Dataset Information

Plasmolysis-deplasmolysis causes changes in endoplasmic reticulum form, movement, flow, and cytoskeletal association.

ABSTRACT: Plasmolysis of hypocotyl cells of transgenic Arabidopsis thaliana and Nicotiana benthamiana diminishes the dynamics of the remodeling of the endoplasmic reticulum (ER) in the central protoplast, namely that withdrawn from the cell wall, and more persistent cisternae are formed, yet little change in the actin network in the protoplast occurs. Also, protein flow within the ER network in the protoplast, as detected with fluorescence recovery after photobleaching (FRAP), is not affected by plasmolysis. After plasmolysis, another network of strictly tubular ER remains attached to the plasma membrane-wall interface and is contained within the Hechtian strands and reticulum. FRAP studies indicate that protein flow within these ER tubules diminishes. Actin is largely absent from the Hechtian reticulum and the ER becomes primarily associated with altered, branched microtubules. The smaller volume of the central protoplast is accompanied by decreased movement rates of tubules, cisternae, and spheroid organelles, but this reduced movement is not readily reversed by the increase in volume that accompanies deplasmolysis.

SUBMITTER: Cheng X

PROVIDER: S-EPMC5853952 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature

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Plasmolysis-deplasmolysis causes changes in endoplasmic reticulum form, movement, flow, and cytoskeletal association.

Cheng Xiaohang X Lang Ingeborg I Adeniji Opeyemi Samson OS Griffing Lawrence L

Journal of experimental botany 20170701 15

Plasmolysis of hypocotyl cells of transgenic Arabidopsis thaliana and Nicotiana benthamiana diminishes the dynamics of the remodeling of the endoplasmic reticulum (ER) in the central protoplast, namely that withdrawn from the cell wall, and more persistent cisternae are formed, yet little change in the actin network in the protoplast occurs. Also, protein flow within the ER network in the protoplast, as detected with fluorescence recovery after photobleaching (FRAP), is not affected by plasmolys ...[more]

PMID: 28922772

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Project description:Aim and experimental set-up: Sequencing of small RNA from total RNA fractions. The aim of this experiment was to compare profiles of miRNA expression between the following genetic backgrounds: Col-0 (wild type), era1-2 (farnesyl transferase mutant), j2-2/j3-2 + pJ3:J3 (j2/j3 double knockout expressing transgenic J3 under the control of the endogenous J3 promoter), j2-2/j3-2 + pJ3:J3C417S (j2/j3 double knockout expressing transgenic J3 mutated in the farnesylation site (C417S) under the control of the endogenous J3 promoter). Seedlings were grown under sterile conditions for 16 days. Two biological replicates of each genotype were harvested, and total RNA was prepared by Trizol extraction. Small RNA libraries were constructed using NEBNext Small RNA Library Prep Set and sequenced on an Illumina platform. Sequencing of small RNA bound to AGO1 in membrane fractions The aim of this experiment was to characterize AGO1-bound small RNAs in membrane fractions, and to answer two specific questions: Can miRNAs be identified whose association with membrane-bound AGO1 is different between the following four genotypes: Col-0 (wild type), era1-2 (farnesyl transferase mutant), j2-2/j3-2 + pJ3:J3 (j2/j3 double knockout expressing transgenic J3 under the control of the endogenous J3 promoter), j2-2/j3-2 + pJ3:J3C417S (j2/j3 double knockout expressing transgenic J3 mutated in the farnesylation site (C417S) under the control of the endogenous J3 promoter) Is the ratio between reads matching miRNA and miRNA* different between the above four genotypes? Seedlings were grown under sterile conditions for 16 days. Two biological replicates of each genotype were harvested. Microsomes were prepared from 2g of seedling tissue, solubilized by 1% deoxycholate and AGO1 was immunoprecipitated with specific antibodies (Agrisera). Immunopurified AGO1 was eluted from Protein A sepharose beads by competitive elution with antigenic AGO1 peptide. Small RNA was extracted by Trizol extraction from eluted AGO1. Small RNA libraries were constructed using NEBNext Small RNA Library Prep Set and sequenced on an Illumina platform.

Dataset Information

Plasmolysis-deplasmolysis causes changes in endoplasmic reticulum form, movement, flow, and cytoskeletal association.

Publications

Plasmolysis-deplasmolysis causes changes in endoplasmic reticulum form, movement, flow, and cytoskeletal association.

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