Imaging mass spectrometry analysis of photosynthetic products in poplar
Imaging mass spectrometry analysis of photosynthetic products in poplar
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杨树光合产物的成像质谱分析
DOI：
10.1093/jmicro/dfv297
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发表时间：
2015
期刊：
影响因子：
1.8
通讯作者：
Mariko Norisada and Akira Isogai
中科院分区：
文献类型：
--
作者：
Miyuki Takeuchi;Mariko Norisada and Akira Isogai
TEM技术非常适合于在高放大率下获取有限区域的图像，但不适合于大面积的图像，如植物组织和大细胞。为了捕获这样的数字图像，我们开发了自动采集TEM系统。我们成功地获得了千兆像素级的高分辨率TEM图像，并使用图像拼接程序进行了合并[1]。为了拍摄拟南芥的千兆像素级TEM照片，使用高压冷冻方法固定根。然后，我们使用自动采集TEM系统采集了数千张重叠的TEM显微照片，并将它们合并[2]。由此获得的图像包括根尖的横向和/或径向纵向截面。使用这些千兆像素的TEM图像，我们发现ER体样纺锤形结构从ER在根尖出芽。为了确定这些结构的分布和作用，将它们标记在根尖切片的千兆像素TEM图像上并计数。这些结构分布在根冠中，尤其是侧根冠。免疫细胞化学分析显示，LRC细胞的内质网小体和空泡中有信号，并融合在一起。拟南芥有三个KDEL尾蛋白酶，AtCEP 1 -3。这些酶在程序性细胞死亡期间在组织和细胞中表达[3]。免疫荧光检测显示AtCEP 2定位于LRC细胞的ER体。这些结果表明，在LRC细胞中的ER体参与KDEL尾蛋白酶，如AtCEP 2到液泡的质量运输。[1]K Toyooka等人，Plant Cell Physiol 55（2014）1544-1555. [2]K Toyooka等人，Plant Morphology 26（2014）3-8. [3]G Hierl等人，Plant Mol. Biol 84（2014）605-620.
TEM techniques are well-suited to the acquisition of images of limited regions under high magnification but not of wide areas such as plant tissues and large cells. To capture such digital images, we developed an auto-acquisition TEM system. We successfully acquired gigapixel-class high-resolution TEM images, which were merged using an image-tiling program [1]. To take gigapixel-class TEM pictures of Arabidopsis, the roots were fixed using a high-pressure freezing method. We then acquired several thousand overlapping TEM micrographs using the auto-acquisition TEM system and merged them [2]. The images thus obtained included transverse and/or radial longitudinal sections of root tips. Using these gigapixel TEM images, we found ER body-like spindle-shaped structures budding from ER in the root tip. To determine the distribution and the role of these structures, they were marked on gigapixel TEM images of the sections of root tips and counted. The structures distributed in the root cap, especially the lateral root cap (LRC). Immunocytochemical analysis for the LRC cells showed that signals were detected in ER bodies and vacuoles, which were fused in places. Arabidopsis has three KDEL-tailed proteases, AtCEP1-3. These enzymes are expressed in tissues and cells during programmed cell death [3]. Immunofluorescence assay showed that AtCEP2 was localized in the ER bodies of LRC cells. These results indicate that ER bodies in the LRC cells are involved in mass transport of KDEL-tailed proteases such as AtCEP2 to the vacuoles.[1] K Toyooka et al, Plant Cell Physiol 55 (2014) 1544-1555.[2] K Toyooka et al, Plant Morphology 26 (2014) 3-8.[3] G Hierl et al, Plant Mol. Biol 84 (2014) 605-620.