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PubMed1988

Genetic analysis of Triticeae shikimate dehydrogenase.

Starch gel electrophoresis and polyacrylamide gel isoelectric focusing (IEF) were used to investigate the genetic control of Triticeae shikimate dehydrogenase-1 (SKDH-1). Studies of wheat-alien species chromosome addition lines established that Skdh-1 of Hordeum vulgare cv. Betzes is located in chromosome 5H, Skdh-V1 of Dasypyrum villosum in 5V, Skdh-R1 of Secale cereale cvs. Dakold and King II in 5R, and Skdh-S1(1) of Triticum longissimum in 5S1S. Also, the chromosomal locations of the genes that encode SKDH-1 in T. aestivum cv. Chinese Spring, T. umbellulatum, and S. cereale cv. Imperial, determined earlier using zone electrophoresis, were reconfirmed using IEF. Zone electrophoresis and IEF do not differ markedly in their ability to detect the expression of alien Skdh-1 genes in wheat-alien species chromosome addition lines. However, IEF may be superior to zone electrophoresis as a technique for detecting and analyzing SKDH-1 genetic variants within Triticeae species; among the species studied, IEF generally resolved two or more isozymes per Skdh-1 allele present, while zone electrophoresis resolved only one.

Alcohol Oxidoreductases↗

PubMed1985

Nucleotide sequence of a gene from chromosome 1D of wheat encoding a HMW-glutenin subunit.

A high molecular weight glutenin gene in hexaploid wheat has been isolated by cloning in bacteriophage lambda and characterized. The gene corresponds to polypeptide 12 encoded by chromosome 1D in the variety "Chinese Spring". The coding sequence predicted contains seven cysteine residues six of which flank a central repetitive region comprising more than 70% of the polypeptide. These findings are related to the role of high molecular weight subunits in the viscoelastic theory of gluten structure.

Amino Acid Sequence↗

PubMed2006

Development of simple sequence repeat markers specific for the Lr34 resistance region of wheat using sequence information from rice and Aegilops tauschii.

Hexaploid wheat (Triticum aestivum L.) originated about 8,000 years ago from the hybridization of tetraploid wheat with diploid Aegilops tauschii Coss. containing the D-genome. Thus, the bread wheat D-genome is evolutionary young and shows a low degree of polymorphism in the bread wheat gene pool. To increase marker density around the durable leaf rust resistance gene Lr34 located on chromosome 7DS, we used molecular information from the orthologous region in rice. Wheat expressed sequence tags (wESTs) were identified by homology with the rice genes in the interval of interest, but were monomorphic in the 'Arina' x 'Forno' mapping population. To derive new polymorphic markers, bacterial artificial chromosome (BAC) clones representing a total physical size of approximately 1 Mb and belonging to four contigs were isolated from Ae. tauschii by hybridization screening with wheat ESTs. Several BAC clones were low-pass sequenced, resulting in a total of approximately 560 kb of sequence. Ten microsatellite sequences were found, and three of them were polymorphic in our population and were genetically mapped close to Lr34. Comparative analysis of marker order revealed a large inversion between the rice genome and the wheat D-genome. The SWM10 microsatellite is closely linked to Lr34 and has the same allele in the three independent sources of Lr34: 'Frontana', 'Chinese Spring', and 'Forno', as well in most of the genotypes containing Lr34. Therefore, SWM10 is a highly useful marker to assist selection for Lr34 in breeding programs worldwide.

Alleles↗

PubMed1998

Influence of high-carbohydrate enriched diets on plasma insulin levels and insulin and IGF-I receptors in trout.

Insulin and insulin growth factor-I (IGF-I) binding to skeletal muscle semipurified receptors were assessed in rainbow trout (Oncorhynchus mykiss) fed with different enriched carbohydrate diets. The animals were fed for 2 months, either in spring, summer or autumn with a control diet (C, commercial diet containing 21% raw carbohydrates) or with two diets supplied with highly digestible carbohydrates (E1, 22% expanded wheat; and E2, 37% expanded wheat). Insulin and IGF-I receptors were semipurified by affinity chromatography (WGA-agarose). Fish fed with a carbohydrate enriched diet did not show lower growth rates than those fed with the control diet. Independently of the season, rainbow trout fed E1 and E2 presented higher insulin and glucose plasma levels as well as higher tissue glycogen reserves than fish fed C. An increase in the number of insulin receptors during the diet adaptation was observed especially in fish fed with E2. No differences in the affinity of receptors were observed. IGF-I specific binding in skeletal muscle was higher than that of insulin in all groups and in all seasons. Furthermore, IGF-I receptors showed the same tendency as insulin receptors, with increases in their number in experimentally fed fish, especially those fed with E2. Insulin and IGF-I receptors TKA increased only slightly, as a consequence of E1 and E2 diet adaptation. In conclusion, rainbow trout can be fed high-carbohydrate levels and show good rates of growth. This adaptation determines increases in circulating glucose and insulin, and muscle insulin receptors, which indicate an adaptation of the fish to higher levels of glucose supply. The response of IGF-I receptors also suggests a possible role in the regulation of metabolism.

Adaptation, Biological↗

PubMed2006

Molecular tagging of stripe rust resistance gene YrZH84 in Chinese wheat line Zhou 8425B.

Stripe rust, caused by Puccinia striiformis f. sp. tritici (PST), is one of the most damaging diseases in common wheat (Triticum aestivum L.). With the objective of identifying and tagging new genes for resistance to stripe rust, F1, F2 and F3 populations from the cross Zhou 8425B/Chinese Spring were inoculated with Chinese PST isolate CYR32 in the greenhouse. A total of 790 SSR primers were used to test the parents and resistant and susceptible bulks. The resulting seven polymorphic markers on chromosome 7BL were used for genotyping F2 and F3 populations. Results indicated that Zhou 8425B carries a single dominant resistance gene, temporarily designated YrZH84, closely linked to SSR markers Xcfa2040-7B and Xbarc32-7B with genetic distances of 1.4 and 4.8 cM, respectively. In a seedling test with 25 PST isolates, the reaction patterns of YrZH84 were different from those of lines carrying Yr2 and Yr6. It was concluded that YrZH84 is probably a new stripe rust resistance gene.

Cloning, Molecular↗

PubMed2005

Expression of fission yeast cdc25 driven by the wheat ADP-glucose pyrophosphorylase large subunit promoter reduces pollen viability and prevents transmission of the transgene in wheat.

Cell number was to be measured in wheat (Triticum aestivum) endosperm expressing Spcdc25 (a fission yeast cell-cycle regulator) controlled by a supposedly endosperm-specific promoter, AGP2 (from the large subunit of ADP glucose pyrophosphorylase). Wheat was transformed by biolistics either with AGP2::GUS or AGP2::Spcdc25. PCR and RT-PCR checked integration and expression of the transgene, respectively. In cv. Chinese Spring, AGP2::GUS was unexpectedly expressed in carpels and pollen, as well as endosperm. In cv. Cadenza, three AGP2::Spcdc25 plants, AGP2::Spcdc25.1, .2 and .3, were generated. Spcdc25 expression was detected in mature leaves of AGP2::Spcdc25.1/.3 which exhibited abnormal spikes, 50% pollen viability and low seed set per plant; both were small compared with the nonexpressing and normal AGP2::Spcdc25.2. Spcdc25 was not transmitted to the T(1) in AGP2::Spcdc25.1 or .3, which developed normally. Spcdc25 was PCR-positive in AGP2::Spcdc25.2, using primers for a central portion, but not with primers for the 5' end, of the ORF, indicating a rearrangement; Spcdc25 was not expressed in either T(0) or T(1). The AGP2 promoter is not tissue-specific and Spcdc25 expression disrupted reproduction.

Gene Expression Regulation, Plant↗

PubMed2000

Direct isolation of differentially expressed genes from a specific chromosome region of common wheat: application of the amplified fragment length polymorphism-based mRNA fingerprinting (AMF) method in combination with a deletion line of wheat.

The amplified restriction fragment length polymorphism (AFLP)-based mRNA fingerprinting (AMF) method makes it possible systematically and conveniently to identify differentially expressed cDNAs with high reproducibility. We have applied the AMF method to the cloning of the Q gene of common wheat, which is located on the long arm of chromosome 5A and pleiotropically controls the spike morphology and the threshing character of seeds. Using the AMF method, we compared the fingerprints of mRNA samples extracted from the young spikes of Triticum aestivum cv. Chinese Spring (CS) carrying the Q gene to those of a chromosome deletion line of CS, namely, q5, which lacks 15% of 5AL including the Q gene. Approximately 12,200 fragments were produced after PCR with 256 primer combinations. Of these, 92 fragments were differentially expressed between CS and q5. Northern and Southern analyses showed that 16 fragments gave specific or relatively stronger transcript signals in CS, and these clones were present in single copy or in low copy numbers in the wheat genome. Four clones were genetically mapped to the region deleted in q5. Subsequently, one clone, pTaQ22, was mapped at the same locus as the Q gene, indicating that pTaQ22 corresponds to the Q gene or is tightly linked to it. DNA sequence data showed that pTaQ22 had no homology to any known genes, thus suggesting a novel function for this gene in flower morphogenesis. This AMF method might provide a straightforward method for isolating genes in the hexaploid background of common wheat.

Base Sequence↗

PubMed2006

H22, a major resistance gene to the Hessian fly (Mayetiola destructor), is mapped to the distal region of wheat chromosome 1DS.

H22 is a major resistance gene conferring high-level of antibiosis to Hessian fly [Mayetiola destructor (Say)] larvae. It was previously assigned to wheat chromosome 1D through monosomic analysis (Raupp et al. in J Hered 84:142-145, 1993). The objective of this study was to identify molecular markers that can be used for marker-assisted selection for wheat breeding, and to further map this gene toward map-based cloning. Forty-five simple sequence repeat (SSR) and sequence-tagged site (STS) markers specific to chromosome 1D were evaluated for linkage to H22 using a segregating population consisting of 192 F(2:3) families, which were derived from the cross Tugela-Dn1 x KS85WGRC01(H22). The STS Xhor2kv and SSR Xgdm33 are two flanking markers that are tightly linked to H22 at genetic distances of 0.3 and 1.0 cM, respectively. Five other SSR markers including Xgpw7082, Xwmc147, Xcfd15, Xwmc432 and Xwmc336 were also linked to H22 at the distance from 0.8 to 20.8 cM. Analysis of Chinese Spring (CS) deletion lines revealed that all the H22-linked markers are located distal to the breakpoint of del 1DS-5, indicating that the H22 gene is located at the distal 30% region on the short arm of wheat chromosome 1D. Genomic comparison suggested that the H22 gene is located in the same or similar chromosomal region as the leaf rust resistance genes Lr21 and Lr40 on 1DS, and orthologous to the H9 gene cluster of 1AS.

Animals↗

PubMed2000

[Background analysis of chromosome controling genetic of water use efficiency of Triticum].

Water use efficiency (WUE) of flag leaf of different genetic material was measured by LCA-3 model photosynthesis apparatus. The results show as follows: The order of flag leaf WUE of different chromosome set is AA > BB > DD > RR. Among twenty Chinese Spring ditelosomic, the flag leaf WUE of A ditelosomic set is the highest, the high WUE genes locate on 1AL, 2AL, 2AS and 7AS chromosome arm. Among seven wheat-S. Cereals addition lines, the high WUE gene locates on 4R chromosome, the flag leaf WUE of 5R chromosome is the lowest. In the end of this paper, the research advance of stress resistance gene location on the fourth chromosome set of Triticum was discussed.

Chromosome Mapping↗

PubMed2004

Similarities of omega gliadins from Triticum urartu to those encoded on chromosome 1A of hexaploid wheat and evidence for their post-translational processing.

The omega-gliadins encoded on chromosome 1 of the A genome were purified from Triticum aestivum L. (2n=6 x=42, AABBDD) cv. Butte86, nullisomic 1D-tetrasomic 1A of cv. Chinese Spring (CS N1DT1A), and the diploid T. urartu (2n=2 x=14, AA ). Reverse-phase high-performance liquid chromatography combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis of gliadin extracts from CS nullisomic-tetrasomic (NT) lines confirmed the assignment to chromosome 1A. The purified omega-gliadins were characterized by mass spectrometry and N-terminal sequencing. The 1A-encoded omega-gliadins were smaller than 1B- or 1D-encoded omega-gliadins. The N-terminal amino acid sequences for 1A omega-gliadin mature peptides were nearly identical to those for the T. urartu omega-gliadins and were more similar to 1D omega-gliadin sequences than to sequences for T. monococum omega-gliadins, barley C-hordeins, or rye omega-secalins. They diverged greatly from the N-terminal sequences for the 1B omega-gliadins. The data suggest that T. urartu is the A-genome donor, and that post-translational cleavage by an asparaginyl endoprotease produces those omega-gliadins with N-terminal sequences beginning with KEL.

Amino Acid Sequence↗

PubMed2002

Characterisation of polymorphic microsatellite markers from Aegilops tauschii and transferability to the D-genome of bread wheat.

Microsatellites were isolated from a Aegilops tauschii (the D-genome donor of bread wheat) library enriched for various motifs. Primers generated from the flanking region of the microsatellites were used successfully to amplify the corresponding loci in the D genome of bread wheat. Additional amplification sometimes also occurred from the A and B genomes. The majority of the microsatellites contained (GA)(n) and (GT)(n) motifs. GA and GT repeats appeared to be both more abundant in this library and more polymorphic than other types of repeats. The allele number for both types of dinucleotide repeats fitted a Poisson distribution. Deviance analysis showed that GA and GT were more polymorphic than other motifs in bread wheat. Within each motif type (di-, tri- and tetra-nucleotide repeats), repeat number has no influence on polymorphism. The microsatellites were mapped using the Triticum aestivum Courtot x Chinese Spring mapping population. A total of 100 markers was developed on this intraspecific map, mainly on the D genome. For polyploid species, isolation of microsatellites from an ancestral diploid donor seems to be an efficient way of developing markers for the corresponding genome in the polyploid plant.

Journal Article↗

PubMed2003

Mapping genes affecting flowering time and frost resistance on chromosome 5B of wheat.

Two populations of single chromosome recombinant lines were used to map genes controlling flowering time on chromosome 5B of wheat, and one of the populations was also used to map a new frost resistance gene. Genetic maps were developed, mainly using microsatellite markers, and QTL analysis was applied to phenotypic data on the performance of each population collected from growth-room tests of flowering time and frost tolerance. Using a recombinant substitution-line mapping population derived from a cross between the substitution-line 'Chinese Spring' ('Cheyenne' 5B) and 'Chinese Spring' (CS), the gene Vrn-B1, affecting vernalization response, an earliness per se locus, Eps-5BL1, and a gene, Fr-B1, affecting frost resistance, were mapped. Using a 'Hobbit Sib' ('Chinese Spring' 5BL) x 'Hobbit Sib' recombinant substitution line mapping population, an earliness per se locus, Eps-5BL2 was mapped. The Vrn-B1 locus was mapped on the distal portion of the long arm of chromosome 5B, to a region syntenous with the segments of chromosomes 5A and 5D containing Vrn-A1 and Vrn-D1 loci, respectively. The two Eps-5BL loci were mapped close to the centromere with a 16-cM distance from each other, one in agreement with the position of a homoeologous locus previously mapped on chromosome 5H of barley, and suggested by the response of 'Chinese Spring' deletion lines. The Fr-B1 gene was mapped on the long arm of chromosome 5B, 40 cM from the centromeric marker. Previous comparative mapping data with rice chromosome 9 would suggest that this gene could be orthologous to the other Fr genes mapped previously by us on chromosomes 5A or 5D of wheat, although in a more proximal position. This study completes the mapping of these homoeoallelic series of vernalization requirement genes and frost resistance genes on the chromosomes of the homoeologous group 5 in wheat.

Acclimatization↗

PubMed2005

Characterization of low-molecular-weight glutenin subunit genes and their protein products in common wheats.

To characterize the low-molecular-weight glutenin subunit (LMW-GS), we developed specific PCR primer sets to distinguish 12 groups of LMW-GS genes of Norin 61 and to decide their loci with nullisomic-tetrasomic lines of Chinese Spring. Three, two, and ten groups were assigned to Glu-A3, Glu-B3, and Glu-D3 loci, respectively. To identify the proteins containing the corresponding amino acid sequences, we determined the N-terminal amino acid sequence of 12 spots of LMW-GSs of Norin 61 separated by two-dimensional gel electrophoresis (2DE). The N-terminal sequences of the LMW-GS spots showed that 10 of 12 groups of LMW-GSs were expressed as protein products, which included LMW-i, LMW-m, and LMW-s types. Four spots were encoded by Glu-A3 (LMW-i). Three spots were encoded by Glu-B3 (LMW-m and LMW-s). Five spots were encoded by Glu-D3 (LMW-m and LMW-s). A minor spot of LMW-m seemed to be encoded by the same Glu-B3 gene as a major spot of LMW-s, but processed at a different site. Comparing among various cultivars, there were polymorphic and non-polymorphic LMW-GSs. Glu-A3 was highly polymorphic, i.e., the a, b, and c alleles showed one spot, the d allele showed four spots, and the e allele had no spot. Insignia used as one of the Glu-A3 null standard cultivars had a LMW-GS encoded by Glu-A3. We also found that Cheyenne had a new Glu-D3 allele. Classification of LMW-GS by a combination of PCR and 2DE will be useful to identify individual LMW-GSs and to study their contribution to flour quality.

Amino Acid Sequence↗

PubMed2004

Characterisation and marker development for low molecular weight glutenin genes from Glu-A3 alleles of bread wheat (Triticum aestivum. L).

PCR was used to amplify low-molecular-weight (LMW) glutenin genes from the Glu-A3 loci of hexaploid wheat cultivars containing different Glu-A3 alleles. The complete coding sequence of one LMW glutenin gene was obtained for each of the seven alleles Glu-A3a to Glu-A3g. Chromosome assignment of PCR products using Chinese Spring nulli-tetrasomic lines confirmed the amplified products were from chromosome 1A. All sequences were classified as LMW-i-type genes based on the presence of an N-terminal isoleucine residue and eight cysteine residues located within the C-terminal domain of the predicted, mature amino acid sequence. All genes contained a single uninterrupted open reading frame, including the sequence from the Glu-A3e allele, for which no protein product has been identified. Comparison of LMW glutenin gene sequences obtained from different alleles showed a wide range of sequence identity between the genes, with between 1 and 37 single nucleotide polymorphisms and between one and five insertion/deletion events between genes from different alleles. Allele-specific PCR markers were designed based on the DNA polymorphisms identified between the LMW glutenin genes, and these markers were validated against a panel of cultivars containing different Glu-A3 alleles. This collection of markers represents a valuable resource for use in marker-assisted breeding to select for specific alleles of this important quality-determining locus in bread wheat.

Alleles↗

PubMedDate not supplied

Preferential flow of bromide, bentazon, and imidacloprid in a Dutch clay soil.

Leaching to ground water and tile drains are important parts of the environmental assessment of pesticides. The aims of the present study were to (i) assess the significance of preferential flow for pesticide leaching under realistic worst-case conditions for Dutch agriculture (soil profile with thick clay layer and high rainfall) and (ii) collect a high-quality data set that is suitable for testing pesticide leaching models. The movement of water, bromide, and the pesticides bentazon [3-isopropyl-1H-2, 1,3-benzothiadiazine-4(3H)-one-2,2-dioxide] and imidacloprid [1-[(6-chloro-3-pyridinyl)-methyl]-N-nitro-2-imidazolidinimine] was monitored in a clay soil for about 1 yr. The 1.2-ha field was located in the central part of the Netherlands (51 degrees 53' N, 5 degrees 43' E). The soil was a Eutric Fluvisol cropped with winter wheat (Triticum aestivum L.). Tile drains were present at a 0.8- to 0.9-m depth and the ground water level fluctuated between a 0.5- and 2-m depth. All chemicals were applied in spring. None of the soil concentration profiles showed bimodal concentration distributions. However, for each substance the highest concentration in drain water was found in the first drainage event after its application, which indicates preferential flow. This preferential flow is probably caused by permanent macropores that were present in the 0.3- to 1.0-m layer. At the time of the first drainage event, the drain water concentration of each substance was about an order of magnitude higher than its ground water concentration. Thus, the flux concentrations in drain water proved to be a more sensitive detector of preferential flow than the resident concentrations in the soil profile and the ground water.

Aluminum Silicates↗

PubMed2026

Genome-Wide Characterization and Salt-Responsive Expression Divergence of Chromosome Group 2 and Group 6 TaBADH Genes in Wheat.

Betaine aldehyde dehydrogenase (BADH) catalyzes the final step in glycine betaine biosynthesis, but the evolutionary divergence and differential salt responsiveness of BADH homeologs in bread wheat remain unclear. We identified six TaBADH genes and analyzed their phylogenetic relationships, conserved motifs, gene structures, promoter cis-acting elements and synteny. RNA-seq and qRT-PCR were used to compare expression in salt-tolerant Jimai 60 and salt-sensitive Chinese Spring under 200 mM NaCl, and BADH activity, glycine betaine, H2O2 and malondialdehyde (MDA) were measured during treatment. The genes separated into chromosome group 2 and group 6 clades with distinct structural and transcriptional patterns. TaBADH-2B encoded a shorter protein and lacked several conserved motifs. Group 6 genes showed stronger salt-responsive expression in Jimai 60, with TaBADH-6D displaying the strongest and most sustained induction. Jimai 60 also showed higher BADH activity and glycine betaine accumulation and lower H2O2 and MDA contents at later time points. Expression of TaBADH-6D improved E. coli growth under 200 mM NaCl. These findings identify homeolog-specific divergence within the BADH wheat family and support TaBADH-6D as a candidate for plant-level functional validation.

TaBADH-6D↗

PubMed1998

Empirie und Dogma in den < > medizinischer Wissenschaft.

Empiric and Dogmatic 'Seasons' in the Progress of Medical ScienceThe current 'postmodern' times are not simply permissive towards any theory whatever. In medicine, the 'external validity' of theories and the exact place of various healing schools has become an important subject. 'Critical appraisal' of the empiric evidence for treatment benefit in all areas of medicine, conventional or unconventional, will separate the wheat from the chaff. Pluralistic evidence-based medicine constitutes the flail for achieving this task. This happens in late summer. Later in winter, the chaff of useless medicine will decay; a new therapeutic spring will only be possible when the current task of identifying effective and beneficial treatments in all areas and of abandoning useless therapies will be fulfilled. The current primacy of the empiric study of medicine, rather than the dogmatic, should not be misunderstood as a pure empirism. Rather, we are in the empiric 'season'. Questions about mechanisms of action will remain, but will retreat to the back of the mind for some time.

Journal Article↗

PubMed2000

The apical membrane glycocalyx of MDCK cells.

The microenvironment near the apical membrane of MDCK cells was studied by quantitation of the fluorescence of wheat germ agglutinin attached to fluorescein (WGA). WGA was shown to bind to sialic acid residues attached to galactose at the alpha-2,3 position in the glycocalyx on the apical membrane. Young MDCK cells (5-8 days after splitting) showed a patchy distribution of WGA at stable sites that returned to the same locations after removal of sialic acid residues by neuraminidase treatment. Other lectins also showed stable binding to patches on the apical membrane of young cells. The ratio of WGA fluorescence emission at two excitation wavelengths was used to measure near-membrane pH. The near-membrane pH was markedly acidic to the pH 7.4 bathing solution in both young and older cells (13-21 days after splitting). Patches on the apical membrane of young cells exhibited a range of near-membrane pH values with a mean +/- SEM of 6.86 +/- 0.04 (n = 121) while the near-membrane pH of older cells was 6.61 +/- 0.04 (n = 120) with a uniform WGA distribution. We conclude that the distribution of lectin binding sites in young cells reflects the underlying nonrandom location of membrane proteins in the apical membrane and that nonuniformities in the pH of patches may indicate regional differences in membrane acid-base transport as well as in the location of charged sugars in the glycocalyx.

Animals↗