Biology subjects

Frank, W.

Publications and source records attributed to Frank, W..

4 recordsLinked to original sources

bioRxiv2025

Expression levels of the Band-7 protein FLOTILLIN modulate salt tolerance, growth and development in the moss Physcomitrium patens

The Band-7 proteins, known as FLOTILLINs (FLOT), are present at the plasma membranes of most land plants. They function in clathrin-independent endocytosis and contribute to nodule formation following symbiotic infections. This study reveals that the single FLOT variant in Physcomitrium patens is located at the thylakoid membranes in chloroplasts, serving an unanticipated function. Phenotypic analysis of knockout and overexpression lines demonstrates that PpFLOT overexpression significantly impairs the high salinity tolerance of P. patens. Additionally, liquid protonema cultures of PpFLOT-OEX lines exhibited a distinct color change due to necrotic events and developed brachycyte-like cells. These changes correlate with the strength of PpFLOT expression and do not occur when these lines are cultivated on solid medium. Our study found that PpFLOT-OEX lines display increased chlorophyll and H2O2 production. We also discovered that PpFLOT is regulated by ABA and light, and its high expression can potentially affect retrograde signaling. Metabolomics and proteomics analyses revealed changes in the pigment and lipid composition as well as differentially accumulated proteins in PpFLOT mutant lines. We also observed changes in the expression of ion-transport related genes, accumulation of lipids crucial during pathogen defense, and differentially accumulated proteins taking part in multiple metabolomic pathways. Consequently, our study suggests a novel role for chloroplastic PpFLOT in plant terrestrialization, as it is putatively involved in Ca2+ and reactive oxygen species (ROS) signaling in response to abiotic and biotic stress, along with the light-dependent regulation of chlorophyll biosynthesis.

plant biology↗

bioRxiv2023

Identification and characterization of DICER-LIKE genes and their roles in Marchantia polymorpha development and stress adaptation

DICER-LIKE (DCL) proteins have a central role in plant small RNA (sRNA) biogenesis. The Marchantia polymorpha genome encodes four DCL proteins: two DCL1 homologs, MpDCL1a and MpDCL1b, MpDCL3 and MpDCL4. While MpDCL1a, MpDCL3 and MpDCL4 show high similarities to their orthologs in Physcomitrium patens and Arabidopsis thaliana, MpDCL1b shares only a limited homology with PpDCL1b, but it is very similar, in terms of functional domains, to orthologs in Anthoceros agrestis and Salvinia cucullata. We generated Mpdclge mutant lines via the CRISPR/Cas9 system and performed comprehensive phenotypic analyses of these mutant lines, under control and salt stress conditions as well as upon exogenous naphthaleneacetic acid (NAA) and abscisic acid (ABA) treatments to gain insights into the respective MpDCL functions. While Mpdcl1age mutants display severe developmental aberrations throughout their development, no adverse phenotypic changes are detectable in Mpdcl1bge and Mpdcl4ge mutants except the development of less and smaller male sexual organs (antheridiophores) when they are cultivated under photoperiod conditions supplemented with far-red light. Mpdcl3ge mutants display rosette-shaped thallus formation and overall faster development, but are not able to form antheridiophores. The rosette-shaped thallus development of Mpdcl3ge can be reverted to a wild-type-like thallus growth upon NAA treatment. Mpdcl1bge mutants can tolerate high levels of salt, whereas Mpdcl4ge mutants show higher salt sensitivity. Moreover, Mpdcl1age and Mpdcl3ge mutants show an ABA-hypersensitive phenotype. It can be concluded that the observed phenotypic alterations, under normal or treatment conditions, are linked to the mutations in the respective MpDCLs and hence to defective or altered sRNA biogenesis pathways in M. polymorpha. In conclusion, MpDCLs and their associated sRNAs regulate development, abiotic stress and phytohormonal response in M. polymorpha.

plant biology↗

bioRxiv2021

The RNA-binding protein RBP45D of Arabidopis plays a role in epigenetic control of flowering time and DCL3-independent RNA-directed DNA methylation

RNA-directed DNA methylation (RdDM) helps to defend plants against invasive nucleic acids. In the canonical form of RdDM, 24-nt small interfering RNAs (siRNAs) are produced by DICER-LIKE 3 (DCL3). Here, we describe the Arabidopsis thaliana prors1 (LUC) transgenic system, in which transcriptional gene silencing (TGS) is independent of DLC3. A forward genetics screen performed with this system identified both known components of RdDM, and the RNA-binding protein RBP45D. RBP45D promotes DNA methylation, and its loss delays flowering, especially at high temperature, presumably mediated by elevated FLC levels. RBP45D is localized to the nucleus, where it is associated with snRNAs and snoRNAs. RBP45D maintains siRNA production originating from the LUC transgene, but does not alter mRNA levels or affect processing of transcripts of known RdDM genes. We suggest that RBPD45 facilitates DCL3-independent siRNA production by stabilising either the precursor RNA or the - as yet unidentified - slicer protein.

plant biology↗

bioRxiv2019

Impact of small RNAs in retrograde signalling pathways in Arabidopsis thaliana

Chloroplast perturbations activate retrograde signalling pathways causing dynamic changes of gene expression. Besides transcriptional control of gene expression different classes of small non-coding RNAs (sRNAs) act in gene expression control, but comprehensive analyses regarding their role in retrograde signalling is lacking. We performed sRNA profiling in response to norflurazon (NF) that provokes retrograde signals in A. thaliana wild type and the two retrograde signalling mutants gun1 and gun5. The RNA samples were also used for mRNA and long non-coding RNA (lncRNA) profiling to link altered sRNA levels to changes of their cognate target RNAs. We identified 122 sRNAs from all known sRNA classes that were responsive to NF in wild type. Strikingly, 140 and 213 sRNAs were found to be differentially regulated in both mutants indicating a retrograde control of these sRNAs. Concomitant with the changes in sRNA expression we detected about 1500 differentially expressed mRNAs in the NF treated wild type and around 900 and 1400 mRNAs that were differentially regulated in the gun1 and gun5 mutant with a high proportion (~30%) of genes encoding plastid proteins. Furthermore, around 20% of predicted miRNA targets code for plastid localised proteins. The analyses of sRNA-target pairs identified pairs with an anticorrelated expression as well pairs showing other expressional relations pointing to a role of sRNAs in balancing transcriptional changes upon retrograde signals. Based on the comprehensive changes in sRNA expression we assume a considerable impact of sRNAs in retrograde-dependent transcriptional changes to adjust plastidic and nuclear gene expression. Significance statementPerturbations of plastid functions trigger retrograde signalling to adjust plastidic and nuclear gene expression, however, the role of small non-coding RNAs acting as regulators in these pathways is not well understood. We analysed small non-coding RNA expression in response to retrograde signals in A. thaliana wild type and two retrograde signalling mutants and identified members of all known small non-coding RNA classes pointing to a functional role of these RNA classes in retrograde pathways.

plant biology↗