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PubMed1998

Dynamics of tandem repetitive Afa-family sequences in Triticeae, wheat-related species.

The Afa-family sequences in wheat-related species, Triticeae, are tandem repetitive sequences of 340 bp. All the analyzed Triticeae species carried the sequences in their genomes, though the copy numbers varied about 100-fold among the species. The nucleotide fragments amplified by PCR were cloned and sequenced, and their behavior in the evolution of Triticeae was analyzed by the neighbor-joining (NJ) method. The sequences in genomes with many copies of this family clustered at independent branches of the phylogenic tree, whereas the sequences in genomes with a few copies did not. This may suggest that Afa-family sequences had amplified several times in the evolution of Triticeae, each using a limited number of different master copies. In addition, the sequences of the A and B genomes of hexaploid common wheat indicated that the Afa-family sequences had not evolved in a concerted manner between the genomes. Furthermore, the sequences of each chromosome of the D genome of this species indicated that the sequences had amplified on all over the D-genome chromosomes in a short period.

Base Sequence↗
PubMed2004

[Effect of the 5R(5A) alien chromosome substitution on the growth habit and winter hardiness of wheat].

The growth habit, ear emergence time, and frost tolerance of wheat/rye substitution lines have been studied in cultivars Rang and Mironovskaya Krupnozernaya whose chromosome 5A is substituted with chromosome 5R of Onkhoyskaya rye. Hybrid analysis has demonstrated that the spring habit of the recipient cultivars Rang and Mironovskaya Krupnozernaya is controlled by dominant gene Vrn-A1 located in chromosome 5A. Onokhoyskaya rye has a dominant gene for the spring habit (Sp1) located in chromosome 5R. It has been found that the resultant 5R(5A) alien-substitution lines have a winter type of development and ears do not emerge during summer in plants sown in spring. The change in growth habit has been shown to be related to the absence of the rye Spl gene expression in the substitution lines. The winter hardiness of winter 5R(5A) alien-substitution lines has been studied under the environmental conditions of Novosibirsk. Testing the lines in the first winter demonstrated that their winter survival is 20-27%. The possible presence of the frost resistance gene homeoallelic to the known genes Fr1 and Fr2 of the common wheat located on chromosomes 5A and 5D, respectively, is discussed.

Adaptation, Physiological↗
PubMed1992

Gamma irradiation induced deletions in an alien chromosome segment of the wheat 'Indis' and their use in gene mapping.

Deletion mutants were produced in a translocated chromosome segment derived from Thinopyrum distichum (Thunb.) Löve. Spikes of the translocation line 'Indis' were irradiated with gamma rays at dosages of 15, 20, and 25 Gy. The irradiated spikes were pollinated with 'Inia 66' pollen and the F2 and F3 generations screened for translocation mutants, using the genes for leaf rust resistance and yellow endosperm pigmentation as markers. Finally, endopeptidase polymorphisms were utilized to select mutant translocation homozygotes within each of 29 families. An investigation of polymorphisms at the alpha-Amy-D2 and Wsp-D1 loci of chromosome arm 7DL revealed that 'Indis' did not produce an alpha-AMY-D2 product, but it did produce a novel WSP-D1 protein. The mutants were characterized for their leaf and stem rust resistances and the presence of WSP-D1 and yellow flour pigments. The stem rust resistance gene could not be accurately mapped. The linear order of the remaining loci on 7DL was centromere - leaf rust resistance - Wsp-D1 and yellow pigment. The data obtained suggested that the 'Indis' translocation has homo(eo)logy to the Lr19 translocation and homoeology to 7DL of common wheat.

Chromosome Deletion↗
PubMed2002

An estimation of the minimum number of SSR alleles needed to reveal genetic relationships in wheat varieties. I. Information from large-scale planted varieties and cornerstone breeding parents in Chinese wheat improvement and production.

The genetic relationships of 43 wheat varieties were analyzed with SSR markers. The materials employed included 14 cornerstone breeding parents used before 1980 and another 29 other large-scale planted varieties currently in use in China. A total of 501 different alleles were amplified, including 166 alleles of the A genome, 174 of the B genome and 161 of the D genome. Data obtained were used to estimate genetic similarity using the DICE coefficient, and dendrograms were constructed using the UPGMA method. The dendrogram with 501 alleles successfully differentiated all the cornerstone breeding parents and the large-scale planted varieties, and the dendogram tree was basically consistent with the pedigrees of these varieties. The correlation coefficient between the genetic distance matrix of 501 alleles and that of 450 was 0.99. Correlation coefficients among random samples of alleles suggested that 350 to 400 alleles were needed to detect genetic relationships among common wheat varieties. Correlation coefficients of a genetic similarity matrix based on 580, and those of 501 and 400, random alleles were 0.96 and 0.94, respectively. However, there were marked differences between the matrix based on the 501 alleles and those based on markers located on the A-, B- or D-genome independently. The correlation coefficients between the genetic distance matrix of 501 alleles and alleles within A, B or D genomes were 0.77, 0.76 and 0.67. The estimation of genetic similarity should be based on data from all genomes rather than any one genome.

China↗
PubMed2003

Chromosomal location of a race-specific resistance gene to Mycosphaerella graminicola in the spring wheat ST6.

Septoria tritici blotch, caused by Mycosphaerella graminicola, is a serious foliar disease of wheat worldwide. Qualitative, race-specific resistance sources have been identified and utilized for resistant cultivar development. However, septoria tritici blotch resistant varieties have succumbed to changes in virulence of M. graminicola on at least three continents. The use of resistance gene pyramids may slow or prevent the breakdown of resistance. A clear understanding of the genetics of resistance and the identification of linked PCR-based markers will facilitate the recovery of wheat lines carrying multiple septoria tritici blotch resistance genes. The resistance gene in ST6 to isolate MG2 of M. graminicola was mapped with microsatellite markers in two populations, ST6/Erik and ST6/Katepwa. Bulk segregant analysis identified a marker on chromosome 4AL putatively linked to the resistance gene. A large linkage group was identified in each population using additional microsatellite markers mapping to chromosome 4AL. The resistance gene in ST6 mapped to the distal end of chromosome 4AL in each mapping population and was designated Stb7. Three of the microsatellite loci, Xwmc313, Xwmc219 and Xgwm160, mapped within 3.5 cM of Stb7; however, none flanked Stb7. Xwmc313 was the closest and mapped 0.3 and 0.5 cM from Stb7 in the crosses ST6/Katepwa and ST6/Erik, respectively. WMC313 will be very useful for marker-assisted selection of Stb7 in Canadian breeding programs because the ST6 allele of Xwmc313 was not identified in any of the Canadian common wheat cultivars tested.

Ascomycota↗
PubMed2005

Differential and coordinated expression of Cbf and Cor/Lea genes during long-term cold acclimation in two wheat cultivars showing distinct levels of freezing tolerance.

The cold acclimation process in plants is primarily regulated through the signal transduction pathways that lead to the induction and enhancement of expression of different sets of Cor/Lea genes. Winter wheat 'Mironovskaya 808' (M808) exhibited a much higher level of freezing tolerance than spring wheat 'Chinese Spring' (CS), and the difference became clearer after the long-term cold acclimation. To understand the molecular basis of this cultivar difference, we isolated two CBF/DREB1 homologs, Wcbf2, which are the candidate gene for a transcription factor of the Cor/Lea genes. Expression of the Wcbf2 gene was induced rapidly by low temperature (LT) and drought but not by abscisic acid (ABA). The gene expression was temporal and at least twice up-regulated by LT. The first up-regulation occurred within 1-4 h, which might correspond to the rapid response to LT, while the second up-regulation occurred during 2-3 weeks of cold acclimation. After the second up-regulation, the amount of Wcbf2 transcript greatly decreased in CS, while it increased again in M808 after 4 weeks until 9 weeks (end of the test period). The maintenance of this high level of the Wcbf2 transcript might represent the long-term effect of cold acclimation. The activation of Cor/Lea genes followed the accumulation of Wcbf2 transcript suggested direct involvement of the Wcbf2 gene in the induction and enhancement of the Cor/Lea gene expression. The cultivar difference in freezing tolerance developed during different stages of cold acclimation can be at least partly explained by the differential and coordinated regulation of the predicted Cor/Lea gene signal transduction pathway that is mediated by the CBF/DREB1 transcription factors in common wheat.

Acclimatization↗
PubMed2005

Intracellular FITC-derivatization with PEG.

In order to investigate the amino acids (AAs) in plant cells, we explore an avenue for intracellular derivatization with FITC. In this method, FITC was used to mark AAs in living protoplasts derived from embryogenic calli of common wheat (Triticum aestivum L. c.v. Jinan 177) mediated by PEG. After FITC-derivatization, the AAs in the lysate were determined by CE. The result reveals that this PEG method can be used to transfer FITC into plant cells efficiently, which provides a good method for AA analysis in plant cells.

Amino Acids↗
PubMed1987

Comparisons of proteins and glycoproteins in neuronal plasma membranes, axolemma, synaptic membranes, and oligodendroglial plasma membranes.

Neuronal membranes are unique in that they consist of several functionally distinct segments: the perikaryal plasma membrane, the axolemma, the synaptic membrane, and the dendritic membrane. Methods are now available to isolate the first three types of membranes as well as to isolate oligodendroglial plasma membranes. The protein and glycoprotein compositions for each set of membranes were analyzed by silver staining after separation by SDS polyacrylamide gradient gel electrophoresis and by radiolabeled lectin binding to glycoproteins transferred to nitrocellulose. Analysis of the composition of each set of membranes reveals that they are all complex structures consisting of heterogeneous mixtures of proteins and glycoproteins, ranging in molecular weights from greater than 200,000 to 15,000. Each membrane fraction presents a unique pattern of staining and of lectin binding. As there were proteins and glycoproteins in common among the membranes, there were also differences. Synaptic membranes and axolemma appeared to have more proteins of higher molecular weight than the other membranes. Neuronal plasma membranes had a major concanavalin A binding glycoprotein at 79 kDa, which was not found in the other membranes. The three neuronal membrane fractions had a common wheat germ agglutinin binding glycoprotein at 82 kDa. The most interesting finding was the intense binding of neuronal plasma membrane glycoproteins to Ulex europaeus, suggesting high levels of fucose-containing glycoproteins.

Animals↗
PubMed1980

Arrangement of chromosomes in the interphase nucleus of plants.

Chromosomal arrangement in the interphase nucleus has two main aspects: (1) arrangement of chromosomes with respect to nuclear polarity and to other nuclear components, and (2) arrangement of chromosomes with respect to one another. The latter aspect consists of two main types of spatial relationships; (1) relationships between different members of one chromosomal set, (b) relationships between different chromosomal sets. Data concerning various aspects of chromosomal arrangement in the interphase nucleus are presented and discussed and the genetic control as well as subcellular mechanisms which are involved in nuclear organization, are elucidated. Evidence is presented indicating that, in common wheat, the gene system that determines the specific pattern of chromosomal arrangement in the nucleus is operating via the microtubular elements of the spindle system. The significance of ordered arrangement of chromosomes in the nucleus for the regularity of genetic activity and chromosomal behavior, is pointed out.

Cell Nucleus↗
PubMed2005

Adult plant and seedling resistance to powdery mildew in a Triticum aestivum x Triticum militinae hybrid line.

In the progeny of a cross between the common wheat cultivar Tähti and Triticum militinae, a member of the timopheevii group of tetraploid wheats, several hybrid lines were selected that are characterized by improved seedling and adult plant resistance (APR) to powdery mildew. An F2 single-seed descendant mapping population segregating for seedling resistance and APR to powdery mildew was analysed for the identification of quantitative trait loci (QTL). The main QTL responsible for APR was detected on the long arm of chromosome 4A tightly linked to the Xgwm160 locus on a T. militinae translocation explaining up to 54% of phenotypic variance. The same translocation influenced seedling resistance to powdery mildew upon inoculation of plants with a synthetic population of Blumeria graminis DC. f. sp. tritici, and explained 28-33% of the phenotypic variance.

Ascomycota↗
PubMed2006

Molecular cytogenetic analyses of hexaploid lines spontaneously appearing in octoploid Triticale.

Genome characterization of 14 hexaploid lines that spontaneously appeared in octoploid Triticales was carried out by sequential genomic in situ hybridization and fluorescence in situ hybridization, high molecular weight glutenin subunits and SSR marker analyses. All of the lines showed a chromosome constitution of complete A and B genomes, and a composite genome consisting of the chromosomes of D and R genomes. The composite genome of the 11 lines consisted of chromosomes 1R, 2D, 3R, 4R, 5R, 6R and 7R, that of the two lines were 1D, 2D, 3R, 4R, 5R, 6R and 7R, and that of one line was 1R, 2D, 3R, 4R, 5R, 6D and 7R. The incompatibility of the D and R genomes in common wheat genetic background, preferential retention of chromosome 2D and importance of these lines for the development of hexaploid Triticale are discussed in this report.

Chromosomes, Plant↗
PubMed2007

Metal ion binding properties of Triticum [corrected] aestivum Ec-1 metallothionein: evidence supporting two separate metal thiolate clusters.

Metallothioneins are ubiquitous low molecular mass, cysteine-rich proteins with an extraordinary high metal ion content. In contrast to the situation for the vertebrate forms, information regarding the properties of members of the plant metallothionein family is still scarce. We present the first spectroscopic investigation aiming to elucidate the metal ion binding properties and metal thiolate cluster formation of the Triticum [corrected] aestivum (common wheat) early cysteine-labeled plant metallothionein (Ec-1). For this, the protein was overexpressed recombinantly in Escherichia coli. Recombinant Ec-1 is able to bind a total of six divalent d10 metal ions in a metal thiolate cluster arrangement. The pH stability of the zinc and cadmium clusters investigated is comparable to stabilities found for mammalian metallothioneins. Using cobalt(II) as a paramagnetic probe, we were able to show the onset of cluster formation taking place with the addition of a fourth metal ion equivalent to the apo protein. Limited proteolytic digestion experiments complemented with mass spectrometry and amino acid analysis provide clear evidence for the presence of two separate metal thiolate clusters. One cluster consists of four metal ions and is made up by a part of the protein containing 11 cysteine residues, comparable to the situation found in the mammalian counterparts. The second cluster features two metal ions coordinated by six cysteine residues. The occurrence of the latter cluster is unprecedented in the metallothionein superfamily so far.

Amino Acid Sequence↗
PubMed2005

Manganese toxicity thresholds for restoration grass species.

Manganese toxicity thresholds for restoration plants have not been established. As a result, ecological risk assessments rely on toxicity thresholds for agronomic species, which may differ from those of restoration species. Our objective was to provide Mn toxicity thresholds for grasses commonly used in restoration. We used a greenhouse screening study where seedlings of redtop, slender wheatgrass, tufted hairgrass, big bluegrass, basin wildrye, and common wheat were grown in sand culture and exposed to increasing concentrations of Mn. The LC50, EC50-plant, EC50-shoot, EC50-root, PT50-shoot, and the PT50-root were then determined. Phytotoxicity thresholds and effective concentrations for the restoration species were generally higher than values reported for agronomic species. Our estimates of PT50-shoot for the five restoration grasses range from 41,528 to 120,082 mg Mn kg(-1). Measures of EC50-plant for these restoration grasses ranged from 877 to >6,000 mg Mn l(-1). These thresholds might be more useful for risk assessors than those based on crop plants that are widely used.

Decontamination↗
PubMed2005

Spelt (Triticum aestivum ssp. spelta) as a source of breadmaking flours and bran naturally enriched in oleic acid and minerals but not phytic acid.

The nutritional value of breadmaking cereal spelt (Triticum aestivum ssp. spelta) is said to be higher than that of common wheat (Triticum aestivum ssp. vulgare), but this traditional view is not substantiated by scientific evidence. In an attempt to clarify this issue, wholemeal and milling fractions (sieved flour, fine bran, and coarse bran) from nine dehulled spelt and five soft winter wheat samples were compared with regard to their lipid, fatty acid, and mineral contents. In addition, tocopherol (a biochemical marker of germ) was measured in all wholemeals, whereas phytic acid and phosphorus levels were determined in fine bran and coarse bran samples after 1 month of storage. Results showed that, on average, spelt wholemeals and milling fractions were higher in lipids and unsaturated fatty acids as compared to wheat, whereas tocopherol content was lower in spelt, suggesting that the higher lipid content of spelt may not be related to a higher germ proportion. Although milling fractionation produced similar proportions of flour and brans in spelt and wheat, it was found that ash, copper, iron, zinc, magnesium, and phosphorus contents were higher in spelt samples, especially in aleurone-rich fine bran and in coarse bran. Even though phosphorus content was higher in spelt than in wheat brans, phytic acid content showed the opposite trend and was 40% lower in spelt versus wheat fine bran, which may suggest that spelt has either a higher endogenous phytase activity or a lower phytic acid content than wheat. The results of this study give important indications on the real nutritional value of spelt compared to wheat. Moreover, they show that the Ca/Fe ratio, combined with that of oleate/palmitate, provides a highly discriminating tool to authenticate spelt from wheat flours and to face the growing issue of spelt flour adulteration. Finally, they suggest that aleurone differences, the nature of which still needs to be investigated, may account for the differential nutrient composition of spelt and wheat.

Bread↗
PubMed2002

In-situ comparative mapping (ISCM) of Glu-1 loci in Triticum and Hordeum.

The ability to detect small low- or single-copy DNA sequences by fluorescence in-situ hybridization (FISH) is an important step towards physical mapping of plant genomes. In this study, the FISH technique was used to physically map the Glu-1 loci controlling high-molecular weight (HMW) glutenin in common wheat (Triticum aestivum cv. 'Chinese Spring') and tritordeum (an amphiploid between T. turgidum cv. durum and Hordeum chilense). The probe used was the single-copy Glu-D1-1d gene coding the 1Dx5 HMW glutenin subunit. Three loci were mapped on chromosomes of wheat homoeologous group 1 (arm 1AL, 1BL and 1DL). The Glu-1 loci were mapped (fraction of the distance from the centromere) at positions 0.76 +/- 0.01, 0.69 +/- 0.01 and 0.76 +/- 0.01, on arms 1AL, 1BL and 1DL, respectively. The Glu-1 loci were also mapped on chromosomes of homoeologous group 1 of tritordeum at positions 0.75 +/- 0.01, 0.70 +/- 0.01 and 0.60 +/- 0.01, on arms 1AL, 1BL and 1HchL, respectively. Chromosomes with positive signals were identified by reprobing chromosome preparations using both the GAA-satellite and pAs1 sequences simultaneously. The application of the FISH technique to study homoeology among different genomes is discussed.

Chromosome Mapping↗
PubMed1995

Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR.

Thermal asymmetric interlaced (TAIL-) PCR is an efficient technique for amplifying insert ends from yeast artificial chromosome (YAC) and P1 clones. Highly specific amplification is achieved without resort to complex manipulations before or after PCR. The adaptation of this method for recovery and mapping of genomic sequences flanking T-DNA insertions in Arabidopsis thaliana is described. Insertion-specific products were amplified from 183 of 190 tested T-DNA insertion lines. Reconstruction experiments indicate that the technique can recover single-copy sequences from genomes as complex as common wheat (1.5 x 10(10) bp). RFLPs were screened using 122 unique flanking sequence probes, and the insertion sites of 26 T-DNA transgenic lines were determined on an RFLP map. These lines, whose mapped T-DNA insertions confer hygromycin resistance, can be used for fine-scale mapping of linked phenotypic loci.

Arabidopsis↗
PubMed2006

Construction of a primary RH panel of Italian ryegrass genome via UV-induced protoplast fusion.

Symmetric and asymmetric somatic hybrids were produced via protoplast fusion between common wheat ( TRITICUM AESTIVUM L.) cv. "Jinan 177" and Italian ryegrass ( LOLIUM MULTIFLORUM Lam.). The ryegrass without or with UV irradiation was used as a donor, providing a small amount of chromatin. In these somatic hybrids, most ryegrass chromosomes have been confirmed preferential elimination and the somatic hybrid calli and plants showed wheat-like morphology. Some of the hybrid lines were used for the analysis of distribution and heredity of donor DNA in the hybrid genome and the possibility of establishing a radiation hybrid (RH) panel of the ryegrass in the present experiment. These hybrids, subcultured for two and three years, retained the ryegrass DNA examined by RFLP and GISH analysis, respectively. Distribution of the ryegrass DNA in the wheat genomes of 20 single-cell individuals, randomly selected from hybrid cell lines produced, were analyzed by 21 ryegrass genome specific SSR markers. The average frequencies of molecular marker retention in symmetric hybrid lines (UV 0), as well as asymmetric hybrid lines from UV 30 s and 1 min were 10.88, 15.48 and 33.86, respectively. It was suggested that the UV dose increased the introgression of donor DNA into wheat genome. The ryegrass SSR fragments in most asymmetric hybrid cell lines remained stable over a period of 2 approximately 3 years. This revealed that those asymmetric somatic hybrids are suitable for the introgression of ryegrass DNA into wheat, and for RH panel and RH mapping.

Base Sequence↗
PubMed2005

Stable barley chromosomes without centromeric repeats.

The satellite sequences (AGGGAG)(n) and Ty3/gypsy-like retrotransposons are known to localize at the barley centromeres. Using a gametocidal system, which induces chromosomal mutations in barley chromosomes added to common wheat, we obtained an isochromosome for the short arm of barley chromosome 7H (7HS) that lacked the barley-specific satellite sequence (AGGGAG)(n). Two telocentric derivatives of the isochromosome arose in the progeny: 7HS* with and 7HS** without the pericentromeric C-band. FISH analysis demonstrated that both telosomes lacked not only the barley-specific centromeric (AGGGAG)(n) repeats and retroelements but also any of the known wheat centromeric tandem repeats, including the 192-bp, 250-bp, and TaiI sequences. Although they lacked these centromeric repeats, 7HS* and 7HS** both showed normal mitotic and meiotic transmission. Translocation of barley centromeric repeats to a wheat chromosome 4A did not generate a dicentric chromosome. Indirect immunostaining revealed that all tested centromere-specific proteins (rice CENH3, maize CENP-C, and putative barley homologues of the yeast kinetochore proteins CBF5 and SKP1) and histone H3 phosphorylated at serines 10 and 28 localized at the centromeric region of 7HS*. We conclude that the barley centromeric repeats are neither sufficient nor obligatory to assemble kinetochores, and we discuss the possible formation of a novel centromere in a barley chromosome.

Azure Stains↗