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PubMed1997

Identification of individual barley chromosomes based on repetitive sequences: conservative distribution of Afa-family repetitive sequences on the chromosomes of barley and wheat.

The Afa-family repetitive sequences were isolated from barley (Hordeum vulgare, 2n = 14) and cloned as pHvA14. This sequence distinguished each barely chromosome by in situ hybridization. Double color fluorescence in situ hybridization using pHvA14 and 5S rDNA or HvRT-family sequence (subtelomeric sequence of barley) allocated individual barley chromosomes showing a specific pattern of pHvA14 to chromosome 1H to 7H. As the case of the D genome chromosomes of Aegilops squarrosa and common wheat (Triticum aestivum) hybridized by its Afa-family sequences, the signals of pHvA14 in barley chromosomes tended to appear in the distal regions that do not carry many chromosome band markers. In the telomeric regions these signals always placed in more proximal portions than those of HvRT-family. Based on the distribution patterns of Afa-family sequences in the chromosomes of barley and D genome chromosomes of wheat, we discuss a possible mechanism of amplification of the repetitive sequences during the evolution of Triticeae. In addition, we show here that HvRT-family also could be used to distinguish individual barley chromosomes from the patterns of in situ hybridization.

Biological Evolution↗
PubMed2004

[Barley chromosome identification using genomic in situ hybridization in the genome of backcrossed progeny of barley-wheat amphiploids [H. geniculatum All. (2n = 28) x T. aestivum L. (2n = 42)] (2n = 70)].

Genomic in situ hybridization (GISH) has been used to study characteristics of the formation of alloplasmic lines detected among self-pollinated backcrossed progeny (BC1F5-BC1F8) of barley--wheat amphiploids [Hordeum geniculatum All. (2n = 28) x Triticum aestivum L. (2n = 42)] (2n = 70). The chromosome material of the wild barley H. geniculatum has been shown to contribute to these lines. For example, fifth-generation plants (BC1F5) had genotypes (2n = 42w + 2g), (2n = 42w + 1g + 1tg), and (2n = 41w + 1g), where w is common wheat chromosomes, g is barley (H. geniculatum) chromosomes, and tg is the telocentric chromosome of wild barley. Beginning from the BC1F6 generation, alloplasmic telocentric addition lines (2n = 42 + 2tg) and (2n = 42 + 1tg) appear. This lines has been found cytogenetically unstable. The progeny of each of these cytological types include not only the (2n = 42 + 2tg) and (2n = 42 + 2tg) addition plants, but also plants with the monosomic (2n = 41 + 1tg) and the disomic (2n = 40 + 2tg) substitutions, as well as the (2n = 41 + 2tg) plants, which lack one wheat chromosome and have two telocentric barley chromosomes. It has been demonstrated that the selection for well-filled grains favors the segregation of telocentric addition lines (2n = 42 = 2tg) and (2n = = 42 + 1tg).

Breeding↗
PubMed2005

Molecular cytogenetic characterization and seed storage protein analysis of 1A/1D translocation lines of durum wheat.

Two durum wheat [Triticum turgidum L. ssp. durum (Desf.) Husn.] lines carrying the high-molecular-weight (HMW) glutenin subunits (GS) 1 D x 5 + 1Dy10 encoded by Glu-D1d, L252 and S99B34, were characterized using fluorescent genomic in-situ hybridization (FGISH) and microsatellite markers. These two durum lines were derived from the crosses in which the common wheat (T. aestivum L.) 'Len' and durum wheat 'Langdon' (LDN) and 'Renville' were involved. FGISH patterns of the mitotic chromosomes indicated that these two durum lines have one pair of 1AS.1AL-1DL translocated chromosomes in which the terminal region of 1AL was replaced by a homoeologous segment of 1DL. The 1DL segment spans approximately 31% of the long arm of the translocated chromosome. Microsatellite marker analysis confirmed the 1AS.1AL-1DL translocation and determined the translocation breakpoint to be distal to Xgwm357 on 1AL. Seed storage proteins (GS and gliadins) were analysed in these two 1AS.1AL-1DL translocation lines and three sib lines (L092, S99B19 and S99B33) using SDS-PAGE and A-PAGE. The SDS-PAGE and A-PAGE profiles demonstrated that the two low yielding lines (L252 and S99B19) had the low-molecular-weight (LMW) -1 GS encoded by Glu-A3k and Glu-B3s and 1B-encoded gliadins from LDN, and the other three lines (L092, S99B33 and S99B34) with higher yield had LMW-2 GS and 1B-encoded gliadins from Renville, suggesting that undesirable genetic components from LDN might limit substantial improvement of yield. Thus, the translocation lines with 1 D x 5 + 1Dy10 and LMW-2, which are associated with good bread-making and pasta qualities, respectively, in a good genetic background will be useful for developing durum cultivars with dual-purpose end-use. Results from this study demonstrate that the D-genome could play an important role in the genetic improvement of durum wheat and evolution of the A- and B-genomes in tetraploid wheat.

Genetic Markers↗
PubMed2002

Production of a new wheat line possessing the 1BL.1RS wheat-rye translocation derived from Korean rye cultivar Paldanghomil.

The 1BL.1RS translocations between wheat ( Triticum aestivum L.) and rye ( Secale cereale L.) are widely used in bread wheat breeding programs, but all modern wheat cultivars with the 1BL.1RS have shown genetic vulnerability due to one rye source - a German cultivar, Petkus. We have developed, a new 1BL.1RS wheat-rye translocation line from the backcross of the F(1) hybrid of wheat cv. Olmil and rye cv. Paldanghomil, both cultivars from Korea. The GISH technique was applied to identify the presence of rye chromatin in 467 BC(1)F(6) lines selected from 77 BC(1)F(5) lines. Only one line, Yw62-11, showed wheat-rye translocated chromosomes, with a somatic chromosome number of 2n=42. C-banding patterns revealed that the translocated chromosome was 1BL.1RS, showing prominent bands in the terminal and sub-terminal regions of the short arm as well as in the centromeric region and terminal region of the long arm. This new 1BL.1RS translocation line formed 21 bivalents like common wheat at meiotic metaphase I, thereby showing complete homology.

Journal Article↗
PubMed2003

[Studies on application of APAGE in male sterile wheat with alloplamic cytoplasm].

Four male sterile wheat with Aeglops kotschyi, Ae. variabilis, Ae. ventricosa and Ae. bicornis cytoplasm, were crossed with some wheat materials, and from the crosses a series of maintenance lines were selected. The APAGE(acid polyacrylamide gel electrophoresis) analysis indicated that most maintenance lines possessed the gliadin marker Gld1B3 of 1BL/1RS translocation chromosome. The root tip chromosomes of this maintenance lines had two satellite chromosomes, but other maintenance lines without Gld1B3 marker had the same four satellite chromosomes as other common wheat. Three male sterile wheats with Aeglops kotschyi, Ae. variabilis, Ae. ventricosa cytoplasm had the same fertility, but the male sterile wheat with Ae. bicornis cytoplasm could produce male sterile. Meantime, the paper discussed application of APAGE technology in breeding of non-1BL/1RS male sterile wheat.

Cytoplasm↗
PubMed2003

[Localization of genes, determining quantitative traits in wheat: amendment to the "catalog of chromosomal mapping of genes in domestic cultivars of wheat"].

An amendment to the catalog of chromosome location of genes in Russian wheat cultivars was constructed with the published data of the recent decade. The results of chromosomal localization were summarized and analyzed by methods of multivariate statistics. Chromosomes critical for 40 quantitative traits under study proved to cluster according to their homeology, i.e., by homeological groups. The hypotheses providing an explanation for this finding are considered. It is suggested that quantitative traits are similarly controlled by genes located on homeological chromosomes in common wheat, making it possible to isolate a limited number of major genes for each particular quantitative trait.

Chromosome Mapping↗
PubMed2001

Effect of spelt wheat flour and kernel on bread composition and nutritional characteristics.

Spelt wheat seeds (Triticum aestivum subsp. spelta cv. Ostro) were used to obtain white spelt flour (64.5% yield), wholemeal spelt flour (100% yield), and scalded spelt wheat kernels. From these materials, white spelt wheat bread (WSB), wholemeal spelt wheat bread (WMSB), and spelt wheat bread with scalded spelt wheat kernels (SSKB) were made and were compared to the reference white wheat bread (WWB). The spelt wheat flours and breads contained more proteins in comparison to wheat flour and bread. Among the samples the highest rate of starch hydrolysis was noticed in WSB. During the first 30 min of incubation this particular bread was shown to have significantly more (P < 0.05) rapidly digestible starch than the WMSB and later on also more starch than in WWB and SSKB, respectively. The WMSB had the lowest hydrolysis index (HI = 95.7). However, the result did not differ significantly from that in the reference common wheat bread. On the other hand, the most refined spelt wheat flour resulted in a bread product (WSB) that was statistically withdrawn (P < 0.05) as one with the highest HI (112.6).

Bread↗
PubMedDate not supplied

[Alpha-amylase as an occupational allergen in baking industry employees].

In a group of 226 bakers and pastry makers and in 88 students of a training school for bakers, we evaluated skin sensitization to the common allergens, wheat and alpha amylase. Skin prick tests were positive to the enzyme in 17 exposed subjects (7.5%) and in one student with previous occupational exposure as a baker; 27 exposed subjects (11.9%) and 2 students were sensitized to wheat. Among the 42 exposed workers who complained of work-related symptoms, 12 (28.6%) cases were skin positive to amylase and 17 (42.9%) to wheat. Among the 17 workers who were positive to amylase, 16 were also sensitized to wheat and/or common allergens, 12 complained of symptoms at work but since in many cases there was a positive response to wheat, too, it is impossible to speculate on the role of each allergen in inducing symptoms. One case, with work-related rhinoconjunctivitis, had skin sensitization only to alpha amylase but no specific IgE in the serum. These findings confirm that bakers are at risk of sensitization not only to wheat allergen but also to amylase from Aspergillus oryzae. The enzyme should be included in the list of substances to be tested among bakers in whom an occupational allergy is suspected, but particular care should be taken in evaluating the cutaneous response, especially if compared to wheat wheals. Further investigations are also needed to identify the source of risk and to better define the characteristics of the enzyme and the relationship between skin reaction to amylase, sensitization to wheat and atopy.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed2004

Aneuploid analyses of three chlorophyll abnormalities in Emmer wheat.

Of the various chlorophyll abnormalities that occur in polyploid wheats, genetic bases of only two types, chlorina and virescence, are known. Here, for the first time, the chromosomal bases of three other chlorophyll abnormalities, striato-virescence, delayed virescence, and albino, which occur in Emmer wheat (2n = 4x = 28, genome constitution AABB) are reported. A set of disomic substitution lines of Langdon durum was used for chromosome identification. All three abnormalities are controlled by two duplicated recessive genes (carrier chromosome in parentheses); striato-virescence by sv1 (3A) and sv2 (2A), delayed virescence by dv1 (2B) and dv2 (supposedly 2A), and albino by abn1 (2A) and abn2 (2B). Genes on the same chromosome are located in different loci and are recombined with each other. The Chinese Spring cultivar of common wheat (2n = 6x = 42, genome constitution AABBDD) carries wild-type homoeoalleles for these abnormalities; Sv3 (2D), Dv3 (2D), and Abn3 (2D).

Aneuploidy↗
PubMed1997

Alloplasmic wheats with Aegilops crassa cytoplasm which express photoperiod-sensitive homeotic transformations of anthers, show alterations in mitochondrial DNA structure and transcription.

Alloplasmic wheat. Triticum aestivum cv. Norin 26, with Aegilops crassa cytoplasm, shows photoperiod-sensitive cytoplasmic male sterility (PCMS). This alloplasmic line expresses pistillody of anthers only when grown in long-day conditions (> 15 h light). To assess the molecular basis of the PCMS, we carried out Southern and Northern hybridization analyses on mitochondrial DNAs and RNAs isolated from seedlings of alloplasmic lines showing various PCMS phenotypes using probes for twelve mitochondrial genes. All RFLP patterns of mitochondrial DNA from alloplasmic lines greatly differed from those of common wheat, and were slightly changed from those of the parental species, i.e., Ae. crassa. This indicates that nuclear substitutions between related plant species induce structural alterations in the mitochondrial genome. Furthermore, RFLP patterns of (cr)-N61 and FR-mutant probed with coxIII and orf25 were identical with each other, but different from those of the other alloplasmic lines, indicating that the nuclei of N61 and FR-mutant harbor some gene(s) that induces structural alterations of the mitochondrial genome in the coxIII and orf25 regions. The transcription patterns of atp6 and cob in Ae. crassa type were different from those of T. aestivum type. Furthermore, the orf25 transcript in alloplasmic wheats was about 300 nucleotides longer than that of euplasmic lines, including the Ae. crassa pure line, suggesting that transcription patterns of orf25 are associated with recovery from the PCMS phenomenon. These data clearly show the mutual cross-talk between the nuclear genome and chondriome. These observations raise the possibility that the dysfunction of mitochondria caused by the failure of a cooperative control of mitochondrial gene(s) expression influences the pathway of flower morphogenesis, especially in the process that determines organ identity.

Blotting, Northern↗
PubMed2006

Multiple genetic pathways for seed shattering in the grasses.

Shattering is an essential seed dispersal mechanism in wild species. It is believed that independent mutations at orthologous loci led to convergent domestication of cereal crops. To investigate genetic relationships of Triticeae shattering genes with those of other grasses, we mapped spike-, barrel- (B-type), and wedge-type (W-type) spikelet disarticulation genes in wheat and its wild relatives. The Br1 gene for W-type disarticulation was mapped to a region delimited by Xpsr598 and Xpsr1196 on the short arm of chromosomes 3A in Triticum timopheevii and 3S in Aegilops speltoides. The spike- and W-type disarticulation genes are allelic at Br1 in Ae. speltoides. The B-type disarticulation gene, designated as Br2, was mapped to an interval of 4.4 cM between Xmwg2013 and Xpsr170 on the long arm of chromosome 3D in Aegilops tauschii, the D-genome donor of common wheat. Therefore, B- and W-type disarticulations are governed by two different orthologous loci on group-3 chromosomes. Based on map position, orthologs of Br1 and Br2 were not detected in barley, maize, rice, and sorghum, indicating multiple genetic pathways for shattering in grasses. The implications of the mapping results are discussed with regard to the evolution of polyploid wheat and domestication of cereals.

Amino Acid Sequence↗
PubMed2000

RFLP-based analysis of three RbcS subfamilies in diploid and polyploid species of wheat.

The RbcS multigene family of hexaploid (bread) wheat, Triticum aestivum (genome BBAADD), which encodes the small subunit of Rubisco, comprises at least 22 genes. Based on their 3' non-coding sequences, these genes have been classified into four subfamilies (SFs), of which three (SF-2, SF-3 and SF-4) are located on chromosomes of homoeologous group 2 and one (SF-1) on homoeologous group 5. In the present study we hybridized three RbcS subfamily-specific probes (for SF-1, SF-2 and SF-3) to total DNA digested with four restriction enzymes and analyzed the RFLP patterns of these subfamilies in eight diploid species of Aegilops and Triticum, and in two tetraploid and one hexaploid species of wheat (the diploid species are the putative progenitors of the polyploid wheats). The three subfamilies varied in their level of polymorphism, with SF-2 being the most polymorphic in all species. In the diploids, the order of polymorphism was SF-2 > SF-3 > SF-1, and in the polyploids SF-2 > SF-1 > SF-3. The RbcS genes of the conserved SF-1 were previously reported to have the highest expression levels in all the wheat tissues studied, indicating a negative correlation between polymorphism and gene expression. Among the diploids, the species with the D and the S genomes were the most polymorphic and the A-genome species were the least polymorphic. The polyploids were less polymorphic than the diploids. Within the polyploids, the A genome was somewhat more polymorphic than the B genome, while the D genome was the most conserved. Among the diploid species with the A genome, the RFLP pattern of T. urartu was closer to that of the A genome of the common wheat cultivar Chinese Spring (CS) than to that of T. monococcum. The pattern in Ae. tauschii was similar to that of the D genome of CS. Only partial resemblance was found between the RFLP patterns of the species with the S genome and the B genome of CS.

Chromosome Mapping↗
PubMed2005

Recent developments in durum wheat chromosome engineering.

Transfer of alien chromosome segments from various Triticeae species into cultivated wheats, commonly referred to as "chromosome engineering", is currently benefiting from the recent, impressive advancements in molecular genetics, cytogenetics and genomics, which are providing new insights into the genetic and physical organization of even complex plant genomes, such as those of the Triticeae. The powerful analytical tools presently available are making the assessment of desired genotypes in the course of chromosome engineering far more precise and effective than in the past, thus giving this transfer strategy renewed and increased potential for meaningful practical achievements. Examples are given here of the application of such tools to the engineering of the durum wheat genome with small alien segments containing genes with beneficial impact on disease resistance and quality traits.

Chromosomes, Plant↗
PubMed2000

[Studies of salt tolerance of alloplasmic wheat].

Using Aegilops crassa cytoplasm wheat as test material, salt tolerance of alloplasmic wheat was identified by tissue culture, nutrient solution culture and artificial saline pond, from calli, seeding stage, recovering stage and mature period. The results showed Ae. crassa cytoplasm lines indicated the difference of salt tolerance in varying degree; the difference of cytoplasm effect among different nucleoplasm combinations showed certain nucleoplasm interaction relation. The salt tolerance of (Ae. crassa)-jian 26 was stronger than that other tested alloplasmic lines. The result demonstrated that at recovering date and at mature period indicated salt tolerance of alloplasmic lines was not only stronger than its nuclear parent but also over control variety. As compared with common wheat, lines having strong salt tolerance were more proportions in tested alloplasmic lines. The study of alloplasmic wheat should be significant for breeding salt tolerance wheat.

Cytoplasm↗
PubMed2004

Molecular cloning and comparative analysis of a y-type inactive HMW glutenin subunit gene from cultivated emmer wheat (Triticum dicoccum L.).

Cultivated emmer (Triticum dicoccum, 2n = 4x = 28, AABB) is closely related to bread wheat and possesses extensive allelic variations in high molecular weight glutenin subunit (HMW-GS) composition. These alleles may be an important genetic resource for wheat quality improvement. To isolate and clone HMW-GS genes from cultivated emmer, two pairs of allele-specific (AS) PCR primers were designed to amplify the coding sequence of y-type HMW-GS genes and their upstream sequences, respectively. The results showed that single bands of strong amplification were obtained through AS-PCR of genomic DNA from emmer. After cloning and sequencing the complete sequence of coding and 5'-flanking regions of a y-type subunit gene at Glu-A1 locus was obtained. Nucleotide and deduced amino acid sequences analysis showed that this gene possessed a similar structure as the previously reported Ay gene from common wheat, and is hence designated as Ay1d. The distinct feature of the Ay1d gene is that its coding region contains four stop codons and its upstream region has a 85-bp deletion in the same position of the Ay gene, which are probably responsible for the silencing of y-type subunit genes at Glu-A1 locus. Phylogenetic analysis of HMW glutenin subunit genes from different Triticum species and genomes were also carried out.

Amino Acid Sequence↗
PubMed2004

Two point mutations identified in emmer wheat generate null Wx-A1 alleles.

In this report, the Wx-A1 mutations carried by a Triticum dicoccoides line from Israel and a Triticum dicoccum line from Yugoslavia are characterized. A single nucleotide insertion in the T. dicoccoides null allele and a single nucleotide deletion in the T. dicoccum null allele each cause frameshift mutations that induce premature termination codons more than 55 nucleotides upstream of the last exon-exon junction. In both mutants, Wx-A1 transcripts were detectable in 10 day post-anthesis endosperm by relative RT-PCR. However, transcript levels of the T. dicoccoides and T. dicoccum null alleles were reduced to approximately 6.5 and 1.5% of wild-type, respectively. Therefore, the lack of Wx-A1 protein in the mutants appears to be largely due to nonsense-mediated mRNA decay. The two mutations described here arose independently, and are not related to either of the Wx-A1 mutations identified in common wheat.

Alleles↗
PubMed2005

Cloning, characterization and expression of wheat EDR1 (enhanced disease resistance) gene.

To investigate if there is an EDR1 pathway in wheat (Triticum aestivum L.), a pair of degenerate primers was designed according to the cDNAs of Arabidopsis thaliana EDR1 gene and its homologs were used to isolate EDR1 gene homologs from wheat. RT-PCR was conducted on the cDNA template synthesized with RNA of wheat leaves. A 627-bp cDNA fragment representing an EDR1 gene (named as TaEDR1) was isolated (GenBank accession number: AY743662). Subsequently, the 3050-bp full-length cDNA sequence of TaEDR1, which encodes a polypeptide consisting of 959 amino acid residues, was obtained by RACE technique. The amino acid sequence of TaEDR1 and that of barley (Hordeum vulgare) EDR1 (signed as HvEDR1) show 92% identity. There is a highly conserved catalytic domain of serine/threonine protein kinases in the C-terminus of TaEDR1. Because this protein has a putative nuclear localization motif, it probably functions in the nucleus. This study provides the first molecular biological evidence of the presence of an EDR1 homolog in common wheat. The transcription pattern of TaEDR1 was investigated in leaves after inoculation with Blumeria graminis (DC.) E.O. Speer f. sp. tritici Em. Marchal (Bgt) through semi-quantitative RT-PCR (semi-QRT-PCR). The result showed that the transcribing of TaEDR1 was enhanced by Bgt. The expression pattern of the TaEDR1 gene in different tissues showed that it expressed in leaves, stems, spikes and roots. This study suggests that the TaEDR1, a MAP kinase kinase kinase, may function in wheat defense responses.

Amino Acid Sequence↗
PubMed1999

[Molecular identification and marker-assisted selection of Pm21 gene conferring resistance to powdery mildew in wheat] [In Process Citation]

RAPD marker (OPH17(1400)) and SCAR markers (SCAR1400 and SCAR1265) as well as fluorescence in situ hybridization (FISH) technique were used to detect wheat powdery mildew resistance gene Pm21 derived from Haynaldia villosa (6AL/6VS) in different genetic background of wheat breeding program. Among a total of 372 RAPD amplification reactions, 28 reactions (7.53%) yield no amplification products, and the amplification products of 21 reactions (5.64%) were difficult to interpret the presence or absence of the OPH17(1400), indicating that RAPD marker OPH17(1400) is less reproducible and reliable, so that its usage in breeding program is limited. However, SCAR markers, SCAR1400 and SCAR1265, were amplified in all 488 PCR reactions, suggesting that SCAR markers are highly reproducible and reliable, and can be used in breeding program. The specific SCAR1265 made it possible to screen a large number of genotypes reliably and rapidly in practical breeding programs for the presence/absence detection between the resistant and susceptible plants. After backcrossed with common wheat lines for several times, no recombination between the 6VS chromosome of Haynaldia villosa and any of the wheat chromosomes was detected by using FISH.

Journal Article↗