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Expression profiles of embryo-defective Arabidopsis thaliana mutants increased size exclusion limit1 (ise1) and ise2

ABSTRACT: We use Arabidopsis thaliana embryogenesis as a model system for studying intercellular transport via plasmodesmata (PD). A forward genetic screen for altered PD transport identified ise1 and ise2 mutants with increased intercellular transport of fluorescent 10-kDa tracers. Both ise1 and ise2 exhibit increased formation of twinned and branched PD. ISE1 encodes a mitochondrial DEAD-box RNA helicase, while ISE2 encodes a DEVH-type RNA helicase. Here we show that ISE2 foci are localized to the chloroplast stroma. Surprisingly, plastid development is defective in both ise1 and ise2 mutant embryos. In an effort to understand how RNA helicases that localize to different organelles have similar impacts on plastid and PD development/function we performed whole genome expression analyses. The most significantly affected class of transcripts in both mutants encodes products that target to and enable plastid function. These results reinforce the importance of plastid-mitochondria-nucleus crosstalk, add PD as a critical player in the plant cell communication network, and thereby illuminate a new signaling pathway, dubbed organelle-nucleus-plasmodesmata signaling (ONPS). Several genes with roles in cell wall synthesis and modification are also differentially expressed in both mutants, providing new targets for investigating PD development and function. Three biological replicates each of either ise1 or ise2 mutant seeds vs. sister wild-type controls

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Patricia Zambryski

PROVIDER: E-GEOD-33558 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Expression profiles of embryo-defective Arabidopsis thaliana mutants increased size exclusion limit1 (ise1) and ise2

Project description:We use Arabidopsis thaliana embryogenesis as a model system for studying intercellular transport via plasmodesmata (PD). A forward genetic screen for altered PD transport identified ise1 and ise2 mutants with increased intercellular transport of fluorescent 10-kDa tracers. Both ise1 and ise2 exhibit increased formation of twinned and branched PD. ISE1 encodes a mitochondrial DEAD-box RNA helicase, while ISE2 encodes a DEVH-type RNA helicase. Here we show that ISE2 foci are localized to the chloroplast stroma. Surprisingly, plastid development is defective in both ise1 and ise2 mutant embryos. In an effort to understand how RNA helicases that localize to different organelles have similar impacts on plastid and PD development/function we performed whole genome expression analyses. The most significantly affected class of transcripts in both mutants encodes products that target to and enable plastid function. These results reinforce the importance of plastid-mitochondria-nucleus crosstalk, add PD as a critical player in the plant cell communication network, and thereby illuminate a new signaling pathway, dubbed organelle-nucleus-plasmodesmata signaling (ONPS). Several genes with roles in cell wall synthesis and modification are also differentially expressed in both mutants, providing new targets for investigating PD development and function.

2011-11-09 | GSE33558 | GEO

Choline transporter-like1 is required for plasmodesmal targeting of Potato leafroll virus movement protein MP17 and long-distance assimilate transport

Project description:Macromolecular trafficking and cell-to-cell communication in plants occurs via plasmodesmata (PD). Currently, little is known about the proteins defining these sophisticated cell-to-cell contacts. To uncover proteins required for PD structure and function we made use of the 17 kDa movement protein (MP17) of the Potato leafroll virus (PLRV). The protein is required for cell-to-cell movement of the virus and localizes specifically to branched PD in source tissues. By forward genetic screening for Arabidopsis mutants with altered PD binding of MP17, several mutants were found. Map-based cloning of one of these mutants revealed a mutation in the choline transporter-like 1 (CHER1) protein. The mutation abolished plasma membrane localization of the protein. As consequence phosphatidylcholine levels and PD binding of MP17 decreased. Transcriptome analysis revealed a down-regulation of defense genes and genes involved in the synthesis of very long chain fatty acids, indicating an impairment of lipid signaling. Furthermore, cher1 mutants showed a reduced assimilate export and stunted growth. These findings highlight the emerging role of phospholipids, especially in the context of PD structure and function as well as potential binding sites for plasma membrane- and PD-associated proteins, which has been neglected for a long time. light exposed source leaves from wild type Col-0, T-DNA insertion line cher1-4 and mutant cher1-5 (10 weeks old). Samples: per pool 2 source leaves from 4 different plants, taken in the afternoon; wildtype (WT): Col-0, cher1-4: T-DNA insertion line SALK-065853 (ecotype Col-0, kanamycin resistance), cher1-5: Col-0 with GGA -->GAA at position 740 of cds

2016-06-01 | E-GEOD-61595 | biostudies-arrayexpress

Assaying effector cell-to-cell mobility in plant tissues identifies hypermobility and indirect manipulation of plasmodesmata

Project description:In plants, plasmodesmata establish cytoplasmic continuity between cells to allow for communication and resource exchange across the cell wall. While plant pathogens use plasmodesmata as a pathway for both molecular and physical invasion, the benefits of molecular invasion (cell-to-cell movement of pathogen effectors) are poorly understood. To establish a methodology for identification and characterization of the cell-to-cell mobility of effectors, we performed a quantitative live imaging-based screen of candidate effectors of the fungal pathogen Colletotrichum higginsianum. We predicted C. higginsianum effectors by their expression profiles, the presence of a secretion signal, and their predicted and in planta localization when fused to GFP. We assayed for cell-to-cell mobility of nucleo-cytosolic effectors and identified 14 that are cell-to-cell mobile. We identified that 3 of these effectors are “hypermobile”, showing cell-to-cell mobility greater than expected for a protein of its size. To explore the mechanism of hypermobility we chose two hypermobile effectors and measured their impact on plasmodesmata function and found that even though they show no direct association with plasmodesmata, each increases the transport capacity of plasmodesmata. Thus, our methods for quantitative analysis of cell-to-cell mobility of candidate microbe-derived36effectors, or any suite of host proteins, can identify cell-to-cell hypermobility and offer greater understanding of how proteins affect plasmodesmal function and intercellular connectivity.

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Choline transporter-like1 is required for plasmodesmal targeting of Potato leafroll virus movement protein MP17 and long-distance assimilate transport

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Comparative proteomics of plasmodesmata

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ELONGATED HYPOCOTYL 5 (HY5) and POPEYE (PYE) Regulate Intercellular Iron Transport in Plants

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