Genome-wide identification and evolution of the tubulin gene family in Camelina sativa.

BACKGROUND: Tubulins play crucial roles in numerous fundamental processes of plant development. In flowering plants, tubulins are grouped into α-, ß- and γ-subfamilies, while α- and ß-tubulins possess a large isotype diversity and gene number variations among different species. This circumstance leads to insufficient recognition of orthologous isotypes and significantly complicates extrapolation of obtained experimental results, and brings difficulties for the identification of particular tubulin isotype function. The aim of this research is to identify and characterize tubulins of an emerging biofuel crop Camelina sativa. RESULTS: We report comprehensive identification and characterization of tubulin gene family in C. sativa, including analyses of exon-intron organization, duplicated genes comparison, proper isotype designation, phylogenetic analysis, and expression patterns in different tissues. 17 α-, 34 ß- and 6 γ-tubulin genes were identified and assigned to a particular isotype. Recognition of orthologous tubulin isotypes was cross-referred, involving data of phylogeny, synteny analyses and genes allocation on reconstructed genomic blocks of Ancestral Crucifer Karyotype. An investigation of expression patterns of tubulin homeologs revealed the predominant role of N6 (A) and N7 (B) subgenomes in tubulin expression at various developmental stages, contrarily to general the dominance of transcripts of H7 (C) subgenome. CONCLUSIONS: For the first time a complete set of tubulin gene family members was identified and characterized for allohexaploid C. sativa species. The study demonstrates the comprehensive approach of precise inferring gene orthology. The applied technique allowed not only identifying C. sativa tubulin orthologs in model Arabidopsis species and tracking tubulin gene evolution, but also uncovered that A. thaliana is missing orthologs for several particular isotypes of α- and ß-tubulins.

Intron length polymorphism of ß-tubulin genes of Aegilops biuncialis Vis.

Intron-specific DNA polymorphism is present among plant ß-tubulin gene family members and is considered to be one of the molecular markers based on the difference of tubulin introns length assayed both separately (TBP: 1st intron) or in combination (h-TBP: 1st and 2nd introns). These two approaches are possibly useful for wheat breeding programs, since TBP and h-TBP help to differentiate between the accessions of Aegilops biuncialis Vis., a wild relative of wheat. PCR-derived polymorphic fragments were resolved by PAGE electrophoresis. The length of amplicons varied significantly (395-3900 bp for TBP and 466-3440 bp for h-TBP), while the numbers of polymorphic bands were 21 for TBP and 23 for h-TBP, respectively. PIC mean value was circa 0.3. Dendrograms constructed on the basis of the Nei and Li coefficient with the high bootstrap support reveal a similar order of hierarchy for the samples analyzed using both methods. Thus, both techniques uncover DNA polymorphism level sufficiently high to distinguish different accessions of Ae. biuncialis Vis.