ACTIN2 is essential for bulge site selection and tip growth during root hair development of Arabidopsis.

Institute of Plant Biology, University of Zurich, 8008 Zurich, Switzerland.
Plant physiology (Impact Factor: 7.39). 09/2002; 129(4):1464-72. DOI: 10.1104/pp.005777
Source: PubMed

ABSTRACT Root hairs develop as long extensions from root epidermal cells. After the formation of an initial bulge at the distal end of the epidermal cell, the root hair structure elongates by tip growth. Because root hairs are not surrounded by other cells, root hair formation provides an excellent system for studying the highly complex process of plant cell growth. Pharmacological experiments with actin filament-interfering drugs have provided evidence that the actin cytoskeleton is an important factor in the establishment of cell polarity and in the maintenance of the tip growth machinery at the apex of the growing root hair. However, there has been no genetic evidence to directly support this assumption. We have isolated an Arabidopsis mutant, deformed root hairs 1 (der1), that is impaired in root hair development. The DER1 locus was cloned by map-based cloning and encodes ACTIN2 (ACT2), a major actin of the vegetative tissue. The three der1 alleles develop the mutant phenotype to different degrees and are all missense mutations, thus providing the means to study the effect of partially functional ACT2. The detailed characterization of the der1 phenotypes revealed that ACT2 is not only involved in root hair tip growth, but is also required for correct selection of the bulge site on the epidermal cell. Thus, the der1 mutants are useful tools to better understand the function of the actin cytoskeleton in the process of root hair formation.

  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Strigolactones (SLs) are plant hormones that regulate shoot and root development in a MAX2-dependent manner. The mechanism underlying SLs' effects on roots is unclear. We used root hair elongation to measure root response to SLs. We examined the effects of GR24 (a synthetic, biologically active SL analog) on localization of the auxin efflux transporter PIN2, endosomal trafficking, and F-actin architecture and dynamics in the plasma membrane (PM) of epidermal cells of the primary root elongation zone in wildtype (WT) Arabidopsis and the SL-insensitive mutant max2. We also recorded the response to GR24 of trafficking (tir3), actin (der1) and PIN2 (eir1) mutants. GR24 increased polar localization of PIN2 in the PM of epidermal cells and accumulation of PIN2-containing brefeldin A (BFA) bodies, increased ARA7-labeled endosomal trafficking, reduced F-actin bundling and enhanced actin dynamics, all in a MAX2-dependent manner. Most of the der1 and tir3 mutant lines also displayed reduced sensitivity to GR24 with respect to root hair elongation. We suggest that SLs increase PIN2 polar localization, PIN2 endocytosis, endosomal trafficking, actin debundling and actin dynamics in a MAX2-dependent fashion. This enhancement might underlie the WT root's response to SLs, and suggests noncell autonomous activity of SLs in roots.
    New Phytologist 02/2014; · 6.37 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Background Small Rab GTPases are important regulators of vesicular trafficking in plants. AtRabA1d, a member of the RabA1 subfamily of small GTPases, was previously found in the vesicle-rich apical dome of growing root hairs suggesting a role during tip growth; however, its specific intracellular localization and role in plants has not been well described.ResultsThe transient expression of 35S::GFP:RabA1d construct in Allium porrum and Nicotiana benthamiana revealed vesicular structures, which were further corroborated in stable transformed Arabidopsis thaliana plants. GFP-RabA1d colocalized with the trans-Golgi network marker mCherry-VTI12 and with early FM4-64-labeled endosomal comparments. Late endosomes and endoplasmic reticulum labeled with FYVE-DsRed and ER-DsRed, respectively, were devoid of GFP-RabA1d. The accumulation of GFP-RabA1d in the core of brefeldin A (BFA)-induced-compartments and the quantitative upregulation of RabA1d protein levels after BFA treatment confirmed the association of RabA1d with early endosomes/TGN and its role in vesicle trafficking. Light-sheet microscopy revealed involvement of RabA1d in root development. In root cells, GFP-RabA1d followed cell plate expansion consistently with cytokinesis-related vesicular trafficking and membrane recycling. GFP-RabA1d accumulated in disc-like structures of nascent cell plates, which progressively evolved to marginal ring-like structures of the growing cell plates. During root hair growth and development, GFP-RabA1d was enriched at root hair bulges and at the apical dome of vigorously elongating root hairs. Importantly, GFP-RabA1d signal intensity exhibited an oscillatory behavior in-phase with tip growth. Progressively, this tip localization dissapeared in mature root hairs suggesting a link between tip localization of RabA1d and root hair elongation. Our results support a RabA1d role in events that require vigorous membrane trafficking.Conclusions RabA1d is located in early endosomes/TGN and is involved in vesicle trafficking. RabA1d participates in both cell plate formation and root hair oscillatory tip growth. The specific GFP-RabA1d subcellular localization confirms a correlation between its specific spatio-temporal accumulation and local vesicle trafficking requirements during cell plate and root hair formation.
    BMC Plant Biology 09/2014; 14(1):252. · 3.94 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: We previously showed that seedlings harboring mutations in genes encoding ARK1, an armadillo repeat-containing kinesin, or AGD1, a class 1 ARF-GAP, have root hairs that exhibit wavy/spiral growth and two tips originating from one initiation site. These root hair defects were accompanied by bundling of endoplasmic microtubules and filamentous actin (F-actin) that extended to the extreme root hair apex. The similar phenotypes of ark1 and agd1 mutants suggest a tight coordination between the cytoskeleton and membrane trafficking in the control of root hair polarity. Indeed, cell biological and genetic studies of the agd1 mutant provided evidence that AGD1's involvement in root hair development involves cross-talk among phosphoinositides (PIs), the actin cytoskeleton and other small GTPases such as ROP2 and RABA4b. Here we show that ark1 root hairs mirror those of agd1 with regard to altered targeting of ROP2 and RABA4b, as well as abnormal tonoplast organization. Furthermore, like agd1, enhanced root hair defects in double mutants in ARK1 and genes encoding a type B phosphatidylinositol-4-phosphate 5-kinase 3 (PIP5K3), a phosphatidylinositol-4-phosphate (PI-4P) phosphatase (RHD4), a phosphatidylinositol transfer protein (COW1), and a vegetative actin isoform (ACT2), were observed. However, root hair shape of some ark1 double mutant combinations, particularly those with act2, pip5k3 and rhd4 (ark1 act2, ark1 pip5k3, ark1 rhd4), differed in some respects from agd1 act2, agd1 pip5k3, and agd1 rhd4. Taken together our results continue to point to commonalities between ARK1 and AGD1 in specifying root hair polarity, but that these two modulators of tip-growth can also regulate root hair development through divergent signaling routes with AGD1 acting predominantly during root hair initiation and ARK1 functioning primarily in sustained tip growth.
    Frontiers in Plant Science 01/2013; 4:528. · 3.64 Impact Factor

Full-text (2 Sources)

Available from
Jul 3, 2014