Journal: Plant Physiology

Article Title: RACK1A interacts and colocalizes with FSD1 in stress granules to regulate salt stress response in Arabidopsis

doi: 10.1093/plphys/kiaf659

Figure Lengend Snippet: Airyscan confocal laser scanning microscopy (ACLSM) of RACK1A-GFP localization in Arabidopsis root. (a) Root tip with a high abundance of RACK1A in the meristematic zone. (b) Root meristematic cells. White arrowhead points to the dividing cell at the metaphase. (c and d) Epidermal cells of root elongation zone—single plane image (c), orthogonal projection of 20 optical planes (d). (e–h) RACK1A-GFP localization in a trichoblast at early (e) and late (f) bulging stage. Fluorescent RACK1A-GFP signal was located in a cytoplasm, particularly at the forming root hair tip (arrowheads in e, f) and in nucleus (n). Unlike that, nucleoli (*) and vacuoles (v) were devoid of RACK1A-GFP. In growing root hairs, RACK1A-GFP was located in cytoplasm, nucleus (n) and growing tip (g), as evidenced by a profile-based measurement of the fluorescence intensity distribution (h) along the line shown in (g). (i) Initiation of lateral root primordia formation. White arrowheads indicate 2 lateral root founder cells with increased RACK1A-GFP fluorescence intensity in cytosol. (j) Lateral root primordia cells after first anticlinal division. White arrowheads indicate 4 lateral root primordia cells with increased RACK1A-GFP fluorescence intensity in cytosol. (k) Initiation of endodermis crossing after periclinal division of primordia cells. White line indicates lateral root primordia cells with increased RACK1A-GFP fluorescence intensity. (l) Cortex/epidermis crossing and emergence of the lateral root. White arrow indicates the lateral root tip. n, nucleus; *, nucleolus; v, vacuole; t, trichoblast; at, atrichoblast. Scale bar = 20 μm.

Article Snippet: Seeds were surface-sterilized by ethanol, dried, and placed on a half-strength Murashige and Skoog (1⁄2 MS) medium and grown at 21 °C and 70% humidity under a 16 h light/8 h darkness photoperiod with a photosynthetic photon flux of 120 μmol·m 2 ·s −1 in an environmental chamber (Weiss Technik, Grand Rapids, MI, United States) provided by cool white fluorescent linear tube light sources (Philips Master TL-D Reflex 36 W, light flow 3350 lm, light efficiency 93 lm·W –1 ) for a maximum of 14 d. Additionally, wild-type Nicotiana benthamiana plants were grown from seeds in pots with soil for 5 weeks under the same conditions as described above.

Techniques: Confocal Laser Scanning Microscopy, Fluorescence

Journal: Plant Physiology

Article Title: RACK1A interacts and colocalizes with FSD1 in stress granules to regulate salt stress response in Arabidopsis

doi: 10.1093/plphys/kiaf659

Figure Lengend Snippet: Analysis of NaCl-induced oxidative stress response in Col-0 (wild type; WT), rack1a-1 , rack1a-5 and fsd1-1rack1a-1 mutants, and RACK1A-GFP line. Five-day-old seedlings were transferred to control and NaCl-containing medium. (a and b) Representative images of seedlings taken 5 d after the transfer to the control (a) and 150 mM NaCl-containing medium (b). (c) Quantification of seedlings with affected viability from (b). Plants with fully bleached cotyledons were considered unviable (mean ± SD; N = 90). (d) Quantification of chlorophylls a and b in control and salt-treated (150 mM NaCl) seedlings (mean ± SD; N = 40). (e) ROS distribution visualized by fluorescent tracker CellRox Deep Red reagent in plasmolyzed root epidermal cells of WT, rack1a-1 , fsd1-1 rack1a-1 lines. ROS accumulation in mock-treated (½ MS) and plasmolyzed root epidermal cells (½ MS with 100 mM NaCl) visualized by fluorescent tracker CellRox Deep Red reagent. The observed regions of root epidermal cells were visualized using transmitted light. Scale bars = 20 µm. (f) Quantification and statistical evaluation of CellRox Deep Red reagent fluorescence intensity (mean ± SD; N = 15). Different italic letters above the columns in (c, d), and (f) indicate a statistically significant difference between the respective lines and treatments at a P < 0.05 as determined by one-way ANOVA with post-hoc Tukey HSD test.

Article Snippet: Seeds were surface-sterilized by ethanol, dried, and placed on a half-strength Murashige and Skoog (1⁄2 MS) medium and grown at 21 °C and 70% humidity under a 16 h light/8 h darkness photoperiod with a photosynthetic photon flux of 120 μmol·m 2 ·s −1 in an environmental chamber (Weiss Technik, Grand Rapids, MI, United States) provided by cool white fluorescent linear tube light sources (Philips Master TL-D Reflex 36 W, light flow 3350 lm, light efficiency 93 lm·W –1 ) for a maximum of 14 d. Additionally, wild-type Nicotiana benthamiana plants were grown from seeds in pots with soil for 5 weeks under the same conditions as described above.

Techniques: Control, Fluorescence