Xu Ahui, Wei Cunxu
1Key Laboratory of Crop Genetics and Physiology of Jiangsu Province, Key Laboratory of Plant Functional Genomics of the Ministry of Education, Yangzhou University, Yangzhou, 225009 China.
2Co-Innovation Center for Modern Production Technology of Grain Crops of Jiangsu Province/Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education, Yangzhou University, Yangzhou, 225009 China.
Plant Methods. 2020 Feb 1;16:8. doi: 10.1186/s13007-020-0558-x. eCollection 2020.
This review summarizes the main applications of different sections and some improved sectioning methods in investigating the microstructure and histochemistry of cereal kernels. Thick sections of developing kernels prepared by free-hand and sliding microtome-aided sectioning method can be employed to elucidate tissue anatomy and histochemistry. The thin sections of mature kernels prepared by ultramicrotome-aided sectioning method can exhibit the micromorphology of starch granules when stained with iodine solution. The paraffin sections of developing kernels can exhibit the tissue anatomy of kernel, the accumulation of storage substances, and the location of protein and gene transcripts with immunohistochemistry and in situ hybridization techniques. The semithin resin sections can clearly exhibit the morphology of cells, starch granules, and protein bodies in kernel, but the sections prepared with different resins have various advantages and disadvantages for research investigating the morphology and histochemistry of cereal kernels. The improved methods of free-hand sectioning and ultramicrotome-aided sectioning of mature kernels are suitable for investigating the morphology of starch granules in a large number of samples in a short time. The modified method for preparing resin sections of whole kernels can be employed to determine the morphology and distribution of cells, starch granules, and storage protein in mature, developing, germinated, and cooked kernels in situ. This review could help researchers choose appropriate sections for investigating the microstructure and histochemistry of cereal kernels according to their study objectives.
本综述总结了不同切片方法及其一些改进方法在研究谷物籽粒微观结构和组织化学方面的主要应用。通过徒手切片和滑走切片法制备的发育中籽粒厚切片可用于阐明组织解剖结构和组织化学。用超薄切片法制备的成熟籽粒薄切片在用碘液染色时可显示淀粉颗粒的微观形态。发育中籽粒的石蜡切片可通过免疫组织化学和原位杂交技术展示籽粒的组织解剖结构、贮藏物质的积累以及蛋白质和基因转录本的定位。半薄树脂切片能清晰显示籽粒中细胞、淀粉颗粒和蛋白体的形态,但不同树脂制备的切片在研究谷物籽粒形态和组织化学方面各有优缺点。成熟籽粒的徒手切片和超薄切片改进方法适用于在短时间内对大量样品的淀粉颗粒形态进行研究。全籽粒树脂切片的改良制备方法可用于原位确定成熟、发育、发芽和煮熟籽粒中细胞、淀粉颗粒和贮藏蛋白的形态及分布。本综述有助于研究人员根据研究目的选择合适的切片方法来研究谷物籽粒的微观结构和组织化学。