Volume 10, Issue 7, Pages (July 2017)

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Volume 10, Issue 7, Pages 1004-1006 (July 2017) Arabidopsis CNGC14 Mediates Calcium Influx Required for Tip Growth in Root Hairs Sisi Zhang, Yajun Pan, Wang Tian, Mengqi Dong, Huifen Zhu, Sheng Luan, Legong Li Molecular Plant Volume 10, Issue 7, Pages 1004-1006 (July 2017) DOI: 10.1016/j.molp.2017.02.007 Copyright © 2017 The Authors Terms and Conditions

Figure 1 Expression Pattern, Phenotypes and Functional Analysis of CNGC14 (A) Expression of CNGC14 in root hairs. Histochemical analysis of CNGC14 promoter activity in plants transformed with a construct containing 1.5-kb genomic DNA upstream of the CNGC14 coding region fused to a GUS reporter gene. (i) 9-day-old plant, (ii) root tip, (iii) root-hypocotyl junction (arrowhead), and (iv) root-hair zone. (B) CNGC14 T-DNA insertion line and microRNA173 transgenic line show the short root-hair phenotype. (i, v) Wild-type, (ii, vi) cngc14-1, (iii, vii) cngc14-miR173 line, and (iv, viii) complemented line CNGC14/cngc14-1 (pCNGC14∷YFP-CNGC14/cngc14-1). Scale bars: (i–iv) 200 μm; (v–viii); 20 μm. Seedlings were grown on 1/2 MS containing 0.6% agar and 1% sucrose for 5 days. (C) CNGC14 is localized to the apex of elongating root hairs (arrowhead) containing proCNGC14∷YFP-CNGC14 construct in cngc14-1 background. Scale bars, 20 μm. (D) Two-electrode voltage-clamp characterization of CNGC14 as a Ca2-permeable channel in Xenopus oocytes. Water-injected oocytes showed background currents, and CNGC14-expressed oocytes showed large inward currents when perfused with 30 mM CaCl2. 300 μM DIDS in the bath solution inhibited the endogenous CaCCs. (E) Steady-state I–V relationships of recordings as in (D) (n = 4). (F) Altered calcium oscillations in cngc14-1 root hairs. (Left) root hairs from wild-type (i) and cngc14-1 (ii) expressing the Ca2+ sensor YC3.6 were imaged every 4 s. Cytosolic Ca2+ levels were calibrated as described in the Supplemental Information and pseudocolor coded according to the scale on the right. Numbers represent time in seconds. Scale bars, 10 μm. (Right) Quantitative analysis of cytosolic Ca2+ oscillations in a representative growing root hair from the wild-type or the cngc14-1 mutant. Increase in the FRET/CFP ratio reflects an increase in cytoplasmic Ca2+ level. Molecular Plant 2017 10, 1004-1006DOI: (10.1016/j.molp.2017.02.007) Copyright © 2017 The Authors Terms and Conditions