Ortiz-Ramírez Carlos, Arevalo Edgar Demesa, Xu Xiaosa, Jackson David P, Birnbaum Kenneth D
Center for Genomics and Systems Biology, Department of Biology, New York University, New York, New York.
Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Curr Protoc Plant Biol. 2018 Sep;3(3):e20072. doi: 10.1002/cppb.20072. Epub 2018 Aug 23.
Maize is one the most widely cultivated crops worldwide and an important model system for the study of genetics and cytogenetics. Although the availability of a genome sequence has enabled new quantitative genomic studies, developing methods to isolate specific types of cells will enable useful approaches for transcriptomic analysis in the crop plant. Fluorescence-activated cell sorting (FACS) is a powerful technique for cell isolation and the study of transcriptional profiles from specific cell populations. The use of FACS on plant cells requires the generation of protoplasts by tissue digestion and cell wall removal. Although some protocols are available, they mainly focus on dicot species and obtaining sufficient protoplasts from inner tissue layers has been challenging in both monocots and dicots. Here, we report a new protocol that dramatically increases protoplast yield from maize for subsequent cell isolation by FACS. This protocol is efficient in generating protoplasts from root and shoot inner layers and can also be applied successfully to Arabidopsis thaliana. © 2018 by John Wiley & Sons, Inc.
玉米是全球种植最广泛的作物之一，也是遗传学和细胞遗传学研究的重要模式系统。尽管基因组序列的可用性推动了新的定量基因组学研究，但开发分离特定类型细胞的方法将为作物的转录组分析提供有用的途径。荧光激活细胞分选（FACS）是一种用于细胞分离和特定细胞群体转录谱研究的强大技术。在植物细胞上使用FACS需要通过组织消化和去除细胞壁来生成原生质体。虽然有一些方案可供使用，但它们主要集中在双子叶植物物种上，并且在单子叶植物和双子叶植物中从内部组织层获得足够的原生质体一直具有挑战性。在这里，我们报告了一种新方案，该方案显著提高了玉米原生质体产量，以便随后通过FACS进行细胞分离。该方案能有效地从根和茎的内层生成原生质体，并且也能成功应用于拟南芥。© 2018 约翰威立父子公司版权所有
