Trethewey R N, ap Rees T
Department of Plant Sciences, University of Cambridge, U.K.
Biochem J. 1994 Jul 15;301 ( Pt 2)(Pt 2):449-54. doi: 10.1042/bj3010449.
At the end of a 12-h day leaves of the mutant of Arabidopsis thaliana L., TC265, contained 4-5 times more starch than those of the wild type. During a subsequent 12-h night the decline in the starch content of the leaves of the mutant was at least 50% of that of the wild-type leaves. Starch labelled in the light in a 30-min pulse in 14CO2 was rapidly broken down in a subsequent 12-h chase in the dark in air in the leaves of both mutant and wild type. Chloroplasts from leaves of the wild type took up [32P]Pi and [U-14C]glucose at 12 and 1.6 mumol/h per mg of chlorophyll respectively; chloroplasts from the mutant showed a similar rate for [32P]Pi but no uptake of [U-14C]glucose. The glucose content of freshly isolated chloroplasts from the mutant was twice that of chloroplasts from the wild type; this difference was accentuated when the isolated chloroplasts were incubated in the dark. SDS/PAGE of preparations of chloroplast envelopes showed that those from the mutant were deficient in a protein band of approximate molecular mass 40 kDa. It is suggested that in mutant TC265 the primary lesion is in a hexose transporter in the chloroplast envelope, and that this transporter moves the products of starch breakdown that are destined for sucrose synthesis from the chloroplast to the cytosol.
在12小时的白昼结束时,拟南芥突变体TC265的叶片中淀粉含量比野生型多4至5倍。在随后的12小时夜间,突变体叶片中淀粉含量的下降幅度至少是野生型叶片的50%。在光照下用14CO2进行30分钟脉冲标记的淀粉,在随后于黑暗空气中进行的12小时追踪过程中,突变体和野生型叶片中的淀粉都迅速分解。野生型叶片的叶绿体分别以每毫克叶绿素12和1.6 μmol/h的速率摄取[32P]Pi和[U-14C]葡萄糖;突变体的叶绿体对[32P]Pi的摄取速率相似,但不摄取[U-14C]葡萄糖。突变体新鲜分离的叶绿体中的葡萄糖含量是野生型叶绿体的两倍;当分离的叶绿体在黑暗中孵育时,这种差异更加明显。叶绿体被膜制剂的SDS/PAGE显示,突变体的叶绿体被膜缺乏一条分子量约为40 kDa的蛋白带。有人认为,在突变体TC265中,主要损伤在于叶绿体被膜中的一种己糖转运蛋白,并且这种转运蛋白将淀粉分解产物从叶绿体转运到细胞质中,用于蔗糖合成。