I. N. Leonova

Microsatellite mapping of the induced sphaerococcoid mutation genes in Triticum aestivum

Abstract The S1, S2 and S3 genes of the induced sphaerococcoid mutation in common wheat (Triticum aestivum) were mapped using three different F2 populations consisting of 71–96 individual plants. Twenty-four microsatellite markers from homeologous group 3 of T. aestivum were used to map the S1, S2 and S3 genes on chromosomes 3D, 3B and 3A, respectively. The S1 locus was found to be closely linked to the centromeric marker Xgwm456 of the long arm (2.9 cM) and mapped not far (8.0 cM) from the Xgdm72 marker of the short arm of chromosome 3D. The S2 gene was tightly linked to 2 centromeric markers (Xgwm566, Xgwm845) of chromosome 3B. S3 was located between Xgwm2 (5.1 cM), the marker of the short arm, and Xgwm720 (6.6 cM), the marker of the long arm, both of chromosome 3A. Mapping the S1, S2 and S3 loci of the induced sphaerococcoid mutation near the centromeric regions supports the hypothesis that the sphaerococcum type may be due to gene duplication resulting from DNA recombination in the centromeric region.

Wheat genome structure: translocations during the course of polyploidization.

The genomic organization of Triticum timopheevii (2n=28, AtAtGG) was compared with hexaploid wheat T. aestivum (2n=42, AABBDD) by comparative mapping using microsatellites derived from bread wheat. Genetic maps for the two crosses T. timopheevii var. timopheevii × T. timopheevii var. typica and T. timopheevii K-38555×T. militinae were constructed. On the first population, 121 loci were mapped, and on the second population 103 loci. The transferability of the wheat markers to T. timopheevii was generally better for the A genome-specific markers (76–78% produced amplification products; 26 and 29% were polymorphic) than for B genome-specific markers (54% produced amplification products; 14 and 16% were polymorphic). Of the D genome-specific markers, one third produced amplification products in T. timopheevii, but only 5 and 2% were polymorphic in the corresponding mapping populations. The maps constructed confirmed the previously described translocation between chromosome arms 6AtS and 1GS and revealed at least two yet unknown rearrangements on chromosomes 4At and 6At. The presence of other translocations and rearrangements between T. timopheevii and T. aestivum was demonstrated by a variety of markers mapping to nonhomoeologous positions.

Detection of quantitative trait loci for leaf rust resistance in wheat--T. timopheevii/T. tauschii introgression lines

Advanced backcross QTL analysis was used to identify QTLs for seedling and adult plant resistance to leaf rust in introgression lines derived from a cross between the spring wheat cultivar ‘Saratovskaya 29’ and a synthetic allopolyploid wheat (T. timopheevii/T. tauschii). F2 mapping populations involving two backcross selections (‘BC5’ and ‘BC9’ lines) were genotyped with microsatellite markers. Two significant QTL for adult plant resistance were identified in line ‘BC5’: one on chromosome 2B, but originating from chromosome 2G, explained 31% of the trait variance. The other, derived from T. tauschii and mapped to the short arm of chromosome 2D explained 19% of the trait variance. In the second line, one major seedling and adult plant resistance QTL was identified on chromosome 2B. Both QTL co-located to the same marker interval. Such introgression lines, resulting from the reconstruction of common wheat genome, are of interest both as initial material for breeding and improvement of current cultivars, and as a resource for the study of the interaction and transformation of genomes.

Molecular Analysis of Leaf Rust-Resistant Introgression Lines Obtained by Crossing of Hexaploid Wheat Triticum aestivum with Tetraploid Wheat Triticum timopheevii

Twenty-four Triticum aestivum×T. timopheevii hybrid lines developed on the basis of five varieties of common wheat and resistant to leaf rust were analyzed by the use of microsatellite markers specific for hexaploid wheat T. aestivum. Investigation of intervarietal polymorphism of the markers showed that the number of alleles per locus ranged from 1 to 4, depending on the marker (2.5 on average). InT. timopheevii, amplification fragments are produced by 80, 55, and 30% of primers specific to the A, B, and D common wheat genomes, respectively. Microsatellite analysis revealed two major areas of introgression of the T. timopheevii genome: chromosomes of homoeological groups 2 and 5. Translocations were detected in the 2A and 2B chromosomes simultaneously in 11 lines of 24. The length of the translocated fragment in the 2B chromosome was virtually identical in all hybrid lines and did not depend on the parental wheat variety. In 15 lines developed on the basis of the Saratovskaya-29, Irtyshanka, and Tselinnaya-20, changes occurred in the telomeric region of the long arm of the 5A chromosome. Analysis with markers specific to the D genome suggested that introgressions of the T. timopheevii genome occurred in chromosomes of the D genome. However, the location of these markers on T. timopheevii chromosomes is unknown. Our data suggest that the genes for leaf rust resistance transferred from T. timopheevii to T. aestivum are located on chromosomes of homoeological group 2.

Molecular analysis of the triticale lines with different Vrn gene systems using microsatellite markers and hybridization in situ

Hexaploid triticale (×Triticosecale Wittmack) lines were examined using molecular markers and the hybridization in situ technique. Triticale lines were generated based on wheat varieties differing by the Vrn gene systems and the earing times. Molecular analysis was performed using Xgwm and Xrms microsatellite markers with the known chromosomal localization in the common wheat Triticum aestivum, and rye Secale cereale genomes. Comparative molecular analysis of triticale lines and their parental forms showed that all lines contained A and B genomes of common wheat and also rye homoeologous chromosomes. In the three lines the presence of D genome markers, mapped to the chromosomes 2D and 7D, was demonstrated. This was probably the consequence of the translocations of homoeologous chromosomes from wheat genomes, which took part during the process of triticale formation. The data obtained by use of genomic in situ hybridization supported the data of molecular genetic analysis. In none of the lines wheat-rye translocations or recombinations were observed. These findings suggest that the change of the period between the seedling appearance and earing time in triticale lines compared to the initial wheat lines, resulted from the inhibitory effect of rye genome on wheat vernalization genes.

Diversification of the Duplicated F3h Genes in Triticeae

The F3h gene encodes the flavonoid synthesis enzyme flavanone 3-hydroxylase. Unlike most plant genomes, the bread wheat (Triticum aestivum L.) B genome has two, rather than just one F3h copy. The paralogous F3h-B2 sequence was isolated by PCR and shown to be transcribed, but its predicted polypeptide differed from the typical F3H sequence at a number of the conserved residues associated with its putative substrate-binding sites. The F3h-B2 promoter region was highly divergent from that of F3h-B1, and the transcriptional profiles of the two genes were distinct. Among a panel of 95 Triticeae accessions, representing 24 species, an F3h-2 copy was only detected within those carrying a B, S, G, or an R genome. Analysis of the coding sequence divergence suggested that a small segmental duplication occurred early in the evolution of the Triticeae tribe. The duplicated F3h copy appears to have acquired a novel function in the progenitor of the closely related B, G, and S genomes, as well as in the R genome. In other Triticeae genomes, the F3h-2 paralog may have been lost following pseudogenization.

Genetic analysis and localization of loci controlling leaf rust resistance of Triticum aestivum × Triticum timopheevii introgression lines

Introgressive lines resulting from crossing common wheat Triticum aestivum with the tetraploid T. timopheevii are characterized by effective resistance to leaf rust caused by Puccinia triticina Eriks. Molecular analysis using 350 specific simple sequence repeat (SSR) markers determined localization of the T. timopheevii genome in chromosomes 1A, 2A, 2B, 5A, 5B, and 6B. A population of F2 offspring of crossing hybrid line 842-2 with common wheat cultivar Skala was obtained for mapping the loci controlling leaf rust resistance. Analysis of association of phenotypic and genotypic data by means of simple interval mapping (SIM) and composite interval mapping (CIM) has shown that the resistance of adult plants is determined by two loci in chromosomes 5B and 2A. The major locus QLr.icg-5B, transferred from T. timopheevii chromosome 5G mapped to the interval of microsatellite loci Xgwm408-Xgwm1257 controls 72% of the phenotypic variance of the trait. The other, minor locus QLr.icg-2A located to chromosome 2A at a distance of 10 cM from Xgwm312 accounts for 7% of the trait expression. Microsatellite markers located near these loci may be used for controlling the transfer of agronomically valuable loci when new lines and cultivars are created.

Molecular markers: Implementation in crop plant breeding for identification, introgression and gene pyramiding

Over the past decades, wide theoretical and practical experience has been obtained in the application of DNA markers for the investigation of plant genetic diversity, the construction of molecular genetic maps, the mapping of genes and quantitative trait loci, and the employment of molecular marker technologies in the development of commercial cultivars and breeding of crop lines. To date, the main practical application of molecular markers is related to germplasm characterization, introgression and the pyramiding of genome fragments associated with agronomically important traits controlled by major genes. The contribution of new technologies to the selection of traits with multigenic inheritance is still insignificant. Despite considerable progress in plant molecular genetics and genomics methods and great interest in new technologies among breeders, there are a large number of constraints affecting the implementation of new technologies in breeding practice. This article considers the potential application of DNA markers in the breeding of crop plants and the benefits and limitations of the use of marker-assisted technologies in comparison with conventional plant breeding methods.

Comparative cytological and molecular analysis of common wheat introgression lines containing genetic material of Triticum timopheevii Zhuk

A total of 40 introgression lines of common wheat (2 n = 42) Triticum aestivum L × T. timopheevii Zhuk., resistant to leaf rust and partly to powdery mildew, were examined. Based on cytological analysis of meiosis in pollen mother cells (PMC), hybrid lines were subdivided into two groups characterized by either stable or unstable meiosis. In cytologically stable lines, chromosome configuration at the MI stage of meiosis was mostly bivalent (21II) with small proportion of defect cells (almost 10%), which at most contained two univalents (20II + 2I). Cytologically unstable group was comprised of the lines, containing high proportions of cells with abnormal chromosome pairing in meiotic PMC, as well as the cells with multivalents, and the lines containing aneuploid plants. Localization of the T. timopheevii fragments performed with the use of SSR markers showed that the lines with unstable meiosis were characterized by higher numbers of introgressions compared to stable lines. The influence of certain chromosomes of T. timopheevii on chromosome pairing stability was also demonstrated. In cytologically unstable lines, the increased frequency of 2A substitutions along with the high frequency of introgression of T. timopheevii genetic material into chromosome 7A was observed. Multivalents were scored in all cases of introgression in chromosome 7A. It was suggested that the reason for the genome instability in hybrid forms lied in insufficient compensating ability of certain T. timopheevii chromosomes and/or their parts, involved into recombination processes.

Preferential Elimination of Chromosome 5R of Rye in the Progeny of 5R5D Dimonosomics

Transmission of chromosome 5R of rye (Secale cereale L.) and chromosome 5D of common wheat (Triticum aestivum L.) through gametes of 5R5D dimonosomics (2n = 42, 20W″ + 5R′ + 5D′) was studied. Chromosome 5R was found to have lower competitiveness as compared to 5D. Gametes with the rye chromosome were two times less often involved in the formation of a progeny. The combined frequency of the karyotypes of wheat (5D5D) and wheat monosomics (5D) was 11.6-fold higher than the frequency of the karyotypes of substitution lines (5R5R) and monosomics for the rye chromosome (5R). The karyotypes of 10.38% of hybrid plants had aberrant 5R chromosomes with different translocations formed as a result of breakages in the centromere and in the proximal region of the long arm. Telocentrics for the short arm t5RS, i5RS isochromosomes, and chromosomes with a terminal deletion T5RS.5RL-del were identified. The absence of amplification of SSR markers mapped on 5RS and the detection of PCR products for a number of 5RL markers (including the genome-specific rye marker Xrms115) permitted nine plants carrying only the long arm of chromosome 5R to be revealed. Since t5RL telocentrics were not detected by the cytological analysis, the results obtained allow us to suggest the presence of small intercalary translocations of the long arm of chromosome 5R in chromosome 5D or in other wheat chromosomes.