Molecular genetic characteristics of the <Emphasis Type=Italic>Wx-B1e</Emphasis> allele from common wheat and applicability of the DNA markers for its identification

Molecular genetic characterization of the Wx-B1e allele identified by the authors of the study in the common wheat cultivar Korotyshka was performed. The 804-bp Wx-B1e fragment was cloned and sequenced. Comparison of the sequence obtained with that for the wild-type allele of common wheat (Wx-B1a) demonstrated that Wx-B1e carried the 34-bp insertion, 8-bp deletion, and 23 nucleotide substitutions. BLAST analysis revealed the highest homology with the nucleotide sequences of Wx genes from Triticum spelta and Triticum durum. The amplification variants of four Wx-B1 molecular markers, applied worldwide for testing the collections for different Wx allelic variants, are demonstrated.     

Identity of the Rht-11 and Rht-B1e reduced plant height genes

The Rht (reduced plant height) genes provided a basis for the ??green revolution??, which led to a substantial increase in wheat productivity. More than 20 Rht genes are now known in wheat, Rht-B1 and Rht-11 being among the most important ones. Analysis of a segregating F2 population and molecular analysis of the common wheat cultivars carrying the Rht-11 gene showed that the Rht-11 and Rht-B1e genes are identical.     

Comparative analysis of <Emphasis Type="Italic">Agropyron intermedium</Emphasis> (Host) Beauv 6Ag<Superscript>i</Superscript> and 6Ag<Superscript>i</Superscript>2 chromosomes in bread wheat cultivars and lines with wheat–wheatgrass substitutions

A comparative study of wheat–wheatgrass substituted cultivars and lines resistant to leaf rust developed by the Agricultural Research Institute for Southeast Regions (Multi 6R, Belyanka, Favorit, Voevoda, Lebedushka) and Samara Agricultural Research Institute (Tulaikovskaya 5, Tulaikovskaya 10, Tulaikovskaya 100, Tulaikovskaya Zolotistaya) breeding was conducted. A complex analysis using molecular cytogenetic (C-differential banding, fluorescent (FISH) and genomic (GISH) in situ hybridization), molecular (PLUG markers), and biochemical (electrophoretic analysis of gliadins) markers demonstrated that they have a substitition of wheat chromosome 6D by the chromosomes 6Agⁱ and 6Agⁱ2 belonging to the J(=E) Agropyron intermedium (Host) Beauv (=Thinopyrum intermedium (Host) Barkworth & D.R. Dewey) subgenome. In spite of the fact that the chromosomes 6Agⁱ and 6Agⁱ2 differ in the C-banding pattern and demonstrated minor differences in the blocks of gliadin components, they had the identical pattern of pSc119.2 and pAs1 probe distribution and conjugated between themselves with insignificant disturbance. Thus, it was demonstrated that 6Agi and 6Agⁱ2 are homologous chromosomes; however, the question about allelism of their leaf rust resistance genes between themselves requires special studies. Nevertheless, using STS and SCAR markers and taking into account the type of reaction to Puccinia triticina, their non-allelism to the Lr9, Lr19, Lr24, Lr29, Lr38, and Lr47 genes was established. It was revealed that the 6Agⁱ and 6Agⁱ2 chromosomes have a different level of transmission in hybrid F₂ populations depending on the hybrid combination gene background.     

Molecular cytogenetic mapping of Humulus lupulus sex chromosomes

Dioecy is relatively rare in plants and sex determination systems vary among such species. A good example of a plant with heteromorphic sex chromosomes is hop (Humulus lupulus). The genotypes carrying XX or XY chromosomes correspond to female and male plants, respectively. Until now no clear cytogenetic markers for the sex chromosomes of hop have been established. Here, for the first time the sex chromosomes of hop are clearly identified and characterized. The high copy sequence of hop (HSR1) has been cloned and localized on chromosomes by fluorescence in situ hybridization. The HSR1 repeat has shown subtelomeric location on autosomes with the same intensity of the signal. The signal has been present in the subtelomeric region of the long arm and in the near-centromeric region but absent in the telomeric region of the short arm of the X chromosome. At the same time the signal has been found in the telomeric region only of the long arm of the Y chromosome. This finding indicates that the sex chromosomes of hop have evolved from a pair of autosomes via ancient translocation or inversion. The observation of the meiotic configuration of the sex bivalents shows the location of a pseudoautosomal region on the long arms of X and Y chromosomes.     

Development of a breeder-friendly functional codominant DNA marker for the I2 locus covering resistance to Fusarium oxysporum f. sp. lycopersici

Fusarium oxysporum f. sp. lycopersici Snyder & Hans. (FOL) is a major soil-borne pathogen and the causal agent of Fusarium wilt of tomato, resulting in significant production yield losses. Resistant cultivars have become the most effective method for controlling this fungal disease, and the most important resistance locus to F. oxysporum f. sp. lycopersici in tomato is I2, conferring resistance to race 2 of the pathogen, and widely used in breeding programs. Although this locus was cloned, a robust codominant DNA marker for the I2 locus is not available to date. The development of such a marker has been hindered by the presence of seven homologous sequences at this locus that tend to amplify, and by the absence of information about the structure of the recessive I2 locus. We performed a comparative analysis of the I2 locus nucleotide sequences of tomato genotypes resistant and susceptible to Fusarium wilt. We developed a breeder-friendly functional codominant cleaved amplified polymorphic sequence marker of I2 based on this analysis that can be used in tomato breeding programs for resistance to FOL race 2.     

Cloning and analysis of the resistance gene fragments from silverleaf sunflower Helianthus agrophyllus

Using a combination of degenerate primers designed from the NBS domains of the resistance genes, amplification and subsequent cloning of the resistance gene fragments from sunflower (Helianthus agrophyllus) was conducted. Sequences of cloned PCR products differed from one another and displayed homology to NBS domain fragments of the already known plant resistance genes, as well as to the analogous genes from different classes. The highest homology was shown to the NBS domain regions of cultivated sunflower and the other members of the family Compositae. Two cloned fragments had open reading frames, while the other sequences carried stop codons and seemed to belong to pseudogenes. Amino acid sequences of Helianthus agrophyllus analyzed contained conservative regions typical of NBS domains of the resistance gene products.     

Synthesis and structure of titanium alkoxides based on tetraphenyl substituted 2,6-dimethanolpyridine moiety

Novel titanocanes and spirobititanocanes based on 2,6-bis[hydroxy(diphenyl)methyl]pyridine (1a) and 2,6-di(hydroxymethyl)pyridine (1b) - [2,6-C₅H₃N(CPh₂O)₂]Ti(O-i-Pr)₂ (2a), [2,6-C₅H₃N(CPh₂O)₂]₂Ti (3a), [2,6-C₅H₃N(CH₂O)₂]₂Ti (3b), [2,6-C₅H₃N(CPh₂O)₂]TiCl₂ (4) - as well as the closely related N-phenyl derivative PhN(CH₂CH₂O)₂Ti(Cl)Cp (5) have been synthesized. Complexes 2-5 were characterized by ¹H and ¹³C NMR spectroscopy and elemental analysis data. The molecular structure of 3a was determined by X-ray structure analysis.     

Synthesis and properties of diadamantyl-containing symmetric diureas as target-oriented inhibitors of human soluble epoxide hydrolase

A series of target-oriented competitive inhibitors of human soluble epoxide hydrolase have been synthesized. The compounds retain the inhibitory properties at concentrations down to 4 nM. Based on the results of molecular modeling, it has been shown that the high inhibitory activity of this series of compounds is achieved by a unique mode of the binding to the active site of the enzyme.     

Cas9-targeted Nanopore sequencing rapidly elucidates the transposition preferences and DNA methylation profiles of mobile elements in plants

Transposable element insertions (TEIs) are an important source of genomic innovation by contributing to plant adaptation, speciation, and the production of new varieties. The often large, complex plant genomes make identifying TEIs from short reads difficult and expensive. Moreover, rare somatic insertions that reflect mobilome dynamics are difficult to track using short reads. To address these challenges, we combined Cas9-targeted Nanopore sequencing (CANS) with the novel pipeline NanoCasTE to trace both genetically inherited and somatic TEIs in plants. We performed CANS of the EVADÉ (EVD) retrotransposon in wild-type Arabidopsis thaliana and rapidly obtained up to 40× sequence coverage. Analysis of hemizygous T-DNA insertion sites and genetically inherited insertions of the EVD transposon in the ddm1 (decrease in DNA methylation 1) genome uncovered the crucial role of DNA methylation in shaping EVD insertion preference. We also investigated somatic transposition events of the ONSEN transposon family, finding that genes that are downregulated during heat stress are preferentially targeted by ONSENs. Finally, we detected hypomethylation of novel somatic insertions for two ONSENs. CANS and NanoCasTE are effective tools for detecting TEIs and exploring mobilome organization in plants in response to stress and in different genetic backgrounds, as well as screening T-DNA insertion mutants and transgenic plants.