Weise Sean E, Weber Andreas P M, Sharkey Thomas D
Department of Botany, University of Wisconsin-Madison, 430 Lincoln Drive, Madison, WI 53706 USA.
Planta. 2004 Jan;218(3):474-82. doi: 10.1007/s00425-003-1128-y. Epub 2003 Oct 18.
Transitory starch is formed in chloroplasts during the day and broken down at night. We investigated carbon export from chloroplasts resulting from transitory-starch breakdown. Starch-filled chloroplasts from spinach ( Spinacia oleracea L. cv. Nordic IV) were isolated 1 h after the beginning of the dark period and incubated for 2.5 h, followed by centrifugation through silicone oil. Exported products were measured in the incubation medium to avoid measuring compounds retained inside the chloroplasts. Maltose and glucose made up 85% of the total exported products and were exported at rates of 626 and 309 nmol C mg(-1) chlorophyll h(-1), respectively. Net export of phosphorylated products was less than 5% and higher maltodextrins were not detected. Maltose levels in leaves of bean ( Phaseolus vulgaris L. cv. Linden), spinach, and Arabidopsis thaliana (L.) Heynh. were low in the light and high in the dark. Maltose levels remained low and unchanged during the light/dark cycle in two starch-deficient Arabidopsis mutants, stf1, deficient in plastid phosphoglucomutase, and pgi, deficient in plastid phosphoglucoisomerase. Through the use of nonaqueous fractionation, we determined that maltose was distributed equally between the chloroplast and cytosolic fractions during darkness. In the light there was approximately 24% more maltose in the cytosol than the chloroplast. Taken together these data indicate that maltose is the major form of carbon exported from the chloroplast at night as a result of starch breakdown. We hypothesize that the hydrolytic pathway for transitory-starch degradation is the primary pathway used when starch is being converted to sucrose and that the phosphorolytic pathway provides carbon for other purposes.
暂存淀粉在白天于叶绿体中形成,并在夜间分解。我们研究了暂存淀粉分解导致的叶绿体碳输出。在黑暗期开始1小时后,从菠菜(Spinacia oleracea L. cv. Nordic IV)中分离出充满淀粉的叶绿体,并孵育2.5小时,随后通过硅油离心。在孵育培养基中测量输出产物,以避免测量保留在叶绿体内的化合物。麦芽糖和葡萄糖占总输出产物的85%,输出速率分别为626和309 nmol C mg(-1)叶绿素h(-1)。磷酸化产物的净输出小于5%,未检测到更高的麦芽糖糊精。菜豆(Phaseolus vulgaris L. cv. Linden)、菠菜和拟南芥(Arabidopsis thaliana (L.) Heynh.)叶片中的麦芽糖水平在光照下较低,在黑暗中较高。在两个淀粉缺陷型拟南芥突变体中,即缺乏质体磷酸葡萄糖变位酶的stf1和缺乏质体磷酸葡萄糖异构酶的pgi,麦芽糖水平在光/暗循环期间保持较低且不变。通过非水相分级分离,我们确定在黑暗期间麦芽糖在叶绿体和细胞质部分之间平均分布。在光照下,细胞质中的麦芽糖比叶绿体中的大约多24%。综合这些数据表明,麦芽糖是夜间淀粉分解导致的叶绿体碳输出的主要形式。我们假设,暂存淀粉降解的水解途径是淀粉转化为蔗糖时使用的主要途径,而磷酸解途径为其他目的提供碳。