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1.
Production and characterization of a transgenic bread wheat line over-expressing a low-molecular-weight glutenin subunit gene 总被引:5,自引:0,他引:5
Masci S. D'Ovidio R. Scossa F. Patacchini C. Lafiandra D. Anderson O.D. Blechl A.E. 《Molecular breeding : new strategies in plant improvement》2003,12(3):209-222
The end-use properties, and thus the value, of wheat flours are determined to a large extent by the proteins that make up the polymeric network called gluten. Low molecular weight glutenin subunits (LMW-GS) are important components of gluten structure. Their relative amounts and/or the presence of specific components can influence dough visco-elasticity, a property that is correlated with the end-use properties of wheat flour. For these reasons, manipulation of gluten dough strength and elasticity is important. We are pursuing this goal by transforming the bread wheat cultivar Bobwhite with a LMW-GS gene driven by its own promoter. Particle bombardment of immature embryos produced several transgenic lines, one of which over-expressed the LMW-GS transgene. Southern blots confirmed that the transgene was integrated into the wheat genome, although segregation analyses showed that its expression was sometimes poorly transmitted to progeny. We have determined that the transgene-encoded LMW-GS accumulates to very high levels in seeds of this line, and that it is incorporated into the glutenin polymer, nearly doubling its overall amount. However, SDS sedimentation test values were lower from the transgenic material compared to a non transgenic flour. These results suggest that the widely accepted correlation between the amount of the glutenin polymers and flour technological properties might not be valid, depending on the components of the polymer. 相似文献
2.
Ikeda TM Nagamine T Fukuoka H Yano H 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2002,104(4):680-687
To clarify the composition of low-molecular-weight glutenin subunits (LMW-GSs) in a soft wheat cultivar, we cloned and characterized
LMW-GS genes from a cDNA library and genomic DNA in Norin 61. Based on alignment of the conserved N- and C- terminal domains
of the deduced amino-acid sequences, these genes are classified into 12 groups. One of these groups (group 5), the corresponding
gene of which has not been reported previously, contains two additional hydrophobic amino-acid clusters interrupting the N-terminal
repetitive domain. Other groups (groups 11 and 12), which were not identified in other cultivars as a protein product, showed
all eight cysteines in the C-terminal conserved domain. With specific primer sets for these groups it was revealed that Glu-D3 and Glu-A3 encoded the former and the latter, respectively. Both groups of genes were expressed in immature seeds. The presence of these
groups of LMW-GSs may affect the dough strength of soft wheat.
Received: 26 March 2001 / Accepted: 16 July 2001 相似文献
3.
Masci S Rovelli L Kasarda DD Vensel WH Lafiandra D 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2002,104(2-3):422-428
Low-molecular-weight glutenin subunits are classically divided into the B, C and D groups. Most attention has been paid to the characterisation of the B and D groups, whereas C subunits, although represented by a large number of protein components, have not been thoroughly characterised, mainly because they tend to separate with the gliadins in many fractionation procedures. Here we describe a procedure for obtaining a fraction strongly enriched in C subunits that has allowed us to determine the chromosomal location of these subunits in the bread wheat cultivar Chinese Spring. This analysis has shown that these subunits are coded on chromosome groups 1 and 6. Comparison between N-terminal amino acid sequencing of B and C subunits has shown that, whereas the former group includes mainly subunits with typical LMW-GS type sequences (76%), the C subunit group is made up almost completely of subunits with gliadin-like sequences (95%), including the alpha-type. These results indicate that the LMW-GSs are likely to be coded not only by the typical Glu-3 loci, but also by loci tightly linked to, and possibly included within, the Gli-1 and Gli-2 loci. 相似文献
4.
根据小麦低分子量谷蛋白基因保守区序列设计引物P1/P2, 采用PCR法对四川小麦地方品种AS1643的基因组DNA进行扩增, 获得1条约900 bp的片段, 分离、纯化后连接到载体pMD18-T上, 对筛选阳性克隆测序, 获得1个低分子量谷蛋白基因LMW-AS1643(GenBank登录号: EF190322), 其编码区长度为909 bp, 可编码302个氨基酸残基组成的成熟蛋白。序列分析结果表明, LMW-AS1643具有典型的低分子量谷蛋白基因的基本结构, 其推导氨基酸序列与其它已知的LMW-GS相比, 最高相似性为93.40%。生物信息学分析表明, 在LMW-AS1643低分子量谷蛋白中, 无规则卷曲含量最高, 为67.90 %, 其次是a-螺旋, 占30.46 %, b-折叠含量最少, 为1.64 %。 相似文献
5.
6.
Isolation of low-molecular-weight glutenin subunit genes from wild emmer wheat (Triticum dicoccoides) 总被引:2,自引:0,他引:2
Three low-molecular-weight glutenin subunit (LMW-GS) genes, designated LMW-Td1, LMW-Td2 and LMW-Td3, were isolated from wild emmer wheat (Triticum dicoccoides), which is the tetraploid progenitor of common wheat (T. aestivum). The complete nucleotide sequence lengths of LMW-Td1, LMW-Td2 and LMW-Td3 are 858, 900 and 1062 bp, respectively. LMW-Td1 and LMW-Td3 can encode proteins with 284 and 352 amino acid residues, respectively, whereas LMW-Td2 is a putative pseudogene due to the presence of 3 inframe stop codons in its C-terminal domain. The deduced protein sequences of the 3 genes share the same typical polypeptide structures with known LMW-GS genes containing 8 cysteines in the mature protein domains. LMW-Td1 was clearly distinguished from all known LMW-GS genes, and considered as a novel LMW-GS gene. Two hydrophobic motifs (i.e. PIIIL and PVIIL) were observed in the repetitive domain of LMW-Td3. Sequence comparison indicates that sequences of the 3 LMW-GS genes from this study are strongly similar to known LMW-GS genes. Our phylogenetic analysis suggests that LMW-Td1 and LMW-Td2 are homologous with genes on chromosome 1A, and LMW-Td3 is closely related to genes on chromosome 1B. 相似文献
7.
S. Masci R. D’Ovidio D. Lafiandra D. D. Kasarda 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2000,100(3-4):396-400
Received: 25 May 1999 / Accepted: 22 June 1999 相似文献
8.
Y.-K. Lee F. Bekes P. Gras M. Ciaffi M. K. Morell R. Appels 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,98(1):149-155
Three genes encoding the low-molecular-weight glutenin subunits (LMW-GSs), LMWG-E2 and LMWG-E4, from A-genome diploid wheat
species, and LMW-16/10 from a D-genome diploid wheat, were expressed in bacteria. The respective proteins were produced on
a relatively large scale and compared with respect to their effects on flour-processing properties such as dough mixing, extensibility
and maximum resistance; these are important features in the end-use of wheat for producing food products. The LMWG-E2 and
LMWG-E4 proteins caused significant increases in peak resistance and mixing time, compared to the control, when incorporated
into dough preparations. The LMWG-16/10 protein was qualitatively less effective in producing these changes. All three proteins
also conferred varying degrees of decrease in dough breakdown. LMWG-E2 and LMWG-E4 caused significant increases in dough extensibility,
and decreases in maximum resistance, relative to the control. LMW-16/10 did not show a significant effect on extensibility
but showed a significant decrease in maximum resistance. The refinement of relating specific features of the structure of
the LMW-GS genes to the functional properties of their respective proteins is discussed.
Received: 24 November 1997 / Accepted: 18 August 1998 相似文献
9.
Scossa F Laudencia-Chingcuanco D Anderson OD Vensel WH Lafiandra D D'Ovidio R Masci S 《Proteomics》2008,8(14):2948-2966
10.
Variation and classification of B low-molecular-weight glutenin subunit alleles in durum wheat 总被引:7,自引:0,他引:7
M. T. Nieto-Taladriz M. Ruiz M. C. Martínez J. F. Vázquez J. M. Carrillo 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,95(7):1155-1160
The B low-molecular-weight (LMW) glutenin subunit composition of a collection of 88 durum wheat cultivars was analyzed. Extensive
variation has been found and 18 different patterns were detected. Each cultivar exhibited 4–8 subunits, and altogether 20
subunits of different mobility were identified. The genetic control of all these subunits was determined through the analysis
of nine F2 populations and one backcross. Five subunits were controlled at the Glu-A3 locus, 14 at Glu-B3 and 1 at Glu-B2. At the Glu-A3 locus each cultivar possessed from zero to three bands and eight alleles were identified. At the Glu-B3 locus each cultivar showed four or five bands and nine alleles were detected. Only one band was encoded by the Glu-B2 locus. A nomenclature for these alleles is proposed and the relationship between them and the commonly used LMW-model nomenclature
is discussed.
Received: 10 February 1997 / Accepted: 25 April 1997 相似文献
11.
Zhang Xiaofei Jin Hui Zhang Yan Liu Dongcheng Li Genying Xia Xianchun He Zhonghu Zhang Aimin 《BMC plant biology》2012,12(1):1-16
Background
The polyphenolic products of the phenylpropanoid pathway, including proanthocyanidins, anthocyanins and flavonols, possess antioxidant properties that may provide health benefits. To investigate the genetic architecture of control of their biosynthesis in apple fruit, various polyphenolic compounds were quantified in progeny from a 'Royal Gala' × 'Braeburn' apple population segregating for antioxidant content, using ultra high performance liquid chromatography of extracts derived from fruit cortex and skin.Results
Construction of genetic maps for 'Royal Gala' and 'Braeburn' enabled detection of 79 quantitative trait loci (QTL) for content of 17 fruit polyphenolic compounds. Seven QTL clusters were stable across two years of harvest and included QTLs for content of flavanols, flavonols, anthocyanins and hydroxycinnamic acids. Alignment of the parental genetic maps with the apple whole genome sequence in silico enabled screening for co-segregation with the QTLs of a range of candidate genes coding for enzymes in the polyphenolic biosynthetic pathway. This co-location was confirmed by genetic mapping of markers derived from the gene sequences. Leucoanthocyanidin reductase (LAR1) co-located with a QTL cluster for the fruit flavanols catechin, epicatechin, procyanidin dimer and five unknown procyanidin oligomers identified near the top of linkage group (LG) 16, while hydroxy cinnamate/quinate transferase (HCT/HQT) co-located with a QTL for chlorogenic acid concentration mapping near the bottom of LG 17.Conclusion
We conclude that LAR1 and HCT/HQT are likely to influence the concentration of these compounds in apple fruit and provide useful allele-specific markers for marker assisted selection of trees bearing fruit with healthy attributes. 相似文献12.
13.
R. B. Gupta K. W. Shepherd 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1987,74(4):459-465
Summary One-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of reduced total protein extracts from the endosperm of hexaploid wheat revealed a new set of faintly-stained bands, having slower electrophoretic mobility than the high-molecular-weight (HMW) glutenin subunits. These new bands have been termed the E group of glutenin subunits. Analysis of aneuploid stocks of Chinese Spring wheat has shown that three of the E bands, in order of increasing electrophoretic mobility, are controlled by genes on the short arms of chromosomes 1B, 1A and 1D, respectively. The E bands are expressed only in the presence of the long arm of chromosome 1B indicating an interaction between two or more genes involved in their production in wheat endosperm. The gene on the short arm of chromosome 1D controlling an E subunit recombined freely with Tri-D1 and the centromere but not at all with Gli-D1, indicating additional complexity at the Gli-DI locus in wheat. 相似文献
14.
A wheat genomic library was screened with two synthetic oligonucleotides (24 and 25 bases in length) complementary to a partial cDNA clone encoding a glutenin gene [Thompson et al. (1983) Theor. Appl. Genet. 67, 87-96]. Glutenins are large molecular weight aggregated proteins of grain endosperm, and major determinants of bread making quality of wheat. Of the two clones obtained one was fully characterized. It contained the sequence of the high molecular weight subunit of glutenin. The amino acid sequence derived from the gene sequence reveals a mature protein (817 amino acids) with a highly repeated structure of two different motifs corresponding to the high glutamine (35.7%), glycine (20.1%) and proline (13.1%) content. The gene does not contain an intron, and possesses a typical eukaryotic promoter; the RNA initiation site is 25-30 bases downstream. 相似文献
15.
Three novel low-molecular-weight glutenin subunit (LMW-GS) genes (designated as Ht1, Ht2, and Ht3) were isolated from the genomic DNA of Hordeum brevisubulatum ssp. turkestanicum by PCR amplification (accession no. Y0695). The coding regions of Ht1, Ht2, and Ht3 were 924, 924, and 903 bp, respectively. The deduced amino acid sequences were 306, 306, and 299 amino acid residues each
with a signal peptide, a central repetitive region rich in proline and glutamine, and N-and C-terminal non-repetitive domains.
A comparison was carried out of these genes with other known B hordein genes from cultivated barley and LMW glutenin genes
from wheat. The results indicated that Ht1, Ht2, and Ht3 had a more similar structure and a higher level of homology with the LMW-GS genes than the B hordein genes. In order to investigate
the evolutionary relationship of the novel genes with the prolamin genes from barley and wheat, the phylogenetic tree was
constructed and the subfamilies of these prolamin genes were identified. The results suggested that the three novel genes
were glutenin-like proteins designated as LMW-m type genes.
The text was submitted by the authors in English. 相似文献
16.
Classification of wheat low-molecular-weight glutenin subunit genes and its chromosome assignment by developing LMW-GS group-specific primers 总被引:9,自引:0,他引:9
Long H Wei YM Yan ZH Baum B Nevo E Zheng YL 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2005,111(7):1251-1259
On the basis of sequence analysis, 69 known low-molecular-weight glutenin subunit (LMW-GS) genes were experimentally classified
into nine groups by the deduced amino acid sequence of the highly conserved N-terminal domain. To clarify the chromosomal
locations of these groups, 11 specific primer sets were designed to carry out polymerase chain reactions (PCR) with the genomic
DNA of group 1 ditelosomic lines of Chinese Spring, among which nine primer sets proved to be LMW-GS group-specific. Each group of LMW-GS genes was specifically assigned on
a single chromosome arm and hence to a specific locus. Therefore, these results provided the possibility to predict the chromosome
location of a new LMW-GS gene based on its deduced N-terminal sequence. The validity of the classification was confirmed by
the amplifications in 27 diploid wheat and Aegilops accessions. The length polymorphisms of LMW-GS genes of groups 1 and 2, and groups 3 and 4.1 were detected in diploid A-genome
and S-genome accessions, respectively. The diploid wheat and Aegilops species could be used as valuable resources of novel allele variations of LMW-GS gene in the improvement of wheat quality.
The nine LMW-GS group-specific primer sets could be utilized to select specific allele variations of LMW-GS genes in the marker-assisted
breeding.
Electronic Supplementary Material Supplementary material is available for this article at
Hai Long and Yu-Ming Wei are the two authors who have contributed equally to this paper 相似文献
17.
18.
Zhang X Liu D Yang W Liu K Sun J Guo X Li Y Wang D Ling H Zhang A 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2011,122(8):1503-1516
Low-molecular-weight glutenin subunits (LMW-GSs) play an important role in determining the bread-making quality of bread wheat.
However, LMW-GSs display high polymorphic protein complexes encoded by multiple genes, and elucidating the complex LMW-GS
gene family in bread wheat remains challenging. In the present study, using conventional polymerase chain reaction (PCR) with
conserved primers and high-resolution capillary electrophoresis, we developed a new molecular marker system for identifying
LMW-GS gene family members. Based on sequence alignment of 13 LMW-GS genes previously identified in the Chinese bread wheat
variety Xiaoyan 54 and other genes available in GenBank, PCR primers were developed and assigned to conserved sequences spanning
the length polymorphism regions of LMW-GS genes. After PCR amplification, 17 DNA fragments in Xiaoyan 54 were detected using
capillary electrophoresis. In total, 13 fragments were identical to previously identified LMW-GS genes, and the other 4 were
derived from unique LMW-GS genes by sequencing. This marker system was also used to identify LMW-GS genes in Chinese Spring
and its group 1 nulli–tetrasomic lines. Among the 17 detected DNA fragments, 4 were located on chromosome 1A, 5 on 1B, and 8 on 1D. The results suggest that this marker system is useful for large-scale identification of LMW-GS genes in bread wheat varieties,
and for the selection of desirable LMW-GS genes to improve the bread-making quality in wheat molecular breeding programmes. 相似文献
19.
O. I. Zaitseva A. A. Burakova A. T. Babkenov S. A. Babkenova M. U. Utebayev V. A. Lemesh 《Cytology and Genetics》2017,51(6):432-440
The composition and quantity of high-molecular-weight glutenin subunits plays an important role in determining the bread-making quality of wheat. Molecular-genetic analysis of allelic composition of high-molecular-weight glutenin genes in 102 bread wheat cultivars and lines from different geographical regions was conducted. Three alleles at the Glu-A1 locus, nine alleles at the Glu-B1 locus, and two alleles at the Glu-D1 locus were identified. Among the investigated cultivars and lines, 21 were characterized by intracultivar polymorphism. High allelic variation of high-molecular-weight glutenin subunit genes was shown for the collection: 21 and 9 combinations were defined in monomorphic and polymorphic cultivars and lines, respectively. However, the major part of the collection (66.7%) contained four allelic combinations: Glu-A1b Glu-B1c Glu-D1d, Glu-A1b Glu-B1c Glu-D1-2a, Glu-A1a Glu-B1c Glu-D1d, and Glu-A1b Glu-B1c Glu-D1d/Glu-D1-2a. Fourteen cultivars of bread wheat were selected, and they were characterized by a favorable allelic composition of Glu-1 loci. 相似文献
20.
McIntire TM Lew EJ Adalsteins AE Blechl A Anderson OD Brant DA Kasarda DD 《Biopolymers》2005,78(2):53-61
The high-molecular-weight glutenin subunits (HMW-GS) of wheat gluten in their native form are incorporated into an intermolecularly disulfide-linked, polymeric system that gives rise to the elasticity of wheat flour doughs. These protein subunits range in molecular weight from about 70 K-90 K and are made up of small N-terminal and C-terminal domains and a large central domain that consists of repeating sequences rich in glutamine, proline, and glycine. The cysteines involved in forming intra- and intermolecular disulfide bonds are found in, or close to, the N- and C-terminal domains. A model has been proposed in which the repeating sequence domain of the HMW-GS forms a rod-like beta-spiral with length near 50 nm and diameter near 2 nm. We have sought to examine this model by using noncontact atomic force microscopy (NCAFM) to image a hybrid HMW-GS in which the N-terminal domain of subunit Dy10 has replaced the N-terminal domain of subunit Dx5. This hybrid subunit, coded by a transgene overexpressed in transgenic wheat, has the unusual characteristic of forming, in vivo, not only polymeric forms, but also a monomer in which a single disulfide bond links the C-terminal domain to the N-terminal domain, replacing the two intermolecular disulfide bonds normally formed by the corresponding cysteine side chains. No such monomeric subunits have been observed in normal wheat lines, only polymeric forms. NCAFM of the native, unreduced 93 K monomer showed fibrils of varying lengths but a length of about 110 nm was particularly noticeable whereas the reduced form showed rod-like structures with a length of about 300 nm or greater. The 110 nm fibrils may represent the length of the disulfide-linked monomer, in which case they would not be in accord with the beta-spiral model, but would favor a more extended conformation for the polypeptide chain, possibly polyproline II. 相似文献