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A high-density genetic map of hexaploid wheat (Triticum aestivum L.) from the cross Chinese Spring x SQ1 and its use to compare QTLs for grain yield across a range of environments.

A population of 96 doubled haploid lines (DHLs) was prepared from F1 plants of the hexaploid wheat cross Chinese Spring x SQ1 (a high abscisic acid-expressing breeding line) and was mapped with 567 RFLP, AFLP, SSR, morphological and biochemical markers covering all 21 chromosomes, with a total map length of 3,522 cM. Although the map lengths for each genome were very similar, the D genome had only half the markers of the other two genomes. The map was used to identify quantitative trait loci (QTLs) for yield and yield components from a combination of 24 site x treatment x year combinations, including nutrient stress, drought stress and salt stress treatments. Although yield QTLs were widely distributed around the genome, 17 clusters of yield QTLs from five or more trials were identified: two on group 1 chromosomes, one each on group 2 and group 3, five on group 4, four on group 5, one on group 6 and three on group 7. The strongest yield QTL effects were on chromosomes 7AL and 7BL, due mainly to variation in grain numbers per ear. Three of the yield QTL clusters were largely site-specific, while four clusters were largely associated with one or other of the stress treatments. Three of the yield QTL clusters were coincident with the dwarfing gene Rht-B1 on 4BS and with the vernalisation genes Vrn-A1 on 5AL and Vrn-D1 on 5DL. Yields of each DHL were calculated for trial mean yields of 6 g plant(-1) and 2 g plant(-1) (equivalent to about 8 t ha(-1) and 2.5 t ha(-1), respectively), representing optimum and moderately stressed conditions. Analyses of these yield estimates using interval mapping confirmed the group-7 effects on yield and, at 2 g plant(-1), identified two additional major yield QTLs on chromosomes 1D and 5A. Many of the yield QTL clusters corresponded with QTLs already reported in wheat and, on the basis of comparative genetics, also in rice. The implications of these results for improving wheat yield stability are discussed.

Biomass↗

Molecular characterization and chromosomal localization of cytochrome P450 genes involved in the biosynthesis of cyclic hydroxamic acids in hexaploid wheat.

The cyclic hydroxamic acids, 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) and 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), are defensive secondary metabolites found in gramineous plants including wheat, maize and rye. cDNAs for five cytochromes P450 (P450s) involved in DIBOA biosynthesis (CYP71C6, CYP71C7v2, CYP71C8v2, CYP71C9v1 and CYP71C9v2) were isolated from seedlings of hexaploid wheat [( Triticum aestivum L. cv. Chinese Spring (2n=6x=42, genomes AABBDD)] by RT-PCR and screening of a cDNA library. CYP71C9v1 and CYP71C9v2 are 97% identical to each other in amino acid and nucleotide sequences. The cloned P450 species showed 76-79% identity at the amino acid level to the corresponding maize P450 species CYP71C1-C4, which are also required for DIBOA biosynthesis. The wheat P450 cDNAs were heterologously expressed in the yeast ( Saccharomyces cerevisiae) strain AH22. Microsome fractions from yeast cells expressing these P450 species catalyzed the same reactions as their maize orthologs. The chromosomes carrying the cyp71C6- C9v1 orthologs were identified by Southern hybridization using aneuploid lines of Chinese Spring wheat. The cyp71C9v1 orthologs were located on the chromosomes of wheat homoeologous group-4. The orthologs of the other P450 genes, cyp71C7v2, cyp71C6 and cyp71C8v2, were located on group-5 chromosomes. The same P450 genes were also present in the three ancestral diploid species of hexaploid wheat, T. monococcum (AA), Aegilops speltoides [BB (approximately SS)] and Ae. squarrosa (DD).

Amino Acid Sequence↗

A family of related sequences associated with (TTTAGGG)n repeats are located in the interstitial regions of wheat chromosomes.

A family of related sequences associated with (TTTAGGG)n repeats has been cloned from the wheat cultivar Chinese Spring. These sequences reveal a high level of polymorphism between wheat varieties when used as restriction fragment length polymorphism (RFLP) probes. Although this family of sequences contains motifs homologous to the repeats in the telomeres of wheat, they are located at interstitial sites on wheat chromosomes.

Blotting, Southern↗

Evidence for a triplicate set of glucosephosphate isomerase structural genes in hexaploid wheat.

The glucosephosphate isomerase (GPI)zymogram phenotypes of 46 aneuploid derivatives of the cultivar Chinese Spring of hexaploid wheat were determined. Variation was observed among the strains in the relative level of expression of three GPI isozymes. The relationships observed between chromosomal constitution and zymogram phenotype support the hypothesis that the three GPI isozymes are dimers composed of protomers encoded by a minimum of three homoeologous structural genes located one each in the short arms of chromosomes 1A, 1B, and 1D. The relative levels of expression per dose of chromosome arm of the products of the three arms differ in a manner consistent with the presence of a two-fold greater quantity of the product of 1BS than of the product of 1AS and of 1DS, indicating that 1BS may contain duplicate GPI structural genes.

Chromosome Mapping↗

Geographically based investigation of prostate cancer mortality in four U.S. Northern Plain states.

BACKGROUND: Historically, prostate cancer mortality rates have been elevated in the U.S. Northern Plains states. The purpose of this study was to investigate possible contributing factors, especially whether there was any association with crop patterns. METHODS: Prostate cancer mortality rates (1950-2000) in four northern plains states (MN, MT, ND, and SD) were compared to rates for 46 other U.S. states. Within the four states, county rates in urban, less urban, and rural areas also were compared. For additional analysis, urban counties and counties with <10% of county area in crops were excluded. The average percent of county area in total cropland 1930-1950 and 1954-1974 was estimated. Using Poisson regression, we investigated whether the average percentage of county area in total cropland, 1930-1950 and 1954-1974, was associated with prostate cancer mortality rates, 1975-2000, respectively. Poisson regression analyses were also used to evaluate associations between rates and major crops, which included spring and durum wheat, winter wheat, corn, and other crops. Population centroids of the Census 2000 block groups were used to estimate the percentage of males aged 35 and older residing in close proximity to small grains crops. RESULTS: Mortality rates were higher in rural compared to urban counties in 1950-2000 (rate ratio [RR]=1.032; 95% CI=1.001-1.063). Rates in 1950-1974 were significantly associated with production of corn and other crops in 1930-1950 (corn: RR per 10% increase=1.033, 95% CI=1.012-1.054; other crops: RR=1.042, 95% CI=1.021-1.063). Mortality rates in 1975-2000 were significantly associated with spring and durum wheat production in 1954-1974 (RR per 10% increase=1.042, 95% CI=1.017-1.067). Prostate cancer mortality rates increased as the percentage of population living within 500 m of small grains crops increased. CONCLUSIONS: Epidemiologic studies to evaluate agricultural practices are warranted to further evaluate the observed associations.

Adult↗

DNase I sensitivity of ribosomal RNA genes in chromatin and nucleolar dominance in wheat.

Ribosomal RNA genes at different nucleolar organizer (NOR) loci in hexaploid wheat are expressed at different levels. The degree of expression of a particular organizer depends on the genetic background, especially on the presence of other NOR loci. For example, when chromosome 1U of Aegilops umbellulata is introduced into the hexaploid wheat cultivar "Chinese Spring" the A. umbellulata NOR accounts for most of the nucleolar activity and seems to suppress the activity of the wheat NOR loci. Even in wild-type "Chinese Spring", the NOR on chromosome 1B is partially dominant to that on chromosome 6B, since the 1B locus is more active in spite of having fewer genes. We have previously shown that these and other examples of nucleolar dominance in wheat are associated with undermethylation of cytosine residues in certain regions of the dominant rDNA. Here, we show that rRNA genes at dominant loci are organized in a chromatin conformation that renders them more sensitive to DNase I digestion than other rRNA genes. In addition, we have mapped several DNase I-hypersensitive sites in the intergenic spacer region of the rDNA repeating unit. Some of these sites are located near the initiation region for the 45 S rRNA precursor, while others are associated with a series of short direct repeats 5' to the 45 S rRNA initiation site. The results are discussed in terms of a model in which repeated sequences in the wheat intergenic DNA are presumed to function as upstream promoters and transcriptional enhancers similar to those in Xenopus.

Binding Sites↗

[Dynamics of soil P pool in a long-term fertilizing experiment of wheat-maize rotation. I. Crop yield effect of fertilizer P and dynamics of soil total P and inorganic P].

The effects of long-term applying fertilizer P and manure on the pools of soil total P and inorganic P and the crop yield in rotation of winter wheat-summer maize-->spring maize were studied. The results showed that the pool of soil total P and inorganic P were increased by applying fertilizer P and manure, and the phosphorus mostly accumulated in soil was inorganic P. The critical amounts of fertilizer P (P2O5) for balancing soil P were 94.7 kg.hm-2 to winter wheat-summer maize and 51.5 kg.hm-2 to spring maize. Based on regression equations, the application rates of fertilizer P (P2O5) for economic optimum and highest yields were 135.8 and 149.8 kg.hm-2 to winter wheat-summer maize, and 88.6 and 95.9 kg.hm-2 to spring maize, respectively.

China↗

Proteome analysis of diploid, tetraploid and hexaploid wheat: towards understanding genome interaction in protein expression.

Hexaploid wheat (Triticum aestivum L.) is derived from a complex hybridization procedure involving three diploid species carrying the A, B and D genomes. The proteome patterns of diploid, tetraploid and hexaploid wheat were analyzed to explore the genome interaction in protein expression. At least two species from each of the diploid and tetraploid were used to compare their proteome maps with a hexaploid wheat cv. Chinese Spring. The ancestral cultivars were selected based on their history of closeness with the cultivated wheat. Proteins were extracted from seed flour and separated by two-dimensional electrophoresis (2-DE) with isoelectric focusing of pH range from 4-10. 2-DE maps of cultivated and ancestral species were analyzed by computer assisted image analyzer. The region of high molecular weight glutenin subunits of hexaploid wheat showed similarity with those of the diploid donors, BB and DD genomes. The omega gliadin, which is controlled by B genome in common wheat, was assumed to have evolved as a result of interaction between AA and BB genomes. The low molecular weight glutenins and alpha and beta gliadin regions were contributed by the three genomes. This result suggests that the function of donor genomes particularly in the expression of proteins in hexaploid wheat is not totally independent; rather it is the product of interactions among the diploid genomes in the hexaploid nuclear constitutions. The expression of nonstorage proteins was affected substantially due to the removal of the D genome from hexaploid constitution. Location of the structural gene controlling one of the alpha amylase inhibitor proteins in the nonstorage protein region was identified in the short arm of chromosome 3D.

Chromosome Aberrations↗

Compensating ability in pollen fertilization between group-6 and -7 homoeologous chromosomes of barley and wheat.

By using alpha-amylase isozymes as markers for chromosomes of homoeologous groups 6 and 7, we analyzed the segregation of chromosome constitution in the progenies from crosses between double-ditelosomic or ditelosomic lines of hexaploid wheat cultivar 'Chinese Spring' (CS) as the female parent and double-monosomic F1 hybrids of CS x wheat-barley substitution lines for barley chromosomes 6H or 7H. From this analysis we estimated the transmission rate via pollen of barley chromosomes 6H and 7H in the double-monosomics and evaluated the compensating ability between barley and wheat chromosomes in homoeologous groups 6 and 7. The results indicated that both 6H and 7H showed their highest compensating ability for their respective homoeologous wheat chromosomes 6A (37.5% transmission rate) and 7A (39.4%), intermediate for 6D (34.1%) and 7D (29.6%), and lowest for 6B (26.6%) and 7B (22.6%) chromosomes.

Chromosomes↗

Dissipation and mobility of the sulfonylaminocarbonyl-triazolinone herbicide propoxycarbazone in the soil of winter wheat crops.

The new sulfonylaminocarbonyltriazolinone herbicide propoxycarbazone has been applied at the rate of 70 g ha-1 post-emergence in the spring on winter wheat fields located at three sites different as to their soil texture and composition. A method has been developed for the analysis of propoxycarbazone in soil by GC and GC-MS, after isolation and transformation of propoxycarbazone. The limit of sensitivity was 1 microgram propoxycarbazone kg-1 dry soil. In the sandy-loam soil at Melle, and in the clay-loam soil at Zevekote, the propoxycarbazone soil half-lives in the 0-10 cm surface soil layer were similar, i.e. about 54 days. In the loam soil at Cortil-Noirmont, the propoxycarbazone soil half-life was 31 days. The difference as to the soil half-lives was related to the organic fertilization practized in the past on the three fields. At Cortil-Noirmont, after the winter wheat harvest at the end of August, the residues of propoxycarbazone in the 0-10 cm surface soil layer were very low, and at the end of September, propoxycarbazone was no more detected. At Melle and Zevekote, at the end of September, the concentrations of propoxycarbazone in the 0-10 cm surface soil layer were very low; at the end of October, propoxycarbazone was no more detected. After its application and until the end of October, propoxycarbazone was not detected in the 10-15 and 15-20 cm surface soil layers.

Belgium↗

Nuclear genes control changes in the organization of the mitochondrial genome in tissue cultures derived from immature embryos of wheat.

Although the mitochondrial genomes of the Chinese Spring and Aquila varieties of wheat are normally similar in organization, this is not so in tissue cultures initiated from their immature embryos where the mitochondrial genomes of both are rearranged and in different, characteristic, ways. However, the mitochondrial genomes of tissue cultures of reciprocal F1 crosses between these varieties were almost identical to one another, showing that nuclear genes control the rearrangement processes. These rearrangements are either due to the appearance of new structures or else result from changes in the relative amounts of subgenomic components. The severe reduction in the amount of certain molecular configurations in tissue cultures from reciprocal crosses is probably due to the presence of dominant information in the Aquila nuclear genome. Data obtained from tissue cultures initiated from F2 embryos of the cross Aquila x Chinese Spring suggest that at least two complementary genes are involved in this control. In contrast, the presence of new molecular arrangements appears to be under the control of a dominant allelic form of a Chinese Spring gene or genes. Thus, this study demonstrates that at least two sets of nuclear genes control the reorganization of the mitochondrial genome which occurs when tissue cultures are initiated from the immature embryos of wheat.

Blotting, Southern↗

Birth malformations and other adverse perinatal outcomes in four U.S. Wheat-producing states.

Chlorophenoxy herbicides are widely used in the United States and Western Europe for broadleaf weed control in grain farming and park maintenance. Most of the spring and durum wheat produced in the United States is grown in Minnesota, Montana, North Dakota, and South Dakota, with more than 85% of the acreage treated with chlorophenoxy herbicides such as 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-chloro-2-methylphenoxyacetic acid (MCPA). Rates of adverse birth outcomes in rural, agricultural counties of these states during 1995-1997 were studied by comparing counties with a high proportion of wheat acreage and those with a lower proportion. Information routinely collected and made available by federal agencies was used for this ecologic study. Significant increases in birth malformations were observed for the circulatory/respiratory category for combined sexes [odds ratio (OR) = 1.65; 95% confidence interval (CI), 1.07-2.55]. A stronger effect was observed for the subcategory, which excluded heart malformations (OR = 2.03; 95% CI, 1.14-3.59). In addition, infants conceived during April-June--the time of herbicide application--had an increased chance of being diagnosed with circulatory/respiratory (excluding heart) malformations compared with births conceived during other months of the year (OR = 1.75; 95% CI, 1.09-2.80). Musculoskeletal/integumental anomalies increased for combined sexes in the high-wheat counties (OR = 1.50; 95% CI, 1.06-2.12). Infant death from congenital anomalies significantly increased in high-wheat counties for males (OR = 2.66; 95% CI, 1.52-4.65) but not for females (OR = 0.48; 95% CI, 0.20-1.15). These results are especially of concern because of widespread use of chlorophenoxy herbicides.

Adolescent↗

Review of methodologies and a protocol for the Agrobacterium-mediated transformation of wheat.

Since the first report of wheat transformation by Agrobacterium tumefaciens in 1997, various factors that influence T-DNA delivery and regeneration in tissue culture have been further investigated and modified. This paper reviews the current methodology literature describing Agrobacterium transformation of wheat and provides a complete protocol that we have developed and used to produce over one hundred transgenic lines in both spring and winter wheat varieties.

Journal Article↗

Low quality roughages for steers grazing wheat pasture. I. Effect on weight gains and bloat.

The effect of feeding low quality roughages (LQR) on live and carcass weight gains and the incidence and severity of bloat of stocker cattle grazed on wheat pasture was evaluated in a 3 yr study. One hundred eighty-five steer calves (172 kg mean initial weight) grazed clean-tilled wheat pasture and were either fed no LQR or had ad libitum access to wheat straw (WS) or sorghum-Sudan hay (SS). Grazing periods were (I) fall grazing, (II) winter grazing, (III) period of lush spring growth of wheat forage and (IV) period of advancing forage maturity and declining quality. Mean dry matter (DM), crude protein and acid detergent fiber (ADF) content (percentage of DM) of wheat forage averaged across years ranged, respectively, from 23.8 to 33.0, 19.8 to 26.4 and 21.5 to 27.7. Mean daily consumption (kg DM/head) of WS and SS by steers ranged from .076 to .100 and .199 to .248, respectively. Live and carcass weight gains of steers during Periods I through III (i.e., the usual wheat pasture grazing period) were not influenced (P greater than .05) by treatments. Carcass weight gains were about 74% of live weight gains. Bloat was observed only during the last 2 wk of Period III of the first year. The incidence (steer days of bloat) and severity (bloat score) of control, WS- and SS-fed steers were 9.5 and 1.2, .5 and .5 and 2.0 and 1.0, and were not different (P greater than .05) among treatments. Intake of WS and SS [g/body weight (BW).75kg] during Periods I to III was, respectively, only about 5 and 12% of roughage intakes (i.e., 37.5 g/BW.75kg) reported in the literature to "effectively control" or aid the prevention of bloat. It seems unlikely that LQR consumed to amounts similar to those of this study would control bloat of stocker cattle on wheat pasture.

Animals↗

Isolation and location of three homoeologous dihydroflavonol-4-reductase (DFR) genes of wheat and their tissue-dependent expression.

DFR is involved in an important step in the flavonoid biosynthesis pathway upstream of anthocyanin, proanthocyanidin, and phlobaphene production, which contributes to the pigmentation of various plant tissues. Full genomic sequences of three DFRs were isolated in hexaploid wheat. Loci of TaDFRs were found in a more proximal region of the long arm of chromosomes of homoeologous group 3 than the R gene for red grain colour of wheat. These DFRs were designated TaDFR-A, TaDFR-B, and TaDFR-D on chromosome 3A, 3B, and 3D, respectively. In the 5' upstream region of DFR genes, two or three combinations of a G box core element and a putative binding site for a Myb-type transcription factor, P, of maize were found. Expression of DFR reached a maximal level in red grain of wheat cv. Chinese Spring (CS) at 5 d post-anthesis (DPA) and decreased gradually in the grain coat tissue from 10 to 20 DPA, in contrast to a very low expression level of DFR in white wheat grain during the same period. These DFRs differed in their expression. TaDFR-B and -D were expressed predominantly in grains. In developing leaves, DFR expression was light-responsive, and TaDFR-B was more up-regulated in leaves and roots than the other two.

Alcohol Oxidoreductases↗

Molecular cloning of the wheat CK2alpha gene and detection of its linkage with Vrn-A1 on chromosome 5A.

The casein kinase CK2 is one of the major multifunctional protein kinases in cells that is expressed ubiquitously and is essential for survival. The alpha-subunit of CK2 is thought to be involved in light-regulated gene expression and rhythmic expression of genes by circadian rhythm in plants. The rice chromosome-3 region containing the photoperiod-response Hd6 gene, an orthologue of the CK2alpha genes of Arabidopsis and maize, is in synteny with the wheat chromosome-5A Vrn-A1 region. This evidence proposes two possibilities, first the wheat Vrn-A1 is an orthologue of the rice CK2alpha, and second the wheat CK2alpha which has not yet been identified is located independently but tightly linked to Vrn-A1. To clarify whether the wheat CK2alpha gene is conserved in the Vrn-A1 region and to elucidate the above two possibilities, we attempted to isolate this gene from the wheat cDNA library and to map it on the chromosome-5A region that is syntenous to the rice Hd6 region. The isolated cDNA clone showed an extremely high homology with the Arabidopsis CK2alpha gene. Using this clone as a probe genomic Southern-blot analyses of the aneuploid lines available in Chinese Spring assigned the wheat homologue of CK2alpha to the long arm of chromosome 5A. Furthermore, a linkage analysis using an F(2) population having recombination in the Vrn-A1 region revealed that the wheat CK2alpha, designated as tck2a, is tightly linked to Vrn-A1 by 1.1 cM

Journal Article↗

Analysis and mapping of randomly chosen bacterial artificial chromosome clones from hexaploid bread wheat.

The current view of wheat genome composition is that genes are compartmentalized into gene-rich and gene-poor regions. This model can be tested by analyzing randomly selected bacterial artificial chromosome (BAC) clones for gene content, followed by placement of these BACs onto physical and genetic maps. Map localization could be difficult for BACs that consist entirely of repeated elements. We therefore developed a technique where repeat junctions are used to generate unique markers. Four BAC clones from hexaploid wheat variety Chinese Spring were randomly selected and sequenced at 4- to 6-fold redundancy. About 50% of the BAC sequences corresponded to previously identified repeats, mainly LTR-retrotransposons, whereas most of the remaining DNA consisted of sequences with unknown origin or function. The average gene content was <1%, although each BAC contained one or two identified genes. Repeat boundaries were amplified and used to map each clone to a chromosome arm. Extrapolation from wheat-rice comparative knowledge suggests that three of the four BAC clones originate from "gene-rich" regions of the wheat genome. Nevertheless, because these BACs carry only a single gene (two BACs) or two genes (one BAC), the predicted gene density is approximately 1 gene per 75 kb, which is considerably lower than previously estimated gene densities (one gene per 5-20 kb) for gene-rich regions in wheat. This analysis of randomly selected wheat BAC clones suggests that genes are more evenly distributed in wheat than previously believed and substantiates the need for large-scale random BAC sequencing to determine wheat genome organization.

Bread↗