DArT Markers Effectively Target Gene Space in the Rye Genome

Large genome size and complexity hamper considerably the genomics research in relevant species. Rye (Secale cereale L.) has one of the largest genomes among cereal crops and repetitive sequences account for over 90% of its length. Diversity Arrays Technology is a high-throughput genotyping method, in which a preferential sampling of gene-rich regions is achieved through the use of methylation sensitive restriction enzymes. We obtained sequences of 6,177 rye DArT markers and following a redundancy analysis assembled them into 3,737 non-redundant sequences, which were then used in homology searches against five Pooideae sequence sets. In total 515 DArT sequences could be incorporated into publicly available rye genome zippers providing a starting point for the integration of DArT- and transcript-based genomics resources in rye. Using Blast2Go pipeline we attributed putative gene functions to 1101 (29.4%) of the non-redundant DArT marker sequences, including 132 sequences with putative disease resistance-related functions, which were found to be preferentially located in the 4RL and 6RL chromosomes. Comparative analysis based on the DArT sequences revealed obvious inconsistencies between two recently published high density consensus maps of rye. Furthermore we demonstrated that DArT marker sequences can be a source of SSR polymorphisms. Obtained data demonstrate that DArT markers effectively target gene space in the large, complex, and repetitive rye genome. Through the annotation of putative gene functions and the alignment of DArT sequences relative to reference genomes we obtained information, that will complement the results of the studies, where DArT genotyping was deployed, by simplifying the gene ontology and microcolinearity based identification of candidate genes

Comparative distribution of markers on consensus reference wheat map and linkage groups from A genome of HM-DH population.

<p>Comparative distribution of markers on consensus reference wheat map and linkage groups from A genome of HM-DH population.</p

Representational Difference Analysis of Transcripts Involved in Jervine Biosynthesis

Veratrum-type steroidal alkaloids (VSA) are the major bioactive ingredients that strongly determine the pharmacological activities of Veratrum nigrum. Biosynthesis of VSA at the molecular and genetic levels is not well understood. Next-generation sequencing of representational difference analysis (RDA) products after elicitation and precursor feeding was applied to identify candidate genes involved in VSA biosynthesis. A total of 12,048 contigs with a median length of 280 bases were received in three RDA libraries obtained after application of methyl jasmonate, squalene and cholesterol. The comparative analysis of annotated sequences was effective in identifying candidate genes. GABAT2 transaminase and hydroxylases active at C-22, C-26, C-11, and C-16 positions in late stages of jervine biosynthesis were selected. Moreover, genes coding pyrroline-5-carboxylate reductase and enzymes from the short-chain dehydrogenases/reductases family (SDR) associated with the reduction reactions of the VSA biosynthesis process were proposed. The data collected contribute to better understanding of jervine biosynthesis and may accelerate implementation of biotechnological methods of VSA biosynthesis

Identification of 1bl/1rs translocation in interspecific hybrids between Aegilops and Triticum.

The aim of studies was testing for presence of 1BL/1RS translocation in 17 lines derived from wide crosses of Aegilops ventricosa and Aegilops juvenalis with common and durum wheats, along with parental forms. PCR analyses detected translocated chromosome in Lanca and CZR1406 wheats and in 4 hybrid lines (JCPC, JCCP, JCC and JCCC). The lack of 1RS arm in lines (VGL and VGLL) derived from crosses with Lanca suggests that another chromosomal rearrangements occurred. We confirmed the usefulness of applied STS-PCR assays for fast and robust identification of 1BL/1RS translocation in breeding materials

Variability in Expression of Male Fertility in Triticale (Xtriticosecale Wittmack) With Triticum Timopheevi Cytoplasm

We present a reliable, visual method for evaluation of the level of male fertility during flowering, that is indispensable for breeding of hybrid cultivars of winter triticale based on the cms-T.timopheevi system. Detailed observations of anther development were performed on 20 F2 and BC1 plants derived from crosses between male-sterile and fertility restoring lines. Variation of anther development within florets, spikelets, spikes, and among spikes of the same plant was examined..