Abstract

The sequencing and comparison of plastid genomes are becoming a standard method in plant genomics, and many researchers are using this approach to infer plant phylogenetic relationships. Due to the widespread availability of next-generation sequencing, plastid genome sequences are being generated at breakneck pace. This trend towards massive sequencing of plastid genomes highlights the need for standardized bioinformatic workflows. In particular, documentation and dissemination of the details of genome assembly, annotation, alignment and phylogenetic tree inference are needed, as these processes are highly sensitive to the choice of software and the precise settings used. Here, we present the procedure and results of sequencing, assembling, annotating and quality-checking of three complete plastid genomes of the aquatic plant genus Cabomba as well as subsequent gene alignment and phylogenetic tree inference. We accompany our findings by a detailed description of the bioinformatic workflow employed. Importantly, we share a total of eleven software scripts for each of these bioinformatic processes, enabling other researchers to evaluate and replicate our analyses step by step. The results of our analyses illustrate that the plastid genomes of Cabomba are highly conserved in both structure and gene content.

Highlights

  • The sequencing of plastid genomes constitutes a reliable and well-tested method in plant genomic research

  • Between three and four contigs with a length equal or greater than 1000 bp were generated by the automatic assembly process, with the largest contig matching the large single-copy (LSC) region in each sample

  • We found that the plastid genomes of Cabomba are highly conserved in both structure and phylogenetic inference process for three complete plastid genomes of the aquatic plant genus and gene content

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Summary

Introduction

The sequencing of plastid genomes constitutes a reliable and well-tested method in plant genomic research. Sequencing plastid genomes for phylogenetic purposes (commonly referred to as “plastid phylogenomics”) has been conducted since the mid-2000s (see [1] for a review). Plastid genome sequences have been used to infer phylogenetic relationships among genera [9,10] and species [11,12,13], to resolve recently radiated plant lineages [14]. Several factors render the comparison of plastid genomes beneficial for phylogenetic analysis. The majority of plastid genomes display strong structural conservation in both gene composition and synteny, enabling the comparison of primarily homologous regions across a wide variety of taxa [16]. Plastid genomes are typically uniparentally (maternally) inherited and display

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