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Chromosomal locations of the genes for histones and a histone gene-binding protein family HBP-1 in common wheat.

The chromosomal locations of the genes in common wheat that encode the five histones and five members of the HBP (histone gene-binding protein)-1 family were determined by hybridizing their cloned DNAs to genomic DNAs of nullitetrasomic and telosomic lines of common wheat, Triticum aestivum cv. Chinese Spring. The H1 and H2a genes are located on different sets of homoeologous chromosomes or chromosome arms, namely, 5A, 5B and 5D, and 2AS, 2BS and 2DS, respectively. Genes for the other histones, H2b, H3 and H4, are found in high copy number and are dispersed among a large number of chromosomes. The genes for all members of the HBP-1 family are present in small copy numbers. Those for HBP-1a(1) are located on six chromosome arms, 3BL, 5AL, 5DL, 6AL, 6BS and 7DL, whereas those for each HBP-1a(c14), 1a(17), 1b(c1), and 1b(c38) are on a single set of homoeologous chromosome arms; 4AS, 4BL, 4DL; 6AS, 6BS, 6DS; 3AL, 3BL, 3DL; and 3AS, 3BS, 3DS, respectively. The genes for histones H1 and H2a, and for all members of the HBP-1 family except HBP-1a(1) are assumed to have different phylogenetic origins. The genes for histone 2a and HBP-1a(17) are located in the RFLP maps of chromosomes 2B and 6A, respectively. Gene symbols are proposed for all genes whose chromosomal locations have been determined.

Aneuploidy

Adenosine diphosphate glucose pyrophosphorylase genes in wheat: differential expression and gene mapping.

A full-length cDNA clone representing the large (shrunken-2) subunit of ADP-glucose pyrophosphorylase (AGP; EC 2.7.7.27) has been isolated from a cDNA library prepared from developing grain of hexaploid wheat (Triticum aestivum L., cv. Chinese Spring). The 2084-bp cDNA insert contains an open reading frame of 1566 nucleotides and primer-extension analysis indicated that the 5' end is 10 nucleotides shorter than the mRNA. The deduced protein contains 522 amino acids (57.8 kDa) and includes a putative transit peptide of 62 amino acids (6.5 kDa). The similarity of the deduced protein to the small subunit of AGP and to other AGP genes from plants and microorganisms is discussed. Northern hybridisation shows that the Agp1 genes (encoding the small subunit in the wheat endosperm) and the Agp2 genes (encoding the large subunit in the wheat endosperm) are differentially expressed in the wheat grain. Transcripts from both gene sets accumulate to high levels in the endosperm during grain development with the majority of the expression in the endopsperm rather than the embryo and pericarp layers. Although enzyme activity is detected in developing grains prior to 10 d post anthesis, only the Agp1 genes are active at this time (the Agp2 genes are not expressed until 10 d post anthesis). The possibility that the enzyme expressed during early grain development is a homotetramer of small subunits is discussed. The Agp1 and Agp2 genes are arranged as triplicate sets of single-copy homoeoloci in wheat. The Agp2 genes are located on the long arms of chromosomes 1A, 1B and 1D, about 80 cM from the centromere. The Agp1 genes have been mapped to a position just distal to the centromere on the long arms of chromosomes 7A, 7B and 7D.

Amino Acid Sequence

Identification of three Wx proteins in wheat (Triticum aestivum L.).

Nullisomic analysis of waxy (Wx) protein of hexaploid wheat (Triticum aestivum L.) cv. "Chinese Spring" using two-dimensional polyacrylamide gel electrophoresis revealed that three Wx loci, Wx-A1, Wx-B1, and Wx-D1, located on chromosome arms 7AS, 4AL, and 7DS, produce three distinct Wx subunit groups, subunit group-A (SGA), SGB, and SGD, respectively. SGA has a higher molecular weight and a more basic isoelectric point (pI) than the other two. SGB and SGD have the same molecular weight but a slightly different pI range. Owing to the detection of these three subunit groups, we were able to identify the expression of three waxy genes in wheat endosperm and to find two types of mutants among Japanese wheat cultivars, one lacking SGA and the others SGB. These results suggest the possibility of breeding a waxy wheat.

Electrophoresis, Polyacrylamide Gel

Abundance, variability and chromosomal location of microsatellites in wheat.

The potential of microsatellite sequences as genetic markers in hexaploid wheat (Triticum aestivum) was investigated with respect to their abundance, variability, chromosomal location and usefulness in related species. By screening a lambda phage library, the total number of (GA)n blocks was estimated to be 3.6 x 10(4) and the number of (GT)n blocks to be 2.3 x 10(4) per haploid wheat genome. This results in an average distance of approximately 270 kb between these two microsatellite types combined. Based on sequence analysis data from 70 isolated microsatellites, it was found that wheat microsatellites are relatively long containing up to 40 dinucleotide repeats. Of the tested primer pairs, 36% resulted in fragments with a size corresponding to the expected length of the sequenced microsatellite clone. The variability of 15 microsatellite markers was investigated on 18 wheat accessions. Significantly, more variation was detected with the microsatellite markers than with RFLP markers with, on average, 4.6 different alleles per microsatellite. The 15 PCR-amplified microsatellites were further localized on chromosome arms using cytogenetic stocks of Chinese Spring. Finally, the primers for the 15 wheat microsatellites were used for PCR amplification with rye (Secale cereale) and barley accessions (Hordeum vulgare, H. spontaneum). Amplified fragments were observed for ten primer pairs with barley DNA and for nine primer pairs with rye DNA as template. A microsatellite was found by dot blot analysis in the PCR products of barley and rye DNA for only one primer pair.

Base Sequence

Restriction fragment length polymorphism (RFLP) analysis in wheat. II. Linkage maps of the RFLP sites in common wheat.

Sixty-six F2 plants from the cross, Triticum aestivum cv. Chinese Spring (abbrev. CS) x T. spelta var. duhamelianum (Spelta), exhibiting the greatest number of RFLPs among eight common wheats, were analyzed for their RFLP genotypes using genomic DNA clones of CS as probes. In total, 204 RFLP loci were identified and their linkage relationships established. By nulli-tetrasomic analyses, all linkage groups were assigned to one another of the 21 wheat chromosomes. In addition, the carrier chromosomes of 228 non-RFLP loci were identified. The linkage maps of these RFLP loci have a total size of 1800 cM and exceed those of the classical genes in both size and locus number. Twenty loci show distorted segregation, four of which are clustered on chromosome 4A and three on the 2D chromosome. The CS alleles on 4A exhibit preferential transmission, while those on 2D exhibit depressed transmission, compared with Spelta alleles. This suggests the influence of gametic factors in those regions. RFLP loci are much fewer in the D genome than in the A and B genomes, but the numbers of non-RFLP loci are nearly the same in these three genomes. This suggests that Spelta wheat originated from a hybridization between T. dicoccum (spelt emmer) and T. aestivum.

Alleles

The effect of enzyme supplementation on the apparent metabolizable energy and nutrient digestibilities of wheat, barley, oats, and rye for the young broiler chick.

The influence of enzyme supplementation on the bioavailable energy (AME(n)) and apparent digestibilities of lipid (ALD) and protein (APD) in young broiler chicks was examined for diets containing either wheat, hulled or hulless barley, naked oats, or spring rye. Dietary AME(n), APD, and ALD values were depressed (P less than or equal to .01) for all test grains (except hulled Bedford barley) as the inclusion rate of the grain replacing wheat increased. The antinutritives, beta-glucans (barley and oats) and pentosans (rye), had the most pronounced effect on ALD. The decreases in ALD were 43, 77, and 67% for chicks fed diets containing 70% Scout barley (hulless), Terra oats, and Gazelle rye, respectively, compared with those fed the control wheat diet. Enzyme supplementation increased (P less than or equal to .01) AME(n), APD, and ALD for all test cereals. The corresponding increases in the AME(n), of the enzyme-supplemented diets containing 70% HY320 wheat, Bedford barley, Scout barley, Terra oats, and Gazelle rye diets were 4, 7, 42, 33, and 14%, respectively, compared with their unsupplemented counterparts. Enzyme treatment also improved (P less than or equal to .01) weight gains and feed conversion efficiencies of chicks fed diets containing each of the cereals. Overall, the results demonstrate that the nutritive value of cereal grains such as wheat, barley, oats, and rye can be improved by the addition of crude fungal extracts to the diet of young chicks.

Animal Feed

Mycotoxin formation in HY-320 wheat during granary storage at 15 and 19% moisture content.

Eleven-kilogram parcels of HY-320 wheat, a cultivar of the new Canada Prairie Spring class, were kept at 15 and 19% initial moisture contents (IMC) in simulated storage in a Manitoba farm granary for 60 weeks to determine biotic and abiotic changes and mycotoxin production. Ochratoxin A reached a maximum of 0.24 ppm by week 20 in the 19% IMC wheat, but was absent in the 15% IMC wheat; no other mycotoxins were detected. Temperature, moisture content, O2 and CO2 levels, fat acidity values, seed germination, microfloral incidence and abundance, and the presence of other mycotoxins were monitored. Principal component analysis of all variables showed that the first principal components accounted for 32-41% of the system variability, and contained the ochratoxin A variable. Ochratoxin A was produced in moist grain that had decreased seed germination and Alternaria activity, and high fungal activity by Penicillium and Aspergillus versicolor. Compared to other stored cereals previously studied, HY-320 wheat would be ranked in a low-risk category for mycotoxin formation, based on the ochratoxin A levels observed.

Carbon Dioxide

Two-dimensional electrophoresis of 1D-encoded B and D glutenin subunits in common wheats with similar omega gliadins.

Gli-D1-encoded omega gliadins of bread wheats show little variation; their electrophoretic patterns can be classified into two main groups which broadly resemble the patterns found in the cultivars Chinese Spring and in Cheyenne. B and D subunits of low molecular weight glutenin encoded by the chromosome 1D loci Glu-D3 and Gli-D1, respectively, also showed little variation. D subunits were found only in bread wheats with "Chinese Spring-type" omega gliadins and they all exhibited the same electrophoretic pattern. This material also showed very similar B subunits. "Cheyenne-type" bread wheats displayed the same electrophoretic distribution of chromosome 1D-encoded B subunits, although they were slightly different from that found in Cheyenne itself.

Electrophoresis, Gel, Two-Dimensional

Escape protein for beef cows: II. Source and level in ammoniated wheat straw-corn silage diets.

Two experiments, using Angus x Hereford spring-calving beef cows in mid- or late lactation nursing Simmental-sired calves, were conducted to evaluate the relative value of a corn gluten meal-blood meal mixture (CGM-BM; 50% of supplemental protein from each source). In Exp. 1 (78 d), cows in late lactation were assigned to one of three treatments: control at 8.2% CP (C), soybean meal at 10.4% CP (SBM), or CGM-BM at 10.3% CP. Diets were calculated to be isocaloric at 55% TDN. In Exp. 2 (65 d), cows in mid-lactation were assigned to four treatments: urea, SBM, low CGM-BM (LM), and high CGM-BM (HM). Diets in Exp. 2 were isonitrogenous (9.5% CP) and isocaloric (55% TDN). Diets in both experiments were based on ammoniated wheat straw and corn silage. Weight gains of cows and cow-calf pairs were greater (P less than .06) when protein was supplemented in Exp. 1. Gains were lower for cows fed urea (P less than .03) in Exp. 2 but were similar when cows were supplemented with SBM vs either the low or the high level of CGM-BM. Performance of calves did not differ among dietary treatments.

Animal Feed

Plant contents of magnesium, calcium and potassium in relation to ruminant nutrition.

Grass tetany and wheat pasture poisoning are metabolic diseases of mature, lactating beef cattle. In some other countries, similar problems occur with dairy cattle. In grass tetany, the animals generally are grazing cool-season forages in which Mg concentration or bioavailability of plant Mg is low. Levels of Mg in the blood serum also generally are low. Grass tetany can occur in beef cattle fed hay made from grass, small grains or alfalfa Medicago sativa L.). In wheat pasture poisoning where animals are grazing small grains forages, blood levels of Ca often are low, and blood Mg also may be low. Both grass tetany and wheat pasture poisoning occur when plants are growing rapidly in the spring, at the time of heavy lactation demand by ruminants for Mg and Ca. When the temperature increases and plants start to grow rapidly in the spring, concentrations of K, N, organic acids, and the ratio of K/(Ca + Mg) all increase, and the percent dry matter decreases. Much research remains to be done to understand all the soil, plant, and animal aspects of grass tetany and wheat pasture poisoning.

Animal Nutritional Physiological Phenomena

Genetic control of shikimate dehydrogenase in hexaploid wheat.

The genetics of shikimate dehydrogenase (SKDH; EC 1.1.1.25) was investigated in Triticum aestivum cv Chinese Spring (2n = 6x = 42; genomic formula ABD) using the zymogram technique. The enzyme occurs in two electrophoretically distinct forms on starch gels. The results of a study of aneuploid derivatives of Chinese Spring indicate that the SKDH isozyme of faster electrophoretic mobility is encoded by a gene, designated Skdh-A1, located in the p (= short) arm of chromosome 5A and that the products of two other genes, designated Skdh-B1 and Skdh-D1, located one each in the p arms of homoeologous chromosomes 5B and 5D, respectively, encode two isozymes of slower and coincident electrophoretic mobility. Additional evidence for this interpretation of the genetic basis of hexaploid wheat SKDH was obtained in studies of the SKDH zymogram phenotypes of various close relatives of hexaploid wheat, including T. monococcum, T. longissimum, T. tauschii, T. turgidum, and T. timopheevii.

Alcohol Oxidoreductases

Molecular analysis of an alcohol dehydrogenase (Adh) gene from chromosome 1 of wheat.

We have cloned and determined the nucleotide sequence of a gene encoding alcohol dehydrogenase (Adh) from Triticum aestivum cv. Millewa. Southern analysis using cv. Chinese Spring nullisomic-tetrasomic and ditelosomic lines established that the cloned gene mapped to the long arm of chromosome 1A and does not correspond to any previously identified wheat Adh locus. Southern analysis also provided evidence for triplicate copies of this Adh gene on the homoeologous group 1 chromosomes, while Northern blots indicated that the homoeologous group 1 Adh genes, like several other plant Adh genes, are transcribed under anaerobic conditions. Sequence analysis indicates that the cloned gene has a structure similar to both monocot and dicot Adh genes with an open reading frame encoding a polypeptide of 379 amino acids. Sequences important for eucaryotic gene expression such as the TATA box, polyadenylation signal, and intron splice sites were found in the expected positions. The open reading frame is interrupted by 8 introns which are in identical positions with 8 of the 9 introns in maize and pea Adh genes, suggesting that during evolution there are processes occurring that result in the loss of introns. Sequence analysis also revealed that the cloned wheat Adh gene shared extensive homology with the barley Adh3 gene not only in the coding region but also in the noncoding regions. However, this homology is discontinuous as a result of a 1.8-kbp insertion (TLM), which is present in the cloned wheat Adh gene and absent in the barley Adh3 gene. Sequence analysis of this insertion reveals features characteristic of the short terminal inverted repeat class of eucaryotic transposable elements. We have no evidence for the transposition of the TLM element. However, Southern blots reveal multiple copies of sequences related to TLM in the wheat genome and in other closely related species, suggesting that transposition may once have played an important role in the evolution of the Gramineae family.

Alcohol Dehydrogenase

A 'zebra' chromosome arising from multiple translocations involving non-homologous chromosomes.

An alloplasmic wheat line carrying a 'zebra' chromosome z5A was isolated from the derivatives of an Elymus trachycaulus x Triticum aestivum cv Chinese Spring hybrid. Chromosome z5A was named zebra because of its striped genomic in situ hybridization pattern. z5A consists of four chromosome segments derived from E. trachycaulus and four chromosome segments, including the centromere, from wheat. The short arm of z5A paired with the telocentric chromosome 1H(t)S of E. trachycaulus and the long arm with the long arm of normal 5A. z5A also carried several genetic markers derived from 1H(t)S. Chromosome 1H(t) was the only E. trachycaulus chromosome found in the sib plants of a previous generation from which z5A was derived. Monosomic 5A and telocentric chromosome 5AL were also found in most of the sib plants. The zebra chromosome most probably originated from spontaneous multiple translocations between chromosomes 5A and 1H(t)S or 5A and 1H(t).

Chromosome Banding

[RAPD markers for wheat chromosomes in Langdon disomic substitution lines].

Optimal conditions were developed for a random amplified polymorphio DNA (RAPD) assay of hexaploid bread wheat and tetraploid durum wheat. AmpliTaq Stoffcl fragment was found to be better than Taq DNA polymerase in generating RAPDs. Studies on chromosome specific RAPD markers of the A- and B- and D-genome were performed using the complete set of Langdon disomic substitution lines and the parental lines (Langdon and Chinese Spring) as templete. Seven out of twelve arbitrary primers (all Operon 10-mer sequences) yielded 13 products that could be assigned to 1.0 chromosomes of A- and B- and D-genome, five of 13 markers for A-genome (2A: J6a and J11b; 3A: D11b; 6A: J17; 7A: J15a), seven for B-genome (1B: J11c; 2B: D5, D11c and J18) and one for D-genome (1D: J11a). Using Chinese Spring ditelosomic lines, four RAPD markers were further mapped to a specific chromosome arm (i.e., J11b-2AL, J17-6AL, D11c-2BL, and J11a-1DL). This study demonstrates that reproduoible RAPID markers can be generated and assigned to wheat chromosomes except 4AL, using Langdon disomic substitution lines and Chinese Spring euploid and aneuploids as malerids.

Base Sequence

Effects of animal and supplement characteristics on average daily gain of grazing beef cattle.

Effects of animal gender and age, use of a growth stimulant, and supplementation with grain alone or grain plus other substances on ADG by growing beef cattle grazing bermudagrass paddocks with sod-seeded rye, wheat, and ryegrass were determined. Two grazing experiments (Exp. 1: late winter through mid-spring; Exp. 2: late spring through mid-summer) were conducted. Experiment 1 used 96, 12- to 13-mo-old Simmental-cross calves (heifers, 240 kg; steers, 272 kg), half of which were implanted with zeranol. Within each implant treatment, cattle received no supplement or .5% BW (DM) of ground corn alone or plus a mix of protein meals, zinc sulfate, thiamin-HCl, or salt. Daily gain was higher (P less than .05) with than without supplementation and was similar (P greater than .10) among supplement treatments. In Exp. 2, 96 crossbred beef steers, approximately 7 (230 kg) or 15 mo old (250 kg), were not supplemented (control) or received .5% BW (DM) of ground corn on d 1 to 84 (C-C), corn plus a protein meal mix on d 1 to 84 (CP-CP), corn on d 43 to 84 (O-C), corn plus the protein meal mix on d 43 to 84 (O-CP), or corn on d 1 to 42 and corn plus the protein meal mix on d 43 to 84 (C-CP). Daily gain on d 1 to 84 was affected (P less than .05) by supplement, age, implant, and the supplement x implant interaction (nonimplanted: .37, .56, .68, .40, .49, and .49; implanted: .37, .62, .54, .49, .70, and .71 kg for control, C-C, CP-CP, O-C, O-CP, and C-CP, respectively).

Adipose Tissue

Nitrate reductase activity (in vivo and in vitro) of ditelosomic stocks of wheat (Triticum aestivum L.).

The nitrate reductase activities (NRA) of 31 ditelosomic stocks were compared with that of the control plant [Chinese Spring (CS) euploid], using in vivo and in vitro assay procedures that had been optimized with respect to the euploid. Fourteen stock exhibited significant differences in in vivo NRA from that of the euploid; the effect of removal of a chromosome arm was always to increase NRA. Eight of these stocks showed similar effects in vitro, although in three, a casein-sensitive factor had to be eliminated before the difference was expressed. Homoeologous group effects were evident among ditelosomics of groups 2, 4, and 7, while for three chromosomes (2D, 7A, and 7B), removal of either are resulted in a similar increase in NRA in vivo and probable in vitro.

Genes

Genomic introgressions from wild relatives in the wheat genome alter meiotic dynamics in inter-varietal hybrids.

The use of wild relatives to introduce original diversity in the genome of bread wheat (Triticum aestivum L.) is an interesting approach to face the challenges of sustainable agriculture and the impact of climate change on wheat production. However, the influence of these wild-species introgressions on meiosis in inter-varietal wheat hybrids remains poorly understood. We analyzed the French wheat variety Renan (Re) carrying Aegilops ventricosa (Aev)-derived 2AS/2NS and 7DL/7DvL introgressions, the reference cultivar Chinese Spring (CS), which lacks these introgressions, and their inter-varietal hybrid Chinese Spring × Renan (CSRe). This analysis combined cytogenetic approaches with the assessment of reproductive performance. Furthermore, we generated a cytological atlas of meiosis in wild tetraploid Aev, quantifying bivalent configurations and chiasma frequency. We observed a reduced pollen viability and a slight decrease in floret fertility in the hybrid CSRe. Exploration of the meiotic behavior showed that CSRe exhibited increased numbers of rod bivalents and univalents, leading to a reduced average chiasma number and frequent chromosome bridges and fragmentations, whereas the parental lines maintained stable chromosome pairing. These rearrangements indicate that homologous chromosome pairing and recombination are affected in CSRe. We applied introgression-specific oligo-Fluorescent In Situ Hybridization to localize alien segments in CSRe, providing a novel strategy to investigate the meiotic behavior of introgressed regions. The 2AS/2NS introgressed segments in CSRe were frequently located on rod bivalents or univalents, while 7DL/7DvL segments consistently formed ring bivalents. Our results provide a foundation for guiding alien gene introgression and for understanding the behavior of chromosomes with introgressions in the wheat genome.

Meiosis

Influence of homoeologous chromosomes on gene-dosage effects in allohexaploid wheat (Triticum aestivum L.).

Gene-dosage responses for a group of six wheat endosperm proteins have been investigated by using compensated nulli-tetrasomic lines of cv. Chinese Spring. Practically linear dosage responses have been observed for all the proteins. However, for two of the proteins (and probably for a third one), the net output of protein, at each dosage of its structural gene, was 30-80% higher when the chromosome carrying an active homoeogene was absent. The possible significance of this effect in connection with the loss of gene redundancy undergone by polyploids is discussed.

Alleles