Li Xiao-li, Luo Liu-bin, Hu Xiao-qian, Lou Bing-gan, He Yong
Guang Pu Xue Yu Guang Pu Fen Xi. 2014 Jun;34(6):1571-6.
Healthy tea and tea infected by anthracnose were first studied by confocal Raman microscopy to illustrate chemical changes of cell wall in the present paper. Firstly, Raman spectra of both healthy and infected sample tissues were collected with spatial resolution at micron-level, and ultrastructure of healthy and infected tea cells was got from scanning electron microscope. These results showed that there were significant changes in Raman shift and Raman intensity between healthy and infected cell walls, indicating that great differences occurred in chemical compositions of cell walls between healthy and infected samples. In details, intensities at many Raman bands which were closely associated with cellulose, pectin, esters were reduced after infection, revealing that the content of chemical compounds such as cellulose, pectin, esters was decreased after infection. Subsequently, chemical imaging of both healthy and infected tea cell walls were realized based on Raman fingerprint spectra of cellulose and microscopic spatial structure. It was found that not only the content of cellulose was reduced greatly after infection, but also the ordered structure of cellulose was destroyed by anthracnose infection. Thus, confocal Raman microscopy was shown to be a powerful tool to detect the chemical changes in cell wall of tea caused by anthracnose without any chemical treatment or staining. This research firstly applied confocal Raman microscopy in phytopathology for the study of interactive relationship between host and pathogen, and it will also open a new way for intensive study of host-pathogen at cellular level.
本文首次利用共聚焦拉曼显微镜对健康茶叶和感染炭疽病的茶叶进行研究,以阐明细胞壁的化学变化。首先,以微米级的空间分辨率采集健康和感染样本组织的拉曼光谱,并通过扫描电子显微镜获得健康和感染茶树细胞的超微结构。这些结果表明,健康和感染细胞壁之间的拉曼位移和拉曼强度存在显著变化,表明健康和感染样本的细胞壁化学成分存在很大差异。具体而言,感染后许多与纤维素、果胶、酯类密切相关的拉曼谱带强度降低,表明感染后纤维素、果胶、酯类等化合物的含量减少。随后,基于纤维素的拉曼指纹光谱和微观空间结构,实现了健康和感染茶树细胞壁的化学成像。研究发现,感染后不仅纤维素含量大幅降低,而且炭疽病感染破坏了纤维素的有序结构。因此,共聚焦拉曼显微镜被证明是一种无需任何化学处理或染色就能检测炭疽病引起的茶树细胞壁化学变化的有力工具。本研究首次将共聚焦拉曼显微镜应用于植物病理学中宿主与病原体相互作用关系的研究,也将为在细胞水平上深入研究宿主-病原体关系开辟一条新途径。