Biology subjects

Palme, K.

Publications and source records attributed to Palme, K..

8 recordsLinked to original sources

bioRxiv2023

FAN, the homolog of mammalian Apoptosis Antagonizing Transcription Factor AATF/Che-1 protein, is involved in safeguarding genome stability through the ATR induced pathway in Arabidopsis

Cellular DNA can be damaged by endogenous or exogenous genotoxins. In plants, reduced genome stability can have a detrimental effect on development. Here, we show the identification of the fan mutant from an ethyl-methanesulfonate (EMS) mutagenized Arabidopsis Col-0 population on the basis of its short root and small leaf phenotype. The causative mutation was identified as a G-to-A transition at the border of the eighth intron and ninth exon of the At5G61330 gene, resulting in a mis-spliced mRNA transcript. FAN is a homolog of the mammalian AATF/Che-1 protein consisting of conserved AATF/Che-1 and TRAUB domains in Arabidopsis. In the fan mutant, under normal conditions, we detected DNA damage and cell death response at the root tip, while hypersensitivity to the exogenously applied hydroxyurea (HU) compared to Col-0, suggesting that FAN plays a role in the DNA damage response (DDR). Furthermore, our results showed that FAN is involved in DDR pathway regulated by ATM/RAD53-RELATED (ATR). Taken together, these suggest that FAN is required for meristem maintenance and the DNA damage response.

plant biology↗

bioRxiv2021

The role of AUX1 during lateral root development in the domestication of the model C4 grass Setaria italica

C4 photosynthesis increases the efficiency of carbon fixation by spatially separating high concentrations of molecular oxygen from rubisco. The specialized leaf anatomy required for this separation evolved independently many times. C4 root systems are highly branched, an adaptation thought to support high rates of photosynthesis; however, little is known about the molecular mechanisms that have driven the evolution of C4 root system architecture (RSA). Using a mutant screen in the C4 model plant Setaria italica, we identify siaux1-1 and siaux1-2 as RSA mutants, and use CRISPR/cas9-mediated genome editing and overexpression to confirm the importance of the locus. As AUX1 is not necessary for lateral root emergence in S. viridis, the species from which S. italica was domesticated, we conducted an analysis of auxin responsive elements in the promoters of auxin-responsive gene families in S. italica, and explore the molecular basis of SiAUX1s role in seedling development using an RNAseq analysis of wild type and siaux1-1 plants. Finally, we use a root coordinate system to compare cell-by-cell meristem structures in siaux1-1 and wild type Setaria plants, observing changes in the distribution of cell volumes in all cell layers and a dependence in the frequency of protophloem and protoxylem strands on siAUX1.

plant biology↗

bioRxiv2021

Estimation of cell cycle kinetics in higher plant root meristem with cellular fate and positional resolution.

Plant root development is a complex spatial-temporal process that originates in the root apical meristem (RAM). To shape the organs structure signaling between the different cells and cell files must be highly coordinated. Thereby, diverging kinetics of chromatin remodeling and cell growth in these files need to be integrated and balanced by differential cell growth and local differences in cell proliferation frequency. Understanding the local differences in cell cycle duration in the RAM and its correlation with chromatin organization is crucial to build a holistic view on the different regulatory processes and requires a quantitative estimation of the chromatin geometry and underlying mitotic cell cycle phases timing at every cell file and every position. Unfortunately, so far precise methods for such analysis are missing. This study presents a robust and straightforward pipeline to determine in parallel the duration of cell cycles key stages in all cell layers of a plants root and their nuclei organization. The methods combine marker-free techniques based on the detection of the nucleus, deep analysis of the chromatin phase transition, incorporation of 5-ethynyl-2'-deoxyuridine (EdU), and mitosis with a deep-resolution plant phenotyping platform to analyze all key cell cycle events kinetics. In the Arabidopsis thaliana L. RAM S-phase duration was found to be as short as 20-30 minutes in all cell files. The subsequent G2-phase duration however depends on the cell type/position and varies from 3.5 hours in the pericycle to more than 4.5 hours in the epidermis. Overall, S+G2+M duration in Arabidopsis under our condition is 4 hours in the pericycle and up to 5.5 hours in the epidermis. Endocycle duration was determined as the time required to achieve 100% EdU index in the transition zone and estimated to be in the range of 3-4 hours. Besides Arabidopsis, we show that the presented technique is applicable also to root tips of other dicot and monocot plants (tobacco (Nicotiana tabacum L.), tomato (Lycopersicon esculentum L.) and wheat (Triticum aestivum L.).

cell biology↗

bioRxiv2020

The auxin transporter PIN1 and the cytokinin transporter AZG1 interact to regulate the root stress response

Root system development is crucial for optimal growth and yield in plants, especially in sub-optimal soil conditions. The architecture of a root system is environmentally responsive, enabling the plant to exploit regions of high nutrient density whilst simultaneously minimizing abiotic stress. Despite the vital contribution of root systems to the growth of both model and crop species, we know little of the mechanisms which regulate their architecture. One factor which is relatively well understood is the transport of auxin, a plant growth regulator which defines the frequency of lateral root (LR) initiation and the rate of LR outgrowth. Here we describe a search for proteins which regulate RSA by interacting directly with a key auxin transporter, PIN1. The native separation of PIN1 identified several co-purifying proteins. Among them, AZG1 was subsequently confirmed as a PIN1 interactor. AZG1-GFP fusions co-localized with PIN1 in procambium cells of the root meristem. Roots of azg1 plants contained less PIN1 and blocking proteolysis restored PIN1 levels, observations which are consistent with PIN1 being stabilized by AZG1 in the plasma membrane. Furthermore, we show that AZG1 is a cytokinin import protein; accordingly, azg1 plants are insensitive to exogenously applied cytokinin. In wild-type plants, the frequency of LRs falls with increasing salt concentration, a response which is not observed in azg1 x azg2 plants, although their drought response is unimpaired. This report therefore identifies a potential point for auxin:cytokinin crosstalk in the environmentally-responsive determination of root system architecture.

plant biology↗

bioRxiv2020

A simple high efficiency and low cost in vitro growth system for phenotypic characterization and seed propagation of Arabidopsis thaliana

BackgroundArabidopsis research relies extensively on the use of in vitro growth for phenotypic analysis of the seedlings and characterization of plant responses to intrinsic and extrinsic cues. For this purpose, stress-free optimal growth conditions should be set up and used as a reference especially in studies aimed at characterizing the plant responses to abiotic and biotic stresses. Currently used standard in vitro protocols for growth and characterization of Arabidopsis thaliana plants often suffer from sub-optimal composition due to an excessively high nutritional content which represents a stress per se and an experimental bias. ResultsWe describe a simple protocol for in vitro growth of Arabidopsis plants in which the phenotypic analysis is based on an optimized and nutritionally balanced culture medium. We show that the protocol is robustly applicable for growth of several Arabidopsis mutants, including mutants lacking the root system. This protocol enables rapid high scale seed production in vitro avoiding soil usage while saving space and time. The optimized in vitro protocol aims at: 1) making in vitro growth as close as possible to natural soil conditions by optimizing nutrient balance in the medium; 2) simplifying phenotypic and molecular investigation of individual plants by standardizing all steps of plant growth; 3) enabling seeds formation also in genotypes with severe defect in the root system; 4) minimizing the amount of waste and space for plant growth by avoiding soil usage. ConclusionsHere we report an optimized protocol for optimal growth of Arabidopsis thaliana plants to avoid biases in phenotypic observation of abiotic/biotic stress experiments. The protocol also enables the completion of the whole life cycle in vitro within 40-45 days and a satisfactory seed set for further propagation with no need for facilities for plant growth in soil and seed sterilisation.

plant biology↗

bioRxiv2020

Retrograde induction of phyB orchestrates ethylene-auxin hierarchy to regulate growth

Exquisitely regulated plastid-to-nucleus communication by retrograde signaling pathways is essential for fine-tuning of responses to the prevailing environmental conditions. The plastidial retrograde signaling metabolite methylerythritol cyclodiphosphate (MEcPP) has emerged as a stress signal transduced into a diverse ensemble of response outputs. Here we demonstrate enhanced phytochrome B protein abundance in red-light grown MEcPP-accumulating mutant (ceh1) plant relative to wild-type seedlings. We further establish MEcPP-mediated coordination of phytochrome B with auxin and ethylene signaling pathways, and uncover differential hypocotyl growth of red-light grown seedlings in response to these phytohormones. Genetic and pharmacological interference with ethylene and auxin pathways outline the hierarchy of responses, placing auxin epistatic to the ethylene signaling pathway. Collectively, our finding establishes the key role of a plastidial retrograde metabolite in orchestrating the transduction of a repertoire of signaling cascades, and positions plastids at the zenith of relaying information coordinating external signals and internal regulatory circuitry to secure organismal integrity. Two sentence summaryThe plastidial retrograde metabolite, MEcPP, orchestrates coordination of light and hormonal signaling cascade through induction of phytochrome B abundance and modulation of auxin and ethylene levels for optimal adaptive responses to light environment.

plant biology↗

bioRxiv2018

Light dynamically regulates growth rate and cellular organisation of the Arabidopsis root meristem.

1Large-scale methods and robust algorithms are needed for a quantitative analysis of cells status/geometry in situ. It allows the understanding the cellular mechanisms that direct organ growth in response to internal and environmental cues. Using advanced whole-stack imaging in combination with pattern analysis, we have developed a new approach to investigate root zonation under different dark/light conditions. This method is based on the determination of 3 different parameters: cell length, cell volume and cell proliferation on the cell-layer level. This method allowed to build a precise quantitative 3D cell atlas of the Arabidopsis root tip. Using this approach we showed that the meristematic (proliferation) zone length differs between cell layers. Considering only the rapid increase of cortex cell length to determine the meristematic zone overestimates of the proliferation zone for epidermis/cortex and underestimates it for pericycle. The use of cell volume instead of cell length to define the meristematic zone correlates better with cell proliferation zone.

plant biology↗

bioRxiv2017

Tissue specific auxin biosynthesis regulates leaf vein patterning

The plant hormone auxin (indole-3-acetic acid, IAA) has a profound influence over plant cell growth and differentiation. Current understanding of vein development in leaves is based on the canalization of auxin into self-reinforcing streams which determine the sites of vascular cell differentiation. However, the role of auxin biosynthesis during leaf development in the context of leaf vein patterning has not been much studied so far. Here we characterize the context specific importance of auxin biosynthesis, auxin transport and mechanical regulations in a growing leaf. We show that domains of auxin biosynthesis predict the positioning of vascular cells. In mutants that have reduced capacity in auxin biosynthesis, leaf vein formation is decreased. While exogenous application of auxin does not compensate the loss of vein formation in auxin biosynthesis mutants, inhibition of polar auxin transport does compensate the vein-less phenotype, suggesting that the site-specific accumulation of auxin, which is likely to be mainly caused by the local auxin biosynthesis, is important for leaf vein formation. Our computational model of midvein development brings forth the interplay of cell stiffness and auxin dependent cell division. We propose that local auxin biosynthesis has the integral role in leaf vascular development.\n\nHighlightsO_LIBuilt spatially and temporally resolved auxin biosynthesis map in growing leaf primordium of Arabidopsis.\nC_LIO_LIExpression domains of auxin biosynthetic enzymes within primordia strongly correlated with leaf vein initiation.\nC_LIO_LIResults show that domains of auxin biosynthesis within primordia drive leaf vein initiation and patterning.\nC_LI\n\nHighlights and eTOC BlurbUsing modelling and a spatiotemporal analysis of auxin biosynthesis and transport, Kneuper et al. show that tissue specific auxin biosynthesis defines places of vein initiation hence underlining the importance of auxin concentration in vein initiation.

plant biology↗