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The Fusarium head blight (FHB) pathogen Fusarium graminearum produces the trichothecene mycotoxin deoxynivalenol and reduces wheat yield and grain quality. Spring wheat ( Triticum aestivum ) genotype CB037 was transformed with constitutive expression (CE) constructs containing sorghum ( Sorghum bicolor ) genes encoding monolignol biosynthetic enzymes caffeoyl coenzyme A (CoA) 3- O -methyltransferase ( SbCCoAOMT ), 4-coumarate-CoA ligase ( Sb4CL ), or coumaroyl shikimate 3-hydroxylase ( SbC3'H ) or monolignol pathway transcriptional activator SbMyb60 . Spring wheats were screened for type I (resistance to initial infection, using spray inoculations) and type II (resistance to spread within the spike, using single-floret inoculations) resistances in the field (spray) and greenhouse (spray and single floret). Following field inoculations, disease index, percentage of Fusarium -damaged kernels (FDK), and deoxynivalenol measurements of CE plants were similar to or greater than those of CB037. For greenhouse inoculations, the area under the disease progress curve (AUDPC) and FDK were determined. Following screens, focus was placed on two each of SbC3'H and SbCCoAOMT CE lines because of trends toward a decreased AUDPC and FDK observed following single-floret inoculations. These four lines were as susceptible as CB037 following spray inoculations. However, single-floret inoculations showed that these CE lines had a significantly reduced AUDPC ( P < 0.01) and FDK ( P ≤ 0.02) compared with CB037, indicating improved type II resistance. None of these CE lines had increased acid detergent lignin compared with CB037, indicating that lignin concentration may not be a major factor in FHB resistance. The SbC3'H and SbCCoAOMT CE lines are valuable for investigating phenylpropanoid-based resistance to FHB., Competing Interests: The author(s) declare no conflict of interest.

Background: Previous research has suggested that proteins and other quality parameters of wheats may have changed over a century of wheat breeding. These changes may affect protein digestibility. The in vitro protein digestibility of breads made with 21 cultivars of wheat introduced or released in the USA between 1870 and 2013 was therefore evaluated., Results: Protein digestibility increased with release year, but was not normally distributed; three older cultivars had significantly lower digestibility than the other cultivars: 42.0 ± 0.3 mol% (primary amino N/total N) versus 34.7 ± 0.7 mol%; P < 0.001. High molecular weight (MW) protein fractions increased and low MW protein fractions decreased with release year, but these changes were not related to protein digestibility. Thus, other differences in protein composition or other flour components may contribute to diminished digestibility of the three older cultivars., Conclusions: This study identified differences in protein digestibility among wheat cultivars that may have important implications for human nutrition. Further investigation is required to determine the specific characteristics that differentiate high- and low-digestibility wheat cultivars. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Wheat streak mosaic virus (WSMV) and triticum mosaic virus (TriMV) are economically important viruses of wheat ( Triticum aestivum L.), causing significant yield losses in the Great Plains region of the United States. These two viruses are transmitted by wheat curl mites, which often leads to mixed infections with synergistic interaction in grower fields that exacerbates yield losses. Development of dual-resistant wheat lines would provide effective control of these two viruses. In this study, a genetic resistance strategy employing an RNA interference (RNAi) approach was implemented by assembling a hairpin element composed of a 202-bp (404-bp in total) stem sequence of the NIb (replicase) gene from each of WSMV and TriMV in tandem and of an intron sequence in the loop. The derived RNAi element was cloned into a binary vector and was used to transform spring wheat genotype CB037. Phenotyping of T 1 lineages across eight independent transgenic events for resistance revealed that i) two of the transgenic events provided resistance to WSMV and TriMV, ii) four events provided resistance to either WSMV or TriMV, and iii) no resistance was found in two other events. T 2 populations derived from the two events classified as dual-resistant were subsequently monitored for stability of the resistance phenotype through the T 4 generation. The resistance phenotype in these events was temperature-dependent, with a complete dual resistance at temperatures ≥25°C and an increasingly susceptible response at temperatures below 25°C. Northern blot hybridization of total RNA from transgenic wheat revealed that virus-specific small RNAs (vsRNAs) accumulated progressively with an increase in temperature, with no detectable levels of vsRNA accumulation at 20°C. Thus, the resistance phenotype of wheat harboring an RNAi element was correlated with accumulation of vsRNAs, and the generation of vsRNAs can be used as a molecular marker for the prediction of resistant phenotypes of transgenic plants at a specific temperature.

The above-mentioned article was published in 2015 with an error in the reverse primer sequence for the PPOA2d1074 marker, which made amplification difficult. The reverse primer was missing a thymine nucleotide at the thirteenth position (GCGGTGCTTCACTTGGT).

Hexaploid waxy wheat ( Triticum aestivum L.) has null mutations in Wx genes and grain lacking amylose with increased digestibility and usability for specialty foods. The waxy cultivar Mattern is susceptible to Fusarium head blight (FHB) caused by Fusarium graminearum species complex, which produces the mycotoxin deoxynivalenol (DON). In experiment 1, conducted during low natural FHB, grain from waxy breeding lines, Mattern, and wild-type breeding lines and cultivars were assessed for Fusarium infection and DON concentration. Nine Fusarium species and species complexes were detected from internally infected (disinfested) grain; F. graminearum infections were not different between waxy and wild-type. Surface- and internally infected grain (nondisinfested) had greater numbers of Fusarium isolates across waxy versus wild-type, but F. graminearum -like infections were similar; however, DON levels were higher in waxy . In experiment 2, conducted during a timely epidemic, disease severity, Fusarium -damaged kernels (FDK), and DON were assessed for waxy breeding lines, Mattern, and wild-type cultivars. Disease severity and FDK were not significantly different from wild-type, but DON was higher in waxy than wild-type lines. Across both experiments, waxy breeding lines, Plant Introductions 677876 and 677877, responded similarly to FHB as moderately resistant wild-type cultivar Overland, showing promise for breeding advanced waxy cultivars with reduced FHB susceptibility.

The development of inexpensive, whole-genome profiling enables a transition to allele-based breeding using genomic prediction models. These models consider alleles shared between lines to predict phenotypes and select new lines based on estimated breeding values. This approach can leverage highly unbalanced datasets that are common to breeding programs. The Southern Regional Performance Nursery (SRPN) is a public nursery established by the USDA-ARS in 1931 to characterize performance and quality of near-release wheat ( L.) varieties from breeding programs in the US Central Plains. New entries are submitted annually and can be re-entered only once. The trial is grown at >30 locations each year and lines are evaluated for grain yield, disease resistance, and agronomic traits. Overall genetic gain is measured across years by including common check cultivars for comparison. We have generated whole-genome profiles via genotyping-by-sequencing (GBS) for 939 SPRN entries dating back to 1992 to explore the potential use of the nursery as a genomic selection (GS) training population (TP). The GS prediction models across years (average = 0.33) outperformed year-to-year phenotypic correlation for yield ( = 0.27) for a majority of the years evaluated, suggesting that genomic selection has the potential to outperform low heritability selection on yield in these highly variable environments. We also examined the predictability of programs using both program-specific and whole-set TPs. Generally, the predictability of a program was similar with both approaches. These results suggest that wheat breeding programs can collaboratively leverage the immense datasets that are generated from regional testing networks., (Copyright © 2018 Crop Science Society of America.)

High Plains wheat mosaic virus (HPWMoV, genus Emaravirus; family Fimoviridae), transmitted by the wheat curl mite (Aceria tosichella Keifer), harbors a monocistronic octapartite single-stranded negative-sense RNA genome. In this study, putative proteins encoded by HPWMoV genomic RNAs 2-8 were screened for potential RNA silencing suppression activity by using a green fluorescent protein-based reporter agroinfiltration assay. We found that proteins encoded by RNAs 7 (P7) and 8 (P8) suppressed silencing induced by single- or double-stranded RNAs and efficiently suppressed the transitive pathway of RNA silencing. Additionally, a Wheat streak mosaic virus (WSMV, genus Tritimovirus; family Potyviridae) mutant lacking the suppressor of RNA silencing (ΔP1) but having either P7 or P8 from HPWMoV restored cell-to-cell and long-distance movement in wheat, thus indicating that P7 or P8 rescued silencing suppressor-deficient WSMV. Furthermore, HPWMoV P7 and P8 substantially enhanced the pathogenicity of Potato virus X in Nicotiana benthamiana. Collectively, these data demonstrate that the octapartite genome of HPWMoV encodes two suppressors of RNA silencing., (Published by Elsevier Inc.)

Previously, we reported that coat protein (CP) of Wheat streak mosaic virus (WSMV) (genus Tritimovirus, family Potyviridae) tolerates deletion of amino acids 36 to 84 for efficient systemic infection of wheat. In this study, we demonstrated that WSMV mutants with deletion of CP amino acids 58 to 84 but not of 36 to 57 induced severe chlorotic streaks and spots, followed by acute chlorosis in wheat, maize, barley, and rye compared with mild to moderate chlorotic streaks and mosaic symptoms by wild-type virus. Deletion of CP amino acids 58 to 84 from the WSMV genome accelerated cell-to-cell movement, with increased accumulation of genomic RNAs and CP, compared with the wild-type virus. Microscopic examination of wheat tissues infected by green fluorescent protein-tagged mutants revealed that infection by mutants lacking CP amino acids 58 to 84 caused degradation of chloroplasts, resulting in acute macroscopic chlorosis. The profile of CP-specific proteins was altered in wheat infected by mutants causing acute chlorosis, compared with mutants eliciting wild-type symptoms. All deletion mutants accumulated CP-specific major protein similarly to that in wild-type virus; however, mutants that elicit acute chlorosis failed to accumulate a 31-kDa minor protein compared with wild-type virus or mutants lacking amino acids 36 to 57. Taken together, these data suggest that deletion of CP amino acids 58 to 84 from the WSMV genome enhanced accumulation of CP and genomic RNA, altered CP-specific protein profiles, and caused severe symptom phenotypes in multiple cereal hosts.

Wheat streak mosaic virus (WSMV) and Triticum mosaic virus (TriMV) are economically important viral pathogens of wheat. Wheat cvs. Mace, carrying the Wsm1 gene, is resistant to WSMV and TriMV, and Snowmass, with Wsm2, is resistant to WSMV. Viral resistance in both cultivars is temperature sensitive and is effective at 18°C or below but not at higher temperatures. The underlying mechanisms of viral resistance of Wsm1 and Wsm2, nonallelic single dominant genes, are not known. In this study, we found that fluorescent protein-tagged WSMV and TriMV elicited foci that were approximately similar in number and size at 18 and 24°C, on inoculated leaves of resistant and susceptible wheat cultivars. These data suggest that resistant wheat cultivars at 18°C facilitated efficient cell-to-cell movement. Additionally, WSMV and TriMV efficiently replicated in inoculated leaves of resistant wheat cultivars at 18°C but failed to establish systemic infection, suggesting that Wsm1- and Wsm2-mediated resistance debilitated viral long-distance transport. Furthermore, we found that neither virus was able to enter the leaf sheaths of inoculated leaves or crowns of resistant wheat cultivars at 18°C but both were able to do so at 24°C. Thus, wheat cvs. Mace and Snowmass provide resistance at the long-distance movement stage by specifically blocking virus entry into the vasculature. Taken together, these data suggest that both Wsm1 and Wsm2 genes similarly confer virus resistance by temperature-dependent impairment of viral long-distance movement.

Pyramiding of alien-derived Wheat streak mosaic virus (WSMV) resistance and resistance enhancing genes in wheat is a cost-effective and environmentally safe strategy for disease control. PCR-based markers and cytogenetic analysis with genomic in situ hybridisation were applied to identify alien chromatin in four genetically diverse populations of wheat (Triticum aestivum) lines incorporating chromosome segments from Thinopyrum intermedium and Secale cereale (rye). Out of 20 experimental lines, 10 carried Th. intermedium chromatin as T4DL*4Ai#2S translocations, while, unexpectedly, 7 lines were positive for alien chromatin (Th. intermedium or rye) on chromosome 1B. The newly described rye 1RS chromatin, transmitted from early in the pedigree, was associated with enhanced WSMV resistance. Under field conditions, the 1RS chromatin alone showed some resistance, while together with the Th. intermedium 4Ai#2S offered superior resistance to that demonstrated by the known resistant cultivar Mace. Most alien wheat lines carry whole chromosome arms, and it is notable that these lines showed intra-arm recombination within the 1BS arm. The translocation breakpoints between 1BS and alien chromatin fell in three categories: (i) at or near to the centromere, (ii) intercalary between markers UL-Thin5 and Xgwm1130 and (iii) towards the telomere between Xgwm0911 and Xbarc194. Labelled genomic Th. intermedium DNA hybridised to the rye 1RS chromatin under high stringency conditions, indicating the presence of shared tandem repeats among the cereals. The novel small alien fragments may explain the difficulty in developing well-adapted lines carrying Wsm1 despite improved tolerance to the virus. The results will facilitate directed chromosome engineering producing agronomically desirable WSMV-resistant germplasm.

An infectious cDNA clone of Triticum mosaic virus (TriMV) (genus Poacevirus; family Potyviridae) was used to establish three independent lineages in wheat to examine intra-host population diversity levels within protein 1 (P1) and coat protein (CP) cistrons over time. Genetic variation was assessed at passages 9, 18 and 24 by single-strand conformation polymorphism, followed by nucleotide sequencing. The founding P1 region genotype was retained at high frequencies in most lineage/passage populations, while the founding CP genotype disappeared after passage 18 in two lineages. We found that rare TriMV genotypes were present only transiently and lineages followed independent evolutionary trajectories, suggesting that genetic drift dominates TriMV evolution. These results further suggest that experimental populations of TriMV exhibit lower mutant frequencies than that of Wheat streak mosaic virus (genus Tritimovirus; family Potyviridae) in wheat. Nevertheless, there was evidence for parallel evolution at a synonymous site in the TriMV CP cistron., (Published by Elsevier Inc.)

Key Message: Hard red wheats can donate genes to hard white wheats for tolerance to preharvest sprouting, the effects are quantitative in nature, and may be tracked with previously described DNA markers., Abstract: Pre-harvest sprouting (PHS) of wheat (Triticum aestivum L.) can negatively impact end-use quality and seed viability at planting. Due to preferences for white over red wheat in international markets, white wheat with PHS tolerance has become increasingly desired for worldwide wheat production. In general, however, red wheat is more tolerant of sprouting than white wheat. The main objective of this study was the identification of PHS tolerance conditioned by genes donated from hard red winter wheat, using markers applicable to the Great Plains hard white wheat gene pool. Three red wheat by white wheat populations, Niobrara/NW99L7068, NE98466/NW99L7068 and Jagalene/NW99L7068 were developed, and white-seeded progenies were analyzed for PHS tolerance and used to identify markers for the trait. In the three populations, marker loci with significant allelic effects were most commonly located on chromosomes of group 2, 3, 4 and 5, though additional markers were detected across the wheat genome. Chromosome 3A was the only chromosome with significant markers in all three populations. Markers were inconsistent across the three populations, and markers linked to tolerance-inducing loci were identified in both tolerant and susceptible parents. Additive effects of marker loci were common. In the present investigation, a wide range of PHS tolerance was observed, even though all lines were fixed for the recently reported positive TaPHS1 allele. PHS tolerance is controlled by additive major gene effects with minor gene effects where variations of minor gene effects were still unclear.

Waxy wheat contains very low concentration (generally <2%) of amylose in endosperm starch, in contrast to conventional wheat whose starch is typically 20% amylose, with the balance being the branched macromolecule, amylopectin. With the release of a commercial hard winter waxy wheat cultivar in the United States, the grain trade, milling, and processing industries seek to have a rapid technique to ensure the purity of identity preserved waxy wheat lots. Near infrared (NIR) reflectance spectroscopy, a technique widely used in the cereals industry for proximate analysis, is a logical candidate for measuring contamination level and thus is the subject of this study. Two sets of wheat samples, harvested, prepared and scanned one year apart, were used to evaluate the NIR concept. One year consisted of nine pairs of conventional:waxy preparations, with each preparation consisting of 29 binary mixtures ranging in conventional wheat fraction (by weight) of 0-100% (261 spectral samples). The second year was prepared in the same fashion, with 12 preparations, thus producing 348 spectral samples. One year's samples were controlled for protein content and moisture level between pair components in order to avoid the basis for the conventional wheat fraction models being caused by something other than spectral differences attributed to waxy and nonwaxy endosperm. Likewise the second year was controlled by selection of conventional wheat for mixture preparation based on either protein content or cluster analysis of principal components of candidate spectra. Partial least squares regression, one and two-term linear regression, and support vector machine regression models were examined. Validation statistics arising from sets within the same year or across years were remarkably similar, as were those among the three regression types. A single wavelength on second derivative transformed spectra, namely 2290 nm, was effective at estimating the mixture level by weight, with standard errors of performance in the 6-9% range. Thus, NIR spectroscopy may be used for measuring conventional hard wheat 'contamination' in waxy wheat at mixture levels above 10% w/w., (Published by Elsevier B.V.)

Triticum mosaic virus (TriMV) (genus Poacevirus, family Potyviridae) is a recently described eriophyid mite-transmitted wheat virus. In vitro RNA transcripts generated from full-length cDNA clones of TriMV proved infectious on wheat. Wheat seedlings inoculated with in vitro transcripts elicited mosaic and mottling symptoms similar to the wild-type virus, and the progeny virus was efficiently transmitted by wheat curl mites, indicating that the cloned virus retained pathogenicity, movement, and wheat curl mite transmission characteristics. A series of TriMV-based expression vectors was constructed by engineering a green fluorescent protein (GFP) or red fluorescent protein (RFP) open reading frame with homologous NIa-Pro cleavage peptides between the P1 and HC-Pro cistrons. We found that GFP-tagged TriMV with seven or nine amino acid cleavage peptides efficiently processed GFP from HC-Pro. TriMV-GFP vectors were stable in wheat for more than 120 days and for six serial passages at 14-day intervals by mechanical inoculation and were transmitted by wheat curl mites similarly to the wild-type virus. Fluorescent protein-tagged TriMV was observed in wheat leaves, stems, and crowns. The availability of fluorescent protein-tagged TriMV will facilitate the examination of virus movement and distribution in cereal hosts and the mechanisms of cross protection and synergistic interactions between TriMV and Wheat streak mosaic virus.

Key Message: Characterized novel mutations present at Ppo loci account for the substantial reduction of the total kernel PPO activity present in a putative null Ppo - A1 genetic background. Wheat (Triticum aestivum) polyphenol oxidase (PPO) contributes to the time-dependent discoloration of Asian noodles. Wheat contains multiple paralogous and orthologous Ppo genes, Ppo-A1, Ppo-D1, Ppo-A2, Ppo-D2, and Ppo-B2, expressed in wheat kernels. To date, wheat noodle color improvement efforts have focused on breeding cultivars containing Ppo-D1 and Ppo-A1 alleles conferring reduced PPO activity. A major impediment to wheat quality improvement is a lack of additional Ppo alleles conferring reduced kernel PPO. In this study, a previously reported very low PPO line, 07OR1074, was found to contain a novel allele at Ppo-A2 and null alleles at the Ppo-A1 and Ppo-D1 loci. To examine the impact of each mutation upon kernel PPO, populations were generated from crosses between 07OR1074 and the hard white spring wheat cultivars Choteau and Vida. Expression analysis using RNA-seq demonstrated no detectable Ppo-A1 transcripts in 07OR1074 while Ppo-D1 transcripts were present at less than 10% of that seen in Choteau and Vida. Novel markers specific for the Ppo-D1 and Ppo-A2 mutations discovered in 07OR1074, along with the Ppo-A1 STS marker, were used to screen segregating populations. Evaluation of lines indicated a substantial genotypic effect on PPO with Ppo-A1 and Ppo-D1 alleles contributing significantly to total PPO in both populations. These results show that the novel mutations in Ppo-A1 and Ppo-D1 present in 07OR1074 are both important to lowering overall wheat seed PPO activity and may be useful to produce more desirable and marketable wheat-based products.

Co-infection of wheat (Triticum aestivum L.) by Wheat streak mosaic virus (WSMV, a Tritimovirus) and Triticum mosaic virus (TriMV, a Poacevirus) of the family Potyviridae causes synergistic interaction. In this study, the effects of the synergistic interaction between WSMV and TriMV on endogenous and virus-derived small interfering RNAs (vsiRNAs) were examined in susceptible ('Arapahoe') and temperature-sensitive resistant ('Mace') wheat cultivars at 18°C and 27°C. Single and double infections in wheat caused a shift in the profile of endogenous small RNAs from 24 nt being the most predominant in healthy plants to 21 nt in infected wheat. Massive amounts of 21 and 22 nt vsiRNAs accumulated in singly and doubly infected Arapahoe at both temperatures and in Mace at 27°C but not 18°C. The plus- and minus-sense vsiRNAs were distributed throughout the genomic RNAs in Arapahoe at both temperature regimens and in Mace at 27°C, although some regions served as hot-spots, spawning an excessive number of vsiRNAs. The vsiRNA peaks were conserved among cultivars, suggesting that the Dicer-like enzymes in susceptible and resistant cultivars similarly accessed the genomic RNAs of WSMV or TriMV. Accumulation of large amounts of vsiRNAs in doubly infected plants suggests that the silencing suppressor proteins encoded by TriMV and WSMV do not prevent the formation of vsiRNAs; thus, the synergistic effect observed is independent from RNA-silencing mediated vsiRNA biogenesis. The high-resolution map of endogenous and vsiRNAs from WSMV- and/or TriMV-infected wheat cultivars may form a foundation for understanding the virus-host interactions, the effect of synergistic interactions on host defense, and virus resistance mechanisms in wheat.

Eriophyid mite-transmitted, multipartite, negative-sense RNA plant viruses with membrane-bound spherical virions are classified in the genus Emaravirus. We report here that the eriophyid mite-transmitted Wheat mosaic virus (WMoV), an Emaravirus, contains eight genomic RNA segments, the most in a known negative-sense RNA plant virus. Remarkably, two RNA 3 consensus sequences, encoding the nucleocapsid protein, were found with 12.5% sequence divergence, while no heterogeneity was observed in the consensus sequences of additional genomic RNA segments. The RNA-dependent RNA polymerase, glycoprotein precursor, nucleocapsid, and P4 proteins of WMoV exhibited limited sequence homology with the orthologous proteins of other emaraviruses, while proteins encoded by additional genomic RNA segments displayed no significant homology with proteins reported in GenBank, suggesting that the genus Emaravirus evolved further with a divergent octapartite genome. Phylogenetic analyses revealed that WMoV formed an evolutionary link between members of the Emaravirus genus and the family Bunyaviridae. Furthermore, genomic-length virus- and virus-complementary (vc)-sense strands of all WMoV genomic RNAs accumulated asymmetrically in infected wheat, with 10- to 20-fold more virus-sense genomic RNAs than vc-sense RNAs. These data further confirm the octapartite negative-sense polarity of the WMoV genome. In WMoV-infected wheat, subgenomic-length mRNAs of vc sense were detected for genomic RNAs 3, 4, 7, and 8 but not for other RNA species, suggesting that the open reading frames present in the complementary sense of genomic RNAs are expressed through subgenomic- or near-genomic-length vc-sense mRNAs. Importance: Wheat mosaic virus (WMoV), an Emaravirus, is the causal agent of High Plains disease of wheat and maize. In this study, we demonstrated that the genome of WMoV comprises eight negative-sense RNA segments with an unusual sequence polymorphism in an RNA encoding the nucleocapsid protein but not in the additional genomic RNA segments. WMoV proteins displayed weak or no homology with reported emaraviruses, suggesting that the genus Emaravirus further evolved with a divergent octapartite genome. The current study also examined the profile of WMoV RNA accumulation in wheat and provided evidence for the synthesis of subgenomic-length mRNAs of virus complementary sense. This is the first report to demonstrate that emaraviruses produce subgenomic-length mRNAs that are most likely utilized for genome expression. Importantly, this study facilitates the examination of gene functions and virus diversity and the development of effective diagnostic methods and management strategies for an economically important but poorly understood virus., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

Triticum mosaic virus (TriMV) and Wheat streak mosaic virus (WSMV) infect winter wheat (Triticum aestivum) in the Great Plains region of the United States. The two viruses are transmitted by wheat curl mites (Aceria tosichella), which also transmit High Plains virus. In a field study conducted in 2011 and 2012, winter wheat cultivars Millennium (WSMV-susceptible) and Mace (WSMV-resistant) were mechanically inoculated with TriMV, WSMV, TriMV+WSMV, or sterile water at the two-leaf growth stage. Chlorophyll meter (soil plant analysis development [SPAD]) readings, area under the SPAD progress curve (AUSPC), grain yield (=yield), yield components (spikes/m 2 , kernels/spike, 1,000-kernel weight), and aerial dry matter were determined. In Millennium, all measured variables were significantly reduced by single or double virus inoculation, with the greatest reductions occurring in the double-inoculated treatment. Among the yield components, the greatest reductions occurred in spikes/m 2 . In Mace, only AUSPC was significantly reduced by the TriMV+WSMV treatment in 2012. There was a significant (P ≤ 0.05), negative linear relationship between SPAD readings and day of year in all inoculation treatments in Millennium and in the TriMV+WSMV treatment in Mace. There were significant (P ≤ 0.05), positive linear relationships between yield and SPAD readings and between yield and aerial dry matter in Millennium but not in Mace. The results from this study indicate that under field conditions, (i) Mace, a WSMV-resistant cultivar, is also resistant to TriMV, and (ii) double inoculation of winter wheat by TriMV and WSMV exacerbates symptom expression and yield loss in a susceptible cultivar.

Waxy mutants, in which endosperm starch contains ~100% amylopectin rather than the wild-type composition of ~70% amylopectin and ~30% amylose, occur in many domesticated cereals. The cultivation of waxy varieties is concentrated in east Asia, where there is a culinary preference for glutinous-textured foods that may have developed from ancient food processing traditions. The waxy phenotype results from mutations in the GBSSI gene, which catalyzes amylose synthesis. Broomcorn or proso millet (Panicum miliaceum L.) is one of the world's oldest cultivated cereals, which spread across Eurasia early in prehistory. Recent phylogeographic analysis has shown strong genetic structuring that likely reflects ancient expansion patterns. Broomcorn millet is highly unusual in being an allotetraploid cereal with fully waxy varieties. Previous work characterized two homeologous GBSSI loci, with multiple alleles at each, but could not determine whether both loci contributed to GBSSI function. We first tested the relative contribution of the two GBSSI loci to amylose synthesis and second tested the association between GBSSI alleles and phylogeographic structure inferred from simple sequence repeats (SSRs). We evaluated the phenotype of all known GBSSI genotypes in broomcorn millet by assaying starch composition and protein function. The results showed that the GBSSI-S locus is the major locus controlling endosperm amylose content, and the GBSSI-L locus has strongly reduced synthesis capacity. We genotyped 178 individuals from landraces from across Eurasia for the 2 GBSSI and 16 SSR loci and analyzed phylogeographic structuring and the geographic and phylogenetic distribution of GBSSI alleles. We found that GBSSI alleles have distinct spatial distributions and strong associations with particular genetic clusters defined by SSRs. The combination of alleles that results in a partially waxy phenotype does not exist in landrace populations. Our data suggest that broomcorn millet is a system in the process of becoming diploidized for the GBSSI locus responsible for grain amylose. Mutant alleles show some exchange between genetic groups, which was favored by selection for the waxy phenotype in particular regions. Partially waxy phenotypes were probably selected against-this unexpected finding shows that better understanding is needed of the human biology of this phenomenon that distinguishes cereal use in eastern and western cultures.

Grain polyphenol oxidase (PPO) activity can cause discoloration of wheat (Triticum aestivum L.) food products. Five crosses (PI 117635/Antelope; Fielder/NW03681; Fielder/Antelope; NW07OR1070/Antelope; NW07OR1066/OR2050272H) were selected to study the genetic inheritance of PPO activity. STS markers, PPO18, PPO29 and STS01, were used to identify lines with putative alleles at the Ppo-A1 and Ppo-D1 loci conditioning low or high PPO activity. ANOVA showed significant genotypic effects on PPO activity (P < 0.0001) in all populations. The generations and generation × genotype effects were not significant in any population. A putative third (null) genotype at Ppo-A1 (no PCR fragments for PPO18) was discovered in NW07OR1066 and NW07OR1070 derived populations, and these had the lowest mean PPO activities. Results demonstrated that both Ppo-A1 and Ppo-D1 loci affect the kernel PPO activity, but the Ppo-A1 has the major effect. In three populations, contrary results were observed to those predicted from previous work with Ppo-D1 alleles, suggesting the markers for Ppo-D1 allele might give erroneous results in some genetic backgrounds or lineages. Results suggest that selection for low or null alleles only at Ppo-A1 might allow development of low PPO wheat cultivars.

Triticum mosaic virus (TriMV) is a recently discovered virus infecting wheat (Triticum aestivum) in the Great Plains region of the United States. It is transmitted by wheat curl mites (Aceria tosichella) which also transmit Wheat streak mosaic virus (WSMV) and Wheat mosaic virus. In a greenhouse study, winter wheat 'Millennium' (WSMV susceptible) and 'Mace' (WSMV resistant) were mechanically inoculated with TriMV, WSMV, TriMV+WSMV, or sterile water at the two-leaf growth stage. At 28 days after inoculation, final chlorophyll meter (soil plant analysis development [SPAD]) readings, area under the SPAD progress curve (AUSPC), the number of tillers per plant, shoot and root weight, and total nitrogen and carbon content were determined. In Millennium, all measured variables were significantly reduced by single or double virus infections, with the greatest reductions occurring in the double-infection treatment. In Mace, only final SPAD readings, AUSPC, and total nitrogen were significantly reduced by single or double virus infections. There was a significant (P ≤ 0.05), positive linear relationship between SPAD readings and shoot weight in Millennium but not in Mace. The relationship between total nitrogen and shoot weight was positive, linear, and significant in both cultivars. The results from this study indicate that Mace, a WSMV-resistant cultivar, is also resistant to TriMV, and double infection of winter wheat by TriMV and WSMV exacerbates symptom expression and loss of biomass in susceptible cultivars.

Wheat (Triticum aestivum L.) breeding programs are currently developing varieties that are free of amylose (waxy wheat), as well as genetically intermediate (partial waxy) types. Successful introduction of waxy wheat varieties into commerce is predicated on a rapid methodology at the commodity point of sale that can test for the waxy condition. Near-infrared (NIR) reflectance spectroscopy, one such technology, was applied to a diverse set of hard winter (hexaploid) wheat breeders' lines representing all eight genotypic combinations of alleles at the wx-A1, wx-B1, and wx-D1 loci. These loci encode granule-bound starch synthase, the enzyme responsible for amylose synthesis. Linear discriminant analysis of principal components scores 1-4 was successful in identifying the fully waxy samples at typically greater than 90% accuracy; however, accuracy was reduced for partial and wild-type genotypes. It is suggested that the spectral sensitivity to waxiness is due to (1) the lipid-amylose complex which diminishes with waxiness, (2) physical differences in endosperm that affect light scatter, or (3) changes in starch crystallinity.

Triticum mosaic virus (TriMV), the type member of the newly proposed Poacevirus genus, and Wheat streak mosaic virus (WSMV), the type member of Tritimovirus genus of the family Potyviridae, infect wheat naturally in the Great Plains and are transmitted by wheat curl mites. In this study, we examined the ability of these viruses to infect selected cereal hosts, and found several differential hosts between TriMV and WSMV. Additionally, we examined the interaction between WSMV and TriMV in three wheat cultivars at two temperature regimens (19 and 20 to 26 degrees C), and quantified the virus concentration in single and double infections by real-time reverse-transcription polymerase chain reaction. Double infections in wheat cvs. Arapahoe and Tomahawk at both temperature regimens induced disease synergism with severe leaf deformation, bleaching, and stunting, with a 2.2- to 7.4-fold increase in accumulation of both viruses over single infections at 14 days postinoculation (dpi). However, at 28 dpi, in double infections at 20 to 26 degrees C, TriMV concentration was increased by 1.4- to 1.8-fold in Arapahoe and Tomahawk but WSMV concentration was decreased to 0.5-fold. WSMV or TriMV replicated poorly in Mace at 19 degrees C with no synergistic interaction whereas both viruses accumulated at moderate levels at 20 to 26 degrees C and induced mild to moderate disease synergism in doubly infected Mace compared with Arapahoe and Tomahawk. Co-infections in Mace at 20 to 26 degrees C caused increased TriMV accumulation at 14 and 28 dpi by 2.6- and 1.4-fold and WSMV accumulated at 0.5- and 1.6-fold over single infections, respectively. Our data suggest that WSMV and TriMV induced cultivar-specific disease synergism in Arapahoe, Tomahawk, and Mace, and these findings could have several implications for management of wheat viruses in the Great Plains.

To realize the full potential of waxy wheat (Triticum aestivum L.), the wet milling properties of waxy wheat flours including their dough-mixing properties were investigated. Flours of six waxy hard wheats, one normal hard wheat ('Karl 92'), and one partial waxy hard wheat ('Trego') were fractionated by the dough-washing (Martin) process, and the yields and recoveries of starch and gluten were compared. When waxy and normal wheat starches each were blended with a wheat gluten to give a mixture containing 14.5% protein, they gave very different mixograms even though the protein was the same in those blends. Waxy wheat starch absorbed more water than normal wheat starch, which apparently retarded hydration of gluten and dough development. Higher water content had to be used for some waxy wheat flours to develop optimum dough. Washing waxy wheat flour dough under a stream of water caused dough to become slack, spread out more on the sieve, and break apart into several pieces, which when thoroughly washed, coalesced into an elastic dough like the controls. By mixing a weak dough with 2% NaCl solution or by adding hemicellulase, stickiness of the dough subsided during the washing step and thereby improved the recovery of the gluten and starch fractions.

Wheat (Triticum aestivum L.) flour properties necessary for optimal tortilla production have not been identified. Transgenic wheats (Triticum aestivum L.) overexpressing high molecular weight glutenin subunit (HMW-GS) 1Dy10 were used to make tortilla and their quality was evaluated. The level of HMW-GS 1Dy10 in flours derived from transgenic wheats was 2.5-5.8-fold greater than in controls. Polymeric proteins in the transgenic samples had a molecular weight distribution shifted toward larger polymers as indicated by increased levels of polymeric proteins present and greater M(w) averages of the largest fractions in the insoluble polymeric proteins. Dough derived from transgenic wheats had greater resistance to extension and lower extensibility than controls. Tortilla quality evaluation revealed that tortillas originated from transgenic wheats had decreased diameter, greater thickness and rupture force, and lower rollability scores and stretchability than controls. The presence of 1RS chromosomal translocations from rye (Secale cereale L.) in transgenic wheat decreased the negative effects of overexpression of HMW-GS 1Dy10, as tortillas made with this flour mostly exhibited quality properties similar to those made from control flour. Results suggested that the negative effects of overexpression of HMW-GS 1Dy10 on tortilla properties were derived from a nonideal gluten matrix formation.

This review focuses on the potential advantages and disadvantages of forages such as switchgrass (Panicum virgatum), and two small grains: sorghum (Sorghum bicolor), and wheat (Triticum aesitvum), as feedstocks for biofuels. It highlights the synergy provided by applying what is known from forage digestibility and wheat and sorghum starch properties studies to the biofuels sector. Opportunities therefore, exist to improve biofuel qualities in these crops via genetics and agronomics. In contrast to cereal crops, switchgrass still retains tremendous exploitable genetic diversity, and can be specifically improved to fit a particular agronomic, management, and conversion platform. Combined with emerging studies on switchgrass genomics, conversion properties and management, the future for genetic modification of this species through conventional and molecular breeding strategies appear to be bright. The presence of brown-midrib mutations in sorghum that alter cell wall composition by reducing lignin and other attributes indicate that sorghum could serve as an important model species for C(4)-grasses. Utilization of the brown-midrib traits could lead to the development of forage and sweet sorghums as novel biomass crops. Additionally, wheat crop residue, and wheat and sorghum with improved starch content and composition represent alternate biofuel sources. However, the use of wheat starch as a biofuel is unlikely but its value as a model to study starch properties on biofuel yields holds significant promise.

Use of near-infrared (NIR) diffuse reflectance on ground wheat meal for prediction of protein content is a well-accepted practice. Although protein content has a strong bearing on the suitability of wheat (Triticum aestivum L.) for processed foods, wheat quality, as largely influenced by the configuration and conformation of the monomeric and polymeric endosperm storage proteins, is also of great importance to the food industry. The measurement of quality by NIR, however, has been much less successful. The present study examines the effects and trends of applying mathematical transformations (pretreatments) to NIR spectral data before partial least-squares (PLS) regression. Running mean smooths, Savitzky-Golay second derivatives, multiplicative scatter correction, and standard normal variate transformation, with and without detrending, were systematically applied to an extensive set of hard red winter wheat and hard white wheat grown over two seasons. The studied properties were protein content, sodium dodecyl sulfate (SDS) sedimentation volume, number of hours during grain fill at temperature <24 degrees C, and number of hours during grain fill at temperature >32 degrees C. The size of the convolution window used to perform a smooth or second derivative was also examined. The results indicate that for easily modeled properties such as protein content, the importance of pretreatment was lessened, whereas for the more difficult-to-model properties, such as SDS sedimentation volume, wide-window (>20 points) smooth or derivative convolutions were important in maximizing calibration performance. By averaging 30 PLS cross-validation trial statistics (standard error) for each property, we were able to ascertain the inherent modeling ability of each wheat property.

A 2RL.2BS wheat-rye translocation, present in the wheat germplasm line Hamlet, carries a gene for resistance to Hessian fly biotype L, one of the most virulent biotypes presently encountered in wheat production environments. Unlike several other wheat-rye chromosome translocations common in wheat breeding programs, 2RL lacks genes encoding storage proteins or other easily selected markers. Oligonucleotide primers synthesized from published sequences derived from the R173 family of moderately repetitive rye DNA were used in the DNA polymerase chain reaction (PCR) to identify specific markers for 2RL. The same primers, when used with DNA extracted from additional wheat-rye translocation lines of importance to the wheat breeding community, gave distinctive PCR products for each genotype. The single primer pair, PAWS5 and PAWS6, may, therefore, have wide applicability for the identification of wheat-rye chromosomal translocations presently encountered in wheat breeding populations.

Wheat (Triticum aestivum L.) breeders world-wide have used rye (Secale cereale L.) as a source of genes for agronomic improvement. The 1BL/1RS wheat-rye chromosomal translocation derived from the Russian cultivars 'Kavkaz' and 'Aurora' has been among the most common means of accessing useful rye genes. Unfortunately, deleterious wheat quality effects are often associated with the presence of 1RS. The identification of genetic backgrounds capable of alleviating the deleterious effects of 1RS is crucial for its continued exploitation. End-use quality parameters and flour protein composition, as measured by size-exclusion high-performance liquid chromatography (SE-HPLC) of 373 wheat lines, derived from seven 1BL/1RS breeding populations, were analyzed. In all populations, significant quality defects were detected in 1BL/1RS lines compared to non-1RS sister lines. The detrimental quality effects resulted from alteration of the ratio of flour protein composition, especially, decreased glutenin concentrations, and increased salt-water soluble protein concentrations. The end-use quality of 1BL/1RS lines, however, was highly dependent on genetic backgrounds. The potential exists for improvement in quality through crosses between 1RS lines with high glutenin, or low salt-water soluble protein concentrations, and non-1RS lines with strong dough properties.

Sequences derived from a rye gamma secalin gene were used as primers in polymerase chain reactions using DNA obtained from a series of wheat and triticale genetic stocks. A 473-bp fragment, the predicted size based on the distance between the selected primers, was found only in rye, triticales, and wheat lines carrying rye chromosome 2RS. Use of triticale lines with various wheat chromosome substitutions confirmed the chromosomal origin of the rye-specific marker. The presence of the 473-bp PCR product was always associated with the production of 75K secalins in grain samples. Thus, the primer sequences, and the clone of origin (pSC503), were both derived from the SEC-2 locus of rye chromosome 2RS.