Abstract

Brassicaceae is a large family of flowering plants, characterized by cruciform corolla, tetradynamous stamen and capsular fruit. In light of the important economic and scientific values of Brassicaceae, many phylogenetic and systematic studies were carried out. One recent and important phylogenetic analysis revealed three major lineages (I, II and III), however, classification at different taxonomic levels (tribe, genus, and species) remained problematic and evolutionary relationships among and within these lineages were still largely unclear. This is partly due to the fact that the past studies lacked information, as they mainly utilized the morphological data, nuclear DNA, partial chloroplast (cp) genes and so on. Nowadays, next generation sequencing (NGS) technology provides the possibility to make use of big data in phylogeny and evolutionary studies. Thus, we sequenced the chloroplast genomes of 80 representative species, using additional 15 reference chloroplast genomes from the NCBI database, and carried out both the phylogenetic reconstruction and the study of protein coding genes evolution in this novel dataset with different methods. Several novel results were obtained. 1 Successful application of NGS technology in chloroplast genome sequencing. During the final assembly, I could reconstruct full chloroplast genomes and the structure maps for 14 out of 80 sampled species, while the remaining were assembled nearly completely with only few gaps remaining. 2 Characterization of chloroplast genome structure. Gene number and order, single sequence repeat (SSR) as well as variety and distribution of large repeat sequence were characterized. 3 The difference of codon usage frequency was calculated between Cardamine resedifolia and Cardamine impatiens. Twelve genes with signatures of positive selection were identified at a family-wide level. 4 Three major lineages (I – III) were confirmed with high support values. Besides, the positions of various tribes were reclassified. Relationships among and within these lineages were highly resolved and supported in the final tree. Most of the tribes in the analyses were inferred to be monophyletic, only Thlaspideae was paraphyletic. Anastaticeae was for the first time classified into position of expanded lineage II, and position of tribe Lepidieae was delimited with relatively low support values in the final phylogenetic tree. This study was a new and successful application of NGS in large-scale Brassicaceae phylogeny and evolution, which offered the chance to look in details of the structural and functional features of the chloroplast genome. These results provided a paradigm on how to proceed towards the full elucidation of the evolutionary relationships among various biological species in the tree of life.