拟南芥静止中心细胞特异性基因表达系统的构建及功能验证
Construction and Functional Assessment of a QC Specific Gene Expression System in Arabidopsis thaliana
DOI: 10.12677/BR.2019.83024, PDF,  被引量    科研立项经费支持
作者: 陈月琴, 严欢欢, 陈玉洁, 陈丽华, 王 甜, 张汉马：重庆师范大学，植物环境适应分子生物学重庆市重点实验室，重庆
关键词: 拟南芥；静止中心；细胞特异性表达；WOX5；启动子；Arabidopsis； QC； Cell Specific Expression； WOX5； Promoter
摘要: 静止中心是高等植物根尖分生组织中一群重要细胞，在根尖生长点功能和干细胞调控中起关键作用。但由于缺少静止中心细胞特异性的基因表达工具，对该细胞群中基因功能的了解非常有限。本研究利用拟南芥WOX5基因的启动子片段和GAL4-VP16-UAS人工合成基因转录激活系统构建了一个可在拟南芥根尖静止中心细胞中特异性地驱动目的基因表达的转基因系统。通过绿色荧光蛋白基因证实了该表达系统的静止中心细胞特异性，并用两个已知根干细胞调控基因WOX5和FEZ检测了该表达系统用于研究静止中心基因功能的可行性及功效。研究结果表明所构建的表达系统具静止中心细胞特异性，可用来研究不同调控基因在该细胞群中的功能。
Abstract: Quiescent center (QC) is an important group of cells in root meristems of higher plants. It plays a key role in the regulation of root meristem function and stem cells. However, due to a lack of cell-specific gene expression tool, there is very limited understanding about gene functions within this group of cells. In this study, we constructed a QC-specific gene expression system using the promoter of Arabidopsis WOX5 gene and the synthetic GAL4-VP16-UAS transcriptional activation system. We confirmed the QC-specificity of this system using GFP as the reporter gene, and also tested its utility and effectiveness for studying gene functions using two known root stem cell regulatory genes, WOX5 and FEZ. Our results demonstrate that the expression system is QC-specific and can be used to study the functions of different genes in QC cells.
文章引用：陈月琴, 严欢欢, 陈玉洁, 陈丽华, 王甜, 张汉马. 拟南芥静止中心细胞特异性基因表达系统的构建及功能验证[J]. 植物学研究, 2019, 8(3): 181-189. https://doi.org/10.12677/BR.2019.83024

参考文献

[1] Dolan, L., Janmaat, K., Willemsen, V., Linstead, P., Poethig, S., Roberts, K. and Scheres, B. (1993) Cellular Organisation of the Arabidopsis thaliana Root. Development, 119, 71-84.
[2] van den Berg, C., Willemsen, V., Hendriks, G., Weisbeek, P. and Scheres, B. (1997) Short-Range Control of Cell Differentiation in the Arabidopsis Root Meristem. Nature, 390, 287-289. [Google Scholar] [CrossRef] [PubMed]
[3] Xu, J., Hofhuis, H., Heidstra, R., Sauer, M., Friml, J. and Scheres, B. (2006) A Molecular Framework for Plant Regeneration. Science, 311, 385-388. [Google Scholar] [CrossRef] [PubMed]
[4] Sarkar, A.K., Luijten, M., Miyashima, S., Lenhard, M., Hashimoto, T., Nakajima. K., Scheres, B., Heidstra, R. and Laux, T. (2007) Conserved Factors Regulate Signalling in Arabidopsis thaliana Shoot and Root Stem Cell Organizers. Nature, 446, 811-814. [Google Scholar] [CrossRef] [PubMed]
[5] Helariutta, Y., Fukaki, H., Wysocka-Diller, J., Nakajima, K., Jung, J., Sena, G., Hauser, M.T. and Benfey, P.N. (2007) The SHORT-ROOT Gene Controls Radial Patterning of the Arabidopsis Root through Radial Signaling. Cell, 101, 555-567. [Google Scholar] [CrossRef] 
[6] Di Laurenzio, L., Wysocka-Diller, J., Malamy, J.E., Pysh, L., Helariutta, Y., Freshour, G., Hahn, M.G., Feldmann, K.A. and Benfey, P.N. (1996) The SCARECROW Gene Regulates an Asymmetric Cell Division that Is Essential for Generating the Radial Organization of the Arabidopsis Root. Cell, 86, 423-433. [Google Scholar] [CrossRef] 
[7] Galinha, C., Hofhuis, H., Luijten, M., Willemsen, V., Blilou, I., Heidstra, R. and Scheres, B. (2007) PLETHORA Proteins as Dosedependent Master Regulators of Arabidopsis Root Development. Nature, 449, 1053-1057. [Google Scholar] [CrossRef] [PubMed]
[8] Willemsen, V., Bauch, M., Bennett, T., Campilho, A., Wolkenfelt, H., Xu, J., Haseloff, J. and Scheres, B. (2008) The NAC Domain Transcription Factors FEZ and SOMBRERO Control the Orientation of Cell Division Plane in Arabidopsis Root Stem Cells. Development Cell, 15, 913-922. [Google Scholar] [CrossRef] [PubMed]
[9] Wildwater, M., Campilho, A., Perez-Perez, J.M., Heidstra, R., Blilou, I., Korthout, H., Chatterjee, J., Mariconti, L., Gruissem, W. and Scheres, B. (2005) The RETINOBLASTOMA-RELATED Gene Regulates Stem Cell Maintenance in Arabidopsis Roots. Cell, 123, 1337-1349. [Google Scholar] [CrossRef] [PubMed]
[10] De Smet, I., Vassileva, V., De Rybel, B., Levesque, M.P., Grunewald, W., Van Damme, D., Van Noorden, G., Naudts, M., Van Isterdael, G., De Clercq, R., Wang, J.Y., Meuli, N., Vanneste, S., Friml, J., Hilson, P., Jürgens, G., Ingram, G.C., Inzé, D., Benfey, P.N. and Beeckman, T. (2008) Receptor Like Kinase ACR4 Restricts Formative Cell Divisions in the Arabidopsis Root. Science, 322, 594-597. [Google Scholar] [CrossRef] [PubMed]
[11] Clough, S.J. and Bent, A.F. (1998). Floral Dip: A Simplified Method for Agrobacterium-Mediated Transformation of Arabidopsis thaliana. The Plant Journal, 16, 735-743.[CrossRef] [PubMed]
[12] Pi, L., Aichinger, E., van der Graaff, E., Llavata-Peris, C.I., Weijers, D., Hennig, L., Groot, E. and Laux, T. (2015) Organizer-Derived WOX5 Signal Maintains Root Columella Stem Cells through Chromatin-Mediated Repression of CDF4 Expression. Development Cell, 33, 576-588. [Google Scholar] [CrossRef] [PubMed]