Photosystem II monomeric antenna CP26 plays a key role in nonphotochemical quenching in <i>Chlamydomonas</i>

Cazzaniga, Stefano; Kim, Minjae; Pivato, Matteo; Perozeni, Federico; Sardar, Samim; D’Andrea, Cosimo; Jin, EonSeon; Ballottari, Matteo

doi:10.1093/plphys/kiad391

Cited by 5 publications

(1 citation statement)

References 61 publications

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“…Four of 96 transformants (4%) showed larger PCR product sizes compared to the parental strain UVM4 (Figure 1C), indicating a potential integration of the aphVII cassette (length 1.631 bp) into the LCYE target site. The observed editing frequency is in line with recent findings for NHEJ-based editing [27,43] but lower compared to recent homologydirected repair strategies (up to 10-60% [28,30,32]).…”

Section: Cas9-mediated Knockout Of Crlcye By Integration Of Selection...supporting

confidence: 91%

CRISPR/Cas9-Mediated Knockout of the Lycopene ε-Cyclase for Efficient Astaxanthin Production in the Green Microalga Chlamydomonas reinhardtii

Kneip,

Kniepkamp,

Jang

et al. 2024

Plants

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Carotenoids are valuable pigments naturally occurring in all photosynthetic plants and microalgae as well as in selected fungi, bacteria, and archaea. Green microalgae developed a complex carotenoid profile suitable for efficient light harvesting and light protection and harbor great capacity for carotenoid production through the substantial power of the endogenous 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway. Previous works established successful genome editing and induced significant changes in the cellular carotenoid content in Chlamydomonas reinhardtii. This study employs a tailored carotenoid pathway for engineered bioproduction of the valuable ketocarotenoid astaxanthin. Functional knockout of lycopene ε-cyclase (LCYE) and non-homologous end joining (NHEJ)-based integration of donor DNA at the target site inhibit the accumulation of α-carotene and consequently lutein and loroxanthin, abundant carotenoids in C. reinhardtii without changes in cellular fitness. PCR-based screening indicated that 4 of 96 regenerated candidate lines carried (partial) integrations of donor DNA and increased ß-carotene as well as derived carotenoid contents. Iterative overexpression of CrBKT, PacrtB, and CrCHYB resulted in a 2.3-fold increase in astaxanthin accumulation in mutant ΔLCYE#3 (1.8 mg/L) compared to the parental strain UVM4, which demonstrates the potential of genome editing for the design of a green cell factory for astaxanthin bioproduction.

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Section: Cas9-mediated Knockout Of Crlcye By Integration Of Selection...supporting

confidence: 91%

CRISPR/Cas9-Mediated Knockout of the Lycopene ε-Cyclase for Efficient Astaxanthin Production in the Green Microalga Chlamydomonas reinhardtii

Kneip,

Kniepkamp,

Jang

et al. 2024

Plants

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Chassis engineering for high light tolerance in microalgae and cyanobacteria

Dou,

Li,

Wang

et al. 2024

Critical Reviews in Biotechnology

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In vivoanalysis of the relationship between CP26 and qE-type NPQ via higher-orderArabidopsis cp26mutants

Walter,

Patel-Tupper,

Lam

et al. 2024

Preprint

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CP26 is a monomeric minor light-harvesting complex of PSII (LHCII) protein that connects major LHCII trimers to the PSII core in photosynthetic thylakoid membranes. Previous studies have proposed that CP26 is not only involved in light harvesting but could also be involved in non-photochemical quenching (NPQ). Here, we analyzed higher-order Arabidopsis cp26 mutants using biophysical and pharmacological approaches to investigate the nature of NPQ and its relationship to known NPQ regulators (PSII subunit S (PsbS), the xanthophyll-converting enzyme VDE and the pH gradient across the thylakoid membrane). Maximum PSII quantum efficiencies (Fv/Fm) and chlorophyll fluorescence lifetimes in the dark were significantly lower in cp26 mutants, confirming that CP26 deficiency leads to a sustained quenched state even in the absence of light. Destabilized PSII-LHCII supercomplexes as observed with native PAGE analysis are the likely cause for this pre-quenched state, without other antenna proteins being able to rescue this phenotype. Further analyses revealed that cp26 mutants exhibit modest (single mutant) to highly significant (double mutants) reductions in overall NPQ capacity, which do not directly rely on PsbS and VDE (although the effect is more pronounced when these qE components are altered) but depend on thylakoid lumen acidification and protonation of protein residues. Together, these results show that the NPQ component lacking in cp26 mutants acts independently of qE and qZ and is induced in a slower phase of NPQ induction that most likely relies on pH-dependent conformational changes.

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Photosystem II monomeric antenna CP26 plays a key role in nonphotochemical quenching in Chlamydomonas

Cited by 5 publications

References 61 publications

CRISPR/Cas9-Mediated Knockout of the Lycopene ε-Cyclase for Efficient Astaxanthin Production in the Green Microalga Chlamydomonas reinhardtii

CRISPR/Cas9-Mediated Knockout of the Lycopene ε-Cyclase for Efficient Astaxanthin Production in the Green Microalga Chlamydomonas reinhardtii

Chassis engineering for high light tolerance in microalgae and cyanobacteria

In vivoanalysis of the relationship between CP26 and qE-type NPQ via higher-orderArabidopsis cp26mutants

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