发布: 2017年07月20日第7卷第14期 DOI: 10.21769/BioProtoc.2408 浏览次数: 8275
评审: Rebecca Van AckerAnonymous reviewer(s)
相关实验方案
基于β-葡萄糖醛酸酶 (GUS) 的细菌竞争测定以评估植物感染期间适应性的细微差异
Julien S. Luneau [...] Alice Boulanger
2022年07月05日 2221 阅读
Abstract
Zygotes of the Fucale species are a powerful model system to study cell polarization and asymmetrical cell division (Bisgrove and Kropf, 2008). The Fucale species of brown algae grow in the intertidal zone where they reproduce by releasing large female eggs and mobile sperm in the surrounding seawater. The gamete release can be induced from sexually mature fronds in the laboratory and thousands of synchronously developing zygotes are easily obtained. In contrast to other eukaryotic models, such as land plants (Brownlee and Berger, 1995), the embryo is free of maternal tissues and therefore readily amenable to pharmacological approaches. The zygotes are relatively large (up to 100 µm in diameter), facilitating manipulations and imaging studies. During the first hours of zygote development, the alignment of the axis to external cues such as light is labile and can be reversed by light gradients from different directions. A few hours before rhizoid emergence, the alignment of the axis and the polarity are fixed and the cells germinate accordingly. At this stage the zygotes are naturally attached to the substratum through the secretion of cell wall adhesive materials (Kropf et al., 1988; Hervé et al., 2016). The first cell division occurs about 24 h after fertilisation and the early embryo is composed of only two cell types that differ in size, shape and developmental fates (i.e., thallus cells and rhizoid cells) (Bouget et al., 1998). The embryo can be successfully cultivated in the laboratory for a few more days (4 weeks maximum) and has an invariant division pattern during the early stages, which allows cell lineages to be traced histologically.
Keywords: Asymmetric cell division (不对称细胞分裂)Background
This protocol provides instructions for the isolation of male and female gametes of Fucus spp. used for in vitro fertilisation and discusses how to monitor the development of the resulting zygotes and early embryos. These instructions are for the use of the dioecious species Fucus serratus, but they can be readily adapted to a monoecious Fucale species.
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文章信息
版权信息
© 2017 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Siméon, A. and Hervé, C. (2017). Isolation of Fucus serratus Gametes and Cultivation of the Zygotes. Bio-protocol 7(14): e2408. DOI: 10.21769/BioProtoc.2408.
分类
植物科学 > 植物细胞生物学 > 细胞分离
细胞生物学 > 细胞分离和培养 > 共培养
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