Establishing of 3D-FISH on frozen section and its applying in chromosome territories analysis in Populus trichocarpa.

IF 5.3 2区 生物学 Q1 PLANT SCIENCES
Yihang Ning, Daxin Shang, Haoyang Xin, Runxin Ni, Ziyue Wang, Yan Zhen, Guangxin Liu, Mengli Xi
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引用次数: 0

Abstract

Key message: Fluorescence in situ hybridization with frozen sections of root tips showed difference of chromosome territories distribution between autosome and sex-chromosome homologous pairs in Populus trichocarpa. The spatial organization of chromatin within the interphase nucleus and the interactions between chromosome territories (CTs) are essential for various biologic processes. Three-dimensional fluorescence in situ hybridization (3D-FISH) is a powerful tool for analyzing CTs, but its application in plants is limited. In this study, we established a 3D-FISH technique using frozen sections of Populus trichocarpa root tips, which was an improvement over the use of paraffin sections and enabled us to acquire good FISH signals. Using chromosome-specific oligo probes, we were able to analyze CTs in interphase nuclei in three dimensions. The distribution of chromosome pairs 17 and 19 in the 3D-preserved nuclei of P. trichocarpa root tip cells were analyzed and showed that the autosome pair 17 associated more often than sex chromosome 19. This research lays a foundation for further study of the spatial position of chromosomes in the nucleus and the relationship between gene expression and spatial localization of chromosomes in poplar.

在冰冻切片上建立 3D-FISH 并将其应用于杨树染色体区系分析。
关键信息根尖冷冻切片荧光原位杂交显示,杨树自体和性染色体同源对的染色体区域分布存在差异。染色质在细胞核间期的空间组织以及染色体区域(CT)之间的相互作用对各种生物过程至关重要。三维荧光原位杂交(3D-FISH)是分析CT的有力工具,但其在植物中的应用还很有限。在这项研究中,我们利用杨树根尖的冷冻切片建立了一种三维荧光原位杂交技术,该技术改进了石蜡切片的使用,使我们能够获得良好的荧光原位杂交信号。利用染色体特异性寡核苷酸探针,我们能够对间期细胞核中的 CT 进行三维分析。我们分析了17号和19号染色体对在三维保存的三叶草根尖细胞核中的分布情况,结果表明17号染色体对比19号染色体对更常见。这项研究为进一步研究杨树细胞核中染色体的空间位置以及基因表达与染色体空间定位之间的关系奠定了基础。
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来源期刊
Plant Cell Reports
Plant Cell Reports 生物-植物科学
CiteScore
10.80
自引率
1.60%
发文量
135
审稿时长
3.2 months
期刊介绍: Plant Cell Reports publishes original, peer-reviewed articles on new advances in all aspects of plant cell science, plant genetics and molecular biology. Papers selected for publication contribute significant new advances to clearly identified technological problems and/or biological questions. The articles will prove relevant beyond the narrow topic of interest to a readership with broad scientific background. The coverage includes such topics as: - genomics and genetics - metabolism - cell biology - abiotic and biotic stress - phytopathology - gene transfer and expression - molecular pharming - systems biology - nanobiotechnology - genome editing - phenomics and synthetic biology The journal also publishes opinion papers, review and focus articles on the latest developments and new advances in research and technology in plant molecular biology and biotechnology.
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