Raffinose in Chloroplasts is Synthesized in the Cytosol and Transported across the Chloroplast Envelope

ISSN： 0032-0781

Source： Plant and Cell Physiology, Vol.50, Iss.12, 2009-12, pp. : 2174-2182

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Abstract

In chloroplasts, several water-soluble carbohydrates have been suggested to act as stress protectants. The trisaccharide raffinose (α-1,6-galactosyl sucrose) is such a carbohydrate but has received little attention. We here demonstrate by compartmentation analysis of leaf mesophyll protoplasts that raffinose is clearly (to about 20%) present in chloroplasts of cold-treated common bugle (Ajuga reptans L.), spinach (Spinacia oleracea L.) and Arabidopsis [Arabidopsis thaliana (L.) Heynh.] plants. The two dedicated enzymes needed for raffinose synthesis, galactinol synthase and raffinose synthase, were found to be extra-chloroplastic (probably cytosolic) in location, suggesting that the chloroplast envelope contains a raffinose transporter. Uptake experiments with isolated Ajuga and Arabidopsis chloroplasts clearly demonstrated that raffinose is indeed transported across the chloroplast envelope by a raffinose transporter, probably actively. Raffinose uptake into Ajuga chloroplasts was a saturable process with apparent K_m and v_max values of 27.8 mM and 3.3 mol mg⁻¹ Chl min⁻¹, respectively.