Three CNGC Family Members, CNGC5, CNGC6, and CNGC9, Are Required for Constitutive Growth of Arabidopsis Root Hairs as Ca2+-Permeable Channels

Abstract

The genetic identities of Ca²⁺ channels in root hair (RH) tips essential for constitutive RH growth have remained elusive for decades. Here, we report the identification and characterization of three cyclic nucleotide-gated channel (CNGC) family members, CNGC5, CNGC6, and CNGC9, as Ca²⁺ channels essential for constitutive RH growth in Arabidopsis. We found that the cngc5-1cngc6-2cngc9-1 triple mutant (designated shrh1) showed significantly shorter and branching RH phenotypes as compared with the wild type. The defective RH growth phenotype of shrh1 could be rescued by either the expression of CNGC5, CNGC6, or CNGC9 single gene or by the supply of high external Ca²⁺, but could not be rescued by external K⁺ supply. Cytosolic Ca²⁺ imaging and patch-clamp data in HEK293T cells showed that these three CNGCs all function as Ca²⁺-permeable channels. Cytosolic Ca²⁺ imaging in growing RHs further showed that the Ca²⁺ gradients and their oscillation in RH tips were dramatically attenuated in shrh1 compared with those in the wild type. Phenotypic analysis revealed that these three CNGCs are Ca²⁺ channels essential for constitutive RH growth, with different roles in RHs from the conditional player CNGC14. Moreover, we found that these three CNGCs are involved in auxin signaling in RHs. Taken together, our study identified CNGC5, CNGC6, and CNGC9 as three key Ca²⁺ channels essential for constitutive RH growth and auxin signaling in Arabidopsis.

Keywords: Arabidopsis; CNGC; Ca2+ channels; polar growth; root hair.