Improving sample preparation to investigate lignin intensity of xylem at the cellular level by confocal Raman microspectroscopy of Populus tomentosa
Improving sample preparation to investigate lignin intensity of xylem at the cellular level by confocal Raman microspectroscopy of Populus tomentosa
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DOI：
10.1007/s11676-020-01244-1
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发表时间：
2020-11
影响因子：
3
通讯作者：
B. Wang;Mei Luo;Yadi Liu;Xiaorui Guo;Xiatong Liu;Chong Zhang;Zhijing Zhao;Di Liu;Hui Li-Hui
中科院分区：
文献类型：
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作者：
B. Wang;Mei Luo;Yadi Liu;Xiaorui Guo;Xiatong Liu;Chong Zhang;Zhijing Zhao;Di Liu;Hui Li-Hui
共焦拉曼显微光谱（CRM）是原位分析细胞壁成分分布的重要工具。在本研究中，我们改进了石蜡包埋切片结合正己烷脱蜡的样品制备方法，以获得高分辨率的拉曼图像。毛白杨木质部纤维细胞的细胞壁成分不同于射线细胞和导管细胞。乙酰溴和CRM方法产生类似的趋势时，木质部区域的木质素强度的差异进行了比较transgenicPtrLac4和野生型P。绒毛草然而，CRM被证明是更有用的，以分析木质素在每个细胞类型中的分布，并区分在细胞水平上的木质素强度的详细差异。因此，CRM被证明是一个有用的原位方法，快速分析木质部木质素含量的空间变异的木本植物。
Confocal Raman microspectroscopy (CRM) is an important tool for analyzing the compositional distribution of cell walls in situ. In this study, we improved the sample preparation method using paraffin-embedded sections combined with hexane dewaxing to obtain high resolution Raman images. We determined that the cell wall components of fiber cells were different from those of ray cells and vessel cells in the xylem ofPopulus tomentosa. Acetyl bromide and CRM methods produced similar trends when the difference in lignin intensity in the xylem region was compared between transgenicPtrLac4and wild-typeP. tomentosa. However, CRM proved more useful to analyze the lignin distribution in each cell type and distinguished the detailed difference in lignin intensity at the cellular level. Thus, CRM proved to be a useful in situ method to rapidly analyze the spatial variation of lignin content in the xylem of woody plants.