AtPHT4;4 is a chloroplast-localized ascorbate transporter in Arabidopsis

• Published in: Nature communications 2015-01, Vol.6 (1), p.5928-5928, Article 5928
• Main Authors: Miyaji, Takaaki, Kuromori, Takashi, Takeuchi, Yu, Yamaji, Naoki, Yokosho, Kengo, Shimazawa, Atsushi, Sugimoto, Eriko, Omote, Hiroshi, Ma, Jian Feng, Shinozaki, Kazuo, Moriyama, Yoshinori
• Format: Article
• Language: English
• Subjects: 631/449/448/1374; 631/449/448/2651; Anion Transport Proteins - genetics; Anion Transport Proteins - metabolism; Arabidopsis; Arabidopsis - genetics; Arabidopsis - metabolism; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Arabidopsis thaliana; Ascorbic Acid - metabolism; Chloroplasts - metabolism; DNA Primers - genetics; DNA, Complementary - genetics; Fluorescence; Gene Knockout Techniques; Humanities and Social Sciences; Immunohistochemistry; In Vitro Techniques; Light; Membrane Transport Proteins - genetics; Membrane Transport Proteins - metabolism; multidisciplinary; Polymerase Chain Reaction; Science; Science (multidisciplinary); Stress, Physiological - genetics
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms6928

Ascorbate is an antioxidant and coenzyme for various metabolic reactions in vivo . In plant chloroplasts, high ascorbate levels are required to overcome photoinhibition caused by strong light. However, ascorbate is synthesized in the mitochondria and the molecular mechanisms underlying ascorbate transport into chloroplasts are unknown. Here we show that AtPHT4;4, a member of the phosphate transporter 4 family of Arabidopsis thaliana , functions as an ascorbate transporter. In vitro analysis shows that proteoliposomes containing the purified AtPHT4;4 protein exhibit membrane potential- and Cl − -dependent ascorbate uptake. The AtPHT4;4 protein is abundantly expressed in the chloroplast envelope membrane. Knockout of AtPHT4;4 results in decreased levels of the reduced form of ascorbate in the leaves and the heat dissipation process of excessive energy during photosynthesis is compromised. Taken together, these observations indicate that the AtPHT4;4 protein is an ascorbate transporter at the chloroplast envelope membrane, which may be required for tolerance to strong light stress. In plants, ascorbate is synthesized in the mitochondria yet plays essential roles as an antioxidant in the chloroplast. Here, Miyaji et al. show that AtPHT4;4 is a chloroplast envelope ascorbate transporter and suggest it is required for dissipation of excess energy under light stress.