共聚焦显微拉曼光谱研究碱处理对稻秸发酵制沼气的影响

碱水解法以其方便高效成为稻秸发酵制沼气中广泛采用的化学预处理方法，但是碱水解对于稻秸细胞壁中高聚物成分及其空间致密交联结构的作用机理有待深入研究。采用共聚焦显微拉曼光谱和透射电镜研究了NaOH碱处理对稻秸厌氧发酵产沼气的影响。首先对未处理稻秸和碱处理稻秸进行微米级空间分辨率的拉曼光谱面扫描，然后对这两类样本进行主成分分析，发现累计贡献率达99%的前两个主成分空间中两类样本分别呈射线状分布，两类样本分界线清楚表明碱处理导致稻叶组织的拉曼光谱响应特性发生了明显变化;进一步分析前两个主成分的载荷图，发现拉曼峰1 739，1 508和1 094 cm-1是影响主成分的重要谱带，而这三个拉曼峰分别归属于半纤维素、木质素和纤维素的拉曼散射效应;结合半纤维素、木质素和纤维素的拉曼特征峰和显微图像信息实现了组织细胞中三种成分的化学成像分析，发现碱处理破坏了稻叶组织中上述三种成分的致密空间结构，并使它们的含量明显减少，尤其是木质素。由此得出结论：共聚焦显微拉曼光谱可实现稻叶组织细胞中半纤维素、木质素和纤维素的无损检测，结合显微空间信息可实现稻秸中三种成分的化学成像分析，该研究有助揭示碱处理促进秸秆厌氧发酵产沼气的作用机理。

Studies on Effect of Alkali Pretreatment on Anaerobic Digestion of Rice Straw with Confocal Raman Microscopy

NaOH pretreatment is a convenient and effective method which is widely used in rice straw anaerobic digestion. But the mechanism of the alkaline (NaOH) hydrolysis of biopolymers compositions and polymeric cross-linked network structures ofrice straw cell wall need further study. This paper firstly studied the effect and mechanism of alkali pretreatment on anaerobic digestion and biogas production of rice straw by using a combination of confocal Raman microscopy and transmission electron microscope. First, the original rice straw and the rice straw pretreated by NaOH were taken for mapping scanning by confocal Raman microscopy withmicron-scale spatial resolution. Then principal component analysis was adoptedto extract main information of Raman spectra, it could be found that the two types of samples were respectively presented with ray-like distribution in the first two principal component space, which were with cumulative contribution of 99%. And there was a clear boundary between the two types of samples without any overlapping, indicating that there was a significant difference of Raman spectralcharacteristic between original rice leaf and rice leaf pretreated by NaOH. Further analysis of the loading weights of the first two principal components showed that the Raman peaks at 1 739, 1 508 and 1 094 cm-1 were the important bands, and these three Raman peaks were attributed to the scattering of hemicellulose, cellulose and lignin respectively. Following, chemical imaging analysisof hemicellulose, cellulose and lignin were achieved by combining these Raman peaks and microscopic image information. It could be found that the NaOH pretreatment resulted in a loss of dense spatial uniformity structure of tissue and great decreases of the contents of these three ingredients, particularly lignin. It can be concluded that it is feasible to non-destructively measure hemicellulose, lignin and cellulose in rice straw tissue by confocal Raman microscopy, and toachieve chemical imaging analysis of the three ingredients in tissue, and this research will be much help for revealing the promotion mechanism of NaOH pretreatment for the rice straw fermentation and biogas production.