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Biology subjects

Maryn, N.

Publications and source records attributed to Maryn, N..

3 recordsLinked to original sources

Natural Variation in Photoprotection: Rapid NPQ Kinetics in Ferns

Land plants perform oxygenic photosynthesis but are unable to use all of the solar radiation that they absorb on a daily basis. To minimize the production of reactive oxygen species in excess light, photosynthetic organisms use non-photochemical quenching (NPQ) mechanisms to dissipate excess excitation energy. However, the on-off transition of these mechanisms is slower than the light fluctuations themselves. In high-to-low light transitions, this can be costly to the overall productivity and carbon gain of the organism across its lifetime, because useful light energy is wasted. Here, we characterize the rapid kinetics of NPQ found in species across the fern lineage. Most of the 23 examined fern species showed faster NPQ induction and faster and more complete NPQ relaxation. Curve fitting suggested that energy-dependent quenching was the dominant contributor to the kinetics. The xerophytic fern Astrolepis windhamii exhibited rapid, dithiothreitol-resistant accumulation of zeaxanthin during NPQ induction, and it maintained low residual NPQ after NPQ relaxation, which however was not associated with rapid re-epoxidation of zeaxanthin. Rapid NPQ kinetics might have been an adaptive trait as ferns radiated in sunflecked forest understories during angiosperm diversification and expansion during the Cretaceous.

plant biology↗

Multiplexed CRISPR/Cas9 mutagenesis of rice PSBS1 non-coding sequences for transgene-free overexpression

Understanding CRISPR/Cas9s capacity to generate native overexpression (OX) alleles would accelerate agronomic gains achievable by gene editing. To generate OX alleles with increased RNA and protein abundance, we leveraged multiplexed CRISPR/Cas9 mutagenesis of non-coding DNA sequences located upstream of the rice PSBS1 gene. We isolated 120 transgene-free, gene-edited alleles with varying NPQ capacity in vivo --ranging from complete knockout to overexpression, using a high-throughput phenotyping and transgene screening pipeline. Overexpression of OsPSBS1 increased protein abundance 2-3-fold, matching fold changes obtained by transgenesis. Increased PsbS protein abundance enhanced non-photochemical quenching capacity and improved water-use efficiency. Across our resolved genetic variation, we identify the role of 5UTR indels and inversions in driving knockout/knockdown and overexpression phenotypes, respectively. Complex structural variants, such as the 252kb duplication/inversion generated in this study, evidence the potential of CRISPR/Cas9 to facilitate significant genomic changes with negligible off-target transcriptomic perturbations. Our results may inform future gene-editing strategies for hypermorphic alleles and have opened the door to the pursuit of gene-edited, non-transgenic rice plants with accelerated relaxation of photoprotection.

plant biology↗

A noncatalytic activity of the H4K20 demethylase DPY-21 regulates condensin DC binding

Condensin is a multi-subunit SMC complex that binds to and compacts chromosomes. Here we addressed the regulation of condensin binding dynamics using C. elegans condensin DC, which represses X chromosomes in hermaphrodites for dosage compensation. We established fluorescence recovery after photobleaching (FRAP) using the SMC4 homolog DPY-27 and showed that a well-characterized ATPase mutation abolishes its binding. Next, we performed FRAP in the background of several chromatin modifier mutants that cause varying degrees of X-chromosome derepression. The greatest effect was in a null mutant of the H4K20me2 demethylase DPY-21, where the mobile fraction of condensin DC reduced from [~]30% to 10%. In contrast, a catalytic mutant of dpy-21 did not regulate condensin DC mobility. Hi-C data in the dpy-21 null mutant showed little change compared to wild type, uncoupling Hi-C measured long-range DNA contacts from transcriptional repression of the X chromosomes. Together, our results indicate that DPY-21 has a non-catalytic role in regulating the dynamics of condensin DC binding, which is important for transcription repression.

cell biology↗