Biology subjects

Zeiner, A.

Publications and source records attributed to Zeiner, A..

5 recordsLinked to original sources

bioRxiv2026

The molecular architecture of the Arabidopsis callose synthase complex

ABSTRACTCallose synthase is responsible for the targeted deposition of the {beta}-1,3-glucan polymer, callose which underlines essential plant developmental processes, including cell division, pathogen defense or cell-cell communication. The architecture of the callose synthase complex (CALSC) as well as the molecular mechanisms of callose synthesis remain unknown. Here we report an integrative characterisation of the Arabidopsis CALS complex, with the most enriched subunits, CALS1, CALS2 and CALS3, forming its core. Structurally, CALSC assembles into a trimer, requiring the plant-specific Bag domain to mediate inter-subunit associations. The biological importance of CALSC assembly is highlighted by the simultaneous loss of CALS1 and CALS3, which abolishes plasmodesmal callose deposition and affects symplastic transport. Site-directed mutagenesis and molecular dynamics simulations depict the topology of the CALS1 active site in detail, including the components of the enzymatic reaction. We pinpoint the translocating tunnel through which the nascent glucan is delivered and mechanistically confirm the role of transmembrane helix 8 in regulating glucan export. Our work provides unprecedented insight into the molecular architecture of the CALSC and the distinct changes from maturation to activity at the plasma membrane, while showcasing the mechanisms involved in callose synthesis at the molecular level.

plant biology↗

bioRxiv2026

Unstructured regions differentially modulate the activation of RBOHD and RBOHH

Reactive oxygen species (ROS) produced by plant NADPH oxidases (RBOHs) must be precisely controlled in their concentration and spatial distribution to support diverse developmental and stress responses. RBOHs are activated by Ca2+ binding and phosphorylation, yet how internal regulatory domains within RBOHs have evolved to translate these inputs into precise levels of ROS production remains unclear. To address this, we performed phylogenetic analyses to define RBOH subfamilies and identify protein regions underlying functional diversification. This analysis revealed that the most variable regions across land-plant RBOHs are two unstructured regions in the N-terminus, UR1 and UR2, which flank the EF-hand Ca2+-binding domain (EFD). We dissected the roles of these regions in Arabidopsis RBOHD, which is central to plant immunity, and in RBOHH, which drives pollen tube elongation and exhibits high Ca2+-induced ROS production. Our analyses revealed that UR1 plays opposing roles in these RBOHs: in RBOHD, UR1 functions as an autoinhibitory module that restrains Ca2+-mediated activation, whereas in RBOHH, UR1 is essential for Ca2+-dependent activation and has coevolved with the EFD to maximize Ca2+-induced ROS production. We further uncovered divergent regulatory roles for UR2. In RBOHD, but not in RBOHH, phosphorylation of UR2 stabilizes an -helical conformation that promotes interaction with the catalytic domain required for enzymatic activation. Furthermore, unlike in RBOHH, the EFD of RBOHD has coevolved with UR2 to maximize phosphorylation-induced activity. Together, our results show how evolution of unstructured regulatory regions adapts a conserved enzymatic core to distinct demands of immune signaling and polarized growth.

plant biology↗

bioRxiv2025

Coordinated action of CRK2 and QSK1 regulate osmotic stress response in Arabidopsis

Precise control of intercellular communication is essential for normal growth and stress responses in all multicellular organisms. In Arabidopsis, two membrane-localized receptor like kinases (RLKs), the Cysteine-rich RLK CRK2 and the Leucine-rich repeat (LRR) RLK QSK1 relocalize from the general plasma membrane (PM) to plasmodesmata (PD) in response to osmotic stress. Both these RLKs regulate callose deposition thereby modulating PD permeability. However, unchecked callose deposition can block the PD and disrupt proper intercellular communication. Here, we show that under normal growth conditions, CRK2 phosphorylates and sequesters QSK1 at the general PM, preventing unnecessary callose deposition at PD. We show that osmotic stress-induced enrichment of QSK1 at PD requires functional CRK2 and establish that phosphorylation of QSK1 in its C-terminal region is inhibitory in this process. We propose that osmotic stress triggers dephosphorylation and release of QSK1 from the CRK2-QSK1 complex, enabling its relocalization from general PM to PD, where it promotes stress-induced callose deposition. Subsequently, CRK2 relocalizes to PD where it negatively influences callose deposition. Our work reveals a tightly coordinated distribution of QSK1 and CRK2 at PM, establishing a dynamic gating mechanism that balances growth and stress responsiveness.

plant biology↗

bioRxiv2025

CYSTEINE-RICH RLK2 regulates development via callose synthase-dependent symplastic transport in Arabidopsis

CYSTEINE-RICH RECEPTOR-LIKE PROTEIN KINASEs (CRKs) play an important role in plant development and stress responses. One of the best described members of the Arabidopsis CRK family is CRK2, which was proposed as a crucial regulator of intercellular transport facilitated by plasmodesmata (PD). As intercellular channels allowing symplastic communication, PD-mediated transport is predominantly regulated by callose synthase (CALS)-mediated callose deposition. This process can impact not just the distribution of molecules between adjacent cells, but also the symplastic loading of vascular tissue, thereby influencing plant stress responses and developmental processes. Here we described the overlapping expression pattern of genes encoding phylogenetically closely related CALS1 and CALS3. Both CALSs were phosphorylated in vitro by CRK2, and the genetic interaction between genes encoding CRK2 and CALS1 or CALS3 revealed their impact on callose deposition, rosette growth, primary root length, and development, represented as a decreased number of true leaves. Importantly, we observed significant accumulation of starch in crk2 mutant plants, especially in developmentally older leaves, which was reverted by the independent introduction of cals1.5 and cals3.1 into the crk2 mutant background. The observed starch accumulation was accompanied by photosynthesis inhibition. We propose that the growth and developmental alterations of crk2 are caused by decreased phloem loading, which resulted in starch accumulation in source organs, and subsequent sink tissue starvation. Our results propose CRK2 as negative regulator of CALS1 and CALS3 regulating source to sink transport, which impacts plant growth and development.

plant biology↗

bioRxiv2025

Pattern-triggered immunity in blue and white seed cultivars of Papaver somniferum

Papaver somniferum (poppy) is a traditional component of Central and Eastern European cuisine and an important oilseed crop in the region. The crucial thread for poppy stable yield is pathogen infection. Thus, we need to understand poppy defence mechanisms in detail. The first robust layer of plant immunity, which plays a crucial role in combat against pathogens, is pattern-triggered immunity (PTI). Here, we provide the first insights into PTI in poppy. We selected four poppy varieties used in the food industry. We investigated poppy response to various peptide elicitors acting as microbe-associated molecular patterns (MAMPs) and damage-associated molecular patterns (DAMPs). Flg22 induced the most robust reactive oxygen species (ROS) burst among all tested peptides. Flg22 also triggered putative mitogen-activated protein kinase (MAPK) phosphorylation and seedling growth inhibition in all tested cultivars. We identified PsWRKY22 and PsPR2 as candidate marker genes suitable for monitoring poppy PTI responses. The tested poppy cultivars have low levels of salicylic acid. Callose accumulation was triggered by wounding but not by flg22. For studying PTI in poppy, wounding is a challenge that needs to be considered as it can obscure potential PTI responses. Our findings highlight conserved aspects of poppy immunity and challenges in studying poppy PTI. The established pipeline facilitates improving our understanding of poppy immunity and has the potential for widespread application in poppy breeding and improving selection for broad-spectrum disease resistance provided by enhanced PTI. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=147 SRC="FIGDIR/small/639761v1_ufig1.gif" ALT="Figure 1"> View larger version (39K): org.highwire.dtl.DTLVardef@1680d52org.highwire.dtl.DTLVardef@42895eorg.highwire.dtl.DTLVardef@488ffeorg.highwire.dtl.DTLVardef@1e5a95d_HPS_FORMAT_FIGEXP M_FIG Graphical abstract The establishment of the methods for studying pattern-triggered immunity (PTI) in Papaver somniferum L. (poppy) was inspired by the knowledge of the model plant Arabidopsis thaliana. The study showed a similarity between Arabidopsis and poppy in response to flg22 but also pointed out the obstacles for PTI analysis in poppy and the differences compared to the model plant. Created with BioRender.com. C_FIG

plant biology↗