Abstract

BackgroundThe plastid acquisition by secondary endosymbiosis is a driving force for the algal evolution, and the comparative genomics was required to examine the genomic change of symbiont. Therefore, we established a pipeline of a de novo assembly of middle-sized genomes at a low cost and with high quality using long and short reads.ResultsWe sequenced symbiotic algae Chlorella variabilis using Oxfofrd Nanopore MinION as the long-read sequencer and Illumina HiSeq 4000 as the short-read sequencer and then assembled the genomes under various conditions. Subsequently, we evaluated these assemblies by the gene model quality and RNA-seq mapping rate. We found that long-read only assembly could not be suitable for the comparative genomics studies, but with short reads, we could obtain the acceptable assembly. On the basis of this result, we established the pipeline of de novo assembly for middle-sized algal genome using MinION.ConclusionsThe genomic change during the early stages of plastid acquisition can now be revealed by sequencing and comparing many algal genomes. Moreover, this pipeline offers a solution for the assembly of various middle-sized eukaryotic genomes with high-quality and ease.

Highlights

  • The plastid acquisition by secondary endosymbiosis is a driving force for the algal evolution, and the comparative genomics was required to examine the genomic change of symbiont

  • Plastid acquisition by secondary endosymbiosis consists of four stages: In the first stage, host organisms prey on algae, and undigested algae temporarily become symbionts

  • We found that the total length was about 10 Mbps shorter than the reference assembly and that the eukaryotic genes conservation rate (ECR) and RNA-seq mapping rates were drastically reduced after the scaffolding step using long reads (Fig. 4b)

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Summary

Introduction

The plastid acquisition by secondary endosymbiosis is a driving force for the algal evolution, and the comparative genomics was required to examine the genomic change of symbiont. Evolutionary research has revealed divergence in photosynthetic eukaryotes with plastid acquisition by secondary endosymbiosis as the driving force [1,2,3]. This phenomenon has occurred many times over the course of evolution. Plastid acquisition by secondary endosymbiosis consists of four stages: In the first stage, host organisms prey on algae, and undigested algae temporarily become symbionts. Curtis et al [5] previously studied the third stage of plastid acquisition by genome and transcriptome analysis of cryptophytes and chlorarachniophytes. Few studies have focused on the early stages of plastid acquisition

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