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利用 (11) C 和 (13) C 联合标记研究大豆(Glycine max)叶片中的碳分配。

Carbon partitioning in soybean (Glycine max) leaves by combined (11) C and (13) C labeling.

机构信息

Department of Chemistry, Washington University, St Louis, MO, 63130, USA.

出版信息

New Phytol. 2012 Dec;196(4):1109-1121. doi: 10.1111/j.1469-8137.2012.04333.x. Epub 2012 Sep 24.

Abstract

We labeled soybean (Glycine max) leaves with 200 and 600 ppm (13) CO(2) spiked with (11) CO(2) and examined the effects of light intensity and water stress on metabolism by using a combination of direct positron imaging and solid-state (13) C nuclear magnetic resonance (NMR) of the same leaf. We first made 60-min movies of the transport of photosynthetically assimilated (11) C labels. The positron imaging identified zones or patches within which variations in metabolism could be probed later by NMR. At the end of each movie, the labeled leaf was frozen in liquid nitrogen to stop metabolism, the leaf was lyophilized, and solid-state NMR was used either on the whole leaf or on various leaf fragments. The NMR analysis determined total (13) C incorporation into sugars, starch, proteins, and protein precursors. The combination of (11) C and (13) C analytical techniques has led to three major conclusions regarding photosynthetically heterogeneous soybean leaves: transient starch deposition is not the temporary storage of sucrose excluded from a saturated sugar-transport system; peptide synthesis is reduced under high-light, high CO(2) conditions; and all glycine from the photorespiratory pathway is routed to proteins within photosynthetically active zones when the leaf is water stressed and under high-light and low CO(2) conditions.

摘要

我们用 200 和 600ppm(13)CO(2)标记大豆(Glycine max)叶片,并掺入(11)CO(2),然后结合直接正电子成像和同一叶片的固态(13)C 核磁共振(NMR),研究了光强和水分胁迫对代谢的影响。我们首先制作了 60 分钟的光合作用同化(11)C 标记物的运输电影。正电子成像识别了代谢可能在以后通过 NMR 进行探测的区域或斑块。在每部电影的结尾,标记的叶片被液氮冷冻以停止代谢,然后将叶片冻干,并对整个叶片或各种叶片碎片进行固态 NMR 分析。NMR 分析确定了总(13)C 掺入到糖、淀粉、蛋白质和蛋白质前体中。(11)C 和(13)C 分析技术的结合得出了关于光合作用不均匀的大豆叶片的三个主要结论:瞬态淀粉沉积不是蔗糖从饱和糖转运系统中排除的临时储存;在高光、高 CO(2)条件下,肽合成减少;当叶片受到水分胁迫和高光、低 CO(2)条件下时,来自光呼吸途径的所有甘氨酸都被导向光合作用活跃区域的蛋白质中。

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