(A) CLSM images of GN-GFP and GNL1-GFP in the epidermis of seedling RAMs. In contrast with GNL1-GFP, GN-GFP localizes to the PM. Numbers in bottom right indicate ratios of RAMs with PM signals. …
Seedlings (A) and adults (B) of gns complemented by GNpro:GN-GFP. No complementation of gns by GNpro:GNL1-GFP (C). (D) Root hair positioning in gns complemented by GNpro:GN-GFP, GNpro:GNfwr-GFP and G…
CLSM (A) and TIRF (B) images of GN-GFP and GNL1-GFP from two transgenic lines analyzed. Scale bars – (A) 10 µm, (B) 2 µm.
TIRF colocalization of GN-GFP with VAN3-mRFP in etiolated hypocotyl epidermis, following a treatment with BFA at 50 µM for 1 hr. The BFA-induced, dense and dynamic punctate signals of GN-GFP do not …
(A) Lateral displacement of individual GN-GFP and GNfwr-GFP structures estimated from kymograph maximum projections of TIRF movies. GN-GFP: 0.555±0.280 µm (mean ± s.d.), n=103; GNfwr-GFP: …
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(A) Eight-day-old seedlings of gns mutants complemented with GNpro:GN-GFP and GNpro:GNfwr-GFP transgenes. Two independent lines of each complemented mutant are shown. (B) Adult gns mutants …
TIRF images (A) and kymographs (B) of GN-GFP and GNfwr-GFP expressed in complemented mutant backgrounds, at basal ends of young trichoblasts. Scale bar – 2 µm.
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TIRF images (A) and kymographs (B) of GN-GFP and CLC-mKO in the epidermis of early elongation zone of seedling roots. The GN-positive structures at the PM are distinct from CCPs. Scale bar – 2 µm. …
TIRF images (A) and kymographs (B) of TPLATE-GFP and AP2A1-TagRFP in the epidermis of early elongation zone of seedling roots. Scale bar – 2 µm. Arrowheads in (A) indicate co-localizing punctae.
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(A) Maximum projections of z-stacks of CLSM images of secRFP in RAMs of wild-type seedlings and in basal ends of gns seedlings. Scale bars - 20 µm. (B) Maximum projection of a z-stack of CLSM images …
(A) CLSM maximum projection of a z-stack of secRFP in RAMs of 2-day-old wild-type seedlings. (B) Maximum projections of z-stacks of CLSM images in gns basal ends with and without secRFP expression. …
(A) CLSM images of GN-GFP in seedling RAM epidermis following a treatment with BFA at 50 µM for 1 hr. After a treatment with BFA, GN-GFP locates to the core of BFA bodies and strongly localizes to …
(A) A CLSM image of GN-GFP in seedling RAM epidermis following a treatment with BFA at 50 µM for 1 hr, demonstrating strong localization of GN-GFP to the cell plates. Scale bar - 10 µm. (B) Control …
(A) Schematic representation of the domain composition of GN, GNL1, and chimeric GBF1-type ARF-GEFs. An LLG chimera was not cloned. (B) No complementation of gns by the expression of GLL-GFP. (C) …
(A) Terminal phenotypes of gns mutants complemented by the expression of LGL-GFP. (B) An additional picture of adult gns mutants complemented by the expression of GLG-GFP. (C) Control images showing …
(A) Seedlings of gns mutants expressing LGG-GFP and GGL-GFP. Expression of LGG-GFP fully complements the mutant phenotype, while seedlings expressing GGL-GFP are characterized by roots growing more …
Col-0 seedlings grown on media supplemented with 1 µM NPA on vertical and tilted plates. NPA causes loss of root waving as exhibited on plates tilted back from the vertical.
Growth on tilted plates showing both general seedling morphology and root waving (A), single and fused cotyledons (B), dark-grown seedlings (C), and apical hooks (D) of indicated lines.
(A) TIRF images of GN-GFP, LGG-GFP, GGL-GFP, GLG-GFP and LGL-GFP in hypocotyls of etiolated seedlings of complemented gns mutants. All complementing chimeras localize to the GN-specific structures …
(A) TIRF images of LGG-GFP, GGL-GFP, GLG-GFP and LGL-GFP in hypocotyls of etiolated seedlings of complemented gns mutants from two transgenic lines tested. Scale bar - 2 µm. (B) CLSM images of …
ARF-GEF | Overall degree of GN function | Adult phenotype | Seedling growth rate | Root waving | Root hair positioning | Apical patterning | Etiolated seedling tropism | Apical hooks |
---|---|---|---|---|---|---|---|---|
GN (GGG) | complete | normal | normal | normal | normal | normal | normal | normal |
LGG | complete | normal | normal | normal | normal | normal | normal | normal |
GGL | very high | normal | normal | decreased | not analyzed | normal | normal | normal |
GLG | moderate | decreased growth and fertility | partially decreased | decreased | not analyzed | ~50% single or fused cotyledons | variable growth directions | pen apical hooks |
LGL | moderate to low | very small, infertile | strongly decreased | absent | not analyzed | ~50% single or fused cotyledons | variable growth directions | pen apical hooks |
GLL | none | - | identical to gn | - | - | identical to gn | - | - |
GNL1 (LLL) | none | - | identical to gn | - | - | identical to gn | - | - |
LLG | not cloned |
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Biological sample (Arabidopsis thaliana) | Col-0 (Columbia) | Nottingham Arabidopsis Stock Centre (NASC) | N1092 | |
Gene (Arabidopsis thaliana) | GNOM | The Arabidopsis Information Resource | AT1G13980 | |
Gene (Arabidopsis thaliana) | GNOM-LIKE1 | The Arabidopsis Information Resource | AT5G39500 | |
Genetic reagent (Arabidopsis thaliana) | gns | SALK T-DNA collection | SALK_103014 |
Additional materials and methods.
(a) Lines generated as part of this study. (b) Primers used in this study. (c) Constructs generated in this study.