Name: shootward movement of cfda tracer loaded in the bottom sink tissues of arabidopsis
Uploaded: 2019-05-11T15:00:00
Description: Watch this Scientific Journal Video about Shootward Movement of CFDA Tracer Loaded in the Bottom Sink Tissues of Arabidopsis at JoVE.com

这项工作由国家自然科学基金 (31671257) 和湖北粮食产业合作创新中心 (LXT-16-16-18) 资助。

1. 多发性能级研究中的垂直生长
<ol><li>层流柜的内部需要在使用前使用30分钟的紫外线和15分钟的气流进行处理。当紫外线亮起时, 请务必关闭玻璃门。在将所有工具和板材放入机柜之前, 请先将其喷洒70% 的乙醇。</li>
	<li>在层流柜下的标准90毫米直径 Petri 培养皿中制备 Murashige 和 Skoog (MS) 介质。 注: MS 介质包含以下组件: 20.6 mM NH 4no3, 18.8 mm kno 3, 1.25 mmKh 2 po 4, 1.5 mm...

<ol>
	<li>Grignon, N., Touraine, B., Durand, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=6(5)Carboxyfluorescein+as+a+tracer+of+phloem+sap+translocation.">6(5)Carboxyfluorescein as a tracer of phloem sap translocation.</a> American Journal of Botany. 76, 871-877 (1989).</li><li>Wright, K. M., Oparka, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+fluorescent+probe+HPTS+as+a+phloem-mobile,+symplastic+tracer:+an+evaluation+using+confocal+laser+scanning+microscopy.">The fluorescent probe HPTS as a phloem-mobile, symplastic tracer: an evaluation using confocal laser scanning microscopy.</a> Journal of Experimental Botany. 47 (3), 439-445 (1996).</li><li>Oparka, K. J., Prior, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Movement+of+Lucifer+Yellow+CH+in+potato+tuber+storage+tissues:+A+comparison+of+symplastic+and+apoplastic+transport.">Movement of Lucifer Yellow CH in potato tuber storage tissues: A comparison of symplastic and apoplastic transport.</a> Planta. 176 (4), 533-540 (1988).</li><li>Knoblauch, M., 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=Multispectral+Phloem-Mobile+Probes:+Properties+and+Applications.">Multispectral Phloem-Mobile Probes: Properties and Applications.</a> Plant Physiology. 167 (4), 1211-1220 (2015).</li><li>Oparka, K. J., Duckett, C. M., Prior, D. A. M., Fisher, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+imaging+of+phloem+unloading+in+the+root+tip+of+Arabidopsis.">Real-time imaging of phloem unloading in the root tip of Arabidopsis.</a> The Plant Journal. 6 (5), 759-766 (1994).</li><li>Viola, R., 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=Tuberization+in+Potato+Involves+a+Switch+from+Apoplastic+to+Symplastic+Phloem+Unloading.">Tuberization in Potato Involves a Switch from Apoplastic to Symplastic Phloem Unloading.</a> The Plant Cell. 13 (2), 385-398 (2001).</li><li>Roberts, A. G., 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=Phloem+Unloading+in+Sink+Leaves+of+Nicotiana+benthamiana:+Comparison+of+a+Fluorescent+Solute+with+a+Fluorescent+Virus.">Phloem Unloading in Sink Leaves of Nicotiana benthamiana: Comparison of a Fluorescent Solute with a Fluorescent Virus.</a> The Plant Cell. 9 (8), 1381-1396 (1997).</li><li>P&#233;ron, T., 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=New+Insights+into+Phloem+Unloading+and+Expression+of+Sucrose+Transporters+in+Vegetative+Sinks+of+the+Parasitic+Plant+Phelipanche+ramosa+L+(Pomel).">New Insights into Phloem Unloading and Expression of Sucrose Transporters in Vegetative Sinks of the Parasitic Plant Phelipanche ramosa L (Pomel).</a> Frontiers in Plant Science. 7 (2048), (2017).</li><li>Spallek, T., 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=Interspecies+hormonal+control+of+host+root+morphology+by+parasitic+plants.">Interspecies hormonal control of host root morphology by parasitic plants.</a> Proceedings of the National Academy of Sciences of the USA. 114 (20), 5283-5288 (2017).</li><li>Complainville, A., 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=Nodule+initiation+involves+the+creation+of+a+new+symplasmic+field+in+specific+root+cells+of+medicago+species.">Nodule initiation involves the creation of a new symplasmic field in specific root cells of medicago species.</a> The Plant Cell. 15 (12), 2778-2791 (2003).</li><li>Bederska, M., Borucki, W., Znojek, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Movement+of+fluorescent+dyes+Lucifer+Yellow+(LYCH)+and+carboxyfluorescein+(CF)+in+Medicago+truncatula+Gaertn.+roots+and+root+nodules.">Movement of fluorescent dyes Lucifer Yellow (LYCH) and carboxyfluorescein (CF) in Medicago truncatula Gaertn. roots and root nodules.</a> Symbiosis. 58 (1-3), 183-190 (2012).</li><li>Robards, A. W., Lucas, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasmodesmata.">Plasmodesmata.</a> Annual Review of Plant Physiology and Plant Molecular Biology. 41 (1), 369-419 (1990).</li><li>Roberts, A. G., Oparka, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasmodesmata+and+the+control+of+symplastic+transport.">Plasmodesmata and the control of symplastic transport.</a> Plant, Cell &amp; Environment. 26 (1), 103-124 (2003).</li><li>Duckett, C. M., Oparka, K. J., Prior, D. A. M., Dolan, L., Roberts, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dye-coupling+in+the+root+epidermis+of+Arabidopsis+is+progressively+reduced+during+development.">Dye-coupling in the root epidermis of Arabidopsis is progressively reduced during development.</a> Development. 120 (11), 3247-3255 (1994).</li><li>Palevitz, B. A., Hepler, P. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+dye+coupling+of+stomatal+cells+of+Allium+and+Commelina+demonstrated+by+microinjection+of+Lucifer+yellow.">Changes in dye coupling of stomatal cells of Allium and Commelina demonstrated by microinjection of Lucifer yellow.</a> Planta. 164 (4), 473-479 (1985).</li><li>van Bel, A. J. E., Kempers, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Symplastic+isolation+of+the+sieve+element-companion+cell+complex+in+the+phloem+of+Ricinus+communis+and+Salix+alba+stems.">Symplastic isolation of the sieve element-companion cell complex in the phloem of Ricinus communis and Salix alba stems.</a> Planta. 183 (1), 69-76 (1991).</li><li>Erwee, M. G., Goodwin, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Symplast+domains+in+extrastelar+tissues+of+Egeria+densa+Planch.">Symplast domains in extrastelar tissues of Egeria densa Planch.</a> Planta. 163 (1), 9-19 (1985).</li><li>Oparka, K. J., Prior, D. A. M., Wright, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Symplastic+communication+between+primary+and+developing+lateral+roots+of+Arabidopsis+thaliana.">Symplastic communication between primary and developing lateral roots of Arabidopsis thaliana.</a> Journal of Experimental Botany. 46 (2), 187-197 (1995).</li><li>Christensen, N. M., Faulkner, C., Oparka, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+for+Unidirectional+Flow+through+Plasmodesmata.">Evidence for Unidirectional Flow through Plasmodesmata.</a> Plant Physiology. 150 (1), 96-104 (2009).</li><li>Wr&#243;bel-Marek, J., Kurczy&#324;ska, E., P&#322;achno, B. J., Kozieradzka-Kiszkurno, M. <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+symplasmic+domains+in+the+embryo+and+seed+of+Sedum+acre+L.+(Crassulaceae).">Identification of symplasmic domains in the embryo and seed of Sedum acre L. (Crassulaceae).</a> Planta. 245 (3), 491-505 (2017).</li><li>Liang, D., White, R. G., Waterhouse, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+silencing+in+Arabidopsis+spreads+from+the+root+to+the+shoot,+through+a+gating+barrier,+by+template-dependent,+non-vascular,+cell+to+cell+movement.">Gene silencing in Arabidopsis spreads from the root to the shoot, through a gating barrier, by template-dependent, non-vascular, cell to cell movement.</a> Plant Physiology. 159 (3), 984-1000 (2012).</li><li>Radford, J. E., White, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+tissue-preparation-induced+callose+synthesis+on+estimates+of+plasmodesma+size+exclusion+limits.">Effects of tissue-preparation-induced callose synthesis on estimates of plasmodesma size exclusion limits.</a> Protoplasma. 216 (1-2), 47-55 (2001).</li><li>Kollist, H., 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=Rapid+Responses+to+Abiotic+Stress:+Priming+the+Landscape+for+the+Signal+Transduction+Network.">Rapid Responses to Abiotic Stress: Priming the Landscape for the Signal Transduction Network.</a> Trends in Plant Science. 24 (1), 25-37 (2019).</li><li>Haupt, S., Duncan, G. H., Holzberg, S., Oparka, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+for+Symplastic+Phloem+Unloading+in+Sink+Leaves+of+Barley.">Evidence for Symplastic Phloem Unloading in Sink Leaves of Barley.</a> Plant Physiology. 125 (1), 209-218 (2001).</li><li>Botha, C. E. J., 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=A+xylem+sap+retrieval+pathway+in+rice+leaf+blades:+evidence+of+a+role+for+endocytosis?.">A xylem sap retrieval pathway in rice leaf blades: evidence of a role for endocytosis?.</a> Journal of Experimental Botany. 59 (11), 2945-2954 (2008).</li></ol>

新出现的研究表明, 植物可以对外部刺激23迅速作出反应, 包括在实验程序中引入的操作22。在我们最初的实验中, 我们对这些知识的监督往往导致染色失败。通过这些教训, 我们建议在进行本实验时应注意以下预防措施: (1) 收获后的种子应存放在低温、低水分的储物柜中;(2) 操纵植物, 特别是暴露在柜子里的空气中, 应保持在最低限度;(3) 实验条件应保持不变, 例如...

许多具有光谱特性的荧光示踪剂, 如 5 (6)-羧荧光素 (CF)1、8-羟基苯甲酸1、3、6-三硫基苯甲酸2、路西奥黄 CH(lsch) 3、ES奎因和 cter4, 已经开发出来,应用于植物中, 以监测共体运动和韧皮部的活性。一般情况下, 共生染料被加载到目标组织中的切割中, 记者连续分散到植物的其他部位将证明细胞间的通信。虽然染料吸收的机制还没有得到充分的理解, 但活细胞内 CF 运动的原理已经得到了广泛的认可。CF (CF 二乙酸酯, CFDA) 的酯形态为非荧光, 但膜渗透性。这种特性允许染料快速地将膜扩散到细胞中。一旦进入活细胞, 细胞内酯酶去除 CFDA 3 ' 和 6 ' 位置的醋酸盐基团, 释放荧光和膜不渗透 CF (图 1, 也参考赖特等人 2);然后, CF 可以通过质粒移动到植物的其他部分。
一个既定的程序与 CFDA 是, 它可以加载到源叶, 并用于监测韧皮部流和韧皮部卸载在许多物种的水槽组织, 例如, cf 卸载在拟南芥根5, 韧皮部卸载马铃薯块茎化过程中6、韧皮部卸除在烟叶中, 叶7, 依此类推。通过类似的加载方法, 其他研究采用这种染料来证明宿主和寄生虫8,9之间的共生联系, 或揭示共生关系10,11.
另一种利用这种染料的方法是通过微注射将其加载到特定的细胞或单个细胞中, 以确定其分布模式。这种复杂的技术极大地促进了我们对质粒介导的细胞间通信的更深入理解, 特别是在共生域 12、13 概念的发展过程中。例如, 在拟南芥子叶细胞中注射 cfda 导致下胚轴表皮的染料耦合模式, 但在底层细胞或根表皮中产生非耦合, 因此下胚轴表皮形成共用域14。类似的领域, 如气孔保护细胞15, 筛片元素-伴生细胞16 , 根毛细胞14和根帽17,18已被确定为微注射技术。最令人惊讶的是, 有些领域允许示踪分子向某一方向移动。以滴管域为例, 微注射荧光探针进入支撑表皮细胞, 导致示踪剂流入滴虫域, 但反向注射并不成立 19.最近的一份报告还在Sedum胚胎20的共生域中发现了类似的情况。因此, 所有上述情况都意味着, 交换加载站点可能会导致对共生通信的新见解。我们以前的实验旨在剖析根-拍摄移动沉默的路径确定了一个新的共生域, 或 HEJ (低音节-表观结合部交界处) 区, 这是进一步验证通过根加载 (非规范的正弦加载) CFDA实验21。在这里, 我们进一步阐述根到拍摄 cf 运动使用一个简单的方法, 并通过移动加载站点恢复一个潜在的符号域。此外, 这一程序可以适应区分改变根对镜头长途运输的遗传背景。

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			<td height="24" style="height:24px;width:223px;">KNO3&#160;&#160;</td>
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			<td>Sinopharm Chemical Reagent</td>
			<td>10017608</td>
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			<td>Sinopharm Chemical Reagent</td>
			<td>10013018</td>
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			<td>Sinopharm Chemical Reagent</td>
			<td>20011160</td>
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			<td height="24" style="height:24px;">MnSO4&#183;H2O&#160;</td>
			<td>Sinopharm Chemical Reagent</td>
			<td>10013418</td>
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			<td>Sinopharm Chemical Reagent</td>
			<td>10019818</td>
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			<td>Sinopharm Chemical Reagent</td>
			<td>10004818</td>
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			<td>Sinopharm Chemical Reagent</td>
			<td>10024018</td>
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			<td>Sinopharm Chemical Reagent</td>
			<td>10008218</td>
			<td></td>
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			<td height="24" style="height:24px;">CoCl2&#183;6H2O</td>
			<td>Sinopharm Chemical Reagent</td>
			<td>10007216</td>
			<td></td>
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			<td height="21" style="height:21px;">KI</td>
			<td>Sinopharm Chemical Reagent</td>
			<td>10017160</td>
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			<td>Sinopharm Chemical Reagent</td>
			<td>10012118</td>
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			<td height="21" style="height:21px;">EDTA&#160;</td>
			<td>Sinopharm Chemical Reagent</td>
			<td>10009717</td>
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			<td>Sinopharm Chemical Reagent</td>
			<td>10019718</td>
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			<td height="21" style="height:21px;">KOH</td>
			<td>Sinopharm Chemical Reagent</td>
			<td>10017018</td>
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			<td height="21" style="height:21px;">Sucrose</td>
			<td>Sinopharm Chemical Reagent</td>
			<td>10021418</td>
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			<td>MACKLIN</td>
			<td>I811835</td>
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			<td>MACKLIN</td>
			<td>N814565</td>
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			<td>MACKLIN</td>
			<td>V820447</td>
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			<td>MACKLIN</td>
			<td>T818865</td>
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			<td height="21" style="height:21px;">Glycine</td>
			<td>MACKLIN</td>
			<td>G800880</td>
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			<td height="21" style="height:21px;">Agar powder</td>
			<td>Novon</td>
			<td>ZZ14022</td>
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			<td height="21" style="height:21px;">Fluorescence Microscope</td>
			<td>Zeiss</td>
			<td>Axio Zoom V16</td>
			<td></td>
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			<td height="21" style="height:21px;">Dissecting microscope</td>
			<td>SDPTOP</td>
			<td>SRE-1030</td>
			<td></td>
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			<td height="21" style="height:21px;">200&#956;l pipette</td>
			<td>Dragon Laboratory Instruments</td>
			<td colspan="2">713111110000-20-200ul</td>
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			<td height="21" style="height:21px;">2.5&#956;l pipette</td>
			<td>Eppendorf</td>
			<td>3120000011</td>
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			<td height="21" style="height:21px;">Fine forceps&#160;</td>
			<td>TWEEZERS</td>
			<td>ST-15</td>
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			<td height="21" style="height:21px;">Parafilm</td>
			<td>PARAFILM</td>
			<td>PM-996</td>
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			<td height="21" style="height:21px;">Stainless steel double-sided blade</td>
			<td>Gillette</td>
			<td colspan="2">Platinum-Plus Double-Edge Blade</td>
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	</tbody>
</table>

shootward movement of cfda tracer loaded in the bottom sink tissues of arabidopsis

对塑性示踪剂 5 (6)-羧基荧光素二乙酸酯 (CFDA) 已广泛应用于活植物中, 以证明细胞间的连接、韧皮部的迁移和血管模式。该方案分别采用根切法和下胚切法显示拟南芥中的底部到顶部羧基荧光素 (cf) 运动。这两个不同的程序导致不同的效率 CF 运动: 约91% 的 CF 出现在芽与下胚轴捏过程, 而只有约70% 的外观 CF 与根切割程序。加载位置的简单变化, 导致这种共生染料的移动效率发生重大变化, 这表明 CF 运动可能受到共生调节的影响, 很可能是由根下胚轴连接。

共变运动往往受到环境波动的影响。植物生长状态的扰动, 甚至组织制备过程都会影响质粒的大小排除极限, 从而影响共生体的迁移 22.为了提高染色效率, 我们将操作限制在温度和湿度受到严格控制的生长室内, 并尽快执行整个过程 (最好是在提升培养皿盖后10-15 内)。在实验中的这些预防措施可以有效地降低不成功的拍摄染色率。
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Watch this Scientific Journal Video about Shootward Movement of CFDA Tracer Loaded in the Bottom Sink Tissues of Arabidopsis at JoVE.com

Shootward Movement of CFDA Tracer Loaded in the Bottom Sink Tissues of Arabidopsis

该协议的目的是展示如何将 CFDA 加载到拟南芥底部的不同部位。然后给出了在芽中产生的 CF 分布模式。

美国食品药品监督管理局追踪者在拟南芥底部水槽组织中的射击运动

The symplastic tracer 5(6)-carboxyfluorescein diacetate (CFDA) has been widely applied in living plants to demonstrate the intercellular connection, ...

广泛使用的探针CFDA，或甲氧氟辛丁二甲酸酯的短，是非荧光的。两个H3组和3'6'位置在这个化合物上使其膜渗透。一旦进入细胞内部，内部细胞醋酸盐将去除醋酸盐组，并释放新的CFDA形式。那是一种碳氧荧光素，或是荧光素的CF。由于这种新形式的膜渗透性较小，重量大，所有细胞都是通过细胞间孔隙或植物中的普拉斯莫德玛塔移动的，这使得这个细胞间在细胞间活动中研究的理想探针。在今天的视频中，我们将分别演示对阿拉伯植物根部和低皮质的CFD加载，我们将展示两种略有不同的程序将如何影响这个荧光探针的从下到上运动。始终如一地获得染色效果，我们将收获的阿拉伯种子储存在4摄氏度，40%的水分柜中。在此条件下储存超过七天的新收获种子很容易用于播种。为了确保每一种植物都生长在培养皿中相对相似的空间中，我们使用种子区等级卡。现在，润湿消毒的尖端，浸入灭菌的种子，并在指定位置一一播种。播种完成后，用胶卷和胶带密封培养皿，把它们放在培养皿上，让它垂直生长。下面是我们将在本实验中使用的工具。请注意，我们最好尽快装上染料。通常，您可以在 10 到 15 分钟内完成整个过程。在执行 CFDA 加载实验之前，我们始终准备新的 CFDA 解决方案。在这里，在立即使用之前，将一毫摩尔CFDA稀释为五微摩尔浓度。将副膜切成三分之三毫米大小的小块。因为Petri盘的盖子和底部有冷凝，我们需要用纸巾把它们清理干净，以避免染料四处流动。一小块胶卷放在根下。提起植物，放在盖子上。切根约5至10毫米以下的低血。把拍摄的小片拍回来。现在，小心地将一个微升 CFDA 涂抹在根的切割端。尽量避免污染所有的原生植物。然后盖上盖子，然后把它们放在生长室两个小时。低氯二苯醚捏合法。如前所述，小副膜片被放在根-低基交界处。我们使用前部轻轻捏紧靠近根组织，并小心地将0.1微升CFDA涂抹在捏边，离开植物两小时。照片从9天到12天后区域染色植物。它们都显示了在科蒂龙和真叶的血管染色模式。只有在极少数情况下，科蒂顿叶才显示出半叶染色的图案。所以基本上，这两种方法在CF染色模式上没有区别，但是如果我们在越来越多的工厂中进行染料装载，我们发现这两种程序在效率上存在显著差异。采用根切法，约70%的植物在拍摄中显示荧光信号。但使用低氯基捏法可将效率提高至91%。我们也尝试了根夹法，但这种方法导致效率更低，大约30%，因此，似乎加载染料的方法没有考虑到染色的差异。区别在于加载站点，这些站点可能由共塑屏障分隔。我们展示了我们如何加载CFDA到阿拉伯根和低皮质。我们使用两个略有不同的程序，这两个略有不同的程序导致在这个底部到顶部探头运动显著差异。我们建议，这种方法可以帮助我们识别根源射射信号的潜在共塑屏障。

Research

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Environment

Shootward Movement of CFDA Tracer Loaded in the Bottom Sink Tissues of Arabidopsis

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