An integrated isotopic labeling and freeze sampling apparatus (ILSA) to support sampling leaf metabolomics at a centi-second scale.
An integrated isotopic labeling and freeze sampling apparatus (ILSA) to support sampling leaf metabolomics at a centi-second scale.
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集成同位素标记和冷冻采样装置 (ILSA)，支持厘秒级叶片代谢组学采样
DOI：
10.1186/s13007-022-00926-7
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发表时间：
2022-07-30
期刊：
影响因子：
5.1
通讯作者：
中科院分区：
文献类型：
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作者：
光合作用与呼吸和氮同化密切相互作用，决定了叶片的光合效率。准确量化光合、呼吸和氮同化的代谢通量有助于提高光合效率的设计。为了准确估计代谢通量，需要在精确控制的环境下收集包括叶片代谢和同位素丰度变化在内的时间序列数据。但对于在特定环境下的同位素标记叶片，人工采样的时间成本通常长于光合代谢中几种中间体的周转时间。在这种情况下，叶片样品的代谢或生理状态会在采样过程中发生变化，代谢组学数据的准确性可能会受到影响。在此基础上，研制了一种同位素标记和冷冻取样一体化装置（ILSA），该装置可在0.05 s内自动完成冷冻取样。ILSA不仅可以用于光合代谢测量的采样，也适用于受控环境（[CO2]和光照）下的叶片同位素标记实验。结合高效液相色谱-质谱联用（HPLC-MS /MS）作为代谢测量方法，研究了：(1)水稻暗积累叶片光合代谢产物池大小的变化规律；(2)水稻与拟南芥光合代谢通量的变化规律。ILSA的开发为光合代谢研究和代谢通量分析提供了支持，为叶片生理研究提供了新的工具。在线版本包含补充资料，下载地址：10.1186/s13007-022-00926-7。
Photosynthesis close interacts with respiration and nitrogen assimilation, which determine the photosynthetic efficiency of a leaf. Accurately quantifying the metabolic fluxes in photosynthesis, respiration and nitrogen assimilation benefit the design of photosynthetic efficiency improvement. To accurately estimate metabolic fluxes, time-series data including leaf metabolism and isotopic abundance changes should be collected under precisely controlled environments. But for isotopic labelled leaves under defined environments the, time cost of manually sampling usually longer than the turnover time of several intermediates in photosynthetic metabolism. In this case, the metabolic or physiological status of leaf sample would change during the sampling, and the accuracy of metabolomics data could be compromised. Here we developed an integrated isotopic labeling and freeze sampling apparatus (ILSA), which could finish freeze sampling automatically in 0.05 s. ILSA can not only be used for sampling of photosynthetic metabolism measurement, but also suit for leaf isotopic labeling experiments under controlled environments ([CO2] and light). Combined with HPLC–MS/MS as the metabolic measurement method, we demonstrated: (1) how pool-size of photosynthetic metabolites change in dark-accumulated rice leaf, and (2) variation in photosynthetic metabolic flux between rice and Arabidopsis thaliana. The development of ILSA supports the photosynthetic research on metabolism and metabolic flux analysis and provides a new tool for the study of leaf physiology. The online version contains supplementary material available at 10.1186/s13007-022-00926-7.
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影响因子：
--
作者：
Kajihata S;Furusawa C;Matsuda F;Shimizu H
通讯作者：
Shimizu H
影响因子：
3.7
作者：
Khan, Naveed;Essemine, Jemaa;Zhu, Xin-Guang
通讯作者：
Zhu, Xin-Guang
影响因子：
6.9
作者：
Arrivault S;Obata T;Szecówka M;Mengin V;Guenther M;Hoehne M;Fernie AR;Stitt M
通讯作者：
Stitt M
影响因子：
7.7
作者：
Mallmann J;Heckmann D;Bräutigam A;Lercher MJ;Weber AP;Westhoff P;Gowik U
通讯作者：
Gowik U
DOI：
10.1007/978-1-4939-7225-8_12
发表时间：
2017-01-01
期刊：
PHOTORESPIRATION: METHODS AND PROTOCOLS
影响因子：
--
作者：
Ma, Fangfang;Jazmin, Lara J.;Allen, Doug K.
通讯作者：
Allen, Doug K.