GhKT2: a novel K+ transporter gene in cotton (Gossypium hirsutum)

doi:10.15302/J-FASE-2017170

Front. Agr. Sci. Eng.

2018, Vol. 5

Issue (2) : 226-235 https://doi.org/10.15302/J-FASE-2017170

RESEARCH ARTICLE

GhKT2: a novel K⁺ transporter gene in cotton (Gossypium hirsutum)

Yiru WANG, Juan XU, Mingcai ZHANG, Xiaoli TIAN(

), Zhaohu LI

Department of Agronomy, College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China

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Abstract

Potassium is an essential nutrient for plant growth and productivity of crops. K⁺ transporters are important for K⁺ uptake and transport in plants. However, information on the function of K⁺ transporters and K⁺ channels in cotton is limited. The KT/KUP/HAK protein family is essential for a variety of physiological processes in plants, including nutrient acquisition and regulation of development. This study, identified a K⁺ transporter gene, GhKT2, expressed in the roots of cotton (Gossypium hirsutum) cv. Liaomian17. The deduced transcript of GhKT2 is highly homologous to Cluster II of KUP/HAK/KT K⁺ transporters and is predicted to contain 11 transmembrane domains. GhKT2 has been localized to the plasma membrane, and its transcripts were detected in roots, stems, leaves and shoot apices of cotton seedlings. Consistently, b-glucuronidase (GUS) expression driven by the GhKT2 promoter could be detected in roots, mesophyll cells, and leaf veins in transgenic Arabidopsis. In addition, the expression of GhKT2 was induced by low K⁺ stress in cotton roots and pGhKT2::GUS-transgenic Arabidopsis seedlings. The GhKT2-overexpression Arabidopsis lines plants were larger and showed greater K⁺ accumulation than the wild type (WT) regardless of K⁺ concentration supplied. The net K⁺ influx rate, measured by the noninvasive micro-test technique, in root meristem zone of GhKT2-transgenic Arabidopsis lines was significantly greater than that of WT. Taken together, this evidence indicates that GhKT2 may participate in K⁺ acquisition from low or high external K⁺, as well as K⁺ transport and distribution in plants.

Keywords cotton GhKT2 potassium transporter uptake

Corresponding Author(s): Xiaoli TIAN

Just Accepted Date: 30 October 2017 Online First Date: 22 November 2017 Issue Date: 28 May 2018

Cite this article:

Yiru WANG,Juan XU,Mingcai ZHANG, et al. GhKT2: a novel K⁺ transporter gene in cotton (Gossypium hirsutum)[J]. Front. Agr. Sci. Eng. , 2018, 5(2): 226-235.

URL:

https://academic.hep.com.cn/fase/EN/10.15302/J-FASE-2017170
https://academic.hep.com.cn/fase/EN/Y2018/V5/I2/226

Fig.1 Phylogenetic analysis of polypeptide GhKT2 sequences

Fig.2 The putative transmembrane structure of GhKT2

Fig.3 Subcellular localization of the GhKT2-GFP fusion protein in transgenic Arabidopsis root cells. (a) Localization of control 35S-GFP fluorescence; (b) localization of 35S-GhKT2-GFP fluorescence; (c) localization of 35S-GhKT2-GFP fluorescence in root tip cells plasmolyzed with 500 mmol·L⁻¹ mannitol.

Fig.4 Real-time relative-quantitative PCR analysis of GhKT2 in various tissues of cotton seedlings. Tissues include roots (R), stems (S), first true leaf (FTL), second true leaf (STL), and third true leaf (TTL) unexpanded leaves (UL), shoot apices (A). The expression of GhKT2 was calculated relative to GhUBQ7 expression level. Data are means±SE (n = 3).

Fig.5 Histochemical staining of pGhKT2::GUS transgenic Arabidopsis seedlings

Fig.6 Relative expression levels of GhKT2 measured by real-time PCR in cotton roots exposed to 0.03 mmol·L^-¹ K⁺. (a) Two-week-old plantlet; (b) leaves at higher magnification; (c) leaf primordia around shoot apex; (d) five-day-old seedling; (e) five-day-old seedling that was transferred to low K⁺ (100 mmol·L⁻¹) MS medium for 1 day. Data are mean±SE for four independent experiments each with three independent seedlings (i.e., n = 12).

Fig.7 Phenotype assays of GhKT2-overexpression Arabidopsis lines. WT is wild type (Columbia). K1, K2 and K3 are three independent transgenic lines with a WT background. (a) Real-time PCR analysis of WT and GhKT2-overexpression lines grown in MS medium for 7 d; (b) phenotype comparison between WT and transgenic plants grown on normal MS medium (left) for 10 d and on LK (low K⁺, 100 mmol·L^-¹) medium (right) for 26 d; (c–h) dry weight, K⁺ content, and accumulation of 11-day-old seedlings (four-day-old seedlings grown on normal MS plates were transferred onto MS or LK medium for 7d). (c, d) Comparison of shoot and root dry weight on MS medium and LK medium, respectively; (e, f) comparison of shoot and root K⁺ content on MS medium and LK medium, respectively; (g, h) comparison of shoot and root K⁺ accumulation on MS medium and LK medium, respectively. Data are means±SE (n = 4). Different lower-case letters indicate significant differences at P<0.05.

Fig.8 Comparison of net K⁺ flux as measured by the noninvasive microtest technique between Arabidopsis wild-type (WT) and GhKT2-overexpression transgenic lines (K1, K2, K3). All genotypes were starved of K⁺ by growing on low K⁺ (100 mmol·L^-¹) medium containing 2 mmol·L⁻¹ CsCl for 3 d, and then transferred to a measuring buffer with 100 mmol·L^-¹ K⁺. Data are means±SE (n = 5 to 7). Different lower-case letters indicate significant differences at P<0.05.

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