Imaging Ca2+ Dynamics in Wild-Type and NADPH Oxidase-Deficient Mutant Pollen Tubes with Yellow Cameleon and Confocal Laser Scanning Microscopy

Franck, Christina Maria; Westermann, Jens; Boisson-Dernier, Aurélien

doi:10.1007/978-1-4939-7286-9_10

Cited by 3 publications

(4 citation statements)

References 20 publications

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“…Pollen germination medium (5 mM KCl, 1 mM MgSO 4 , 0.01% H 3 BO 3 [w/v], 5 mM CaCl 2 , and 10% sucrose [w/v], dissolved in double-distilled water and adjusted to pH 7.5) was prepared as previously described (Boavida and McCormick, 2007). Pollen germination assays were conducted as described by Franck et al (2017) and imaged using a Leica DM5500 fluorescence microscope equipped with differential interference contrast optics. Subcellular localization of AUN1-YFP and AUN2-YFP was investigated using a confocal laser scanning microscope (Leica TSC SP8).…”

Section: In Vitro Pollen Germination Assaysmentioning

confidence: 99%

The Protein Phosphatases ATUNIS1 and ATUNIS2 Regulate Cell Wall Integrity in Tip-Growing Cells

et al. 2018

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Fast tip-growing plant cells such as pollen tubes (PTs) and root hairs (RHs) require a robust coordination between their internal growth machinery and modifications of their extracellular rigid, yet extensible, cell wall (CW). Part of this essential coordination is governed by members of the receptor-like kinase1-like (RLK1L) subfamily of RLKs with FERONIA (FER) and its closest homologs, ANXUR1 (ANX1) and ANX2, controlling CW integrity during RH and PT growth, respectively. Recently, Leucine-Rich Repeat Extensin 8 (LRX8) to LRX11 were also shown to be important for CW integrity in PTs. We previously reported an suppressor screen in that revealed MARIS (MRI) as a positive regulator of both FER- and ANX1/2-dependent CW integrity pathways. Here, we characterize a suppressor that exhibits a weak rescue of the PT bursting phenotype and a short RH phenotype. The corresponding suppressor mutation causes a D94N substitution in a Type One Protein Phosphatase we named ATUNIS1 (AUN1). We show that AUN1 and its closest homolog, AUN2, are nucleocytoplasmic negative regulators of tip growth. Moreover, we demonstrate that AUN1 and AUN1 harboring mutations in key amino acids of the conserved catalytic site of phosphoprotein phosphatases function as dominant amorphic variants that repress PT growth. Finally, genetic interaction studies using the hypermorph MRI and amorph AUN1 dominant variants indicate that LRX8-11 and ANX1/2 function in distinct but converging pathways to fine-tune CW integrity during tip growth.

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Section: In Vitro Pollen Germination Assaysmentioning

confidence: 99%

The Protein Phosphatases ATUNIS1 and ATUNIS2 Regulate Cell Wall Integrity in Tip-Growing Cells

et al. 2018

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“…BacMam system [113] pathway signal transduction isolated cell transfection FRET Biosensors [166][167][168][169][170][171] pathway signal transduction organ infection (HSV) Biosensors [179,180] The best improvements were obtained through the introduction of side chains bearing phosphonates or hydroxyl groups and substitution of the lateral aromatic ring (Figure 13A). On the other hand, rigidification of the amino moieties improves fluorescence by preventing the formation of twisted low-emitting states [242].…”

Section: Development Of Organic Probes For Sted Nanoscopymentioning

confidence: 99%

“…With the development of lifetime imaging systems and new fluorescent proteins, simultaneous use of two FLIM-FRET biosensors within one sample is allowed [169,170]. The biosensor strategy may bypass difficulties like probe penetration and autofluorescence in plants or microalgae [171]. In addition, photoactivation-induced specific expression of biosensors in a group of cells, in cells, and in subcellular compartments offers a new perspective in cell biology studies [172].…”

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confidence: 99%

“Probe, Sample, and Instrument (PSI)”: The Hat-Trick for Fluorescence Live Cell Imaging

et al. 2018

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Cell Imaging Platforms (CIPs) are research infrastructures offering support to a number of scientific projects including the choice of adapted fluorescent probes for live cell imaging. What to detect in what type of sample and for how long is a major issue with fluorescent probes and, for this, the “hat-trick” “Probe–Sample–Instrument” (PSI) has to be considered. We propose here to deal with key points usually discussed in CIPs including the properties of fluorescent organic probes, the modality of cell labeling, and the best equipment to obtain appropriate spectral, spatial, and temporal resolution. New strategies in organic synthesis and click chemistry for accessing probes with enhanced photophysical characteristics and targeting abilities will also be addressed. Finally, methods for image processing will be described to optimize exploitation of fluorescence signals.

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“…Pollen is amenable to immuno-and dye-based labeling (e.g., used in [10,[31][32][33]), transgenics (e.g., used in [15,29,34]), availability of intracellular sensors (mainly exploited in Arabidopsis thaliana), and setups for efficient live-cell imaging (e.g., [35][36][37]). 4.…”

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confidence: 99%

Two Is Company, but Four Is a Party—Challenges of Tetraploidization for Cell Wall Dynamics and Efficient Tip-Growth in Pollen

Westermann

2021

Plants

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Some cells grow by an intricately coordinated process called tip-growth, which allows the formation of long tubular structures by a remarkable increase in cell surface-to-volume ratio and cell expansion across vast distances. On a broad evolutionary scale, tip-growth has been extraordinarily successful, as indicated by its recurrent ‘re-discovery’ throughout evolutionary time in all major land plant taxa which allowed for the functional diversification of tip-growing cell types across gametophytic and sporophytic life-phases. All major land plant lineages have experienced (recurrent) polyploidization events and subsequent re-diploidization that may have positively contributed to plant adaptive evolutionary processes. How individual cells respond to genome-doubling on a shorter evolutionary scale has not been addressed as elaborately. Nevertheless, it is clear that when polyploids first form, they face numerous important challenges that must be overcome for lineages to persist. Evidence in the literature suggests that tip-growth is one of those processes. Here, I discuss the literature to present hypotheses about how polyploidization events may challenge efficient tip-growth and strategies which may overcome them: I first review the complex and multi-layered processes by which tip-growing cells maintain their cell wall integrity and steady growth. I will then discuss how they may be affected by the cellular changes that accompany genome-doubling. Finally, I will depict possible mechanisms polyploid plants may evolve to compensate for the effects caused by genome-doubling to regain diploid-like growth, particularly focusing on cell wall dynamics and the subcellular machinery they are controlled by.

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Imaging Ca2+ Dynamics in Wild-Type and NADPH Oxidase-Deficient Mutant Pollen Tubes with Yellow Cameleon and Confocal Laser Scanning Microscopy

Cited by 3 publications

References 20 publications

The Protein Phosphatases ATUNIS1 and ATUNIS2 Regulate Cell Wall Integrity in Tip-Growing Cells

The Protein Phosphatases ATUNIS1 and ATUNIS2 Regulate Cell Wall Integrity in Tip-Growing Cells

“Probe, Sample, and Instrument (PSI)”: The Hat-Trick for Fluorescence Live Cell Imaging

Two Is Company, but Four Is a Party—Challenges of Tetraploidization for Cell Wall Dynamics and Efficient Tip-Growth in Pollen

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