CASP microdomain formation requires cross cell wall stabilization of domains and non-cell autonomous action of LOTR1.
Kolbeck, A., Marhavý, P., De Bellis, D., Li, B., Kamiya, T., Fujiwara, T., Kalmbach, L., & Geldner, N.
eLife, 11: e69602. January 2022.
![CASP microdomain formation requires cross cell wall stabilization of domains and non-cell autonomous action of LOTR1 [link]](//bibbase.org/img/filetypes/link.svg "Paper")
Paper
doi
link
bibtex
abstract
@article{kolbeck_casp_2022,
title = {{CASP} microdomain formation requires cross cell wall stabilization of domains and non-cell autonomous action of {LOTR1}},
volume = {11},
issn = {2050-084X},
url = {https://doi.org/10.7554/eLife.69602},
doi = {10/gpjfdm},
abstract = {Efficient uptake of nutrients in both animal and plant cells requires tissue-spanning diffusion barriers separating inner tissues from the outer lumen/soil. However, we poorly understand how such contiguous three-dimensional superstructures are formed in plants. Here, we show that correct establishment of the plant Casparian Strip (CS) network relies on local neighbor communication. We show that positioning of Casparian Strip membrane domains (CSDs) is tightly coordinated between neighbors in wild-type and that restriction of domain formation involves the putative extracellular protease LOTR1. Impaired domain restriction in lotr1 leads to fully functional CSDs at ectopic positions, forming ‘half strips’. LOTR1 action in the endodermis requires its expression in the stele. LOTR1 endodermal expression cannot complement, while cortex expression causes a dominant-negative phenotype. Our findings establish LOTR1 as a crucial player in CSD positioning acting in a directional, non-cell-autonomous manner to restrict and coordinate CS positioning.},
urldate = {2022-02-17},
journal = {eLife},
author = {Kolbeck, Andreas and Marhavý, Peter and De Bellis, Damien and Li, Baohai and Kamiya, Takehiro and Fujiwara, Toru and Kalmbach, Lothar and Geldner, Niko},
editor = {Benitez-Alfonso, Yoselin and Kleine-Vehn, Jürgen and Jallais, Yvon and Somssich, Marc},
month = jan,
year = {2022},
keywords = {arabidopsis, casparian strip, endodermis, microdomains, neprosin, network},
pages = {e69602},
}
Efficient uptake of nutrients in both animal and plant cells requires tissue-spanning diffusion barriers separating inner tissues from the outer lumen/soil. However, we poorly understand how such contiguous three-dimensional superstructures are formed in plants. Here, we show that correct establishment of the plant Casparian Strip (CS) network relies on local neighbor communication. We show that positioning of Casparian Strip membrane domains (CSDs) is tightly coordinated between neighbors in wild-type and that restriction of domain formation involves the putative extracellular protease LOTR1. Impaired domain restriction in lotr1 leads to fully functional CSDs at ectopic positions, forming ‘half strips’. LOTR1 action in the endodermis requires its expression in the stele. LOTR1 endodermal expression cannot complement, while cortex expression causes a dominant-negative phenotype. Our findings establish LOTR1 as a crucial player in CSD positioning acting in a directional, non-cell-autonomous manner to restrict and coordinate CS positioning.
RNA Isolation from Nematode-Induced Feeding Sites in Arabidopsis RootsRoots Using Laser Capture Microdissection.
Anjam, M. S., Siddique, S., & Marhavý, P.
In Duque, P., & Szakonyi, D., editor(s),
Environmental Responses in Plants: Methods and Protocols, of Methods in Molecular Biology, pages 313–324. Springer US, New York, NY, 2022.
Paper
link
bibtex
abstract
@incollection{anjam_rna_2022,
address = {New York, NY},
series = {Methods in {Molecular} {Biology}},
title = {{RNA} {Isolation} from {Nematode}-{Induced} {Feeding} {Sites} in {Arabidopsis} {RootsRoots} {Using} {Laser} {Capture} {Microdissection}},
isbn = {978-1-07-162297-1},
url = {https://doi.org/10.1007/978-1-0716-2297-1_22},
abstract = {Nematodes are diverse multicellular organisms that are most abundantly found in the soil. Most nematodes are free-living and feed on a range of organisms. Based on their feeding habits, soil nematodes can be classified into four groups: bacterial, omnivorous, fungal, and plant-feeding. Plant-parasitic nematodes (PPNs) are a serious threat to global food security, causing substantial losses to the agricultural sector. Root-knot and cyst nematodes are the most important of PPNs, significantly limiting the yield of commercial crops such as sugar beet, mustard, and cauliflower. The life cycle of these nematodes consists of four molting stages (J1–J4) that precede adulthood. Nonetheless, only second-stage juveniles (J2), which hatch from eggs, are infective worms that can parasitize the host’s roots. The freshly hatched juveniles (J2) of beet cyst nematode, Heterodera schachtii, establish a permanent feeding site inside the roots of the host plant. A cocktail of proteinaceous secretions is injected into a selected cell which later develops into a syncytium via local cell wall dissolution of several hundred neighboring cells. The formation of syncytium is accompanied by massive transcriptional, metabolic, and proteomic changes inside the host tissues. It creates a metabolic sink in which solutes are translocated to feed the nematodes throughout their life cycle. Deciphering the molecular signaling cascades during syncytium establishment is thus essential in studying the plant-nematode interactions and ensuring sustainability in agricultural practices. However, isolating RNA, protein, and metabolites from syncytial cells remains challenging. Extensive use of laser capture microdissection (LCM) in animal and human tissues has shown this approach to be a powerful technique for isolating a single cell from complex tissues. Here, we describe a simplified protocol for Arabidopsis-Heterodera schachtii infection assays, which is routinely applied in several plant-nematode laboratories. Next, we provide a detailed protocol for isolating high-quality RNA from syncytial cells induced by Heterodera schachtii in the roots of Arabidopsis thaliana plants.},
language = {en},
urldate = {2022-04-29},
booktitle = {Environmental {Responses} in {Plants}: {Methods} and {Protocols}},
publisher = {Springer US},
author = {Anjam, Muhammad Shahzad and Siddique, Shahid and Marhavý, Peter},
editor = {Duque, Paula and Szakonyi, Dóra},
year = {2022},
keywords = {Arabidopsis root dissection, Laser capture dissection, Plant-nematode infection, RNA extraction, Syncytial cell isolation},
pages = {313--324},
}
Nematodes are diverse multicellular organisms that are most abundantly found in the soil. Most nematodes are free-living and feed on a range of organisms. Based on their feeding habits, soil nematodes can be classified into four groups: bacterial, omnivorous, fungal, and plant-feeding. Plant-parasitic nematodes (PPNs) are a serious threat to global food security, causing substantial losses to the agricultural sector. Root-knot and cyst nematodes are the most important of PPNs, significantly limiting the yield of commercial crops such as sugar beet, mustard, and cauliflower. The life cycle of these nematodes consists of four molting stages (J1–J4) that precede adulthood. Nonetheless, only second-stage juveniles (J2), which hatch from eggs, are infective worms that can parasitize the host’s roots. The freshly hatched juveniles (J2) of beet cyst nematode, Heterodera schachtii, establish a permanent feeding site inside the roots of the host plant. A cocktail of proteinaceous secretions is injected into a selected cell which later develops into a syncytium via local cell wall dissolution of several hundred neighboring cells. The formation of syncytium is accompanied by massive transcriptional, metabolic, and proteomic changes inside the host tissues. It creates a metabolic sink in which solutes are translocated to feed the nematodes throughout their life cycle. Deciphering the molecular signaling cascades during syncytium establishment is thus essential in studying the plant-nematode interactions and ensuring sustainability in agricultural practices. However, isolating RNA, protein, and metabolites from syncytial cells remains challenging. Extensive use of laser capture microdissection (LCM) in animal and human tissues has shown this approach to be a powerful technique for isolating a single cell from complex tissues. Here, we describe a simplified protocol for Arabidopsis-Heterodera schachtii infection assays, which is routinely applied in several plant-nematode laboratories. Next, we provide a detailed protocol for isolating high-quality RNA from syncytial cells induced by Heterodera schachtii in the roots of Arabidopsis thaliana plants.
Photo: Fredrik Larsson
![A proxitome-RNA-capture approach reveals that processing bodies repress coregulated hub genes [link]](http://bibbase.org/img/filetypes/link.svg "Paper")