Isoda Minako, Oyama Tokitaka
Department of Botany, Graduate School of Science, Kyoto University, Kitashirakawa-oiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan.
Plant Biotechnol (Tokyo). 2018 Dec 25;35(4):387-391. doi: 10.5511/plantbiotechnology.18.0721a.
We developed a new model system to analyze physiological behavior at the single-cell level in whole plants. is a species of rootless duckweed, which has a thin and very small structure and can grow rapidly on the surface of culture medium. Epidermal and mesophyll cells were transfected with a reporter gene using particle bombardment and were observed at the single-cell level in the whole living plant. An EM-CCD camera system with a macro zoom microscope was used to capture time-lapse images of bioluminescence, and we successfully detected circadian rhythms in individual cells that expressed a luciferase gene under the control of a circadian promoter. We also detected individual -phase cells in meristematic tissues of intact plants by using a 5-ethynyl-2'-deoxyuridine (EdU)-labeling assay. Our observations indicated that low-molecular-weight compounds could access the inside of the plant body. Thus, showed the experimental characteristics suitable for single-cell analyses that could be combined with whole-plant observations and/or pharmacological analyses/chemical biology.
我们开发了一种新的模型系统,用于分析全株植物单细胞水平的生理行为。无根浮萍是一种无根浮萍属植物,其结构薄且非常小,能在培养基表面快速生长。利用粒子轰击将报告基因转染到表皮细胞和叶肉细胞中,并在全株活体植物的单细胞水平上进行观察。使用配备宏观变焦显微镜的EM-CCD相机系统来捕获生物发光的延时图像,我们成功地在受昼夜节律启动子控制下表达荧光素酶基因的单个细胞中检测到了昼夜节律。我们还通过使用5-乙炔基-2'-脱氧尿苷(EdU)标记测定法,在完整无根浮萍植物的分生组织中检测到了单个G1期细胞。我们的观察结果表明,低分子量化合物可以进入植物体内。因此,无根浮萍显示出适合单细胞分析的实验特性,可与全株观察和/或药理分析/化学生物学相结合。