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Hexaploid Wheat (hexaploid + wheat)

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Life Sciences	100%

Selected Abstracts

Efficient cloning of plant genomes into bacterial artificial chromosome (BAC) libraries with larger and more uniform insert size

PLANT BIOTECHNOLOGY JOURNAL, Issue 3 2004
Boulos Chalhoub
Summary The construction of bacterial artificial chromosome (BAC) libraries remains relatively complex and laborious, such that any technological improvement is considered to be highly advantageous. In this study, we addressed several aspects that improved the quality and efficiency of cloning of plant genomes into BACs. We set the ,single tube vector' preparation method with no precipitation or gel electrophoresis steps, which resulted in less vector DNA damage and a remarkable two- to threefold higher transformation efficiency compared with other known vector preparation methods. We used a reduced amount of DNA for partial digestion (up to 5 µg), which resulted in less BAC clones with small inserts. We performed electrophoresis in 0.25 × TBE (Tris, boric acid, ethylenediaminetetraacetic acid) buffer instead of 0.5 × TBE, which resulted in larger and more uniformly sized BAC inserts and, surprisingly, a two- to threefold higher transformation efficiency, probably due to less contamination with borate ions. We adopted a triple size selection that resulted in an increased mean insert size of up to 70 kb and a transformation efficiency comparable with that of double size selection. Overall, the improved protocol presented in this study resulted in a five- to sixfold higher cloning efficiency and larger and more uniformly sized BAC inserts. BAC libraries with the desired mean insert size (up to 200 kb) were constructed from several plant species, including hexaploid wheat. The improved protocol will render the construction of BAC libraries more available in plants and will greatly enhance genome analysis, gene mapping and cloning. [source]

Wheat leaf rust resistance gene Lr59 derived from Aegilops peregrina

PLANT BREEDING, Issue 4 2008
G. F. Marais
Abstract An Aegilops peregrina (Hackel in J. Fraser) Maire & Weiller accession that showed resistance to mixed leaf rust (Puccinia triticina Eriks.) inoculum was crossed with, and backcrossed to, hexaploid wheat (Triticum aestivum L.). During backcrossing a chromosome segment containing a leaf rust resistance gene (here designated Lr59) was spontaneously translocated to wheat chromosome 1A. Meiotic, monosomic and microsatellite analyses suggested that the translocated segment replaced most of, or the complete, 1AL arm, and probably resulted from centromeric breaks and fusion. The translocation, of which hexaploid wheat line 0306 is the appropriate source material, provided seedling leaf rust resistance against a wide range of South African and Canadian pathotypes. [source]

Molecular cytogenetic analysis of a durum wheat ×Thinopyrum distichum hybrid used as a new source of resistance to Fusarium head blight in the greenhouse

PLANT BREEDING, Issue 5 2001
Q. Chen
Abstract Fusarium head blight (FHB, scab), caused by Fusarium graminearum Schwabe, is a serious and damaging disease of wheat. Although some hexaploid wheat lines express a good level of resistance to FHB, the resistance available in hexaploid wheat has not yet been transferred to durum wheat. A germplasm collection of Triticum durum× alien hybrid lines was tested as a potential source of resistance to FHB under controlled conditions. Their FHB reaction was evaluated in three tests against conidial suspensions of three strains of F. graminearum at the flowering stage. Two T. durum×Thinopyrum distichum hybrid lines, ,AFR4' and ,AFR5,, expressed a significantly higher level of resistance to the spread of FHB than other durum-alien hybrid lines and a resistant common wheat line ,Nyu-Bay'. Genomic in situ hybridization using total genomic DNA from alien grass species demonstrated that ,AFR5' had 13 or 14 alien genome chromosomes plus 27 or 28 wheat chromosomes, while ,AFR4' had 22 alien genome and 28 wheat chromosomes. All of the alien chromosomes present in these two lines belonged to the J genome. ,AFR5' is likely to be more useful as a source of FHB resistance than ,AFR4' because of its relatively normal meiotic behaviour, high fertility and fewer number of alien chromosomes. ,AFR5' shows good potential as a source for transferring FHB resistance gene into wheat. The development of T. durum addition lines carrying resistance genes from ,AFR5' is underway. [source]

A new DNA extraction method for high-throughput marker analysis in a large-genome species such as Triticum aestivum

PLANT BREEDING, Issue 4 2001
N. Stein
Abstract Gene mapping and marker-assisted selection in complex, polyploid genomes still relies strongly on restriction fragment length polymorphism (RFLP) analysis, as conversion of RFLP to polymerase chain reaction (PCR) markers can be very difficult. DNA extraction in amounts suitable for RFLP analysis represents the most time-consuming and labour-intensive step in molecular marker analysis of plant populations. In this paper, a new flexible method for plant DNA extraction is presented. It allows a high-throughput of samples in a short time without the need for freezing or lyophilizing the plant material. The method allows the isolation of genomic DNA with a yield of 100 ,g for a minimal amount of 200 mg of leaf material. This is sufficient for work with large-genome plant species such as hexaploid wheat, where 20 ,g of genomic DNA are required for a single RFLP analysis. [source]

A microsatellite marker linked to leaf rust resistance transferred from Aegilops triuncialis into hexaploid wheat

PLANT BREEDING, Issue 3 2001
M. Aghaee-Sarbarzeh
Abstract Aegilops triuncialis (UUCC) is an excellent source of resistance to various wheat diseases, including leaf rust. Leaf rust-resistant derivatives from a cross of a highly susceptible Triticum aestivum cv.,WL711' as the recurrent parent and Ae. triuncialis Ace.3549 as the donor and with and without a pair of acrocentric chromosomes were used for molecular tagging. The use of a set of sequence tagged microsatellite (STMS) markers already mapped to different wheat chromosomes unequivocally indicated that STMS marker gwm368 of chromosome 4BS was tightly linked to the Ae. triuncialis leaf rust resistance gene transferred to wheat. The presence of the Ae. Triuncialis -specific STMS gwm368 homoeoallele along with the non-polymorphic 4BS allele in the rust-resistant derivatives with and without the acrocentric chromosome indicates that the resistance has been transferred from the acrocentric chromosome to either the A or the D genome of wheat. This alien leaf rust resistance gene has been temporarily named as LrTr. [source]

Transcriptional profiling of hexaploid wheat (Triticum aestivum L.) roots identifies novel, dehydration-responsive genes

PLANT CELL & ENVIRONMENT, Issue 5 2007
MOHSEN MOHAMMADI
ABSTRACT We used a long-oligonucleotide microarray to identify transcripts that increased or decreased in abundance in roots of dehydration-tolerant hexaploid bread wheat, in response to withholding of water. We observed that the major classes of dehydration-responsive genes (e.g. osmoprotectants, compatible solutes, proteases, glycosyltransferases/hydrolases, signal transducers components, ion transporters) were generally similar to those observed previously in other species and osmotic stresses. More specifically, we highlighted increases in transcript expression for specific genes including those putatively related to the synthesis of asparagine, trehalose, oligopeptide transporters, metal-binding proteins, the gamma-aminobutyric acid (GABA) shunt and transcription factors. Conversely, we noted a decrease in transcript abundance for diverse classes of glutathione and sulphur-related enzymes, specific amino acids, as well as MATE-efflux carrier proteins. From these data, we identified a novel, dehydration-induced putative AP2/ERF transcription factor, which we predict to function as a transcriptional repressor. We also identified a dehydration-induced ,little protein' (LitP; predicted mass: 8 kDa) that is highly conserved across spermatophytes. Using qRT-PCR, we compared the expression patterns of selected genes between two related wheat genotypes that differed in their susceptibility to dehydration, and confirmed that these novel genes were highly inducible by water limitation in both genotypes, although the magnitude of induction differed. [source]

Differential seedling resistance to the eyespot pathogens, Oculimacula yallundae and Oculimacula acuformis, conferred by Pch2 in wheat and among accessions of Triticum monococcum

PLANT PATHOLOGY, Issue 5 2010
C. Burt
Eyespot is an economically important stem-base disease of wheat caused by two fungal species: Oculimacula yallundae and Oculimacula acuformis. This study investigated the efficacy of two sources of resistance, viz. the genes Pch1, introgressed into hexaploid wheat from Aegilops ventricosa, and Pch2, identified in wheat cv. Cappelle Desprez, against O. yallundae and O. acuformis separately. In a series of seedling bioassays Pch1 was found to be highly effective against both species. Although Pch2 was found to confer resistance against both pathogen species, it was significantly less effective against penetration from O. yallundae than O. acuformis. Furthermore, a quantitative trait locus (QTL) analysis was not able to locate any resistance to O. yallundae on chromosome 7A of Cappelle Desprez. This has important implications for the use of Pch2 in commercial cultivars as it is necessary to have genes that confer resistance to both pathogens for effective eyespot control. In addition, a set of 22 T. monococcum accessions was screened for resistance to both O. yallundae and O. acuformis to identify potentially novel resistances and to assess the accessions for evidence of differential resistance to the eyespot species. Significant differences in resistance to the two pathogens were identified in four of these lines, providing evidence for differential resistance in T. monococcum. This study demonstrates that future screening for novel sources of eyespot resistance should investigate both pathogen species. [source]

Two different CC-NBS-LRR genes are required for Lr10 -mediated leaf rust resistance in tetraploid and hexaploid wheat

THE PLANT JOURNAL, Issue 6 2009
Caroline Loutre
Summary Comparative study of disease resistance genes in crop plants and their relatives provides insight on resistance gene function, evolution and diversity. Here, we studied the allelic diversity of the Lr10 leaf rust resistance gene, a CC-NBS-LRR coding gene originally isolated from hexaploid wheat, in 20 diploid and tetraploid wheat lines. Besides a gene in the tetraploid wheat variety ,Altar' that is identical to the hexaploid wheat Lr10, two additional, functional resistance alleles showing sequence diversity were identified by virus-induced gene silencing in tetraploid wheat lines. In contrast to most described NBS-LRR proteins, the N-terminal CC domain of LR10 was found to be under strong diversifying selection. A second NBS-LRR gene at the Lr10 locus, RGA2, was shown through silencing to be essential for Lr10 function. Interestingly, RGA2 showed much less sequence diversity than Lr10. These data demonstrate allelic diversity of functional genes at the Lr10 locus in tetraploid wheat, and these new genes can now be analyzed for agronomic relevance. Lr10 -based resistance is highly unusual both in its dependence on two, only distantly, related CC-NBS-LRR proteins, as well as in the pattern of diversifying selection in the N-terminal domain. This indicates a new and complex molecular mechanism of pathogen detection and signal transduction. [source]

Detection of HMW glutenin subunit variations among 205 cultivated emmer accessions (Triticum turgidum ssp. dicoccum)

PLANT BREEDING, Issue 2 2006
Q. Y. Li
Abstract The high molecular weight glutenin subunits (HMW-GS) encoded by Glu-1 loci among 205 accessions of cultivated emmer wheat (Triticum turgidum ssp. dicoccum Schrank) collected from different regions of Europe and China were separated and characterized by SDS-PAGE in combination with two-dimensional gel electrophoresis (A-PAGE × SDS-PAGE) and acidic capillary electrophoresis. High genetic polymorphisms in HMW-GS compositions were found. A total of 40 alleles (6 for Glu-A1 and 34 for Glu-B1) and 62 subunit combinations (genotypes) were detected, some of which were not previously described. At Glu-A1 locus, two novel alleles, designated Glu-A1x coding for the subunit 1A × 1.1 and Glu-A1y coding for the subunit 1A × 2.1, were found while seven new subunits (1B × 17*, 1B × 6,, 1B × 13,, 1B × 20*, 1By9*, 1By14.1 and 1By8.1) and 20 novel alleles at Glu-B1 locus were detected. In particular, some additional protein components were detected, which probably were 1Ay subunits encoded by Glu-A1 locus. The introduction of both Ax and Ay subunits from tetraploid wheats into hexaploid wheats may increase the genetic variability of gluten genes and consequently improve flour technological properties. [source]