Comparative analysis of early divergent land plants and construction of DNA tools for hyper-expression in Marchantia chloroplasts

ABSTRACT Chloroplast genes are present at high ploidy in plants, and capable of driving very high levels of gene expression if mRNA production and stability are properly regulated. Marchantia polymorpha is a simple model plant that allows rapid transformation studies, however post-transcriptional re...

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Published inbioRxiv
Main Authors Frangedakis, Eftychios, Guzman-Chavez, Fernando, Rebmann, Marius, Markel, Kasey, Yu, Ying, Perraki, Artemis, Tse, Sze Wai, Liu, Yang, Rever, Jenna, Sauret-Gueto, Susanna, Goffinet, Bernard, Schneider, Harald, Haseloff, Jim
Format Paper
LanguageEnglish
Published Cold Spring Harbor Cold Spring Harbor Laboratory Press 27.11.2020
Subjects
Binding sites
Chloroplasts
Comparative analysis
Deoxyribonucleic acid
Divergence
DNA
Gene expression
Gene regulation
Genetic transformation
Marchantia
mRNA stability
Plastids
Ploidy
Post-transcription
Proteins
Reporter gene
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Summary:ABSTRACT Chloroplast genes are present at high ploidy in plants, and capable of driving very high levels of gene expression if mRNA production and stability are properly regulated. Marchantia polymorpha is a simple model plant that allows rapid transformation studies, however post-transcriptional regulation in plastids is poorly characterized in this liverwort. We have mapped patterns of transcription in Marchantia chloroplasts. Furthermore, we have obtained and compared sequences from 51 early-divergent plant species, and identified putative sites for pentatricopeptide repeat protein binding that are thought to play important roles in mRNA stabilisation. Candidate binding sites were tested for their ability to confer high levels of reporter gene expression in Marchantia chloroplasts, and levels of protein production and effects on growth were measured in homoplasmic transformed plants. We have produced novel DNA tools for protein hyper-expression in a facile plant system that is a test-bed for chloroplast engineering. Competing Interest Statement The authors have declared no competing interest.
DOI:10.1101/2020.11.27.401802