Integrated Effects of Genetic Resistance and Prothioconazole + Tebuconazole Application Timing on Fusarium Head Blight in Wheat.

Integrated Fusarium head blight (FHB) management programs consisting of different combinations of cultivar resistance class and an application of the fungicide prothioconazole + tebuconazole at or after 50% early anthesis were evaluated for efficacy against FHB incidence (INC; percentage of diseased spikes), index (IND; percentage of diseased spikelets per spike), Fusarium damaged kernel (FDK), deoxynivalenol (DON) toxin contamination, grain yield, and test weight (TW) in inoculated field trials conducted in 11 U.S. states in 2014 and 2015. Mean log response ratios and corresponding percent control values for INC, IND, FDK, and DON, and mean differences in yield and TW relative to a nontreated, inoculated susceptible check (S_CK), were estimated through network meta-analyses as measures of efficacy. Results from the analyses were then used to estimate the economic benefit of each management program for a range of grain prices and fungicide applications costs. Management programs consisting of a moderately resistant (MR) cultivar treated with the fungicide were the most efficacious, reducing INC by 60 to 69%, IND by 71 to 76%, FDK by 66 to 72%, and DON by 60 to 64% relative to S_CK, compared with 56 to 62% for INC, 68 to 72% for IND, 66 to 68% for FDK, and 58 to 61% for DON for programs with a moderately susceptible (MS) cultivar. The least efficacious programs were those with a fungicide application to a susceptible (S) cultivar, with less than a 45% reduction of INC, IND, FDK, or DON. All programs were more efficacious under conditions favorable for FHB compared with less favorable conditions, with applications made at 50% early anthesis being of comparable efficacy to those made 2 to 7 days later. Programs with an MS cultivar resulted in the highest mean yield increases relative to S_CK (541 to 753 kg/ha), followed by programs with an S cultivar (386 to 498 kg/ha) and programs with an MR cultivar (250 to 337 kg/ha). Integrated management programs with an MS or MR cultivar treated with the fungicide at or after 50% early anthesis were the most likely to result in a 50 or 75% control of IND, FDK, or DON in a future trial. At a fixed fungicide application cost, these programs were $4 to $319/MT more economically beneficial than corresponding fungicide-only programs, depending on the cultivar and grain price. These findings demonstrate the benefits of combining genetic resistance with a prothioconazole + tebuconazole treatment to manage FHB, even if that treatment is applied a few days after 50% early anthesis.

Meta-Analysis of the Effects of QoI and DMI Fungicide Combinations on Fusarium Head Blight and Deoxynivalenol in Wheat.

Field trials were conducted in 17 U.S. states to evaluate the effects of quinone outside inhibitor (QoI) and demethylation inhibitor (DMI) fungicide programs on Fusarium head blight index (IND) and deoxynivalenol (DON) toxin in wheat. Four DMI-only treatments applied at Feekes 10.5.1, five QoI-only treatments applied between Feekes 9 or Feekes 10.5, three QoI+DMI mixtures applied at Feekes 10.5, and three treatments consisting of a QoI at Feekes 9 followed by a DMI at Feekes 10.5.1 were evaluated. Network meta-analytical models were fitted to log-transformed mean IND and DON data and estimated contrasts of log means were used to obtain estimates of mean percent controls relative to the nontreated check as measures of efficacy. Results from the meta-analyses were also used to assess the risk of DON increase in future trials. DMI at Feekes 10.5.1 were the most effective programs against IND and DON and the least likely to increase DON in future trials. QoI-only programs increased mean DON over the nontreated checks and were the most likely to do so in future trials, particularly when applied at Feekes 10.5. The effects of QoI+DMI combinations depended on the active ingredients and whether the two were applied as a mixture at heading or sequentially. Following a Feekes 9 QoI application with a Feekes 10.5.1 application of a DMI reduced the negative effect of the QoI on DON but was not sufficient to achieve the efficacy of the Feekes 10.5.1 DMI-only treatments. Our results suggest that one must be prudent when using QoI treatments under moderate to high risk of FHB, particularly where the QoI is used without an effective DMI applied in combination or in sequence.

Efficacy and Stability of Integrating Fungicide and Cultivar Resistance to Manage Fusarium Head Blight and Deoxynivalenol in Wheat.

Integration of host resistance and prothioconazole + tebuconazole fungicide application at anthesis to manage Fusarium head blight (FHB) and deoxynivalenol (DON) in wheat was evaluated using data from over 40 trials in 12 U.S. states. Means of FHB index (index) and DON from up to six resistance class-fungicide management combinations per trial (susceptible treated [S_TR] and untreated [S_UT]; moderately susceptible treated [MS_TR] and untreated [MS_UT]; moderately resistant treated [MR_TR] and untreated [MR_UT]) were used in multivariate meta-analyses, and mean log response ratios across trials were estimated and transformed to estimate mean percent control ( ) due to the management combinations relative to S_UT. All combinations led to a significant reduction in index and DON (P < 0.001). MR_TR was the most effective combination, with a of 76% for index and 71% for DON, followed by MS_TR (71 and 58%, respectively), MR_UT (54 and 51%, respectively), S_TR (53 and 39%, respectively), and MS_UT (43 and 30%, respectively). Calculations based on the principle of treatment independence showed that the combination of fungicide application and resistance was additive in terms of percent control for index and DON. Management combinations were ranked based on percent control relative to S_UT within each trial, and nonparametric analyses were performed to determine management combination stability across environments (trials) using the Kendall coefficient of concordance (W). There was a significant concordance of management combinations for both index and DON (P < 0.001), indicating a nonrandom ranking across environments and relatively low variability in the within-environment ranking of management combinations. MR_TR had the highest mean rank (best control relative to S_UT) and was one of the most stable management combinations across environments, with low rank stability variance (0.99 for index and 0.67 for DON). MS_UT had the lowest mean rank (poorest control) but was also one of the most stable management combinations. Based on Piepho's nonparametric rank-based variance homogeneity U test, there was an interaction of management combination and environment for index (P = 0.011) but not for DON (P = 0.147), indicating that the rank ordering for index depended somewhat on environment. In conclusion, although the magnitude of percent control will likely vary among environments, integrating a single tebuconazole + prothioconazole application at anthesis with cultivar resistance will be a more effective and stable management practice for both index and DON than either approach used alone.

Mapping net form net blotch and septoria speckled leaf blotch resistance Loci in barley.

Septoria speckled leaf blotch (SSLB), caused by Septoria passerinii Sacc., and net form net blotch (NB), caused by Pyrenophora teres f. teres Drechsler, are fungal diseases that decrease the yields of barley in the Upper Midwest. An effective way to manage these diseases is to plant resistant cultivars. To characterize the genetics of resistance to both pathogens, two advanced barley breeding lines, one resistant to NB (M120) and another resistant to SSLB (Sep2-72), were crossed, creating a population of 115 recombinant inbred lines. The two parents and the population were evaluated in three greenhouse seedling assays for each pathogen and for simple-sequence repeat and diversity arrays technology markers. Composite interval mapping revealed two major quantitative trait loci (QTL) associated with NB on chromosome 6H, located in bins 2 and 6. The QTL located in bin 6 explained 19 to 48% of the phenotypic variation and the QTL located in bin 2 explained 25 to 44% of the phenotypic variation. A new locus for resistance to SSLB, Rsp4, was identified on chromosome 6H, located in bins 3 to 4. Mapping these genes in elite breeding germplasm will accelerate the development and utilization of marker-assisted selection to enhance resistance to these diseases.

Colonization of the Residues of Diverse Plant Species by Gibberella zeae and Their Contribution to Fusarium Head Blight Inoculum.

The presence of Fusarium spp. was examined in the residues of wheat, barley, corn, sunflower, pasture, and gramineous weed species common in wheat and barley cropping systems collected from no-tillage and reduced-tillage plots from February 2001 to March 2003 in Uruguay. Gibberella zeae was recovered from residues of wheat, barley, corn, sunflower, fescue, and the gramineous weeds Digitaria sanguinalis, Setaria spp., Lolium multiflorum, and Cynodon dactylon, except from birdsfoot trefoil or white clover. Of the Fusarium spp. obtained, G. zeae was the most frequently recovered from wheat and barley residues, while other species were more common in other crops. G. zeae declined over time in all residues examined. Wheat and barley residues produced more ascospores of G. zeae than corn or other gramineous residues. Sunflower residue did not support ascospore production, indicating that it probably did not contribute to primary inoculum. Wheat and barley residues supported G. zeae colonization longer in no-till than in reduced-tillage production systems and, thus, may represent major contributors to Fusarium head blight (FHB) inoculum in Uruguay. The presence of G. zeae in the gramineous components of pastures, weed species, and sunflower should be considered when implementing control strategies for FHB.

Effect of Extended Irrigation and Host Resistance on Deoxynivalenol Accumulation in Fusarium-Infected Wheat.

Deoxynivalenol (DON) levels are not easily predicted from visual disease assessment, and it is thought likely that environmental conditions such as temperature and moisture influence DON accumulation. This field study examined the influence of environmental moisture on DON accumulation in Fusarium-infected wheat (Triticum aestivum). The effect of extended mist irrigation applied from inoculation (at anthesis) until harvest was compared with mist irrigation applied from inoculation until disease assessment (at early dough), as is generally applied in screening nurseries used for germplasm selection and cultivar improvement. DON concentrations were quantified in kernels at early dough, hard dough, kernel hard, and maturity. Kernels from plots with extended mist irrigation generally had lower DON concentrations than those from plots where mist irrigation was not applied following disease assessment. DON concentrations tended to decrease from disease assessment until harvest, regardless of the irrigation treatment. DON concentrations in the cultivars moderately resistant to Fusarium head blight were lower than those in the susceptible cultivar. Environmental moisture is an important factor determining the DON content of Fusarium-infected wheat.

Host genetic effect on deoxynivalenol accumulation in fusarium head blight of barley.

ABSTRACT One of the major concerns with Fusarium head blight (FHB) of barley is the potential health risks to livestock and humans through the accumulation of the mycotoxin deoxynivalenol (DON) in infected grain. To define the role of the host in DON accumulation during the early stages of disease development, we conducted a series of greenhouse experiments. We inoculated single spikelets of greenhouse-grown plants with Fusarium graminearum, moved the plants to a dew chamber, and harvested the inoculated spikelets after 72 h for DON analysis. We conducted a quantitative trait locus (QTL) analysis using a genetic mapping population, constructed with the parents Stander and Frederickson, that segregated for DON accumulation after single-spikelet inoculation in two experiments. A single QTL on chromosome 3 explained 18 and 35% of the phenotypic variation in the two experiments. To validate this QTL for DON accumulation, we used a DNA marker to select near-isogenic lines from a family from the mapping population that was segregating at this QTL. Disease symptom development was similar between the nearisogenic lines; however, the mean DON concentration of the lines homozygous for the allele from the high DON parent was 2.5-fold more than the lines homozygous for the alternate allele. A time course experiment showed that this effect on toxin accumulation was observed at 10 days post inoculation. The near-isogenic lines developed in this study should prove useful for further exploration of the role of DON in FHB.

Survival and Inoculum Production of Gibberella zeae in Wheat Residue.

Survival and inoculum production of Gibberella zeae (Schwein.) Petch (anamorph Fusarium graminearum (Schwabe)), the causal agent of Fusarium head blight of wheat and barley, was related to the rate of wheat (Triticum aestivum L.) residue decomposition. Infested wheat residue, comprising intact nodes, internodes, and leaf sheaths, was placed in fiberglass mesh bags on the soil surface and at 7.5- to 10-cm and 15- to 20-cm depths in chisel-plowed plots and 15 to 20 cm deep in moldboard-plowed plots in October 1997. Residue was sampled monthly from April through November during 1998 and every 2 months through April to October 1999. Buried residue decomposed faster than residue placed on the soil surface. Less than 2% of the dry-matter residue remained in buried treatments after 24 months in the field, while 25% of the residue remained in the soil-surface treatment. Survival of G. zeae on node tissues was inversely related to the residue decomposition rate. Surface residue provided a substrate for G. zeae for a longer period of time than buried residue. Twenty-four months after the initiation of the trial, the level of colonization of nodes in buried residue was half the level of colonization of residue on the soil surface. Colonization of node tissues by G. zeae decreased over time, but increased for other Fusarium spp. Ascospores of G. zeae were still produced on residue pieces after 23 months, and these spores were capable of inducing disease. Data from this research may assist in developing effective management strategies for residues infested with G. zeae.

Engineering deoxynivalenol metabolism in wheat through the expression of a fungal trichothecene acetyltransferase gene.

Fusarium head blight occurs in cereals throughout the world and is especially important in humid growing regions. Fusarium head blight (FHB) has re-emerged as a major disease of wheat and barley in the U.S. and Canada since 1993. The primary causal agents of FHB, Fusarium graminearum and Fusarium culmorum, can produce deoxynivalenol (DON), a trichothecene mycotoxin that enhances disease severity and poses a health hazard to humans and monogastric animals. To reduce the effects of DON on wheat, we have introduced FsTRI101, a Fusarium sporotrichioides gene formerly known as TriR, into the regenerable cultivar Bobwhite. TRI101 encodes an enzyme that transfers an acetyl moiety to the C3 hydroxyl group of trichothecenes. Four different transgenic plants carrying the FsTRI101 gene were identified. Although expression levels varied among the four lines, all of them accumulated FsTRI101 transcripts in endosperm and glume. TRI101-encoded acetyltransferase activity was detected in endosperm extracts of a single plant that accumulated FsTRI101 mRNA. Greenhouse resistance tests indicated that the accumulation of FsTRI101-encoded acetyltransferase in this plant confers partial protection against the spread of F. graminearum in inoculated wheat heads (spikes).

Biosynthesis of Deoxynivalenol in Spikelets of Barley Inoculated with Macroconidia of Fusarium graminearum.

This research examined the biosynthesis of deoxynivalenol (DON) and 15-acetyldeoxynivalenol (15-ADON) in barley spikelets inoculated with macroconidia of Fusarium graminearum (Group-II). Investigations were conducted to determine if these toxins were present in macroconidia of the pathogen prior to inoculating barley spikelets. Extracts of macroconidia cultured from mung bean agar were analyzed using gas chromatography-mass spectrometry. Neither DON or 15-ADON was detected in three isolates' macroconidia when compared with macroconidia-DON-matrix standards adjusted to 100, 200, 300, and 400 ng/g with a detection limit of 100 ng/g. Mean recovery of DON that was added to macroconidia was 89.5%. The same isolates were pathogenic on barley cultivars Robust (moderately susceptible) and Chevron (moderately resistant) and produced DON (0 to 3.69 ng/g) and 15-ADON (detected but not quantified) when grown in rice culture. Greenhouse experiments were performed to determine when DON and 15-ADON were detectable after inoculation and to quantify their amount in inoculated barley spikelets. The three isolates of F. graminearum were separately inoculated to a central spikelet on heads of barley cultivars Robust and Chevron. Both toxins were detected in spikelets 48 h postinoculation (PI). DON increased dramatically after 72 h and did not diminish thereafter. Accumulation of 15-ADON peaked at 72 to 120 h and decreased by 240 h PI. There were no statistical differences between cultivars or among fungal isolates for accumulation of either toxin when averaged over the time intervals. Differences of toxin accumulation at each sampling interval were significant (P < 0.0001) when averaged over isolates and cultivars. Spikelets of six cultivars and lines were sampled at inoculation and 18, 36, 54, 72, and 90 h PI. DON and 15-ADON were detected at 36 h PI, but differences among the cultivars and lines were not significant. Yield of DON in inoculated spikelets of 31 barley cultivars and lines at 72 h PI ranged from 0.14 to 1.26 µg per spikelet, and differences among the cultivars and lines were significant (P < 0.002). The data demonstrate a useful range of variability for toxin accumulation in inoculated spikelets among germ plasm in the Minnesota breeding program. Macroconidia with no detectable DON or 15-ADON could be used for in vitro studies of toxin biosynthesis. Establishing when DON and 15-ADON are synthesized facilitates studying the effects of promising fungicides, biocontrol organisms, and new or novel genetic resistance mechanisms and if or how they may prevent or delay the biosynthesis of toxins.

The Effect of Stand Density on the Development of Puccinia graminis f. sp. tritici in Barley.

The progress of stem rust epidemics caused by race Pgt-QCCJ of Puccinia graminis f. sp. tritici was examined in field plots of the six-rowed barley cvs. Robust and Steptoe at four stand densities, and in short rows and hill plots. Cv. Robust carries Rpg1, which confers resistance to most races of P. graminis f. sp. tritici, other than Pgt-QCCJ, while cv. Steptoe lacks any reported genes for resistance to P. graminis f. sp. tritici. The stand-density treatments established were approximately double, the equivalent of, half of, and one quarter of the planting rate recommended for commercial barley. Stem rust caused by race Pgt-QCCJ spread more rapidly in cv. Steptoe than in cv. Robust, irrespective of the stand-density treatment. The effect of stand density on rust severity was, however, greater than the effect of cultivar. Rust severity was 18 to 36% greater in sparse stands than in dense stands of both cultivars. Reductions in kernel size were most severe in sparse stands where rust development was greatest. Knowledge that reduced stand densities may promote the development of stem rust in barley may be useful in refining procedures for screening breeding material for resistance to stem rust, and in allaying concern over the high levels of stem rust occasionally reported in lightly seeded seed increases and in commercial fields where sparse stands are encountered.

The Effect of Previous Crop Residues and Tillage on Fusarium Head Blight of Wheat.

Effects of previous crop residues and tillage practices on Fusarium head blight (FHB) of wheat were examined. Fusarium head blight was monitored in plots of the FHB-susceptible spring wheat cultivar Norm following crops of corn, wheat, and soybeans in 1995, 1996, and 1997. Moldboard plow, chisel plow, and no-till treatments were imposed perpendicular to crop strips to establish a range of residue levels in each of the previous crop residues. Fusarium head blight incidence and severity were greatest when wheat followed corn and least when wheat followed soybeans. Incidence and severity were lower in moldboard plowed plots than in either chisel plowed or no-till plots, although differences among chisel plow and no-till treatments were not apparent. Yields of wheat were approximately 15% lower in plots where wheat followed corn or wheat than in wheat following soybeans and were 10% greater in moldboard plowed plots than in either chisel plowed or no-till treatments. The deoxynivalenol (DON) content of harvested grain was significantly correlated with FHB incidence and severity. The DON level in wheat following soybeans, averaged across tillage treatments, was 25% lower than in wheat following wheat and 50% of the level in wheat following corn. These findings suggest that changes in regional tillage practices, principally the move toward conservation tillage and reduced-till systems, contributed to the recent FHB epidemics in the Upper Midwest. Because differences in the type and quantity of crop residues in small plots affected disease development, it is likely that local sources of inoculum, such as those within a grower's field, contribute directly to the inoculum load and disease potential. The implication of these findings is that selection of cultural practices aimed to reduce inoculum-borne residues will assist in the control of FHB.

Agronomic performance of lines derived from anther culture, maize pollination and single-seed descent in a spring wheat cross.

Anther culture and maize hybridization are two frequently used techniques for doubled haploid production in wheat (Triticum aestivum L.). Information on the field performance of lines derived from these techniques is limited. This study was conducted to compare the performance of F(4:6) lines obtained by single-seed descent with lines obtained by anther culture and maize (Zea mays L.) pollination from the same cross of spring wheat, 'Chris'/MN 7529. Thirty-three lines derived from each of those techniques were evaluated in six environments for grain yield, protein content, test weight, heading date, kernel weight and plant height. Mean performance of the single-seed descent lines exceeded performance of the anther culture lines for grain yield, kernel weight and plant height with no apparent differences for grain protein content, test weight and heading date. No differences between trait means for the single-seed descent and maize pollination lines were found except for plant height. The best 5 lines from each method for grain yield, protein content and test weight were similar in performance except that the protein content was higher for the maize pollination lines than for the single-seed descent lines. Acceptable levels of agronomic performance could be found among lines from each method. Wide acceptance of the doubled haploid technique for pure line production in breeding programs may, however, be limited by the often poor efficiency of doubled haploid line production, resulting in smaller population sizes for selection of desirable traits in comparison to the single-seed descent method.

Quantitative trait loci associated with resistance to Fusarium head blight and kernel discoloration in barley.

Resistance to Fusarium head blight (FHB), deoxynivalenol (DON) accumulation, and kernel discoloration (KD) in barley are difficult traits to introgress into elite varieties because current screening methods are laborious and disease levels are strongly influenced by environment. To improve breeding strategies directed toward enhancing these traits, we identified genomic regions containing quantitative trait loci (QTLs) associated with resistance to FHB, DON accumulation, and KD in a breeding population of F(4:7) lines using restriction fragment length polymorphic (RFLP) markers. We evaluated 101 F(4:7) lines, derived from a cross between the cultivar Chevron and an elite breeding line, M69, for each of the traits in three or four environments. We used 94 previously mapped RFLP markers to create a linkage map. Using composite interval mapping, we identified 10, 11, and 4 QTLs associated with resistance to FHB, DON accumulation, and KD, respectively. Markers flanking these QTLs should be useful for introgressing resistance to FHB, DON accumulation, and KD into elite barley cultivars.

Daily Inoculum Levels of Gibberella zeae on Wheat Spikes.

The inoculum level of Gibberella zeae on wheat spikes was measured during 1995 and 1996 in nine locations of Canada and the United States prone to Fusarium head blight of wheat. Spikes were exposed after exsertion and until kernel milk or soft dough stage in fields with wheat or corn residue as a source of inoculum; other spikes were exposed in a location remote from any obvious inoculum source; and in 1995 only, control plants remained in a greenhouse. After 24 h, spikes were excised and vigorously shaken in water to remove inoculum. Propagules were enumerated on selective medium and identified as G. zeae from subcultures. Significantly more inoculum was detected from fields in epidemic areas than from remote sites in an epidemic and from fields in nonepidemic areas. The median inoculum level was 20 CFU of G. zeae per spike per day in fields experiencing an epidemic, 4 CFU in locations remote from epidemic fields, 2 CFU in nonepidemic fields, and 1 CFU in locations remote from a source of inoculum in non-epidemic areas. In an epidemic region, inoculum levels near corn stubble reached up to 587 CFU of G. zeae per spike per day, and the median inoculum level of 126 CFU was significantly higher than the median of 13 CFU found near wheat residue. Inoculum was not detected or occurred sporadically during extended dry periods. While inoculum increased during rainy periods, timing of increased levels was variable. Fusarium head blight epidemics were associated with multiple inoculation episodes and coincident wet periods.

The Effect of Race QCCJ of Puccinia graminis f. sp. tritici on Yield and Quality in Barley.

The progress of epidemics of race QCCJ of Puccinia graminis f. sp. tritici was examined in field plots of the barley cultivars Robust, Excel, Stander, Azure, Stark, Larker, Bonanza, and Steptoe. In addition, a potential source of resistance (cv. Diamond), and a susceptible line (ND11075) were included. Rust-free treatments were maintained to provide comparisons of grain yield and quality. Barleys with Rpg1, which confers resistance to races of P. graminis f. sp. tritici other than Pgt-QCCJ, were generally more resistant than cultivars or lines that lacked this gene. However the severity of losses indicated that under high disease pressure the resistance conferred by Rpg1 would not be effective. Yield reductions of up to 29% were recorded and were generally associated with reduced grain size and weight. The continued presence of Pgt-QCCJ as a component of the P. graminis population in the United States, since it was first detected in 1989, indicates that Pgt-QCCJ remains a threat to commercial barley production.