling blight of wheat caused by Fusarium graminearum and related species using the bacterial endophyte Bacillus mojavensis

Fusarium-infected wheat seed decreases germination, seedling emergence, and causes post emergence seedling death, and can contribute to wheat scab and ear rot of maize, with consequent production of mycotoxins such as deoxynivalenol and zearalenone. Current seed treatments have proved ineffective in controlling seedling blight and scab. A patented endophytic bacterial strain, Bacillus mojavensis RRC 101, and several other strains of this species were studied to determine in vitro antagonism to some Fusarium species and to assess the potential of this bacterium to serve as an endophytic biocontrol for seedling blight of wheat produced by species within the F. graminearum complex, as well as other species of Fusarium. Seedling emergence and seed germination were two tests used as indicators of seedling blight. These tests were conducted in growth rooms with two wheat cultivars highly susceptible to scab, Norm and Pioneer 2552, and other cultivars with varying resistance to scab. The results indicated that all strains of this bacterium were antagonistic in vitro to the strains of F. graminearum and its seven related species, as well as four strains of F. pseudograminearum and the two strains of F. verticillioides. Germination of the highly scab susceptible cultivar 2552 was increased from 77 to 97% when planted in soil containing a mixed inoculum of F. graminearum and related species. Seedling emergence in the very susceptible wheat cultivar Norm increased from 20 to 82% when treated with the bacterium. The data indicated that inoculating wheat kernels with B. mojavensis reduced seedling blight of wheat produced by F. graminearum and related Fusarium species indicating the potential for this bacterium as a biocontrol under field condition.     

Genotype x strain interactions for resistance to Fusarium head blight caused by Fusarium culmorum in winter wheat

Summary In 3 consecutive years, a set of 17 winter wheat genotypes, representing a wide range of Fusarium head blight resistance, was inoculated with four strains of Fusarium culmorum. Fusarium head blight ratings were analyzed. The interaction between genotypes, strains, and years was described using a Finlay-Wilkinson model and an Additive Main effects and Multiplicative Interaction effects (AMMI) model. The interaction consisted primarily of a divergence of genotypical responses with increasing disease pressure, modified by genotype specific reactions in certain years. The divergence was mainly caused by one very pathogenic strain. The Fusarium head blight resistance in this study can be described as horizontal resistance in terms of Vanderplank, with the exception of three genotypes selected from one particular cross that showed a strain-year combination dependent resistance which was ineffective in 1 year.     

Fusarium head blight biological control with Lysobacter enzymogenes strain C3

Fusarium head blight (FHB), caused by Fusarium graminearum (= Gibberella zeae), is a destructive disease of wheat for which biological controls are needed. Lysobacter enzymogenes strain C3, a bacterial antagonist of fungal pathogens via lytic enzymes and induced resistance, was evaluated in this study for control of FHB. In greenhouse experiments, chitin broth cultures of C3 reduced FHB severity to <10% infected spikelets as compared to >80% severity in the controls in some experiments. C3 broth cultures heated to inactivate cells and lytic enzymes, but retaining the elicitor factor for induced resistance, also were effective in reducing FHB severity, suggesting induced resistance is one mechanism of action. C3 broth cultures also were effective when applied in highly diluted form and when applied 1 week prior to pathogen inoculation. When applied to 8 cultivars of hard red spring wheat in the greenhouse, C3 treatments reduced FHB in 5 cultivars but not in the others. These findings also are consistent with induced resistance. Protection offered by C3 treatments, however, was not systemic and required that C3 be applied uniformly to all susceptible florets. Field tests were conducted in South Dakota and Nebraska to evaluate the efficacy of C3 chitin broth cultures in spring and winter wheat, respectively. In experiments involving two hard red spring wheat cultivars, treatment with C3 reduced FHB severity in ‘Russ’ but not in ‘Ingot’. In three other field experiments comparing C3, the fungicide tebuconazole, and the combination of C3 and tebuconazole, treatments with the bacterial culture alone and the fungicide alone were inconsistent across experiments, each treatment being ineffective in controlling FHB in one experiment. The biocontrol agent–fungicide combination was more consistently effective, reducing FHB incidence or severity in all three experiments. Thus, the potential for using L. enzymogenes C3 as a biological control agent for FHB was demonstrated along with a number of factors that might affect control efficacy in the field.     

FcRav2, a gene with a ROGDI domain involved in Fusarium head blight and crown rot on durum wheat caused by Fusarium culmorum

Fusarium culmorum is a soil‐borne fungal pathogen which causes foot and root rot and Fusarium head blight on small‐grain cereals, in particular wheat and barley. It causes significant yield and quality losses and results in the contamination of kernels with type B trichothecene mycotoxins. Our knowledge of the pathogenicity factors of this fungus is still limited. A transposon tagging approach based on the mimp1/impala double‐component system has allowed us to select a mutant altered in multiple metabolic and morphological processes, trichothecene production and virulence. The flanking regions of mimp1 were used to seek homologies in the F. culmorum genome, and revealed that mimp1 had reinserted within the last exon of a gene encoding a hypothetical protein of 318 amino acids which contains a ROGDI‐like leucine zipper domain, supposedly playing a protein–protein interaction or regulatory role. By functional complementation and bioinformatic analysis, we characterized the gene as the yeast Rav2 homologue, confirming the high level of divergence in multicellular fungi. Deletion of FcRav2 or its orthologous gene in F. graminearum highlighted its ability to influence a number of functions, including virulence, trichothecene type B biosynthesis, resistance to azoles and resistance to osmotic and oxidative stress. Our results indicate that the FcRav2 protein (and possibly the RAVE complex as a whole) may become a suitable target for new antifungal drug development or the plant‐mediated resistance response in filamentous fungi of agricultural interest.     

Comparative pathogenicity of Fusarium graminearum isolates from China revealed by wheat coleoptile and floret inoculations

Fusarium head blight (FHB) or scab caused by Fusarium species is an economically important disease on small grain cereal crops worldwide. Accurate assessments of the pathogenicity of fungal isolates is a key obstacle toward a better understanding of the Fusarium-wheat scab system. In this study, a new laboratory method for inoculation of wheat coleoptiles was developed, which consists of cutting off the coleoptile apex, covering the cut apex with a piece of filter paper soaked in conidial suspension, and measuring the lengths of brown lesions 7 days post inoculation. After coleoptile inoculation, distinct brown lesions in the diseased stems were observed, in which the presence of the fungus was verified by PCR amplification with F.␣graminearum Schwable-specific primers. Coleoptile inoculation of six wheat varieties indicated that a highly susceptible wheat variety was more suitable as a differentiating host for the pathogenicity assay. Analysis of the coleoptiles inoculated with a set of 58 different isolates of F. graminearum showed a significant difference in the lengths of the lesions, forming the basis by which pathogenicity of the isolates was assessed. Field inoculation of florets of three wheat varieties over 2 years revealed significant differences in pathogenicity among the 58 isolates, and that the highly resistant and highly susceptible wheat varieties were more appropriate and stable for pathogenicity assessment in field trials. Comparative analyses of eight inoculation experiments of wheat with 58 F. graminearum isolates showed significant direct linear correlations (P<0.001) between coleoptile and floret inoculations. These results indicate that the wheat coleoptile inoculation is a simple, rapid and reliable method for pathogenicity studies of F.␣graminearum in wheat.     

Genetic dissection of a major Fusarium head blight QTL in tetraploid wheat

The devastating effect of Fusarium head blight (FHB) caused by Fusarium graminearum has led to significant financial losses across the Upper Midwest of the USA. These losses have spurred the need for research in biological, chemical, and genetic control methods for this disease. To date, most of the research on FHB resistance has concentrated on hexaploid wheat (Triticum aestivum L.) lines originating from China. Other sources of resistance to FHB would be desirable. One other source of resistance for both hexaploid wheat and tetraploid durum wheat (T. turgidum L. var. durum) is the wild tetraploid, T. turgidum L. var. dicoccoides (T. dicoccoides). Previous analysis of the `Langdon'-T. dicoccoides chromosome substitution lines, LDN(Dic), indicated that the chromosome 3A substitution line expresses moderate levels of resistance to FHB. LDN(Dic-3A) recombinant inbred chromosome lines (RICL) were used to generate a linkage map of chromosome 3A with 19 molecular markers spanning a distance of 155.2 cM. The individual RICL and controls were screened for their FHB phenotype in two greenhouse seasons. Analysis of 83 RICL identified a single major quantitative trait locus, Qfhs.ndsu-3AS, that explains 37% of the phenotypic or 55% of the genetic variation for FHB resistance. A microsatellite locus, Xgwm2, is tightly linked to the highest point of the QTL peak. A region of the LDN (Dic-3A) chromosome associated with the QTL for FHB resistance encompasses a 29.3 cM region from Xmwg14 to Xbcd828.     

Overexpression of defense response genes in transgenic wheat enhances resistance to Fusarium head blight

Fusarium head blight (FHB) of wheat, caused by Fusarium graminearum and other Fusarium species, is a major disease problem for wheat production worldwide. To combat this problem, large-scale breeding efforts have been established. Although progress has been made through standard breeding approaches, the level of resistance attained is insufficient to withstand epidemic conditions. Genetic engineering provides an alternative approach to enhance the level of resistance. Many defense response genes are induced in wheat during F. graminearum infection and may play a role in reducing FHB. The objectives of this study were (1) to develop transgenic wheat overexpressing the defense response genes α-1-purothionin, thaumatin-like protein 1 (tlp-1), and β-1,3-glucanase; and (2) to test the resultant transgenic wheat lines against F. graminearum infection under greenhouse and field conditions. Using the wheat cultivar Bobwhite, we developed one, two, and four lines carrying the α-1-purothionin, tlp-1, and β-1,3-glucanase transgenes, respectively, that had statistically significant reductions in FHB severity in greenhouse evaluations. We tested these seven transgenic lines under field conditions for percent FHB disease severity, deoxynivalenol (DON) mycotoxin accumulation, and percent visually scabby kernels (VSK). Six of the seven lines differed from the nontransgenic parental Bobwhite line for at least one of the disease traits. A β-1,3-glucanase transgenic line had enhanced resistance, showing lower FHB severity, DON concentration, and percent VSK compared to Bobwhite. Taken together, the results showed that overexpression of defense response genes in wheat could enhance the FHB resistance in both greenhouse and field conditions.     

Suppression of wheat Fusarium head blight by novel amphiphilic aminoglycoside fungicide K20

K20 is a novel amphiphilic aminoglycoside capable of inhibiting many fungal species. K20's capabilities to inhibit Fusarium graminearum the causal agent wheat Fusarium head blight (FHB) and to this disease were examined. K20 inhibited the growth of F. graminearum (minimum inhibitory concentrations, 7.8–15.6 mg L^?1) and exhibited synergistic activity when combined with triazole and strobilurin fungicides. Application of K20 up to 720 mg L^?1 to wheat heads in the greenhouse showed no phytotoxic effects. Spraying wheat heads in the greenhouse with K20 alone at 360 mg L^?1 lowered FHB severity below controls while combining K20 with half–label rates of Headline (pyraclostrobin) improved its disease control efficacy. In field trials, spraying K20 at 180 mg L^?1 and 360 mg L^?1 combined with half-label rates of Headline, Proline 480 SC (prothioconazole), Prosaro 421 SC (prothioconazole + tebuconazole), and Caramba (metconazole) reduced FHB indices synergistically. In addition, the K20 plus Proline 480 SC combination reduced levels of the mycotoxin deoxinivalenol by 75 % compared to the control. These data suggest that K20 may be useful as a fungicide against plant diseases such as FHB particularly when combined with commercial fungicides applied at below recommended rates.     

Back-cross reciprocal monosomic analysis of Fusarium head blight resistance in wheat (Triticum aestivum L.)

 Fusarium head blight (FHB or scab) caused by Fusarium spp. is a widespread disease of cereals causing yield and quality losses and contaminating cereal products with mycotoxins. The breeding of resistant varieties is the method of choice for controlling the disease. Unfortunately, the genetic basis of scab resistance is still poorly understood. We present the results of a back-cross reciprocal monosomic analysis of FHB resistance using the highly resistant Hungarian winter wheat line ‘U-136.1’ and the highly susceptible cultivar ‘Hobbit-sib’. Resistance testing was performed in a field trial artificially inoculated with a Fusarium culmorum conidial suspension. Five hemizygous families containing ‘U-136.1’ chromosomes 6B, 5A, 6D, 1B, and 4B had a visually reduced spread of infection compared to lines having the ‘Hobbit-sib’ chromosome. Chromosome 2B from ‘U-136.1’ had an increased spread of infection. The critical chromosomes controlling seed weight were 6D, 3B, 5A, and 6B while those controlling deoxynivalenol (DON) content were homoeologous groups 2 and 6, although the latter effects were not significant due to a high coefficient of variation. Results from this and other studies show that chromosomes 6D, 6B, 5A, 4D, and 7A have frequently been associated with scab resistance in a number of wheat cultivars. Research groups now attempting to map scab resistance in wheat using markers should pay special attention to the above-mentioned chromosomes. Received: 31 March 1998 / Accepted: 14 July 1998     

Selection and evaluation of the potential of choline-metabolizing microbial strains to reduce Fusarium head blight

Choline and betaine are found in wheat flower tissues and have been implicated in stimulating hyphal growth of the primary causal agent of Fusarium head blight (FHB), Gibberella zeae. Choline metabolizing strains (CMS) from wheat anthers may therefore be a useful source of antagonists of G. zeae. One-hundred twenty-three of 738 microbial strains that were recovered from wheat anthers collected from plants grown in Illinois and Ohio were CMS as determined by growth in a liquid medium containing choline as a sole carbon and nitrogen source and a colorimetric, choline oxidase-based assay of culture filtrate. Thirty-one out of 123 CMS reduced FHB disease severity by at least 25% in greenhouse tests on wheat and 17 reduced FHB severity by at least 50%. All five CMS selected for field testing in 2003 reduced disease severity compared to the untreated check at both field locations on moderately resistant cultivar Freedom. Freedom wheat treated with Pseudomonas sp. AS 64.4 had 63% and 46% less FHB severity than untreated wheat at the two sites. Three of five CMS reduced severity at both locations on susceptible cultivar Pioneer Brand 2545. Disease control was comparable to that obtained using the fungicide Folicur 3.6F. Selection of wheat anther colonists for ability to utilize choline as a sole carbon and nitrogen source has utility as a screening tool in the search for efficacious antagonists of G. zeae although choline utilization does not insure that an isolate will be an effective biocontrol agent against Fusarium head blight.     

Genetic architecture is more complex for resistance to Septoria tritici blotch than to Fusarium head blight in Central European winter wheat

Background

Fusarium head blight (FHB) and Septoria tritici blotch (STB) severely impair wheat production. With the aim to further elucidate the genetic architecture underlying FHB and STB resistance, we phenotyped 1604 European wheat hybrids and their 135 parental lines for FHB and STB disease severities and determined genotypes at 17,372 single-nucleotide polymorphic loci.

Results

Cross-validated association mapping revealed the absence of large effect QTL for both traits. Genomic selection showed a three times higher prediction accuracy for FHB than STB disease severity for test sets largely unrelated to the training sets.

Conclusions

Our findings suggest that the genetic architecture is less complex and, hence, can be more properly tackled to perform accurate prediction for FHB than STB disease severity. Consequently, FHB disease severity is an interesting model trait to fine-tune genomic selection models exploiting beyond relatedness also knowledge of the genetic architecture.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1628-8) contains supplementary material, which is available to authorized users.