CamelliA-based simultaneous imaging of Ca2+ dynamics in subcellular compartments

Guo, Jingzhe; He, Jiangman; Dehesh, Katayoon; Cui, Xinping; Yang, Zhenbiao

doi:10.1093/plphys/kiac020

Cited by 13 publications

(4 citation statements)

References 120 publications

(189 reference statements)

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“…However, substantial oscillations of cytosolic pH in pollen tubes, guard cells or mesophyll cells had no visible effects on cytosolic R-GECO1 (Li et al, 2021). Measurements of cytosolic Ca 2+ during pH oscillations in Arabidopsis pollen tubes showed that jR-GECO1a gave similar results to the pH-insensitive Ca 2+ sensor yellow cameleon YC3.6 (Guo et al, 2022). R-GECO1 based Ca 2+ indicators have previously been used to measure Ca 2+ dynamics in organelles, such as the mitochondria of animal cells, although they exhibited some sensitivity to large increases in pH induced by 30 mM NH4Cl (Hou et al, 2017;Kanemaru et al, 2020).…”

Section: Introductionmentioning

confidence: 79%

Diatoms exhibit dynamic chloroplast calcium signals in response to high light and oxidative stress

Flori,

Dickenson,

Gaikwad

et al. 2023

Preprint

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Diatoms are a group of silicified algae that play a major role in marine and freshwater ecosystems. Diatom chloroplasts were acquired by secondary endosymbiosis and exhibit important structural and functional differences from the primary plastids of land plants and green algae. Many functions of primary plastids, including photoacclimation and inorganic carbon acquisition, are regulated by calcium-dependent signalling processes. Calcium signalling has also been implicated in the photoprotective responses of diatoms, although the nature of calcium elevations in diatom chloroplasts and their wider role in cell signalling remains unknown. Using genetically encoded calcium indicators, we find that the diatom Phaeodactylum tricornutum exhibits dynamic chloroplast calcium elevations that are distinct from those found in land plants. Chloroplast calcium ([Ca2+]chl) acts independently from the cytosol and is not elevated by stimuli that induce large cytosolic calcium ([Ca2+]cyt) elevations. In contrast, high light and exogenous hydrogen peroxide (H2O2) induce large, sustained calcium elevations in the chloroplast stroma that are not replicated in the cytosol. Measurements using the fluorescent H2O2 sensor roGFP2-Orp1 indicate that [Ca2+]chl elevations induced by these stimuli correspond to the accumulation of H2O2 in the chloroplast. [Ca2+]chl elevations were also induced by the addition of methyl viologen, which acts to generate superoxide within the chloroplast, and by treatments that disrupt non-photochemical quenching (NPQ). The findings indicate that diatoms generate specific calcium elevations in the chloroplast in response to high light and oxidative stress that likely modulate the activity of calcium-sensitive components in photoprotection and other regulatory pathways.

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Section: Introductionmentioning

confidence: 79%

Diatoms exhibit dynamic chloroplast calcium signals in response to high light and oxidative stress

Flori,

Dickenson,

Gaikwad

et al. 2023

Preprint

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“…Primers used for amplification and assembly of these constructs are listed in Supplemental Table S3 . The ERN1 native promoter–driven complementation construct (ERN1pro::ERN1-mEGFP) was generated in a single reaction, using the NEBuilder HiFi DNA Assembly Master Mix (NEB, E2621), to assemble 3 fragments into a modified pCAMBIA1300-MCS vector that lacks a promoter ( Guo et al 2022 ) ( Supplemental Table S3 ). The promoter and coding sequence of ERN1 without the stop codon were amplified from genomic DNA of WT Arabidopsis plants.…”

Section: Methodsmentioning

confidence: 99%

A G-type lectin receptor kinase negatively regulates Arabidopsis immunity against root-knot nematodes

Zhou

Godinez-Vidal

et al. 2023

Plant Physiology

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Root-knot nematodes (Meloidogyne spp., RKN) are responsible for extensive crop losses worldwide. During infection, they penetrate plant roots, migrate between plant cells, and establish feeding sites, known as giant cells, near the root vasculature. Previously, we found that nematode perception and early responses in plants were similar to those of microbial pathogens and required the BRI1-ASSOCIATED KINASE1/SOMATIC EMBRYOGENESIS RECEPTOR KINASE3 (BAK1/SERK3) coreceptor in Arabidopsis (Arabidopsis thaliana) and tomato (Solanum lycopersicum). Here, we implemented a reverse genetic screen for resistance or sensitivity to RKN using Arabidopsis T-DNA alleles of genes encoding transmembrane receptor-like kinases to identify additional receptors involved in this process. This screen identified a pair of allelic mutations with enhanced resistance to RKN in a gene we named ENHANCED RESISTANCE TO NEMATODES1 (ERN1). ERN1 encodes a G-type lectin receptor kinase (G-LecRK) with a single-pass transmembrane domain. Further characterization showed that ern1 mutants displayed stronger activation of MAP kinases, elevated levels of the defense marker MYB51, and enhanced H2O2 accumulation in roots upon RKN elicitor treatments. Elevated MYB51 expression and ROS bursts were also observed in leaves of ern1 mutants upon flg22 treatment. Complementation of ern1.1 with 35S- or native promotor-driven ERN1 rescued the RKN infection and enhanced defense phenotypes. Our results indicate that ERN1 is an important negative regulator of immunity.

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“…Wild-type (Col-0) expressing GCaMP6 (ref. 44) driven by the pollenspecific LAT52 promoter 48,49,72 ] i in pollen grains were carried out as described in previous studies with modifications 6,25,56,72,[78][79][80][81][82][83] . Pollen grains from freshly opened mature (stage 12-13) flowers were dispersed on the solid germination medium with indicated osmolarity.…”

Section: Imaging Of [Ca 2+ ] I In Pollen Grainsmentioning

confidence: 99%

Osmosensor-mediated control of Ca2+ spiking in pollen germination

Pei,

Tao,

et al. 2024

Nature

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Higher plants survive terrestrial water deficiency and fluctuation by arresting cellular activities (dehydration) and resuscitating processes (rehydration). However, how plants monitor water availability during rehydration is unknown. Although increases in hypo-osmolarity-induced cytosolic Ca2+ concentration (HOSCA) have long been postulated to be the mechanism for sensing hypo-osmolarity in rehydration1,2, the molecular basis remains unknown. Because osmolarity triggers membrane tension and the osmosensing specificity of osmosensing channels can only be determined in vivo3–5, these channels have been classified as a subtype of mechanosensors. Here we identify bona fide cell surface hypo-osmosensors in Arabidopsis and find that pollen Ca2+ spiking is controlled directly by water through these hypo-osmosensors—that is, Ca2+ spiking is the second messenger for water status. We developed a functional expression screen in Escherichia coli for hypo-osmosensitive channels and identified OSCA2.1, a member of the hyperosmolarity-gated calcium-permeable channel (OSCA) family of proteins6. We screened single and high-order OSCA mutants, and observed that the osca2.1/osca2.2 double-knockout mutant was impaired in pollen germination and HOSCA. OSCA2.1 and OSCA2.2 function as hypo-osmosensitive Ca2+-permeable channels in planta and in HEK293 cells. Decreasing osmolarity of the medium enhanced pollen Ca2+ oscillations, which were mediated by OSCA2.1 and OSCA2.2 and required for germination. OSCA2.1 and OSCA2.2 convert extracellular water status into Ca2+ spiking in pollen and may serve as essential hypo-osmosensors for tracking rehydration in plants.

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CamelliA-based simultaneous imaging of Ca2+ dynamics in subcellular compartments

Cited by 13 publications

References 120 publications

Diatoms exhibit dynamic chloroplast calcium signals in response to high light and oxidative stress

Diatoms exhibit dynamic chloroplast calcium signals in response to high light and oxidative stress

A G-type lectin receptor kinase negatively regulates Arabidopsis immunity against root-knot nematodes

Osmosensor-mediated control of Ca2+ spiking in pollen germination

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