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Reliable reference genes and abiotic stress marker genes in Klebsormidium nitens.

ABSTRACT: Microalgae have recently emerged as a key research topic, especially as biological models. Among them, the green alga Klebsormidium nitens, thanks to its particular adaptation to environmental stresses, represents an interesting photosynthetic eukaryote for studying the transition stages leading to the colonization of terrestrial life. The tolerance to different stresses is manifested by changes in gene expression, which can be monitored by quantifying the amounts of transcripts by RT-qPCR. The identification of optimal reference genes for experiment normalization was therefore necessary. In this study, using four statistical algorithms followed by the RankAggreg package, we determined the best reference gene pairs suitable for normalizing RT-qPCR data in K. nitens in response to three abiotic stresses: high salinity, PEG-induced dehydration and heat shock. Based on these reference genes, we were able to identify marker genes in response to the three abiotic stresses in K. nitens.

SUBMITTER: Chatelain P

PROVIDER: S-EPMC9643330 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature

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Reliable reference genes and abiotic stress marker genes in Klebsormidium nitens.

Chatelain Pauline P Blanchard Cécile C Astier Jeremy J Klinguer Agnès A Wendehenne David D Jeandroz Sylvain S Rosnoblet Claire C

Scientific reports 20221108 1

Microalgae have recently emerged as a key research topic, especially as biological models. Among them, the green alga Klebsormidium nitens, thanks to its particular adaptation to environmental stresses, represents an interesting photosynthetic eukaryote for studying the transition stages leading to the colonization of terrestrial life. The tolerance to different stresses is manifested by changes in gene expression, which can be monitored by quantifying the amounts of transcripts by RT-qPCR. The ...[more]

PMID: 36348043

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Project description:During the evolution of life on Earth, the conquest of land by plants played a pivotal role producing a boost in land biomass, a substantial drop in atmospheric CO2, an increase in oxygen and the emergence of new terrestrial habitats facilitating land colonization by animals. Therefore, the characterization of the molecular mechanisms that allowed plant terrestralization is a cornerstone in evolutionary studies. Viridiplantae or the green lineage is divided into two clades Chlorophyta or green microalgae and Streptophyta that in turn splits into Embryophyta or land plants and Charophyta. The latest are mainly considered aquatic algae although some facultative terrestrial species has been identified. Charophyta are generally accepted as the extant algal species most closely related to current land plants and, therefore, they are used in evolutionary studies on plant terrestralization. High light irradiance was one of the major stressors that ancestral charophytic algae needed to overcome during the transition from aquatic to terrestrial environments. In this study, we have chosen the facultative terrestrial early charophytic alga Klebsormidium nitens to perform an integrative transcriptomic and metabolomic analysis under high light in order to unveil key mechanisms involved in the early steps of plants terrestralization. We found a fast chloroplast retrograde signaling possibly mediated by reactive oxygen species and the inositol polyphosphate 1-phosphatase (SAL1) and 3′-phosphoadenosine-5′-phosphate (PAP) pathways inducing gene expression and accumulation of specific metabolites. Systems used by both Chlorophyta and Embryophyta were activated such as the xanthophyll cycle with an accumulation of zeaxanthin and protein folding and repair mechanisms constituted by NADPH-dependent thioredoxin reductases, thioredoxin-disulfide reductases and peroxiredoxins. Similarly, cyclic electron flow, specifically the pathway dependent on Proton Gradient Regulation 5, was strongly activated under high light. We detected a simultaneous co-activation of the non-photochemical quenching mechanisms based on LHC-like Stress Related protein and the photosystem II subunit S that are specific to Chlorophyta and Embryophyta respectively. Exclusive Embryophyta systems for the synthesis, sensing and response to the phytohormone auxin were also activated under high light in Klebsormidium leading to an increase in auxin content with the concomitant accumulation of amino acids such as tryptophan, histidine and phenylalanine.

Project description:The characterization of the molecular mechanisms, such as high light irradiance resistance, that allowed plant terrestralization is a cornerstone in evolutionary studies since the conquest of land by plants played a pivotal role in life evolution on Earth. Viridiplantae or the green lineage is divided into two clades, Chlorophyta and Streptophyta, that in turn splits into Embryophyta or land plants and Charophyta. Charophyta are used in evolutionary studies on plant terrestralization since they are generally accepted as the extant algal species most closely related to current land plants. In this study, we have chosen the facultative terrestrial early charophyte alga Klebsormidium nitens to perform an integrative transcriptomic and metabolomic analysis under high light in order to unveil key mechanisms involved in the early steps of plants terrestralization. We found a fast chloroplast retrograde signaling possibly mediated by reactive oxygen species and the inositol polyphosphate 1-phosphatase (SAL1) and 3'-phosphoadenosine-5'-phosphate (PAP) pathways inducing gene expression and accumulation of specific metabolites. Systems used by both Chlorophyta and Embryophyta were activated such as the xanthophyll cycle with an accumulation of zeaxanthin and protein folding and repair mechanisms constituted by NADPH-dependent thioredoxin reductases, thioredoxin-disulfide reductases, and peroxiredoxins. Similarly, cyclic electron flow, specifically the pathway dependent on proton gradient regulation 5, was strongly activated under high light. We detected a simultaneous co-activation of the non-photochemical quenching mechanisms based on LHC-like stress related (LHCSR) protein and the photosystem II subunit S that are specific to Chlorophyta and Embryophyta, respectively. Exclusive Embryophyta systems for the synthesis, sensing, and response to the phytohormone auxin were also activated under high light in K. nitens leading to an increase in auxin content with the concomitant accumulation of amino acids such as tryptophan, histidine, and phenylalanine.

Dataset Information

Reliable reference genes and abiotic stress marker genes in Klebsormidium nitens.

Publications

Reliable reference genes and abiotic stress marker genes in Klebsormidium nitens.

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