Hybrid Label-Free Molecular Microscopies for Simultaneous Visualization of Changes in Cell Wall Polysaccharides of Peach at Single- and Multiple-Cell Levels during Postharvest Storage
Weinan Huang,
Yating Nie,
Nan Zhu,
Yifan Yang,
Changqing Zhu,
Minbiao Ji,
Di Wu,
Kunsong Chen
Affiliations
Weinan Huang
College of Agriculture & Biotechnology/Zhejiang Provincial Key Laboratory of Horticultural Plant Integrative Biology/The State Agriculture Ministry Laboratory of Horticultural Plant Growth, Development and Quality Improvement, Zhejiang University, Zijingang Campus, Hangzhou 310058, China
Yating Nie
College of Agriculture & Biotechnology/Zhejiang Provincial Key Laboratory of Horticultural Plant Integrative Biology/The State Agriculture Ministry Laboratory of Horticultural Plant Growth, Development and Quality Improvement, Zhejiang University, Zijingang Campus, Hangzhou 310058, China
Nan Zhu
College of Agriculture & Biotechnology/Zhejiang Provincial Key Laboratory of Horticultural Plant Integrative Biology/The State Agriculture Ministry Laboratory of Horticultural Plant Growth, Development and Quality Improvement, Zhejiang University, Zijingang Campus, Hangzhou 310058, China
Yifan Yang
State Key Laboratory of Surface Physics and Department of Physics, Human Phenome Institute, Multiscale Research Institute of Complex Systems, Key Laboratory of Micro and Nano Photonic Structures (Ministry of Education), Fudan University, Shanghai 200433, China
Changqing Zhu
College of Agriculture & Biotechnology/Zhejiang Provincial Key Laboratory of Horticultural Plant Integrative Biology/The State Agriculture Ministry Laboratory of Horticultural Plant Growth, Development and Quality Improvement, Zhejiang University, Zijingang Campus, Hangzhou 310058, China
Minbiao Ji
State Key Laboratory of Surface Physics and Department of Physics, Human Phenome Institute, Multiscale Research Institute of Complex Systems, Key Laboratory of Micro and Nano Photonic Structures (Ministry of Education), Fudan University, Shanghai 200433, China
Di Wu
College of Agriculture & Biotechnology/Zhejiang Provincial Key Laboratory of Horticultural Plant Integrative Biology/The State Agriculture Ministry Laboratory of Horticultural Plant Growth, Development and Quality Improvement, Zhejiang University, Zijingang Campus, Hangzhou 310058, China
Kunsong Chen
College of Agriculture & Biotechnology/Zhejiang Provincial Key Laboratory of Horticultural Plant Integrative Biology/The State Agriculture Ministry Laboratory of Horticultural Plant Growth, Development and Quality Improvement, Zhejiang University, Zijingang Campus, Hangzhou 310058, China
Softening of fruit during the postharvest storage, which is mainly associated with both compositional and spatial changes of polysaccharides within cell wall, affects the texture and quality of fruit. Current research on the fruit softening mechanism lacks an understanding of the overall softening at the cell level. The objective of this work was to investigate the change in the spatial distribution of cell wall polysaccharides in peach flesh cells at both single- and multiple-cell levels in a label-free way during the postharvest storage. Nonmelting peaches (Prunus persica L. Batsch cv.”Zhonghuashoutao”) at commercial maturity were stored at 0 °C and 20 °C. Firmness measurement and chemical analysis were performed at each storage time. In addition, three molecular imaging techniques, namely confocal Raman microspectroscopy (CRM), Fourier transform infrared microspectroscopy (FTIRM), and stimulated Raman scattering microscopy (SRS) were used to visualize changes in the spatial distribution of cell wall polysaccharides of peach fruit in a label-free way during the postharvest storage. The combination of CRM and FTIRM provided complementary spectral information to visualize the spatial changes of cellulose, hemicellulose, and pectin in the cell wall of peach flesh during softening at the single-cell level, and found that the cell wall polysaccharides tended to be concentrated in the cell corner of parenchymal cells at the late stage. Furthermore, SRS, which is an ultrafast Raman imaging technique (approximately three or four orders of magnitude faster than CRM), was used for high-throughput cell wall phenotypes measurement. Different degradation degrees of parenchymal cells during fruit softening were found based on the gray-scale statistical analysis of SRS data. In general, cell wall polysaccharides decreased during softening and tended to be concentrated in the cell corner for most parenchymal cells at the late stage, but there were also some cells not in line with the whole softening trends. The results show that there were differences in the content and spatial changes of cell wall polysaccharides among parenchymal cells of peach fruit during the softening process, and the hybrid use of CRM, FTIRM, and SRS is a promising method for simultaneous visualization of changes in cell wall polysaccharides of peach.
