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 β- 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 β-tubulins.
结果:我们报告了紫花苜蓿中微管蛋白基因家族的全面鉴定和表征,包括外显子-内含子组织的分析,重复的基因比较,正确的同种型指定,系统发育分析,以及在不同组织中的表达模式。17α-,鉴定了34个β-和6个γ-微管蛋白基因,并将其分配给特定的同种型。交叉引用了直系同源微管蛋白同种型的识别,涉及系统发育数据,祖先十字花科动物核型重建基因组块的同种学分析和基因分配。对微管蛋白同源物表达模式的研究揭示了N6(A)和N7(B)亚基因组在各个发育阶段的微管蛋白表达中的主要作用,与H7(C)亚基因组转录本的一般优势相反。
结论:首次鉴定了一套完整的微管蛋白基因家族成员,并对其进行了表征。该研究证明了精确推断基因正交的综合方法。所应用的技术不仅可以识别拟南芥模型物种中的苜蓿微管蛋白直向同源物,还可以跟踪微管蛋白基因进化,但也发现拟南芥缺少α-和β-微管蛋白的几种特定同种型的直向同源物。