ADP-ribosylation factor D1 modulates Golgi morphology, cell plate formation, and plant growth in Arabidopsis

Abstract

ADP-ribosylation factor (ARF) family proteins, one type of small guanine-nucleotide-binding (G) proteins, play a central role in regulating vesicular traffic and organelle structures in eukaryotes. The Arabidopsis (Arabidopsis thaliana) genome contains more than 21 ARF proteins, but relatively little is known about the functional heterogeneity of ARF homologs in plants. Here, we characterized the function of a unique ARF protein, ARFD1B, in Arabidopsis. ARFD1B exhibited both cytosol and punctate localization patterns, colocalizing with a Golgi marker in protoplasts and transgenic plants. Distinct from other ARF1 homologs, overexpression of a dominant-negative mutant form of ARFD1B did not alter the localization of the Golgi marker mannosidase I (ManI)-RFP in Arabidopsis cells. Interestingly, the ARFD1 artificial microRNA knock-down mutant arfd1 displayed a deleterious growth phenotype, while this phenotype was restored in complemented plants. Further, confocal imaging and transmission electron microscopy analyses of the arfd1 mutant revealed defective cell plate formation and abnormal Golgi morphology. Pull-down and liquid chromatography-tandem mass spectrometry analyses identified Coat Protein I (COPI) components as interacting partners of ARFD1B, and subsequent bimolecular fluorescence complementation, yeast (Saccharomyces cerevisiae) two-hybrid, and co-immunoprecipitation assays further confirmed these interactions. These results demonstrate that ARFD1 is required for cell plate formation, maintenance of Golgi morphology, and plant growth in Arabidopsis.

Role of TRDMT1/DNMT2 in stress adaptation and its influence on transcriptome and proteome dynamics under osmotic stress in Physcomitrium patens

“…ADP‐ribosylation factor4‐like proteins that encode small GTP binding proteins that catalyse post translational modifications (ADP ribosylation) were also observed to be reduced. These play crucial roles in vesicle assembly and intracellular trafficking necessary for isotropic and polarised growth of filamentous cells, maintenance of golgi morphology and cell‐plate formation (Lee & Shaw, 2008; Niu et al, 2022). DnaJ/Hsp40 family proteins that help in folding, unfolding, translation or degradation of proteins by acting as co‐chaperons and binding to Hsp70s to promote their ATP hydrolysis activity were also down‐regulated (Qiu et al, 2006).…”

“…were also observed to be reduced. These play crucial roles in vesicle assembly and intracellular trafficking necessary for isotropic and polarised growth of filamentous cells, maintenance of golgi morphology and cell-plate formation (Lee & Shaw, 2008;Niu et al, 2022).…”

BrrA02.LMI1 Encodes a Homeobox Protein That Affects Leaf Margin Development in Brassica rapa

“…RNA extraction, first-strand cDNA synthesis, and qRT-PCR were performed as described previously [ 36 ]. The B. rapa GAPDH [ 51 ] and ACTIN2 (At3g18780) [ 52 ] genes in Arabidopsis were used as reference genes to normalize the expression level of the candidate gene, and the 2 −∆∆Ct method was used to calculate the relative expression of the candidate gene. Three biological replicates and technical replicates were used for each PCR.…”

WRKY6 transcription factor modulates root potassium acquisition through promoting expression of AKT1 in Arabidopsis

“…Arabidopsis root protoplasts prepared from suspensioncultured cells of Plant System Biology Dark-type were used for transient expression experiments (Miao & Jiang, 2007;Niu et al, 2022). In brief, Arabidopsis root cells cultured for 5 days were harvested and treated with an enzyme solution (1% cellulose, 0.05% pectinase, 0.2% driselase) on a shaker for 2-3 h. Afterward, protoplasts were gathered and washed with an electroporation buffer (2.4 g/L HEPES, 0.4 M sucrose, 600 mg/L CaCl 2 , 6 g/L KCl, pH 7.2) twice.…”