Understanding the Key Border: Structure, Function, and Dynamics of the Plant Nuclear Envelope

The nuclear lamina (NL) is a complex network of nuclear lamins and lamina-associated nuclear membrane proteins, which scaffold the nucleus to maintain structural integrity. In animals, type V intermediate filaments are the main constituents of NL. Plant genomes do not encode any homologs of these intermediate filaments, yet plant nuclei contain lamina-like structures that are present in their nuclei. In Arabidopsis thaliana, CROWDED NUCLEI (CRWN), which are required for maintaining structural integrity of the nucleus and specific perinuclear chromatin anchoring, are strong candidates for plant lamin proteins. Recent studies revealed additional roles of Arabidopsis Nuclear Matrix Constituent Proteins (NMCPs) in modulating plants’ response to pathogen and abiotic stresses. However, detailed analyses of Arabidopsis NMCP activities are challenging due to the presence of multiple homologs and their functional redundancy. In this study, we investigated the sole NMCP gene in the liverwort Marchantia polymorpha (MpNMCP). We found that MpNMCP proteins preferentially were localized to the nuclear periphery. Using CRISPR/Cas9 techniques, we generated an MpNMCP loss-of-function mutant, which displayed reduced growth rate and curly thallus lobes. At an organelle level, MpNMCP mutants did not show any alteration in nuclear morphology. Transcriptome analyses indicated that MpNMCP was involved in regulating biotic and abiotic stress responses. Additionally, a highly repetitive genomic region on the male sex chromosome, which was preferentially tethered at the nuclear periphery in wild-type thalli, decondensed in the MpNMCP mutants and located in the nuclear interior. This perinuclear chromatin anchoring, however, was not directly controlled by MpNMCP. Altogether, our results unveiled that NMCP in plants have conserved functions in modulating stress responses.

Abscisic acid (ABA) induces stomatal closure by utilizing complex signaling mechanisms, allowing for sessile plants to respond rapidly to ever-changing environmental conditions. ABA regulates the activity of plasma membrane ion channels and calcium-dependent protein kinases, Ca²⁺ oscillations, and reactive oxygen species (ROS) concentrations. Throughout ABA-induced stomatal closure, the cytoskeleton undergoes dramatic changes that appear important for efficient closure. However, the precise role of this cytoskeletal reorganization in stomatal closure and the nature of its regulation are unknown. We have recently shown that the plant KASH proteins SINE1 and SINE2 are connected to actin organization during ABA-induced stomatal closure but their role in microtubule (MT) organization remains to be investigated. We show here that depolymerizing MTs using oryzalin can restore ABA-induced stomatal closure deficits in sine1-1 and sine2-1 mutants. GFP-MAP4-visualized MT organization is compromised in sine1-1 and sine2-1 mutants during ABA-induced stomatal closure. Loss of SINE1 or SINE2 results in loss of radially organized MT patterning in open guard cells, aberrant MT organization during stomatal closure, and an overall decrease in the number of MT filaments or bundles. Thus, SINE1 and SINE2 are necessary for establishing MT patterning and mediating changes in MT rearrangement, which is required for ABA-induced stomatal closure.