共查询到20条相似文献,搜索用时 15 毫秒
1.
Y.-K. Lee M. Ciaffi R. Appels M. K. Morell 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,98(1):126-134
Three accessions of T. boeoticum were selected for the cloning and sequencing of novel low-molecular-weight glutenin subunit (LMW-GS) genes, based on the
results of SDS-PAGE and PCR analyses of the LMW-GS diversity in A-genome wheat (Lee et al. 1998 a). A comparison of the nucleotide
and deduced amino-acid sequences of three cloned genes, LMWG-E2, LMWG-E4 and LMWG-AQ1, both to each other and to other known
LMW-GS genes was carried out. The N-terminal domains showed one variable position; GAG (coding for a glutamic acid) for the E-type, and GAT (coding for an aspartic acid) for the Q-type. The comparisons of the LMW-GSs in the literature and this paper define three
different types of N-terminal sequences; METSCIPGLERPW and MDTSCIPGLERPW from the durum and A-genome wheats, and METRCIPGLERPW from the hexaploid and D-genome wheats. The repetitive domains were AC-rich at the nucleotide level and coded for
a large number of glutamine residues; this region showed 16 variable positions changing 12 amino-acid residues, three triple
nucleotide deletions/additions, a large deletion of 18 nucleotides in LMWG-E4 and a deletion of 12 nucleotides in LMWG-E2.
In the C-terminal domains 26 variable positions were found and 12 of these mutations changed amino-acid residues; no deletions/
additions were present in this region. It was shown that the LMWG-E2 and LMWG-E4 genes could be expressed in bacteria and
this allowed the respective protein products to be related back to the proteins defined as LMW-GSs in vivo.
Received: 24 November 1997 / Accepted: 18 August 1998 相似文献
2.
Sequence similarity between allelic Glu-B3 genes related to quality properties of durum wheat 总被引:14,自引:0,他引:14
R. D’Ovidio C. Marchitelli L. Ercoli Cardelli E. Porceddu 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,98(3-4):455-461
Low-molecular-weight glutenin subunits (LMW-GSs) are wheat endosperm proteins mostly encoded by genes located at the Glu-3 loci. These proteins are of particular interest in durum wheat because a correlation between LMW-GSs encoded by genes at
the Glu-B3 locus and the pasta-making quality of durum wheat semolina has been shown. We isolated and characterized two allelic lmw-gs genes located at the Glu-B3 locus and present in durum wheat lines displaying different qualitative properties. The clones pLMW1CL and λLMW3.1 were found
to contain allelic sequences encoding LMW-GSs belonging to the good and poor quality-related groups named LMW-2 and LMW-1,
respectively. The LMW-GSs specified by these genes have very large repetitive domains which are composed of repeats regularly
distributed along the domain. The main difference between these two proteins is an insertion of 13 amino acids within the
repetitive domain which, by itself, seems insufficient to explain the qualitative differences between LMW-2 and LMW-1. These
results further support the hypothesis that the greater amount of LMW-2, rather than sequence peculiarities, accounts for
the better quality observed in durum wheat cultivars possessing these subunits. The characterization of the complete primary
structure of these alleles, other than providing information for an understanding of the structure-function relationship among
LMW-GSs and furnishing basic material for wheat engineering, should also assist in our understanding of the evolutionary relationship
between the different lmw-gs genes.
Received: 8 May 1998 / Accepted: 5 August 1998 相似文献
3.
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
4.
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
5.
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
6.
Y.-K. Lee F. Bekes R. Gupta R. Appels M. K. Morell 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,98(1):119-125
A Tris-Tricine gel-electrophoresis system (Schaegger and von Jagow 1987), combined with a gradient gel, has been employed
to provide an improved resolution of the B and C low-molecular-weight glutenin subunits (LMW-GSs) found in the endosperm of
wheat grain. The gel system was used to document the variation in the gluten subunit proteins present in A-genome diploid
wheats. The majority of LMW-GSs found in the A-genome diploid wheats were not present in normal bread wheats; the data suggest
that they represent a rich source of new variation for the LMW-GSs which are considered to be very important in modulating
wheat flour-processing properties. The analysis of variation in the nature of the LMW-GS genes, using PCR, demonstrated that
the subclass of C-subunits assayed by primers from a previously published sequence did not show as much variation as the proteins.
However, the data collected suggest that sufficient variation may exist in the LMW-GS genes of A-genome diploid wheats to
use them as a source of genes for altering the flour-processing properties of hexaploid wheat.
Received: 24 November 1997 / Accepted: 18 August 1998 相似文献
7.
8.
Zhang X Liu D Jiang W Guo X Yang W Sun J Ling H Zhang A 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2011,123(8):1293-1305
Low-molecular-weight glutenin subunits (LMW-GSs) are encoded by a multi-gene family and are essential for determining the
quality of wheat flour products, such as bread and noodles. However, the exact role or contribution of individual LMW-GS genes
to wheat quality remains unclear. This is, at least in part, due to the difficulty in characterizing complete sequences of
all LMW-GS gene family members in bread wheat. To identify full-length LMW-GS genes, a polymerase chain reaction (PCR)-based
method was established, consisting of newly designed conserved primers and the previously developed LMW-GS gene molecular
marker system. Using the PCR-based method, 17 LMW-GS genes were identified and characterized in Xiaoyan 54, of which 12 contained
full-length sequences. Sequence alignments showed that 13 LMW-GS genes were identical to those found in Xiaoyan 54 using the
genomic DNA library screening, and the other four full-length LMW-GS genes were first isolated from Xiaoyan 54. In Chinese
Spring, 16 unique LMW-GS genes were isolated, and 13 of them contained full-length coding sequences. Additionally, 16 and
17 LMW-GS genes in Dongnong 101 and Lvhan 328 (chosen from the micro-core collections of Chinese germplasm), respectively,
were also identified. Sequence alignments revealed that at least 15 LMW-GS genes were common in the four wheat varieties,
and allelic variants of each gene shared high sequence identities (>95%) but exhibited length polymorphism in repetitive regions.
This study provides a PCR-based method for efficiently identifying LMW-GS genes in bread wheat, which will improve the characterization
of complex members of the LMW-GS gene family and facilitate the understanding of their contributions to wheat quality. 相似文献
9.
Hai Long Yuming Wei Zehong Yan Bernard Baum Eviatar Nevo Youliang Zheng 《中国科学C辑(英文版)》2006,49(4):322-331
The thirty-three 5′ flanking conserved sequences of the known low-molecular-weight subunit (LMW-GS) genes have been divided
into eight clusters, which was in agreement with the classification based on the deduced N-terminal protein sequences. The
DNA polymorphism between the eight clusters was obtained by sequence alignment, and a total of 34 polymorphic positions were
observed in the approximately 200 bp regions, among which 18 polymorphic positions were candidate SNPs. Seven cluster-specific
primer sets were designed for seven out of eight clusters containing cluster-specific bases, with which the genomic DNA of
the ditelosomic lines of group 1 chromosomes of a wheat variety ‘Chinese Spring’ was employed to carry out chromosome assignment.
The subsequent cloning and DNA sequencing of PCR fragments validated the sequences specificity of the 5′ flanking conserved
sequences between LMW-GS gene groups in different genomes. These results suggested that the coding and 5′ flanking regions
of LMW-GS genes are likely to have evolved in a concerted fashion. The seven primer sets developed in this study could be
used to isolate the complete ORFs of seven groups of LMW-GS genes, respectively, and therefore possess great value for further
research in the contributions of a single LMW-GS gene to wheat quality in the complex genetic background and the efficient
selections of quality-related components in breeding programs. 相似文献
10.
Hai Long Yuming Wei Zehong Yan Bernard Baum Eviatar Nevo Youliang Zheng 《中国科学:生命科学英文版》2006,49(4):322-331
The thirty-three 5′ flanking conserved sequences of the known low-molecular-weight subunit (LMW-GS) genes have been divided into eight clusters, which was in agreement with the classification based on the deduced N-terminal protein sequences. The DNA polymorphism between the eight clusters was obtained by sequence alignment, and a total of 34 polymorphic positions were observed in the approximately 200 bp regions, among which 18 polymorphic positions were candidate SNPs. Seven cluster-specific primer sets were designed for seven out of eight clusters containing cluster-specific bases, with which the genomic DNA of the ditelosomic lines of group 1 chromosomes of a wheat variety ‘Chinese Spring’ was employed to carry out chromosome assignment. The subsequent cloning and DNA sequencing of PCR fragments validated the sequences specificity of the 5′ flanking conserved sequences between LMW-GS gene groups in different genomes. These results suggested that the coding and 5′ flanking regions of LMW-GS genes are likely to have evolved in a concerted fashion. The seven primer sets developed in this study could be used to isolate the complete ORFs of seven groups of LMW-GS genes, respectively, and therefore possess great value for further research in the contributions of a single LMW-GS gene to wheat quality in the complex genetic background and the efficient selections of quality-related components in breeding programs. 相似文献
11.
Isolation and characterization of wheat ω-gliadin genes 总被引:1,自引:0,他引:1
C. C. Hsia O. D. Anderson 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2001,103(1):37-44
The DNA sequences of two full-length wheat ω-gliadin prolamin genes (ωF20b and ωG3) containing significant 5′ and 3′ flanking
DNA sequences are reported. The ωF20b DNA sequence contains an open reading frame encoding a 30,460-Dalton protein, whereas
the ωG3 sequence would encode a putative 39,210-Dalton protein except for a stop codon at amino-acid residue position 165.
These two ω-gliadin genes are closely related and are of the ARQ-/ARE-variant type as categorized by the derived N-terminal
amino-acid sequences and amino-acid compositions. The ω-gliadins were believed be related to the ω-secalins of rye and the
C-hordeins of barley, and analyses of these complete ω-gliadin sequences confirm this close relationship. Although the ω-type
sequences from all three species are closely related, in this analysis the rye and barley ω-type sequences are the most similar
in a pairwise comparison. A comparison of ω-gliadin flanking sequences with respect to that of their orthologs and with respect
to wheat gliadin genes suggests the conservation of flanking DNA necessary for gene function. Sequence data for members of
all major wheat prolamin families are now available.
Received: 24 August 2000 / Accepted: 15 December 2000 相似文献
12.
Huang XQ Cloutier S 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2008,116(7):953-966
In this study, we report on the molecular characterization and genomic organization of the low molecular weight glutenin subunit
(LMW-GS) gene family in hexaploid wheat (Triticum aestivum L.). Eighty-two positive BAC clones were identified to contain LMW-GS genes from the hexaploid wheat ‘Glenlea’ BAC library
via filter hybridization and PCR validation. Twelve unique LMW glutenin genes and seven pseudogenes were isolated from these
positive BAC clones by primer-template mismatch PCR and subsequent primer walking using hemi-nested touchdown PCR. These genes
were sequenced and each consisted of a single-open reading frame (ORF) and untranslated 5′ and 3′ flanking regions. All 12
LMW glutenin subunits contained eight cysteine residues. The LMW-m-type subunits are the most abundant in hexaploid wheat.
Of the 12 LMW-GS, 1, 2 and 9 are i-type, s-type and m-type, respectively. The phylogenetic analysis suggested that the LMW-i
type gene showed greater differences to LMW-s and LMW-m-type genes, which, in turn, were more closely related to one another.
On the basis of their N-terminal sequences, they were classified into nine groups. Fingerprinting of the 82 BAC clones indicated
30 BAC clones assembled into eight contigs, while the remaining clones were singletons. BAC end sequencing of the 82 clones
revealed that long terminal repeat (LTR) retrotransposons were abundant in the Glu-3 regions. The average physical distance between two adjacent LMW-GS genes was estimated to be 81 kb. Most of LMW-GS genes
are located in the d-genome, suggesting that the Glu-D3 locus is much larger than the Glu-B3 locus and Glu-A3 locus. Alignments of sequences indicated that the same type (starting with the same N-terminal sequence) LMW-GS genes were
highly conserved in the homologous genomes between hexaploid wheat and its donors such as durum wheat and T. tauschii.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
13.
小麦新品种“川麦42”低分子量谷蛋白亚基新基因的分子克隆 总被引:5,自引:2,他引:3
采用PCR方法从小麦(Triticum aestivum L.)新品种“川麦42”中克隆得到一个低分子量谷蛋白亚基(LMW-GS)新基因,暂命名为LMWCM42-1。该基因编码区全长846 bp,编码281个氨基酸,具有LMW-GS基因的典型结构特征。推导氨基酸序列比较显示,尽管LMWCM42-1与已知LMW-GS高度相似,但在N-末端重复区部分重复单元和C-末端区中仍存在明显差异。聚类分析表明,LMWCM42-1可能是由Glu-D3位点编码的。 相似文献
14.
15.
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
16.
Gliadins account for about 40–50% of the total proteins in wheat seeds and play an important role in the nutritional and processing
quality of flour. Usually, gliadins can be divided into α-(α/β), γ-, and ω-groups, whereas the low-molecular-weight (LMW)
gliadins are novel seed storage proteins. The low-molecular-weight glutenin subunits (LMW-GSs) are also designated as gliadins
in a few publications. The genes encoding gliadins are mainly located on the short arms of group 6 and group 1 chromosomes,
and not evenly distributed. Repetitive sequences cover most of the uncoding regions, which attributed greatly to the evolution
of wheat genome. The primary structure of each gliadin is divided into several domains, and the long repetitive domains consist
of peptide motifs. Conserved cysteine residues mainly form intramolecular disulfide bonds. The rare potential intermolecular
disulfide bonds and the long repetitive domains play an important role in the quality of wheat flour. There is a general idea
that gliadin genes, even prolamin genes, have a common origin and subsequent divergence leads to gene polymorphism. The γ-gliadins
are considered to be the most ancient of the wheat prolamin family. Several elements in the 5′-flanking (e.g., CAAT and TATA
box) and the 3′-flanking sequences have been detected, which has been shown to be necessary for the proper expression of gliadins.
Published in Russian in Molekulyarnaya Biologiya, 2006, Vol. 40, No. 5, pp. 796–807.
The text was submitted by the authors in English. 相似文献
17.
Characterization of low-molecular-weight glutenin subunit genes and their protein products in common wheats 总被引:10,自引:0,他引:10
Ikeda TM Araki E Fujita Y Yano H 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2006,112(2):327-334
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. 相似文献
18.
19.
A BamHI DNA fragment of 301 bp corresponding to the main repeating unit of 5S rRNA was isolated from barley genomic DNA. The primary nucleotide sequence of this fragment was determined and a high level of homology was found between coding sequences of 5S rRNA genes of barley, wheat and rye. At the same time, spacer's nucleotide sequences of different species of cereals were changed dramatically. At least two types of 5S rRNA tandem repeats of 301 and 450 bp were found in barley genome. Polymorphism for restriction fragment length in 5S rRNA repeats allowed to discriminate between all barley varieties used in this work. 相似文献
20.
Molecular characterization of a LMW-GS gene located on chromosome 1B and the development of primers specific for the Glu-B3 complex locus in durum wheat 总被引:13,自引:0,他引:13
R. D’Ovidio M. Simeone S. Masci E. Porceddu 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,95(7):1119-1126
Low-molecular-weight glutenin subunits (LMW-GS) represent a specific class of wheat storage proteins encoded at the Glu-3 loci. Particularly interesting are the LMW-GS encoded at the Glu-B3 locus because they have been shown to play an important role in determining the pasta-making properties of durum wheat. Genes
encoding LMW-GS have been characterized but only a few of them have been assigned to specific loci. Notably, no complete LMW-GS
gene encoded at the Glu-B3 locus has yet been described. The present paper reports the isolation and characterization of a lmw-gs gene located at the Glu-B3 locus. The clone involved, designated pLDNLMW1B, contains the entire coding region and 524 bp of the 5′ upstream region.
A nucleotide comparison between the pLDNLMW1B clone and other LMW-GS genes showed the presence of some peculiar structural
characteristics, such as short insertions in the promoter region, the presence of a cysteine codon in the repetitive domain,
and a more regular structure of this region, which could be important for its tissue-specific expression and for the functional
properties of the encoded protein, respectively.
Received : 30 May 1997 / Accepted : 29 July 1997 相似文献