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1.
Common non-waxy (Wx) rice cultivars contain two different alleles at the waxy locus, designated Wx a and Wx b, which encode different levels of granule-bound starch synthases and are hence involved in the control of endosperm amylose content. The Wx a allele was predominant in non-waxy indica cultivars, whereas the Wx b allele was common to the non-waxy japonica variety. Recently, some of the molecular mechanisms underlying the differentiation of Wx a from Wx b have been characterized. One structural difference between these two alleles was shown to be due to alternative splicing caused by a single-base substitution (AGGT to AGTT) at a donor site of the first intron within the Wx gene. In the case of waxy (wx) rice, it was not possible to distinguish whether the each wx allele was derived from Wx a or Wx b alleles by phenotypic analysis. However, we succeeded in developing a derived cleaved amplified polymorphic sequence (dCAPS) marker for the detection of the one-base splicing mutation without the need for sequencing. A mismatch primer was used to generate a restriction site in the Wx a allele (AGGT) but not in the Wx b allele (AGTT). Three hundred fifty-three waxy rice strains that are widely found in Asia were then employed for analysis using this dCAPS marker. Our findings suggested that waxy rice strains have both Wx a- and Wx b-derived alleles, but that the Wx b-derived allele was predominant, and its distribution was independent of indica-japonica differentiation. The wild relatives of cultivated rice all possessed the AGGT allele. It was concluded that the waxy mutations, and the corresponding rice cultivation, originated from japonica during the evolution and domestication process of rice and was preferentially selected by most Asian peoples.Communicated by J. Heslop-Harrison  相似文献   

2.
Four pairs of specific PCR primers have been designed on the basis of the sequence of the granule-bound starch synthase gene (GBSS; dominant non-waxy gene Wx) and used to amplify its homologous sequence from thirteen waxy and two non-waxy inbred lines. Results from electrophoresis indicated that the recessive waxy gene was wx, derived from the dominant non-waxy gene Wx by mutation at its 3′ end. The sequence of the mutated 3′ end was amplified by the TAIL-PCR technique. Sequence alignment showed that the mutation of the wx gene was caused by transposition of the aldehyde dehydrogenase gene rf2. Two pairs of specific primers were designed on the basis of the sequence difference between the dominant gene Wx and its mutated recessive allele wx and used as intragenic selection markers to identify individual plants of genotypes WxWx, Wxwx, and wxwx by PCR amplification from the segregating population of the F2 generation crossed between waxy and non-waxy inbred lines. Iodine solution staining and starch component assay showed that all the 35 F2 plants identified as genotype WxWx produced non-waxy kernels of the F3 generation and that all 33 F2 plants identified as genotype wxwx produced waxy kernels of the F3 generation. This result can be used to improve the selection efficiency of waxy maize breeding and for selection of other single genes and major polygenes.  相似文献   

3.
The Wx gene encodes the granule-bound starch synthase I or waxy protein, which is the sole enzyme responsible for amylose synthesis in wheat seeds. Triticum urartu and einkorn (T. monococcum L. ssp. monococcum), which are related to the A genome of bread wheat, could be important sources of variation for this gene. This study evaluated the Wx gene variability in 52 accessions of these species and compared their nucleotide sequences with the Wx-A1a allele of bread wheat. The level of polymorphism found was high, although not distributed equally between the two species. Five different alleles were found in T. urartu, of which four were novel (Wx-A u 1b, -A u 1c, -A u 1d and -A u 1e). All einkorn accessions had the same allele, which was also novel and was named Wx-A m 1a. A comparison between the proteins deduced from the novel alleles and the Wx-A1a protein showed that there were up to 33 amino acid changes in both the transit peptide and the mature protein. These results showed that these species, especially T. urartu, are a potential source of novel waxy variants.  相似文献   

4.
To examine continuous variation of amylose levels in Asian rice (Oryza sativa) landraces, the five putative alleles (Wx a , Wx in , Wx b , Wx op , and wx) at the wx locus were investigated in near-isogenic lines (NILs). Apparent amylose levels ranged from 0.5 to 29.9% in the NILs, showing a positive relation with the levels of Wx gene product, granule-bound starch synthase (GBSS) as well as the enzymatic activity per milligram starch granule. Only opaque (Wx op ) accessions had an enzymatic activity per GBSS that was reduced to half the level of the others. Nucleotide sequences in the Wx gene were compared among 18 accessions harboring the five different alleles. Each of the Wx alleles had a unique replacement, frame-shift or splice donor site mutation, suggesting that these nucleotide changes could be reflected in phenotype alterations. A molecular phylogenetic tree constructed using the Wx gene indicated that ssp. japonica forms a distinct clade, whereas ssp. indica forms different clades together with the wild progenitor. Unexpectedly, the wx allele of 160 (indica from Taiwan) joined the japonica lineage; however, comparisons using linked genes for two Taiwanese accessions revealed that the wx gene was the product of gene flow from japonica to indica. Therefore, the japonica lineage frequently included Wx in , Wx b and wx, while Wx a and Wx op were found in the other lineages, strongly suggesting that allelic diversification occurred after divergence of the two subspecies. The present results were discussed in relation to the maintenance of agronomically valuable genes in various landraces.  相似文献   

5.
Post-Domestication Selection in the Maize Starch Pathway   总被引:1,自引:0,他引:1  
Modern crops have usually experienced domestication selection and subsequent genetic improvement (post-domestication selection). Chinese waxy maize, which originated from non-glutinous domesticated maize (Zea mays ssp. mays), provides a unique model for investigating the post-domestication selection of maize. In this study, the genetic diversity of six key genes in the starch pathway was investigated in a glutinous population that included 55 Chinese waxy accessions, and a selective bottleneck that resulted in apparent reductions in diversity in Chinese waxy maize was observed. Significant positive selection in waxy (wx) but not amylose extender1 (ae1) was detected in the glutinous population, in complete contrast to the findings in non-glutinous maize, which indicated a shift in the selection target from ae1 to wx during the improvement of Chinese waxy maize. Our results suggest that an agronomic trait can be quickly improved into a target trait with changes in the selection target among genes in a crop pathway.  相似文献   

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7.
The waxy (Wx) locus of Zea mays was cloned from strains carrying the wild-type and wxm-8 mutant alleles. The receptor component of the Suppressor-Mutator (Spm) controlling element system in the wxm-8 allele was shown to be a 2 kb long insertion within the transcribed region of the Wx gene. The insertion, termed Spm-I8, is excised during somatic reversion events induced by the autonomous controlling element Enhancer (En), which is an equivalent to Spm. Integration of Spm-I8 into the Wx gene generates a 3-bp target site duplication. Spm-I8 has a 13 bp long inverted repeat at its termini. The ends of the element can be further folded to build a large double-stranded structure consisting of five perfectly matching double-stranded regions of 9–13 bp in length, interrupted by single-stranded loops. A comparison of the wild-type and wxm-8 alleles revealed two additional insertions 6 (insert-1) and 0.25 (insert-2) kb in length. No En-induced excision of insert-1 and insert-2 could be detected so far. There is remarkable structure and sequence homology between Spm-I8 and the transposable elements Tam1 and Tam2 of Antirrhinum majus at their termini, reflecting a possible evolutionary and/or functional relationship between transposons in different plant species.  相似文献   

8.
Nelson OE 《Genetics》1975,79(1):31-44
The effect of heterozygosity for structural rearrangements on recombination between two wx heteroalleles (C and 90) and the pattern of flanking markers in the resultant Wx gametes has been examined. The rearrangements are Tp9, an insertional translocation in which a segment of chromosome 3 has been inserted into the short arm of chromosome 9 close to the wx locus; In9a, a long pericentric inversion with wx in the inverted segment; and Rearr 9, a complex rearrangement of chromosome 9. Heterozygosity for rearrangements decreases the frequency of Wx gametes to varying degrees.—Heterozygosity for Tp9 enhances the proportion of Wx gametes that are apparent convertants and allows the conclusion that such gametes do not normally arise from an exchange in the wx locus plus a second exchange distal to wx. Heterozygosity for In9a markedly decreases the frequency of Wx gametes that are recombinant for outside markers but does not decrease the frequency of convertants.—Heterozygosity for Rearr 9 permits a low frequency of Wx gametes, all of which are apparent convertants.—A high proportion of the convertants have the flanking markers that entered the cross with C so recombination is polarized in normal homologs and in heterozygotes for all rearrangements.  相似文献   

9.
Emmer wheat is a neglected crop that could be used in the breeding of modern durum wheat for quality, one important aspect of which is the starch composition that is related to the waxy proteins. A collection of 87 accessions of Spanish emmer wheat was analysed for waxy protein composition by SDS?CPAGE. No polymorphism was found for the Wx-A1 gene. However, for the Wx-B1 gene, three alleles were detected, two of them new. The whole gene sequence of these alleles was amplified by PCR in three fragments, which were digested with several endonucleases to determine internal differences in the sequence. These variants were also compared with the Wx alleles present in durum wheat. Differences in size and restriction sites were detected. DNA sequence analysis confirmed that the alleles found in emmer wheat are different from those in durum wheat. The first data suggested that these alleles showed a different influence on the amylose content of these lines. The variation found could be used to enlarge the gene pool of durum and emmer wheat, and design new materials with different amylose content.  相似文献   

10.
Differential regulation of waxy gene expression in rice endosperm   总被引:36,自引:0,他引:36  
Summary In order to examine the effects of different alleles on the gene expression at the waxy locus, the Wx gene product which controls the synthesis of amylose was isolated from endosperm starch of rice plants and analysed by electrophoretic techniques. The major protein bound to starch granules was absent in most of waxy strains and increased with the number of Wx alleles in triploid endosperms, suggesting that the major protein is the Wx gene product. In addition to wx alleles which result in the absence or drastic reduction of the Wx gene product and amylose, differentiation of Wx alleles seemed to have occurred among nonwaxy rice strains. At least two Wx alleles with different efficiencies in the production of the major protein as well as amylose were detected. These alleles are discussed in relation to regulation of the gene expression.  相似文献   

11.
Development of high-yielding cereal crops could meet increasing global demands for food, feed and bio-fuels. Wheat is one of the world??s most important cereal crops. The biosynthesis of starch is the major determinant of yield in wheat. Two starch biosynthesis genes, the waxy (Wx) genes and the starch synthase IIa (SSIIa) genes, were amplified and sequenced in 92 diverse wheat genotypes using genome-specific primers. Nucleotide diversity, haplotype analysis and association mapping were performed. The first exon (5??-UTR) and the first intron of the three homoeologous Wx genes were isolated using expressed sequence tag sequences. The Wx genes contained 12 exons separated by 11 introns. SNP (single nucleotide polymorphism) frequency ranged from 1 SNP/3,648?bp for Wx-D1 to 1 SNP/135?bp for SSIIa-A1, with an average of 1 SNP/230?bp. The average SNP frequencies in exon and intron regions were 1 SNP/322?bp and 1 SNP/228?bp, respectively. Thirty, 23 and 5 SNPs were identified and formed five, six and five haplotypes for SSIIa-A1, SSIIa-B1 and SSIIa-D1, respectively. However, no association was found between these SNPs and seven yield-related traits. Twenty-two, 15 and 1 SNPs were detected and formed nine, five and two haplotypes for Wx-A1, Wx-B1 and Wx-D1, respectively. Three unique nucleotides C+A+T at SNP5, SNP6 and SNP12 formed Wx-B1-H3, which was significantly associated with increased grain weight, thousand kernel weight, and total starch content in three spring wheat genotypes and five winter wheat genotypes. Cost-effective and co-dominant SNP markers were developed using temperature-switch (TS)-PCR and are being used for marker-assisted selection of doubled haploid lines with enhanced grain yield and starch content in winter wheat breeding programs.  相似文献   

12.
Apparent amylose content (AAC) is a key determinant of eating and cooking quality in rice and it is mainly controlled by the Wx gene which encodes a granule-bound starch synthase (GBSS). In this study, sixteen single-segment substitution lines harboring the Wx gene from 16 different donors and their recipient HJX74 were used to detect the naturally occurring allelic variation at the Wx locus. The AAC in the materials varied widely and could be grouped into glutinous, low, intermediate, and two high AAC sub-classes, high I (24.36?C25.20%) and high II (25.81?C26.19%), under different experimental environments, which showed a positive correlation with the enzymatic activity of GBSS. One insertion/deletion (InDel) and three single nucleotide polymorphisms in the Wx gene were detected and their combinations resulted in the variation of five classes of AAC. Based on the results of AAC phenotypes, GBSS activities and cDNA sequences, five Wx alleles, wx, Wx t, Wx g1, Wx g2, and Wx g3, were identified, two of which, Wx g2 and Wx g3, are separated for the first time in this study. Under different cropping seasons, the AAC differed significantly for the Wx t and Wx g1 alleles, with higher AAC in the fall season than in the spring season, but did not differ significantly for the wx, Wx g2, and Wx g3 alleles. In conclusion, the present results might contribute to our understanding of the naturally occurring allelic variation at the Wx locus and will facilitate the improvement of rice quality by marker-assisted selection.  相似文献   

13.
Perisperm starch granules of the dicotyledonous plant Amaranthus hypochondriacus L. were prepared from two homozygous lines (WxWx and wxwx) and their hybrid (Wxwx). The hybrid line was obtained by natural hybridization. By Sephadex G-75 column chromatography of isoamylase-debranched starches, the amylose content of WxWx starch was 16.9%, that of Wxwx was 10.7, and wxwx was zero. SDS-polyacrylamide gel electrophoresis showed that starch granules from two genotypes (WxWx and Wxwx) contained a Wx protein (MW = 68,000) which was supposed to be a starch granule-bound starch synthase and was associated with amylose synthesis, as observed in nonwaxy maize. The intensities of the stained protein bands were apparently correlated with the number of the Wx alleles. The Wx protein was not detected in the wxwx starch. These findings suggest that the Wx allele produces the Wx protein and amylose in the perisperm of A. hypochondriacus, with incomplete dominance over the wx allele. The Wx allele did not affect the fine structure of amylopectin and had little if any effect on susceptibility to glucoamylase and pasting properties of starch granules from these genotypes.  相似文献   

14.
With the use of chromosome interchanges, the waxy (wx) locus on chromosome 9 has been relocated to various positions in the maize genome. Four wx alleles, wx C, wx B, wx 90, and wx H21, were crossed to six chromosome translocation stocks (four with break points proximal to wx and, two distal to wx). Of the 26 possible homozygous translocation heteroallelic combinations, the results of eight are available in this report. In most instances, the frequencies of wx intragenic recombination of the rearranged chromosomes were lower than that of the control. A significant difference in degree of reduction in recombination values is found for different heteroallelic combinations at the same location and in one instance for the same heteroallelic combination at a different chromosome position. The linear order of the 4 wx mutants within the wx cistron is wx C-wx H21-wx 90 (wx B). Additional effects from both genetic background and seasonal factors of the different plantings also are observed.Journal Paper No. J-6906 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Project No. 1335.  相似文献   

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17.
Waxy maize was first reported in China in 1909 and is mainly used in food production in Asia. The evidence for strong domestication selection in the Waxy locus of rice and a selective sweep around its genomic region make us to wonder whether there has been similar selection in Waxy in glutinous maize. To address this issue, DNA sequences of Waxy, three flanking genes and an unlinked gene (Adh1) of 30 accessions sampled from Chinese waxy maize accessions, including representative landraces and inbred lines, were determined in this study. Sharp reduction of nucleotide diversity and significant neutrality tests (Tajima’s D and Fu and Li’s F*) were observed in the Waxy locus in Chinese waxy maize but not in nonglutinous maize; comparison with the unlinked gene confirmed that this pattern was different to Waxy. Sequence analysis across a 143 kb genomic segment centered on the Waxy locus revealed patterns consistent with a selective sweep in the upstream region of Waxy. The selective sweep detected based on current limited genomic sequences exceeded over 50 kb, indicating strong selection in this or a bigger region. However, No sweep effect was detected in the repetitive downstream region of Waxy. Phylogenetic analysis indicated that Chinese waxy maize was domesticated from the cultivated flint maize (Zea mays ssp. mays) that was introduced from the new world. At least two independent deletions in exon 7 (30 bp) and 10 (15 bp) were identified in the Chinese accessions respectively. These findings demonstrate a similar pattern of domestication selection in the Waxy genomic region in both glutinous maize and rice, suggesting that this pattern in the rise of glutinous phenotype is likely in other cereal crops.  相似文献   

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19.
Plants form their gametes late in somatic development and, as a result, often pass somatic mutations on to their progeny. Classic examples of this process are the germinal revertants of unstable, Ac/Ds transposon-induced kernel mutations in maize: frequent and early reversion events during somatic development are generally correlated with a high frequency of revertant gametes. We have characterized a Ds allele of the maize waxy(wx) gene, wx-m5:CS7, for which the correlation between somatic and germinal reversion frequencies no longer holds. The ability of wx-m5:CS7 (CS7) to produce revertant gametes is suppressed ~100-fold in comparison with a second Ds allele, wx-m5:CS8 (CS8), which has an identical insertion at Wx and the same frequent and early somatic reversion pattern in endosperm. The excision of Ds from wx is not reduced 100-fold in the somatic tissues of CS7 plants as compared with CS8 plants. Suppressed formation of CS7 revertant gametes is independent of the Ac transposase source and is heritably passed to the embryos of progeny kernels; however, frequent and early somatic reversion is observed again in endosperms of these progeny kernels. This suppression appears to be caused by a dominant mutation in a trans-acting product that can suppress the germinal reversion of other Ds-induced alleles as well; the mutation is tightly linked to Wx but is not in the CS7 Ds itself. Taken together, the data suggest a novel mode of developmental control of Ac/Ds elements by the host plant, suppressing element excision in the shoot meristem.  相似文献   

20.
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