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用于植原体体内成像的4',6-二脒基-2-苯基吲哚(DAPI)和共聚焦激光扫描显微镜技术

DAPI and Confocal Laser-Scanning Microscopy for In Vivo Imaging of Phytoplasmas.

作者信息

Musetti Rita, Buxa Stefanie Vera

机构信息

Department of Agricultural, Food, Environmental and Animal Sciences, University of Udine, Udine, Italy.

Centre for BioSystems, Land Use and Nutrition, Institute of Phytopathology, Justus Liebig University Giessen, Giessen, Germany.

出版信息

Methods Mol Biol. 2019;1875:301-306. doi: 10.1007/978-1-4939-8837-2_22.

Abstract

As phytoplasmas are located inside the phloem tissue, always surrounded by numerous layers of other cells, they can result difficult candidates for microscopical investigations. Moreover, the necessity to kill the plant tissues for microscopy observations causes instantaneous and irreversible modifications in the sieve elements, leading to misleading information and erroneous interpretations. Phytoplasmas were here investigated in intact Vicia faba host plants using DAPI as fluorescent probe and confocal laser scanning microscopy. The described nondestructive technique may be applied for the imaging of phytoplasmas and of different pathogen-related responses in planta.

摘要

由于植原体位于韧皮部组织内部,总是被多层其他细胞包围,因此它们可能很难成为显微镜检查的对象。此外,为进行显微镜观察而杀死植物组织的必要性会导致筛管分子立即发生不可逆转的变化,从而产生误导性信息和错误解读。本文使用4',6-二脒基-2-苯基吲哚(DAPI)作为荧光探针,通过共聚焦激光扫描显微镜对完整的蚕豆寄主植物中的植原体进行了研究。所描述的非破坏性技术可用于对植物中的植原体以及不同的病原体相关反应进行成像。

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