da Silveira Sylvia Rodrigues, Koehler Andréa Dias, Mendes Gomes Ana Cristina M, Cabral Glaucia Barbosa, de Campos Carneiro Vera Tavares, de Alencar Dusi Diva Maria, Martinelli Adriana Pinheiro
Plant Biotechnology Lab, Center of Nuclear Energy in Agriculture, University of São Paulo, Piracicaba, SP, Brazil.
Brazilian Agricultural Research Corporation (Embrapa), Embrapa Genetic Resources and Biotechnology, Brasilia, DF, Brazil.
Methods Mol Biol. 2022;2527:247-263. doi: 10.1007/978-1-0716-2485-2_18.
In situ hybridization with mRNA probes enables the detection and localization of gene expression in plant somatic embryogenesis samples. BbrizSERK is a gene that is expressed in embryogenic cells and tissues of Brachiaria. Here we describe methods used for in situ hybridization to localize BbrizSERK transcripts during somatic embryogenesis of Brachiaria brizantha according to the plant material and observations intended, using paraffin or butyl methyl methacrylate resin-embedded samples, as well as a method for whole-mount preparation applicable for the analysis of other genes involved in embryogenic processes, along with other in vitro processes.
使用mRNA探针进行原位杂交能够检测和定位植物体细胞胚胎发生样本中的基因表达。BbrizSERK是一个在臂形草的胚性细胞和组织中表达的基因。在此，我们描述了根据植物材料和预期观察结果，使用石蜡或丁基甲基丙烯酸甲酯树脂包埋样本，在臂形草体细胞胚胎发生过程中对BbrizSERK转录本进行定位的原位杂交方法，以及一种适用于分析参与胚胎发生过程的其他基因以及其他体外过程的整装制备方法。
