Fast forward genetics to identify mutations causing a high light tolerant phenotype in Chlamydomonas reinhardtii by whole-genome-sequencing

BMC Genomics
Lisa SchierenbeckOlaf Kruse

Abstract

High light tolerance of microalgae is a desired phenotype for efficient cultivation in large scale production systems under fluctuating outdoor conditions. Outdoor cultivation requires the use of either wild-type or non-GMO derived mutant strains due to safety concerns. The identification and molecular characterization of such mutants derived from untagged forward genetics approaches was limited previously by the tedious and time-consuming methods involving techniques such as classical meiotic mapping. The combination of mapping with next generation sequencing technologies offers alternative strategies to identify genes involved in high light adaptation in untagged mutants. We used the model alga Chlamydomonas reinhardtii in a non-GMO mutation strategy without any preceding crossing step or pooled progeny to identify genes involved in the regulatory processes of high light adaptation. To generate high light tolerant mutants, wildtype cells were mutagenized only to a low extent, followed by a stringent selection. We performed whole-genome sequencing of two independent mutants hit1 and hit2 and the parental wildtype. The availability of a reference genome sequence and the removal of shared bakground variants between the wildtype ...Continue Reading

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Datasets Mentioned

BETA
SRP037721
SRS557198
SRS558641
SRS558642

Methods Mentioned

BETA
nucleotide exchange
PCR
RNA-seq
ubiquitination

Software Mentioned

SAMtools
Genome Analysis Toolkit
SAMtools mpileup
- PdbViewer
GATK Haplotype Caller
TASSER
awk
Trimmomatic Aligner ( BWA )
BWA Aligner
Swiss

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