An in vitro grafting method to quantify mechanical forces of adhering tissues
An in vitro grafting method to quantify mechanical forces of adhering tissues
复制标题
DOI：
10.5511/plantbiotechnology.20.0925a
复制
发表时间：
2020-12-01
影响因子：
1.6
通讯作者：
Notaguchi, Michitaka
中科院分区：
文献类型：
--
作者：
Kawakatsu, Yaichi;Sawai, Yu;Notaguchi, Michitaka
嫁接是繁殖有用树种并同时获得两个品种/物种（砧木和接穗）有益性状的不可缺少的农业技术。最近对嫁接过程中分子事件的研究揭示了动态的生理和转录组变化。需要专注于特定移植步骤的策略，以进一步将每个生理和分子事件与这些步骤关联起来。在这项研究中，我们通过改进人工体外嫁接系统，开发了一种研究组织粘附事件（早期嫁接步骤）的方法，该系统将两片 1.5 毫米厚的本塞姆氏烟草切茎切片组合并在培养基上培养。我们准备了一块含有五个特殊切口的硅胶片，用于粘附切好的茎片。我们使用测力计定量测量了这些接枝界面的粘合力，发现接枝物在接枝后2天开始粘合，随着时间的推移粘合力逐渐增加。在通过该方法确认生长素对移植的积极作用后，我们测试了纤维素酶处理的效果，并观察到移植组织粘附的显着增强。与单独添加生长素或纤维素酶相比，同时添加生长素和纤维素酶时的粘附力更强。因此，本研究开发的体外移植方法可用于检查移植物粘附过程。
Grafting is an indispensable agricultural technology for propagating useful tree varieties and obtaining beneficial traits of two varieties/species-as stock and scion-at the same time. Recent studies of molecular events during grafting have revealed dynamic physiological and transcriptomic changes. Strategies focused on specific grafting steps are needed to further associate each physiological and molecular event with those steps. In this study, we developed a method to investigate the tissue adhesion event, an early grafting step, by improving an artificial in vitro grafting system in which two pieces of 1.5-mm thick Nicotiana benthamiana cut stem sections were combined and cultured on medium. We prepared a silicone sheet containing five special cutouts for adhesion of cut stem slices. We quantitatively measured the adhesive force at these grafting interfaces using a force gauge and found that graft adhesion started 2 days after grafting, with the adhesive force gradually increasing over time. After confirming the positive effect of auxin on grafting by this method, we tested the effect of cellulase treatment and observed significant enhancement of graft tissue adhesion. Compared with the addition of auxin or cellulase individually, the adhesive force was stronger when both auxin and cellulase were added simultaneously. The in vitro grafting method developed in this study is thus useful for examining the process of graft adhesion.