细胞生物学

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Rice Root Hair Phenotypes Imaged by Cryo-SEM

水稻根毛表型的低温扫描电镜成像

HY Haiting Yan YW Yue Wang JZ Jingrong Zhang XC Xinru Cui JW Jiasong Wu JZ Jie Zhou YC Yuan Chen JL Jia Lu RG Ruiyang Guo MO Maggie Ou HL Hongxu Lai

Zhiming Yu*

0 Q&A 3712 Views Jun 5, 2021

Cryo-scanning electron microscopy (cryo-SEM) was first introduced for scientific use in the 1980s. Since then, cryo-SEM has become a routine technique for studying the surfaces and internal structures of biological samples with a high water content. In contrast to traditional SEM, cryo-SEM requires no sample pretreatment processes; thus, we can obtain the most authentic images of the sample shape and structure. Cryo-SEM has two main steps: cryoprocessing of samples and scanning electron microscopy (SEM) observation. The cryoprocessing step includes preparation of the cooled slushing station, cooling of the preparation chamber, sample preparation, and sputtering. The sample is then transferred to an SEM cold stage for observation. We used cryo-SEM to study rice root hair tissues, but the methods and protocols can be applied to other root systems. This protocol optimizes the two key operation steps of reducing the humidity in the growth chamber and previewing the samples before sputtering and can more quickly obtain high-quality images.

Cell Wall Compositional Analysis of Rice Culms

水稻茎杆的细胞壁组成分析

LZ Lanjun Zhang BZ Baocai Zhang* YZ Yihua Zhou*

0 Q&A 3880 Views Oct 20, 2019

The plant cell wall is a complicated network that is mainly constituted of polysaccharides, such as cellulose, hemicellulose and pectin. Many noncellulosic polysaccharides are further acetylated, which confers these polymers flexible physicochemical properties. Due to the significance of cell wall in plant growth and development, the analytic platform has been the focus for a long time. Here, we use internodes/culms, an important organ to provide mechanical support for rice plants, as an experimental sample to explore the method for cell wall composition analysis. The method includes preparation of cell wall residues, sequential extraction of polysaccharides, and measurement of cellulose. The procedure for acetate examination is also described. This method is applicable to determine the composition of individual cell wall polymers and the modifier acetates, and is suitable to identify cell wall relevant mutants based on the advantages in high throughput, precision and repeatability.