Biology subjects

Vogler, H.

Publications and source records attributed to Vogler, H..

5 recordsLinked to original sources

bioRxiv2021

Cell Wall Biochemistry Drives Pollen Tube Mechanics and Affects Growth Rate

Pollen tubes maintain cell wall integrity as they rapidly grow towards the ovule, yet need to rupture at a precise moment to release the sperm cells. This biomechanical balance is critical for fertilization and relies on the interplay between turgor pressure and cell wall rigidity. How cell wall composition affects its mechanical properties is, however, not well understood. In this study, we combine experimental and simulation techniques to determine key mechanical parameters using Arabidopsis cell wall mutants. We integrated cellular force microscopy with a Finite Element Method-based model to predict growth rates of different mutant pollen tubes. The Finite Element Method-based model allowed us to quantify the effects of cell wall mutations on the time-independent turgor pressure and cell wall elasticity, while cellular force microscopy enabled determination of time-dependent viscoelastic properties of the cell wall. This novel approach can be applied across biological systems and advances mechanical studies of cell and tissue morphogenesis.

systems biology↗

bioRxiv2021

Fast and flexible processing of large FRET image stacks using the FRET-IBRA toolkit

Ratiometric time-lapse FRET analysis requires a robust and accurate processing pipeline to eliminate bias in intensity measurements on fluorescent images before further quantitative analysis can be conducted. This level of robustness can only be achieved by supplementing automated tools with built-in flexibility for manual ad-hoc adjustments. FRET-IBRA is a modular and fully parallelized configuration file-based tool written in Python. It simplifies the FRET processing pipeline to achieve accurate, registered, and unified ratio image stacks. The flexibility of this tool to handle discontinuous image frame sequences with tailored configuration parameters further streamlines the processing of outliers and time-varying effects in the original microscopy images. FRET-IBRA offers cluster-based channel background subtraction, photobleaching correction, and ratio image construction in an all-in-one solution without the need for multiple applications, image format conversions, and/or plug-ins. The package accepts a variety of input formats and outputs TIFF image stacks along with performance measures to detect both the quality and failure of the background subtraction algorithm on a per frame basis. Furthermore, FRET-IBRA outputs images with superior signal-to-noise ratio and accuracy in comparison to existing background subtraction solutions, whilst maintaining a fast runtime. The FRET-IBRA package has been extensively used in quantifying the spatial distribution of calcium ions during pollen tube growth under mechanical constraints. Benchmarks against existing tools clearly demonstrate the need for FRET-IBRA in extracting reliable insights from FRET microscopy images of dynamic physiological processes at high spatial and temporal resolution. The source code for Linux and Mac operating systems is released under the BSD license and, along with installation instructions, test images, example configuration files, and a step-by-step tutorial, is freely available at github.com/gmunglani/fret-ibra. Author SummaryFRET is a fundamental imaging technique used to generate fluorescence signals sensitive to molecular conformations and interactions. Despite its wide use and the large body of literature on the theoretical steps required to process images generated from this procedure, we were unable to locate a tool that contained the entire processing workflow, whilst allowing the user the flexibility to adjust parameters for maximum accuracy and runtime efficiency. FRET-IBRA was thus created to be an all-in-one, open-source, parallel solution to process FRET images, while eliminating complications arising from repeated image format conversions. Besides enhancing the background subtraction algorithm for FRET images, several additional options were implemented for the user to extract the cleanest signal possible for their specific use case. FRET-IBRA is primarily built for flexibility when handling large image stacks by supporting sequences of image frames to be treated independently, greatly reducing time spent on splitting and concatenating image stacks. In accuracy and speed benchmarks against more general background subtraction packages, FRET-IBRA was able to provide the cleanest results with a fast runtime, leading to reliable analysis without additional tuning.

bioinformatics↗

bioRxiv2019

The mechanical basis for snapping of the Venus flytrap, Darwin’s ‘most wonderful plant in the world’

The carnivorous Venus flytrap catches prey by an ingenious snapping mechanism. Based on work over the past 190 years, it has become generally accepted that two touches of the traps sensory hairs within 30 seconds, each one generating an action potential, are required to trigger closure of the trap. We developed an electromechanical model which, however, suggests that under certain circumstances one touch is sufficient to generate two action potentials. Using a force-sensing microrobotics system, we precisely quantified the sensory hair deflection parameters necessary to trigger trap closure, and correlated them with the elicited action potentials in vivo. Our results confirm the models predictions, suggesting that the Venus flytrap may be adapted to a wider range of prey movement than previously assumed.

plant biology↗

bioRxiv2018

Feeling the force: how pollen tubes deal with obstacles

HighlightPollen tubes literally feel their way through their environment to avoid obstacles as they deliver male gametes to the ovule. We measured their force sensitivity to understand this remarkable behavior.\n\nAbstractPhysical forces are involved in the regulation of plant development and morphogenesis by translating mechanical stress into the modification of physiological processes, which, in turn, can affect cellular growth. Pollen tubes are tip-growing cells that provide an ideal system to study processes induced by exposure to mechanical stress. We combined a lab-on-a-chip device with cellular force microscopy to mimic and quantify the forces that are involved in pollen tube navigation upon confronting mechanical obstacles. Several stages of obstacle avoidance were identified, including force perception, growth adjustment, and penetration. We have experimentally determined the perceptive force, which is the force threshold at which the pollen tube senses the obstacle, for Lilium longiflorum and Arabidopsis thaliana. In addition, we provide evidence that pollen tubes are capable of penetrating narrow gaps by increasing turgor pressure. Taken together, our data indicate that pollen tubes sense physical barriers and actively adjust their growth behavior to overcome them.

plant biology↗

bioRxiv2017

LRX Proteins play a crucial role in pollen grain and pollen tube cell wall development

Leucine-rich repeat extensins (LRXs) are chimeric proteins containing an N-terminal leucine-rich repeat (LRR) and a C-terminal extensin domain. LRXs are involved in cell wall formation in vegetative tissues and required for plant growth. However, the nature of their role in these cellular processes remains to be elucidated. Here, we used a combination of molecular techniques, light microscopy, and transmission electron microscopy to characterize mutants of pollen-expressed LRXs in Arabidopsis thaliana. Mutations in multiple pollen-expressed lrx genes causes severe defects in pollen germination and pollen tube (PT) growth, resulting in a reduced seed set. Physiological experiments demonstrate that manipulating Ca2+ availability partially suppresses the PT growth defects, suggesting that LRX proteins influence Ca2+-related processes. Furthermore, we show that LRX protein localizes to the cell wall, and its LRR-domain (which likely mediates protein-protein interactions) is associated with the plasma membrane. Mechanical analyses by cellular force microscopy and finite element method-based modelling revealed significant changes in the material properties of the cell wall and the fine-tuning of cellular biophysical parameters in the mutants compared to the wild type. The results indicate that LRX proteins might play a role in cell wall-plasma membrane communication, influencing cell wall formation and cellular mechanics.

plant biology↗