Dataset Information

SNP Discovery for mapping alien introgressions in wheat.

ABSTRACT: BACKGROUND: Monitoring alien introgressions in crop plants is difficult due to the lack of genetic and molecular mapping information on the wild crop relatives. The tertiary gene pool of wheat is a very important source of genetic variability for wheat improvement against biotic and abiotic stresses. By exploring the 5Mg short arm (5MgS) of Aegilops geniculata, we can apply chromosome genomics for the discovery of SNP markers and their use for monitoring alien introgressions in wheat (Triticum aestivum L). RESULTS: The short arm of chromosome 5Mg of Ae. geniculata Roth (syn. Ae. ovata L.; 2n = 4x = 28, UgUgMgMg) was flow-sorted from a wheat line in which it is maintained as a telocentric chromosome. DNA of the sorted arm was amplified and sequenced using an Illumina Hiseq 2000 with ~45x coverage. The sequence data was used for SNP discovery against wheat homoeologous group-5 assemblies. A total of 2,178 unique, 5MgS-specific SNPs were discovered. Randomly selected samples of 59 5MgS-specific SNPs were tested (44 by KASPar assay and 15 by Sanger sequencing) and 84% were validated. Of the selected SNPs, 97% mapped to a chromosome 5Mg addition to wheat (the source of t5MgS), and 94% to 5Mg introgressed from a different accession of Ae. geniculata substituting for chromosome 5D of wheat. The validated SNPs also identified chromosome segments of 5MgS origin in a set of T5D-5Mg translocation lines; eight SNPs (25%) mapped to TA5601 [T5DL?·?5DS-5MgS(0.75)] and three (8%) to TA5602 [T5DL?·?5DS-5MgS (0.95)]. SNPs (gsnp_5ms83 and gsnp_5ms94), tagging chromosome T5DL?·?5DS-5MgS(0.95) with the smallest introgression carrying resistance to leaf rust (Lr57) and stripe rust (Yr40), were validated in two released germplasm lines with Lr57 and Yr40 genes. CONCLUSION: This approach should be widely applicable for the identification of species/genome-specific SNPs. The development of a large number of SNP markers will facilitate the precise introgression and monitoring of alien segments in crop breeding programs and further enable mapping and cloning novel genes from the wild relatives of crop plants.

SUBMITTER: Tiwari VK

PROVIDER: S-EPMC4051138 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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SNP Discovery for mapping alien introgressions in wheat.

Tiwari Vijay K VK Wang Shichen S Sehgal Sunish S Vrána Jan J Friebe Bernd B Kubaláková Marie M Chhuneja Praveen P Doležel Jaroslav J Akhunov Eduard E Kalia Bhanu B Sabir Jamal J Gill Bikram S BS

BMC genomics 20140410

<h4>Background</h4>Monitoring alien introgressions in crop plants is difficult due to the lack of genetic and molecular mapping information on the wild crop relatives. The tertiary gene pool of wheat is a very important source of genetic variability for wheat improvement against biotic and abiotic stresses. By exploring the 5Mg short arm (5MgS) of Aegilops geniculata, we can apply chromosome genomics for the discovery of SNP markers and their use for monitoring alien introgressions in wheat (Tri ...[more]

PMID: 24716476

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Project description:BackgroundTriticum kiharae (AtAtGGDD, 2n?=?42) is of interest for the improvement of bread wheat as a source of high grain protein and gluten content, as well as resistance to many diseases. The use of T. kiharae for the improvement of T. aestivum L. is complicated by the fact that the homology degree of their genomes is low and this leads to an unbalanced set of chromosomes in the gametes of its first generations and the elimination of some genotypes. The aim of this study was to analyze the nature of alien introgressions and their effect on the cytological stability of hybrids obtained from crossing of bread wheat varieties with T. kiharae.ResultsUsing C-banding, the presence of entire chromosomes of T. kiharae in the karyotypes of hybrid lines (intergenomic substitution 2G/2B), chromosome arms (centric translocation ?2AtS:2AL) and large inserts in the form of terminal translocations involving chromosomes of 1st, 3rd and 5th homoeologous groups of B- and G-genomes were found. Molecular markers revealed short introgression of T. kiharae into the genome of common wheat varieties. The highest introgression frequency was shown for 1A, 1B, 2A, 5B, and 6A chromosomes, while no foreign chromatin was detected in 4A and 4B chromosomes. A high level of cytological stability (a meiotic index of 88.18-93.0%) was noted for the majority of introgression lines. An exception was found for the lines containing the structural reorganization of chromosome 5B, affecting the main genes of chromosome synapsis in terms of their functioning.ConclusionsDuring the stabilization of hybrid karyotypes, the introgression of genetic material from T. kiharae into the genome of T. aestivum occurs in the form of short fragments detectable only by molecular markers and in the form of whole chromosomes (intergenomic substitution) and their large fragments (centric and terminal translocations). The level of cytological stability achieved in F10 by the majority of introgression lines ensures the formation of functional gametes sufficient for the successful reproduction of the obtained hybrids.

Project description:Genome-wide introgressions of Thinopyrum bessarabicum into wheat resulted in 12 recombinant lines. Cytological and molecular techniques allowed mapping of 1150 SNP markers across all seven chromosomes of the J genome. Thinopyrum bessarabicum (2n = 2x = 14, JJ) is an important source for new genetic variation for wheat improvement due to its salinity tolerance and disease resistance. Its practical utilisation in wheat improvement can be facilitated through development of genome-wide introgressions leading to a variety of different wheat-Th . bessarabicum translocation lines. In this study, we report the generation of 12 such wheat-Th . bessarabicum recombinant lines, through two different crossing strategies, which were characterized using sequential single colour and multi-colour genomic in situ hybridization (sc-GISH and mc-GISH), multi-colour fluorescent in situ hybridization (mc-FISH) and single nucleotide polymorphic (SNP) DNA markers. We also detected 13 lines containing different Th. bessarabicum chromosome aberrations through sc-GISH. Through a combination of molecular and cytological analysis of all the 25 lines containing Th. bessarabicum recombinants and chromosome aberrations we were able to physically map 1150 SNP markers onto seven Th. bessarabicum J chromosomes which were divided into 36 segmental blocks. Comparative analysis of the physical map of Th. bessarabicum and the wheat genome showed that synteny between the two species is highly conserved at the macro-level and confirmed that Th. bessarabicum contains the 4/5 translocation also present in the A genome of wheat. These wheat-Th . bessarabicum recombinant lines and SNP markers provide a useful genetic resource for wheat improvement with the latter having a wider impact as a tool for detection of introgressions from other Thinopyrum species containing the J or a closely-related genome such as Thinopyrum intermedium (JrJrJvsJvsStSt) and Thinopyrum elongatum (EeEe), respectively.

Project description:BACKGROUND: Wheat (Triticum aestivum L.) is a staple food crop worldwide. The wheat genome has not yet been sequenced due to its huge genome size (approximately 17,000 Mb) and high levels of repetitive sequences; the whole genome sequence may not be expected in the near future. Available linkage maps have low marker density due to limitation in available markers; therefore new technologies that detect genome-wide polymorphisms are still needed to discover a large number of new markers for construction of high-resolution maps. A high-resolution map is a critical tool for gene isolation, molecular breeding and genomic research. Single feature polymorphism (SFP) is a new microarray-based type of marker that is detected by hybridization of DNA or cRNA to oligonucleotide probes. This study was conducted to explore the feasibility of using the Affymetrix GeneChip to discover and map SFPs in the large hexaploid wheat genome. RESULTS: Six wheat varieties of diverse origins (Ning 7840, Clark, Jagger, Encruzilhada, Chinese Spring, and Opata 85) were analyzed for significant probe by variety interactions and 396 probe sets with SFPs were identified. A subset of 164 unigenes was sequenced and 54% showed polymorphism within probes. Microarray analysis of 71 recombinant inbred lines from the cross Ning 7840/Clark identified 955 SFPs and 877 of them were mapped together with 269 simple sequence repeat markers. The SFPs were randomly distributed within a chromosome but were unevenly distributed among different genomes. The B genome had the most SFPs, and the D genome had the least. Map positions of a selected set of SFPs were validated by mapping single nucleotide polymorphism using SNaPshot and comparing with expressed sequence tags mapping data. CONCLUSION: The Affymetrix array is a cost-effective platform for SFP discovery and SFP mapping in wheat. The new high-density map constructed in this study will be a useful tool for genetic and genomic research in wheat.

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SNP Discovery for mapping alien introgressions in wheat.

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SNP Discovery for mapping alien introgressions in wheat.

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