OsEXPA8基因对水稻悬浮细胞的周期和大小的影响

发布时间：2018-01-16 16:18

本文关键词：OsEXPA8基因对水稻悬浮细胞的周期和大小的影响　出处：《农业生物技术学报》2016年01期 　论文类型：期刊论文

【摘要】：膨胀素是植物细胞特有的一种细胞壁松弛蛋白,在很大程度上决定着植物细胞的生长速率。α-膨胀素(α-expansin,EXPA)和β-膨胀素(β-expansin,EXPB)是植物膨胀素家族中两个较大的亚家族。水稻(Oryza sativa)中至少有34个α-膨胀素基因。本研究通过转基因技术,研究了一个水稻根特异性表达的α-膨胀素基因Os EXPA8对水稻悬浮细胞的细胞周期和细胞大小的影响。利用流式细胞仪,分析不同培养时期野生型和转基因型细胞周期的差异,结果发现,35S::Os EXPA8(过表达)转基因悬浮细胞大部分处于G2/M期,而Os EXPA8-RNAi(knock-down)转基因悬浮细胞多处在G1期;利用血球计数板测定细胞数量,结果表明,35S::Os EXPA8转基因细胞数量野生型细胞数量Os EXPA8-RNAi转基因细胞数量;利用显微镜测定不同培养时期的细胞大小,发现35S::Os EXPA8转基因细胞大小野生型细胞大小Os EXPA8-RNAi转基因细胞大小;测定愈伤组织块大小,发现35S::Os EXPA8转基因愈伤组织块大小野生型愈伤组织块大小Os EXPA8-RNAi转基因愈伤组织块大小。本研究结果表明,Os EXPA8能够促进水稻悬浮细胞的分裂和生长,从而增加细胞数量,增大细胞体积,对水稻悬浮细胞的生长具有调控作用。该研究将有助于更好地了解植物细胞壁的松弛过程,从而深入地认识植物的生长发育进程。同时,膨胀素作为一种细胞壁松弛因子在农业上具有潜在的生物工程应用价值。
[Abstract]:Expansin is a cell wall loosening proteins in plant cell specific, largely determines the growth rate of plant cells. Alpha expansin (alpha -expansin, EXPA) and beta expansin (beta -expansin, EXPB) is a plant expansion in two large family of rice (Oryza sativa subfamily.) at least 34 alpha expansin genes. This study by transgenic technology, studies on an expression of rice root specific alpha expansion effect of Os gene on cell cycle of EXPA8 in rice suspension cell and cell size. Using flow cytometry, analysis of differences in different culture period of wild type and transgenic type the cell cycle was found: 35S: Os EXPA8 (expression) of most transgenic suspension cells in G2/M phase, and Os EXPA8-RNAi (knock-down) transgenic suspension cells in G1 phase; Determination of cell number using hemocytometer. The results showed that: 35S: Os EXPA The number of cell number 8 transgenic wild type cell number of Os transgenic EXPA8-RNAi cells; Determination of different incubation times with microscope cell size, 35S:: Os EXPA8 transgenic cell size of wild type cell size Os EXPA8-RNAi transgenic cell size; Determination of callus size, block size: 35S: block found in wild type callus Os EXPA8 transgenic callus block size Os EXPA8-RNAi transgenic callus block size. The results of this study show that Os EXPA8 can promote the proliferation and growth of rice suspension cells, thereby increasing the number of cells, increase in cell volume, has effects on rice suspension cell growth. The study will contribute to a better understanding of the relaxation process of plant cell wall, thus in-depth understanding the process of growth and development of plants. At the same time, expansion as a cell wall relaxation factor is a potential student in agriculture The application value of the material engineering.

【作者单位】：重庆大学生物工程学院;
【基金】：国家自然科学基金项目(No.31271685) 重庆市自然科学基金项目(No.cstc2014jcyja80017)
【分类号】：S511
【正文快照】： 植物生长过程中,位于细胞壁上的膨胀素使细胞壁膨胀,促进细胞伸长(Mcqueen-Mason et al.,1992)。膨胀素最初在黄瓜(Cucumis sativus)下胚轴细胞的胞壁中发现(Cosgrove et al.,1997;Lee et al.,2001)。随后,膨胀素基因也相继在水稻、拟南芥(Arabidopsis thaliana)、大豆(Glycin

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