Biology subjects

Dall'Osto, L.

Publications and source records attributed to Dall'Osto, L..

5 recordsLinked to original sources

bioRxiv2025

Structural determinants for red-shifted absorption in higher-plants Photosystem I

- Higher plants Photosystem I absorbs near-infrared light through long-wavelength chlorophylls, enriched under vegetation canopies, to enhance photon capture. Far-red absorption originates from chlorophylls pairs within the Lhca3 and Lhca4 subunits of the LHCI antenna, known as the "red cluster" composed of chlorophylls a603 and a609. - We used reverse genetics to produce an Arabidopsis mutant devoid of red-shifted absorption, and we obtained high-resolution cryo-EM structures from purified PSI-LHCI complexes in both wild-type and mutant plants. - Computed excitonic coupling values suggested a possible contribution of additional nearby pigment molecules, namely chlorophyll a615 and violaxanthin in L2 site, to far-red absorption. Therefore, we investigated the structural determinants of far-red absorption and analyzed the spectroscopic effects of these additional pigments by producing further Arabidopsis transgenic lines. The two experimental structures were used for quantum mechanics calculations, revealing that excitonic interactions alone cannot explain far-red absorption, while charge transfer states were needed for accurate spectral simulations. - Our findings demonstrate that the molecular mechanisms of light-harvesting under shaded conditions rely on very precise tuning of chromophore interactions, an understanding of which is crucial for designing light-harvesting complexes with engineered absorption spectra

plant biology↗

bioRxiv2025

The minor antennae of photosystem II contribute to qH-energy dissipation in Arabidopsis

Photosynthesis is a biological process that converts light energy into chemical energy. Excessive light can damage the photosynthetic machinery, so plants have evolved photoprotective mechanisms such as non-photochemical quenching (NPQ). Among the NPQ mechanisms, qH is a form of sustained quenching, dependent on LIPOCALIN IN THE PLASTID (LCNP) and repressed by SUPPRESSOR OF QUENCHING 1 (SOQ1), protecting against abiotic stress. Recently, we showed in Arabidopsis thaliana that qH can occur in the major light-harvesting complexes (Lhcb1, Lhcb2, Lhcb3) but independently of any specific major antenna. Interestingly, in mutants with little or no accumulation of major antennae (koLHCII, lhcb1, cpsrp43), qH can still be induced. Here, we show that the minor antennae can be quenched by qH and remain quenched once isolated. To investigate the role of minor antennae in qH, we combined the soq1 mutant, which displays high qH, with mutations in each minor antenna type (Lhcb4, Lhcb5, or Lhcb6), or with a mutant lacking all minor antennae. None are strictly required for qH to occur. Still, the absence of Lhcb6 decreases qH induction likely due to an indirect effect from the slower electron transport rate and/or a different macro-organization of photosynthetic complexes in the thylakoids. Overall, this work demonstrates that the minor antennae are a secondary target for qH and could serve as an additional safety valve for photoprotective energy dissipation during prolonged stress.

plant biology↗

bioRxiv2022

Thylakoid grana stacking revealed by multiplex genome editing of LHCII encoding genes

Land plant chloroplasts differ from algal ones for their thylakoid membranes being organized in grana: piles of vesicles paired by their stromal surface, forming domains including Photosystem (PS) II and its antenna while excluding PS I and ATPase to stroma membranes, connecting grana stacks. The molecular basis of grana stacking remain unclear. We obtained genotypes lacking the trimeric antenna complex (Lhcb1-2-3), the monomeric Lhcb4-5-6, or both. Full deletion caused loss of grana, while either monomers or trimers support 50% stacking. The expression of Lhcb5 alone restored stacking at 50%, while Lhcb2 alone produced huge grana which broke down upon light exposure. Cyclic electron transport was maintained in the lack of stacking, while excitation energy balance between photosystems and the repair efficiency of damaged Photosystem II were affected. We conclude that grana evolved for need of regulating energy balance between photosystems under terrestrial canopy involving rapid changes in photon spectral distribution.

plant biology↗

bioRxiv2021

Supramolecular assembly of chloroplast NAD(P)H dehydrogenase-like complex with photosystem I from Arabidopsis thaliana

Cyclic electron transport/flow (CET/CEF) in chloroplasts is a regulatory mechanism crucial for optimization of plant photosynthetic efficiency. CET is catalyzed by a membrane-embedded NAD(P)H dehydrogenase-like (NDH) complex containing at least 29 protein subunits and associating with photosystem I (PSI) to form the NDH-PSI supercomplex. Here we report the 3.9 [A] resolution structure of Arabidopsis thaliana NDH-PSI (AtNDH-PSI) supercomplex. We have constructed structural models for 26 AtNDH subunits, among which 11 subunits are unique to chloroplast and stabilize the core part of NDH complex. In the supercomplex, one NDH can bind up to two PSI-LHCI complexes at both sides of its membrane arm. Two minor LHCIs, Lhca5 and Lhca6, each present in one PSI-LHCI, interact with NDH and contribute to the supercomplex formation and stabilization. Our results showed structural details of the supercomplex assembly and provide molecular basis for further investigation of the regulatory mechanism of CEF in plants.

plant biology↗

bioRxiv2018

Optimized Cas9 expression systems for highly efficient Arabidopsis genome editing facilitate isolation of complex alleles in a single generation

Genetic resources for the model plant Arabidopsis comprise mutant lines defective in almost any single gene in reference accession Columbia. However, gene redundancy and/or close linkage often render it extremely laborious or even impossible to isolate a desired line lacking a specific function or set of genes from segregating populations. Therefore, we here evaluated strategies and efficiencies for the inactivation of multiple genes by Cas9-based nucleases and multiplexing. In first attempts, we succeeded in isolating a mutant line carrying a 70 kb deletion, which occurred at a frequency of ~1.6% in the T2 generation, through PCR-based screening of numerous individuals. However, we failed to isolate a line lacking Lhcb1 genes, which are present in five copies organized at two loci in the Arabidopsis genome. To improve efficiency of our Cas9-based nuclease system, regulatory sequences controlling Cas9 expression levels and timing were systematically compared. Indeed, use of DD45 and RPS5a promoters improved efficiency of our genome editing system by approximately 25-30-fold in comparison to the previous ubiquitin promoter. Using an optimized genome editing system with RPS5a promoter-driven Cas9, putatively quintuple mutant lines lacking detectable amounts of Lhcb1 protein represented approximately 30% of T1 transformants. These results show how improved genome editing systems facilitate the isolation of complex mutant alleles, previously considered impossible to generate, at high frequency even in a single (T1) generation.

plant biology↗