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Genome-wide functional analysis in Arabidopsis with the mechanical properties as a measure

以机械特性为衡量标准的拟南芥全基因组功能分析

基本信息

批准号：
17510159
负责人：
HOSON Takayuki
金额：
$ 2.43万
依托单位：
Osaka City University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17510159/
关键词：
Gene Genome Plant Arabidoosis Mechanical orooerties

项目摘要

To identify the function of every gene in Arabidopsis, it is required to develop the systematic genome-wide screening method with a suitable phenotype as a measure. For this purpose, we established the efficient procedures and conditions for analyzing the mechanical properties of Arabidopsis hypocotyls. To accurately measure the mechanical properties of tiny hypocotyls, we introduced a new tensile tester and developed feasible software and interface. The newly developed rheological method, in which the load-strain analysis and the stress-relaxation analysis are successively applied to a common sample, enabled the efficient analysis of the mechanical properties of hypocotyls. We also determined cultivation conditions of Arabidopsis seedlings suitable for effective sample preparation.We applied the method established in such a way to the analysis of the mechanical properties of Arabidopsis mutants defective in formation, metabolism, or function of the cell wall, the plasma membrane, and cortical microtubules. Out of various mutants examined, the mechanical properties of the knockout and over-expressed mutants in certain xyloglucan endotransglucosylase/hydrolase (XTH) genes were shown to be modified, the degree of which matched with the degrees of modifications in growth phenotype or xyloglucan metabolism. Moreover, α- and β-tubulin, MAPs (mor1, spr1, spr2, MAP65, katanin) and hmg mutants showed disordered growth pattern, such as dwarfism and helical growth, and also the corresponding changes in the mechanical properties. These results indicate the effectiveness of the method established in the present study for genome-wide functional analysis in Arabidopsis.

为了明确拟南芥中每个基因的功能，需要建立一种以合适的表型为指标的系统的全基因组筛选方法。为此，我们建立了有效的程序和条件，分析拟南芥下胚轴的力学特性。为了准确测量微小下胚轴的力学性能，研制了一种新型的拉伸试验机，并开发了可行的软件和界面。新开发的流变学方法，其中的载荷-应变分析和应力松弛分析连续施加到一个共同的样品，使下胚轴的力学性能的有效分析。此外，还确定了适合于有效制样的拟南芥幼苗培养条件，并将该方法应用于拟南芥细胞壁、质膜、皮层微管的形成、代谢或功能缺陷突变体的力学特性分析。在所检查的各种突变体中，某些木葡聚糖内切转葡糖基酶/水解酶（XTH）基因中的敲除和过表达突变体的机械性质显示出被修饰，其程度与生长表型或木葡聚糖代谢中的修饰程度相匹配。此外，α-和β-微管蛋白、MAPs（mor 1、spr 1、spr 2、MAP 65、katanin）和hmg突变体的生长模式也发生了紊乱，如矮化和螺旋生长，力学性能也发生了相应的变化。这些结果表明，在本研究中建立的方法在拟南芥全基因组功能分析的有效性。