Name: a labor-saving and repeatable touch-force signaling mutant screen protocol for the study of thigmomorphogenesis of a model plant arabidopsis thaliana
Uploaded: 2019-08-06T13:00:00
Description: Watch this Scientific Journal Video about A Labor-saving and Repeatable Touch-force Signaling Mutant Screen Protocol for the Study of Thigmomorphogenesis of a Model Plant Arabidopsis thaliana at JoVE.

这项研究得到以下拨款的支持:31370315、31570187、31870231(中国国家科学基金会)、16100318、661613、16101114、16103615、16103817、AoE/M-403/16(香港RGC)。作者感谢巨峰精密自动化技术有限公司(中国深圳)提供图1所示的几张原理图。
作者们还要感谢张国宝和李华华为触摸力装载机的发展所做出的贡献。

1. 种子制备
注:野生型(Col-0)以及mkk1和mkk2所用功能丧失突变体的种子从阿拉伯生物多样性生物资源中心(ABRC、https://www.arabidopsis.org、哥伦布、OH)购买。
<ol><li>计算将用每个基因型的植物个体数量进行可靠的统计分析。根据每行的发芽率准备足够数量的种子,通常比实验所需的种子多4-5倍。确保有足够的健康且均匀大小的植物可用于触摸响应测定。根据该协?...

<ol>
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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+unstable+transcripts+in+Arabidopsis+by+cDNA+microarray+analysis:+rapid+decay+is+associated+with+a+group+of+touch-and+specific+clock-controlled+genes.">Identification of unstable transcripts in Arabidopsis by cDNA microarray analysis: rapid decay is associated with a group of touch-and specific clock-controlled genes.</a> Proceedings of the National Academy of Sciences of the United States of America. 99, 11513-11518 (2002).</li><li>Lee, D., Polisensky, D. H., Braam, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genome-wide+identification+of+touch-and+darkness-regulated+Arabidopsis+genes:+a+focus+on+calmodulin-like+and+XTH+genes.">Genome-wide identification of touch-and darkness-regulated Arabidopsis genes: a focus on calmodulin-like and XTH genes.</a> New Phytologist. 165, 429-444 (2005).</li><li>Wang, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+and+functional+posttranslational+modification+proteomics+reveals+that+TREPH1+plays+a+role+in+plant+touch-delayed+bolting.">Quantitative and functional posttranslational modification proteomics reveals that TREPH1 plays a role in plant touch-delayed bolting.</a> Proceedings of the National Academy of Sciences United States of America. 115, 10265-10274 (2018).</li><li>Hamilton, E. S., Schlegel, A. M., Haswell, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=United+in+diversity:+mechanosensitive+ion+channels+in+plants.">United in diversity: mechanosensitive ion channels in plants.</a> Annual Review of Plant Biology. 66, 113-137 (2015).</li><li>Knight, M. R., Campbell, A. K., Smith, S. M., Trewavas, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenic+plant+aequorin+reports+the+effects+of+touch+and+cold-shock+and+elicitors+on+cytoplasmic+calcium.">Transgenic plant aequorin reports the effects of touch and cold-shock and elicitors on cytoplasmic calcium.</a> Nature. 352, 524 (1991).</li><li>Toyota, M., Furuichi, T., Tatsumi, H., Sokabe, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytoplasmic+calcium+increases+in+response+to+changes+in+the+gravity+vector+in+hypocotyls+and+petioles+of+Arabidopsis+seedlings.">Cytoplasmic calcium increases in response to changes in the gravity vector in hypocotyls and petioles of Arabidopsis seedlings.</a> Plant Physiology. 146, 505-514 (2008).</li><li>Knight, M. R., Smith, S. M., Trewavas, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wind-induced+plant+motion+immediately+increases+cytosolic+calcium.">Wind-induced plant motion immediately increases cytosolic calcium.</a> Proceedings of the National Academy of Sciences of the United States of America. 89, 4967-4971 (1992).</li><li>Braam, J., Davis, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rain-,+wind-,+and+touch-induced+expression+of+calmodulin+and+calmodulin-related+genes+in+Arabidopsis.">Rain-, wind-, and touch-induced expression of calmodulin and calmodulin-related genes in Arabidopsis.</a> Cell. 60, 357-364 (1990).</li><li>Chehab, E. W., Yao, C., Henderson, Z., Kim, S., Braam, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arabidopsis+touch-induced+morphogenesis+is+jasmonate+mediated+and+protects+against+pests.">Arabidopsis touch-induced morphogenesis is jasmonate mediated and protects against pests.</a> Current Biology. 22, 701-706 (2012).</li><li>Telewski, F. W., Pruyn, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thigmomorphogenesis:+a+dose+response+to+flexing+in+Ulmus+americana+seedlings.">Thigmomorphogenesis: a dose response to flexing in Ulmus americana seedlings.</a> Tree Physiology. 18, 65-68 (1998).</li><li>De Vylder, J., Vandenbussche, F. J., Hu, Y., Philips, W., Van Der Straeten, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rosette+tracker:+an+open+source+image+analysis+tool+for+automatic+quantification+of+genotype+effects.">Rosette tracker: an open source image analysis tool for automatic quantification of genotype effects.</a> Plant Physiology. , (2012).</li><li>Clark, T., Bradburn, M., Love, S., Altman, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Survival+analysis+part+I:+basic+concepts+and+first+analyses.">Survival analysis part I: basic concepts and first analyses.</a> British Journal of Cancer. 89, 232 (2003).</li><li>Bradburn, M. J., Clark, T. G., Love, S., Altman, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Survival+analysis+part+II:+multivariate+data+analysis&#8211;an+introduction+to+concepts+and+methods.">Survival analysis part II: multivariate data analysis&#8211;an introduction to concepts and methods.</a> British Journal of Cancer. 89, 431 (2003).</li><li>Jaffe, M., Forbes, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thigmomorphogenesis:+the+effect+of+mechanical+perturbation+on+plants.">Thigmomorphogenesis: the effect of mechanical perturbation on plants.</a> Plant Growth Regulation. 12, 313-324 (1993).</li><li>Kutschera, U., Niklas, K. 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三毛磷发生是一种复杂的植物生长反应对机械扰动,这涉及到细胞信号和植物激素的作用网络。它是植物在不良环境条件下生存的适应性进化的结果25、26。机械触摸,特别是人的手指触摸和手持棉签触摸,已被选定来研究这种形态变化在以前的三元遗传学研究14,20。这种简化的触摸力加载版本可触发工厂触摸响应...

植物异质性是1973年由Jaffe,MJ所创建的术语。它是一种植物对流,但不同于众所周知的光动或重力引起的阳光或重力刺激2,3。它描述了与周期性机械刺激相关的表型变化,在早期4,5中,植物学家经常观察到这些改变。雨滴、风、植物、动物和人类接触,甚至动物叮咬,都被认为是不同类型的梅查诺刺激,触发植物4,5的力信号。植物异质性的特征包括螺栓的延迟、较短的茎、草本植物中较小的玫瑰花/叶大小,以及木本植物6、7、8的较厚茎。这与在米莫萨植物或其他对美茶敏感的藤蔓中经常发现的异体或地皮反应不同,在这些快速接触反应中更容易观察到1、9、10。另一方面,由于生长反应缓慢,因此相对难以观察到三角性发生。在连续的力载刺激后,通常观察三重磷发生。植物触摸反应的这种独特特性使得使用人手触摸刺激来分离抗触摸信号突变体的正向遗传屏幕变得困难。
为了阐明强力信号转导途径和分子机制,在过去6年进行了分子和细胞生物学实验。12,13,14.这些研究提出,植物力信号受体主要由机械敏感电通道(MSC)和系绳MSC复合物组成,由膜跨度蛋白11、14的多体复合体组成。,15.细胞质Ca2+瞬态尖峰在初始接触后几秒钟内生成。风、雨或重力刺激可能与下游钙传感器相互作用,将力信号转用于核事件14、16、17、18。除了分子和细胞研究外,用手动手指触摸植物的正向遗传屏幕发现,植物激素和继发代谢物与随后的接触诱导(TCH)基因表达有关。触摸力加载13,19 。例如,到目前为止,从基因研究中已经发现了aos和opr320突变体。然而,在血性发生研究中应用前向遗传学的主要问题仍然是定量触摸反应水平和接触大量基因突变所需的大量劳动。个别植物。耗时的问题也仍然存在于手触摸为基础的突变屏幕14,20。例如,要完成一轮触摸力刺激,一个人需要触摸单个植物30-60次(每秒一次)。为了有足够的植物进行统计表型分析,触摸力加载过程通常需要20-50个相同基因型的单个植物。这种触摸力加载机制意味着一个人需要重复执行600-3,000次触摸选择一个基因型。这种类型的触摸通常需要每天重复3至5轮,这相当于大约1,800-15,000指或棉拭子触摸每天每个基因型的植物。训练有素的人通常需要在一天的多轮重复中,在理想的范围内保持多重接触的强度和力量,以避免力量和力量的很大变化。众所周知,血性发生是一个可饱和和剂量依赖的过程6,21,触摸力/强度成为成功触发植物触摸反应的关键。
为了消除与人相关的触摸力载荷,并将机械应用保持在可接受的误差范围14内,我们设计了一个自动触摸力装载机,以取代手工操作的触摸。机器有4个移动臂,每个都配有一个人发刷。此版本名为模型 K1,以指定其人发触摸力加载功能。如果在一台机器下定量测量4个基因型的血性发生或触摸反应,可以测量每个基因型40-48个个体。每轮触摸重复(每株触摸不到 60 次)使用移动速度可调机械臂持续不到 5 分钟。因此,K1 型触摸机上的工厂可以机械地刺激一天多轮,或者具有恒定的触摸力负载或最初编程的不同强度级别。
阿拉伯植物有机体模型,因此被选为测试全自动头发触摸力装载机应用的目标植物物种。由于有几个大型种子库可用于检索突变体的各种种质和开花的大小,阿拉伯兰多普西斯非常适合安装在使用型号 K1 触摸机的生长架上可用的空间。
K1 型自动触摸机由三个主要部件组成:(1) 由两个皮带驱动的线性执行器组成的 H 形金属机架,(2) 配备发刷的机械金属臂,以及 (3) 控制器。对于定制的 K1 型触摸机,每个 X/Y 轴模块由一个皮带驱动导轨、两个滑动块(红色)和一个 57 步进电机(预安装和拆卸)组成(图 1A,B)。上部水平执行器允许机械臂左右水平移动,下部垂直皮带驱动的线性执行器允许机械臂垂直上下移动(图 1B,图 2A).垂直执行器上安装了四个可拆卸的机械臂(图1C,图2B)。四个人类发刷被绑定到四个机械臂上,分别(图1C,图2B)。图1C中标有下面用粗体字构建K1型触摸机的所有机械部件(另见材料表)。

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			<td height="21" style="height:21px;width:375px;">4 hair brushes</td>
			<td style="width:393px;"></td>
			<td style="width:149px;">customized</td>
			<td style="width:224px;"></td>
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			<td height="21" style="height:21px;width:375px;">4 robot arms with one holder</td>
			<td style="width:393px;"></td>
			<td style="width:149px;">customized</td>
			<td>1000 mm length holder and 560 mm length robot arm</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:375px;">57 stepper motor</td>
			<td style="width:393px;"></td>
			<td style="width:149px;">57HS22-A</td>
			<td></td>
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			<td height="21" style="height:21px;width:375px;">All purpose potting soil</td>
			<td style="width:393px;">Plantmate, Hong Kong</td>
			<td style="width:149px;"></td>
			<td></td>
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			<td height="21" style="height:21px;width:375px;">Arabidopsis plant seeds</td>
			<td style="width:393px;">Arabidopsis Biological Resource Centers, Columbus, OH</td>
			<td style="width:149px;"></td>
			<td>For arabidopsis seed purchase</td>
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			<td height="21" style="height:21px;width:375px;">BIO-MIX potting substratum</td>
			<td style="width:393px;">Jiffy Products International BV, the Netherlands</td>
			<td style="width:149px;">1000682050</td>
			<td>Two soils were mixed together to grow Arabidopsis. The ratio of All purpos potting soil and&#160; BIO-MIX is 1:2</td>
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			<td height="21" style="height:21px;width:375px;">IL 1700 research radiometer</td>
			<td style="width:393px;">International Light, Newburyport, MA</td>
			<td style="width:149px;"></td>
			<td>The light intensity of both full-wavelength and photosynthetic active radiation can be measured.</td>
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			<td height="21" style="height:21px;width:375px;">ImageJ</td>
			<td style="width:393px;">https://imagej.nih.gov/ij/download.html</td>
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			<td>Free downloaded software</td>
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			<td height="21" style="height:21px;width:375px;">Ju Feng Precision and Automation Technology Limited</td>
			<td style="width:393px;">Shenzhen, China</td>
			<td style="width:149px;"></td>
			<td>For belt-driven linear actuators and other mechanical modules purchase</td>
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			<td height="21" style="height:21px;width:375px;">Junction plate of the slide block</td>
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			<td>To fix the Y guide-rail module or Y auxiliary girder onto backs of slide blocks</td>
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			<td height="21" style="height:21px;width:375px;">Junction plate of the X axis module</td>
			<td style="width:393px;"></td>
			<td style="width:149px;">customized</td>
			<td>To connect the X guide-rail module and X auxiliary girder</td>
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			<td height="21" style="height:21px;width:375px;">Slide block</td>
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			<td height="21" style="height:21px;width:375px;">WDT4045 X axis guide-rail module</td>
			<td style="width:393px;"></td>
			<td style="width:149px;">843 mm, customized</td>
			<td>Pre-installed with two slide blocks and one 57 stepper motor</td>
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			<td height="42" style="height:42px;width:375px;">WDT4045 Y axis guide-rail module</td>
			<td style="width:393px;"></td>
			<td style="width:149px;">1038 mm, customized</td>
			<td>Pre-installed with two slide blocks and one 57 stepper motor</td>
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			<td height="21" style="height:21px;width:375px;">X axis auxiliary girder</td>
			<td style="width:393px;"></td>
			<td style="width:149px;">843 mm, customized</td>
			<td>Pre-installed with two slide blocks</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:375px;">Y axis auxiliary girder</td>
			<td style="width:393px;"></td>
			<td style="width:149px;">1038 mm, customized</td>
			<td>Pre-installed with two slide blocks</td>
		</tr>
	</tbody>
</table>

a labor-saving and repeatable touch-force signaling mutant screen protocol for the study of thigmomorphogenesis of a model plant arabidopsis thaliana

植物对细胞内和细胞外机械刺激(或力信号)作出反应,并产生特殊的形态变化,称为血性发生。在过去的几十年中,已经发现并报告了一些信号成分,这些成分参与了干扰(例如,钙离子结合蛋白和jasmonic酸生物合成酶)。然而,在力信号或异位磷发生研究中研究相对缓慢,主要归因于两个原因:需要人工操纵的触摸感应的异质性发生和力强度误差与人们的手触摸相关。为了提高植物有机体上外力装载的效率,建立了自动触摸力装载机。这款机械臂驱动式毛刷触感可提供省力且易于重复的触摸力模拟、无限轮的触摸重复和可调节的触摸强度。这种头发触摸力装载机可用于触摸力信号突变体的大规模筛选和植物造影的鸡学研究。此外,触摸材料,如人的头发,可以替代其他天然材料,如动物的头发,丝线和棉纤维。机器上的自动移动臂可配备喷水喷嘴和空气鼓风机,分别模拟雨滴和风的自然力。通过结合手工棉签触摸,结合这种自动发触摸力装载机,我们研究了两种力信号突变体MAP KINASE KINASE 1 (MKK1) 和 MKK2 工厂的触摸响应.对触摸力加载的野生型植物和两种突变体的现象进行了统计评价。他们在触摸响应方面表现出显著差异。

自动发触摸力装载机 为了观察植物的形态变化,可重复的生长条件和治疗方法都是获得可重复结果的关键。这种高通量和自动触摸力信号突变筛选是由新建成的头发触摸力加载机,型号K1(图1,图2)实现的。这些发刷可以同时触摸最多 4 个托盘的植物。在托盘中放置了24个杯子,在一组中放置了12个植物,作为控制和处理过的植物(...

Watch this Scientific Journal Video about A Labor-saving and Repeatable Touch-force Signaling Mutant Screen Protocol for the Study of Thigmomorphogenesis of a Model Plant Arabidopsis thaliana at JoVE.

A Labor-saving and Repeatable Touch-force Signaling Mutant Screen Protocol for the Study of Thigmomorphogenesis of a Model Plant Arabidopsis thaliana

一种省力和可重复的触摸力信号突变屏幕协议,用于研究阿拉伯植物示范植物的三角性发生

一个温和的触摸力装载机是由人的头发刷,机械臂和控制器。发刷由安装在机器上的机械臂驱动,并定期移动,对植物施加触摸力。机器驱动的头发触摸强度可与手动应用的触摸强度相媲美。

Plants responding to both intracellular and extracellular mechanical stimulations (or force signals) and develop special morphological changes, a called ...

我们制造一台自动触摸力机来观察植物信号形态。我安装的机械臂与人的头发刷，以应用温柔的触摸力植物。与人手指触摸相比，自动人发触摸机可以提供更多的人力节省，均匀的触摸力加载。一轮强制装载所需时间可以在几分钟内限制。与几个小时的手指触摸和棉签触摸相比，机器人机器可以在很大程度上改善植物商业反应研究。通过提高处理植物的效率和数量，我们预计自动触摸机将应用于四个信号突变屏幕和植物和动物的商反应研究。首先，在用MS培养基在8%的阿加盘上培育种子后，测试每种基因型的发芽率7天。根据发芽率，准备种子比所需的植物数量多四到五倍。将种子浸入冷水中，在1.5毫升塑料管中，用铝箔覆盖，以保持在黑暗中。储存在摄氏四度的种子。在形成后五到七天，选择适合植物生长的土壤。避免大团块，并使用叉子将它们均匀地混合。准备 24 个 207 毫升塑料杯，上缘直径为 7.4 厘米。在杯子底部钻三个圆孔，用于灌溉。用混合土壤填充这些塑料杯。让土壤堆积到比杯边高一厘米左右，用尺子轻轻地压平堆积的土壤表面。将 24 个杯子转移到塑料托盘中。将生长室的光照强度设置为每平方米180至240微爱因斯坦/秒，高于光合作用活性辐射，使植物快速生长。将光条件设置为 24 小时常数。将生长室的温度设置为 23.5 正负 1.5 摄氏度，湿度在 35 至 45%之间，将 2.5 升水加入每个托盘。让土壤从杯底的孔中吸收水分，等待土壤表面下降到杯边缘水平。两小时后，将三到四个种子播种到一个点上，在一个杯子里播下四个均匀分布的点。在每个托盘上方放置一个透明的塑料盖，让种子发芽一周。然后，取下盖子，让幼苗再生长一个星期。种子发芽后每隔一天用1.5升水灌溉植物。种子播种后9至10天，通过变薄去除额外的芽。使用小剪刀切割滑道，避免将整个植物直接从土壤中拉出。每个杯子有三到四个大小相似的植物个体。要将触摸发刷安装到机械臂上，首先使用胶水将一层人的头发均匀地固定到 330 毫米长的钢尺上。头发的长度是126毫米。使用两个金属夹，将钢制尺子连接到机械臂上。在控制面板中，通过按 Jog F 加来提升和 Jog R 减压机械臂和刷子，沿垂直尺寸设置机器臂的高度。保持头发刷的尖端比杯子边缘低5厘米。按 Inc F 加或 R 减号预运行机器一到两个周期，以确保所有工厂人员被触摸。在正式实验之前，使用电子刻度测量触摸力。调整机器臂的高度，以保持一到两毫微吨之间的接触力。接下来，手动沿水平尺寸设置机器臂的起始位置。让发刷挂在每个托盘的边缘。按 Jog F 加或 Jog R 减号，一点一点地水平移动机器臂，以确保在接触实验开始之前没有接触任何植物。按下行驶按钮，将水平尺寸中的头发刷行驶距离设置为 365 毫米。按 Inc F 加或 R 减号移动机器臂以获得完整的行驶距离，并确保接触所有经过处理的植物。然后，按自动速度按钮，将机器臂 x 轴的移动速度设置为每分钟 5000 毫米。按下次要循环按钮，将触摸时间设置为 20 次试验，等于每轮 40 次触摸。按下主要周期按钮，使头发刷每天接触植物三轮，间隔时间为八小时，将触摸圆的重复间隔设置为每天 480 分钟。当倒计时变为零时，机器将自动开始新一轮的触摸。将主要周期设置为 12 次试验，使机器在四天内接触工厂 12 圈。按"启动"按钮启动预设程序。在整个接触期间，每天调整和校准刷子和发头到相同的高度。要跟踪每个植物在触摸实验中的螺栓日，每天观察每个植物的第一个注入茎。测量长度并记录生长时间，当它达到一厘米，这意味着它已过渡到植物阶段繁殖阶段。玫瑰花环半径的测量，从顶部拍摄整个托盘的照片。使用图像处理软件测量从玫瑰花环中心到长叶尖的距离，杯轮作为尺子。要测量玫瑰花环面积，首先，去除花序，而不影响其他玫瑰花环器官。从每个植物的顶部拍摄照片，并放置一个比例尺放在附近。在图像处理软件中，测量玫瑰花叶的水平二维表面积。本实验中棉签手动触摸处理与机器驱动式头发触摸处理野型控制哥伦比亚零植物进行比较。连续棉签触摸处理后，螺栓延迟 1.7 天。同样，对于自动机器驱动的头发触摸，观察到延迟螺栓。单变量考克斯危害分析表明，与对照组中的植物相比，接触组中植物的螺栓化风险分别为31%和52%。然而，在tDNA插入突变体、NKK一和NKK二中未观察到这种螺栓延迟。单变量考克斯比例危害模型显示，只有野生类型哥伦比亚零表现出控制植物和接触植物之间的显著差异，估计HR为41。对于形态变化，野生类型哥伦比亚零植物显示明显较小的玫瑰花环半径和较短的叶长度后，三天的恒定和重复的自动机器驱动头发触摸。经过13天的接触，预计的玫瑰花环面积从20平方厘米改为16平方厘米。为确保触摸力负载不会导致伤害，需要仔细设置适当的触摸力。需要一套适当的生长条件来生产一种合适的阿拉伯植物，用于触摸反应测定。例如，24 小时的光强为 180 到 240 微爱因斯坦/平方米/秒，需要观察信号形态生成。为了扩展到其他形式的机械力装载，机器还可以在将来分别使用喷水喷嘴或风吹风机进行改装，以模拟雨或风效应。

Research

JoVE Journal

Biology

A Labor-saving and Repeatable Touch-force Signaling Mutant Screen Protocol for the Study of Thigmomorphogenesis of a Model Plant Arabidopsis thaliana

Computer-Generated Animal Model Stimuli

计算机生成的动物模型的刺激

Communication between animals is diverse and complex. Animals may communicate using auditory, seismic, chemosensory, electrical, or visual signals. In ...

科研

生物学

Choice and No-Choice Assays for Testing the Resistance of A. thaliana to Chewing Insects

测试拟南芥耐咀嚼昆虫的选择和非选择性检测

Larvae of the small white cabbage butterfly are a pest in agricultural settings. This caterpillar species feeds from plants in the cabbage family, which ...

Efficient and Rapid Isolation of Early-stage Embryos from Arabidopsis thaliana Seeds

Efficient and Rapid Isolation of Early-stage Embryos from Arabidopsis thaliana Seeds

高效，快速的早期胚胎分离<em&gt;拟南芥</em&gt;种子

In flowering plants, the embryo develops within a nourishing tissue - the endosperm - surrounded by the maternal seed integuments (or seed coat). As a ...

Metabolic Labeling and Membrane Fractionation for Comparative Proteomic Analysis of Arabidopsis thaliana Suspension Cell Cultures

Metabolic Labeling and Membrane Fractionation for Comparative Proteomic Analysis of Arabidopsis thaliana Suspension Cell Cultures

代谢标记和膜分馏的比较蛋白质组学分析<em&gt;拟南芥</em&gt;悬浮细胞培养

Plasma membrane microdomains are features based on the physical properties of the lipid and sterol environment and have particular roles in signaling ...

Imaging Spatial Reorganization of a MAPK Signaling Pathway Using the Tobacco Transient Expression System

成像使用烟草瞬时表达系统MAPK信号通路的空间重组

Visualization of dynamic signaling events in live cells has been a challenge. We expanded an established transient expression system, the biomolecular ...

Rapid Analysis of Circadian Phenotypes in Arabidopsis Protoplasts Transfected with a Luminescent Clock Reporter

在拟南芥原生质体转染昼夜的表型有荧光时钟记者的快速分析

The plant circadian clock allows the anticipation of daily changes to the environment. This anticipation aids the responses to temporally predictable ...

A Hydroponic Co-cultivation System for Simultaneous and Systematic Analysis of Plant/Microbe Molecular Interactions and Signaling

用于植物/微生物分子相互作用和信号的同时系统分析的水培共培养系统

An experimental design mimicking natural plant-microbe interactions is very important to delineate the complex plant-microbe signaling processes. ...

Standardized Method for High-throughput Sterilization of Arabidopsis Seeds

拟南芥种子高通量灭菌的标准化方法

Arabidopsis thaliana (Arabidopsis) seedlings often need to be grown on sterile media. This requires prior seed sterilization to prevent the growth of ...

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A (EYFP-CENP-A)

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A EYFP-CENP-A

质谱分析，用于识别 EYFP 标记 CENP-A （EYFP-CENP-A） 的泛化

Studying the structure and the dynamics of kinetochores and centromeres is important in understanding chromosomal instability (CIN) and cancer ...

Forward Genetic Screen Using Transgenic Calcium Reporter Aequorin to Identify Novel Targets in Calcium Signaling

使用转基因钙处理器 Aequorin 前向基因屏幕，以识别钙信号中的新目标

Forward genetic screens have been important tools in the unbiased identification of genetic components involved in several biological pathways. The basis ...