Name: an optimized rhizobox protocol to visualize root growth and responsiveness to localized nutrients
Uploaded: 2018-10-22T15:00:00
Description: Watch this Scientific Journal Video about An Optimized Rhizobox Protocol to Visualize Root Growth and Responsiveness to Localized Nutrients at JoVE.com

作者希望感谢匿名审阅者对他们的反馈意见, 以及 J.C. 卡希尔和谭宝在开发 rhizobox 协议方面的初步指导。资金由粮食和农业研究基金会、美国农业部 (USDA) 国家粮食和农业研究所、农业试验站项目 CA-D-PLS-2332-H、司法部和加州大学戴维斯分校提供。科学通过 J.S. 的奖学金

1. 前、后面板和垫片的准备工作
<ol><li>准备前面板和背板。<ol>
<li>将两块透明0.635 厘米厚的亚克力切成40.5 厘米宽61厘米每盒或购买预切割件 (见材料表)。</li>
		<li>使用专为亚克力设计的钻头孔, 从顶部的2.5、19、38和53.3 厘米的侧面边缘0.635 厘米直径1.3 厘米的钻孔。钻孔1.3 厘米从底部边缘在 2.5, 20.3 和38厘米从左侧 (图 1)。 注意: 一次使用钻床进行六到十页的?...

<ol>
	<li>Hodge, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Roots:+The+Acquisition+of+Water+and+Nutrients+from+the+Heterogeneous+Soil+Environment.">Roots: The Acquisition of Water and Nutrients from the Heterogeneous Soil Environment.</a> Progress in Botany 71. , 307-337 (2010).</li><li>Grossman, J. D., Rice, 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=Evolution+of+root+plasticity+responses+to+variation+in+soil+nutrient+distribution+and+concentration.">Evolution of root plasticity responses to variation in soil nutrient distribution and concentration.</a> Evolutionary Applications. 5 (8), 850-857 (2012).</li><li>Zhang, H., Forde, B. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+Arabidopsis+MADS+box+gene+that+controls+nutrient-induced+changes+in+root+architecture.">An Arabidopsis MADS box gene that controls nutrient-induced changes in root architecture.</a> Science. 279 (5349), 407-409 (1998).</li><li>Hodge, A., Stewart, J., Robinson, D., Griffiths, B. S., Fitter, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Competition+between+roots+and+soil+micro-organisms+for+nutrients+from+nitrogen-rich+patches+of+varying+complexity.">Competition between roots and soil micro-organisms for nutrients from nitrogen-rich patches of varying complexity.</a> Journal of Ecology. 88 (1), 150-164 (2000).</li><li>Trachsel, S., Kaeppler, S. M., Brown, K. M., Lynch, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shovelomics:+high+throughput+phenotyping+of+maize+(Zea+mays+L.)+root+architecture+in+the+field.">Shovelomics: high throughput phenotyping of maize (Zea mays L.) root architecture in the field.</a> Plant and Soil. 341 (1-2), 75-87 (2011).</li><li>Rogers, E. D., Monaenkova, D., Mijar, M., Nori, A., Goldman, D. I., Benfey, P. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=X-ray+computed+tomography+reveals+the+response+of+root+system+architecture+to+soil+texture.">X-ray computed tomography reveals the response of root system architecture to soil texture.</a> Plant Physiology. , (2016).</li><li>Groleau-Renaud, V., Plantureux, S., Guckert, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+mechanical+constraint+on+nodal+and+seminal+root+system+of+maize+plants.">Effect of mechanical constraint on nodal and seminal root system of maize plants.</a> Comptes Rendus De L Academie Des Sciences Serie Iii-Sciences De La Vie-Life Sciences. 321 (1), 63-71 (1998).</li><li>Lin, Y., Allen, H. E., Di Toro, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Barley+root+hair+growth+and+morphology+in+soil,+sand,+and+water+solution+media+and+relationship+with+nickel+toxicity.">Barley root hair growth and morphology in soil, sand, and water solution media and relationship with nickel toxicity.</a> Environmental Toxicology and Chemistry. 35 (8), 2125-2133 (2016).</li><li>Wenzel, W. W., Wieshammer, G., Fitz, W. J., Puschenreiter, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+rhizobox+design+to+assess+rhizosphere+characteristics+at+high+spatial+resolution.">Novel rhizobox design to assess rhizosphere characteristics at high spatial resolution.</a> Plant and Soil. 237 (1), 37-45 (2001).</li><li>Spohn, M., Carminati, A., Kuzyakov, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Soil+zymography+-+A+novel+in+situ+method+for+mapping+distribution+of+enzyme+activity+in+soil.">Soil zymography - A novel in situ method for mapping distribution of enzyme activity in soil.</a> Soil Biology and Biochemistry. 58, 275-280 (2013).</li><li>Vollsnes, A. V., Futsaether, C. M., Bengough, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantifying+rhizosphere+particle+movement+around+mutant+maize+roots+using+time-lapse+imaging+and+particle+image+velocimetry.">Quantifying rhizosphere particle movement around mutant maize roots using time-lapse imaging and particle image velocimetry.</a> European Journal of Soil Science. 61 (6), 926-939 (2010).</li><li>Hewitt, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Sand and Water Culture Methods Used in the Study of Plant Nutrition. , (1966).</li><li>Choudhary, M. I., Shalaby, A. A., Al-Omran, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Water+holding+capacity+and+evaporation+of+calcareous+soils+as+affected+by+four+synthetic+polymers.">Water holding capacity and evaporation of calcareous soils as affected by four synthetic polymers.</a> Communications in Soil Science and Plant Analysis. 26 (13-14), 2205-2215 (1995).</li><li>Bakker, P. A. H. M., Berendsen, R. L., Doornbos, R. F., Wintermans, P. C. A., Pieterse, C. M. 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+rhizosphere+revisited:+root+microbiomics.">The rhizosphere revisited: root microbiomics.</a> Frontiers in Plant Science. 4, 2013 (2013).</li><li>McNear, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Rhizosphere+-+Roots,+Soil,+and+Everything+In+Between.">The Rhizosphere - Roots, Soil, and Everything In Between.</a> Nature Education Knowledge. 4 (3), 1 (2013).</li><li>Ortas, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+the+extent+of+rhizosphere+soil.">Determination of the extent of rhizosphere soil.</a> Communications in Soil Science and Plant Analysis. 28 (19-20), 1767-1776 (1997).</li><li>. Carbon (13C) and Nitrogen (15N) Sample Preparation Available from: <a target="_blank" href="https://stableisotopefacility.ucdavis.edu/13cand15nsamplepreparation.html">https://stableisotopefacility.ucdavis.edu/13cand15nsamplepreparation.html</a> (2018)</li><li>Barraclough, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=15N+isotope+dilution+techniques+to+study+soil+nitrogen+transformations+and+plant+uptake.">15N isotope dilution techniques to study soil nitrogen transformations and plant uptake.</a> Fertilizer research. 42 (1-3), 185-192 (1995).</li><li>Belter, P. R., Cahill, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disentangling+root+system+responses+to+neighbours:+identification+of+novel+root+behavioural+strategies.">Disentangling root system responses to neighbours: identification of novel root behavioural strategies.</a> AoB PLANTS. 7, (2015).</li><li>Nagel, K. 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=GROWSCREEN-Rhizo+is+a+novel+phenotyping+robot+enabling+simultaneous+measurements+of+root+and+shoot+growth+for+plants+grown+in+soil-filled+rhizotrons.">GROWSCREEN-Rhizo is a novel phenotyping robot enabling simultaneous measurements of root and shoot growth for plants grown in soil-filled rhizotrons.</a> Functional Plant Biology. 39 (11), 891-904 (2012).</li><li>Adu, M. O., Yawson, D. O., Bennett, M. J., Broadley, M. R., Dupuy, L. X., White, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+scanner-based+rhizobox+system+enabling+the+quantification+of+root+system+development+and+response+of+Brassica+rapa+seedlings+to+external+P+availability.">A scanner-based rhizobox system enabling the quantification of root system development and response of Brassica rapa seedlings to external P availability.</a> Plant Root. 11, 16-32 (2017).</li><li>Neumann, G., George, T. S., Plassard, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strategies+and+methods+for+studying+the+rhizosphere-the+plant+science+toolbox.">Strategies and methods for studying the rhizosphere-the plant science toolbox.</a> Plant and Soil. 321 (1-2), 431-456 (2009).</li><li>Bodner, G., Alsalem, M., Nakhforoosh, A., Arnold, T., Leitner, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RGB+and+Spectral+Root+Imaging+for+Plant+Phenotyping+and+Physiological+Research:+Experimental+Setup+and+Imaging+Protocols.">RGB and Spectral Root Imaging for Plant Phenotyping and Physiological Research: Experimental Setup and Imaging Protocols.</a> JoVE (Journal of Visualized Experiments). (126), e56251-e56251 (2017).</li><li>Kuchenbuch, R. O., Ingram, K. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Image+analysis+for+non-destructive+and+non-invasive+quantification+of+root+growth+and+soil+water+content+in+rhizotrons.">Image analysis for non-destructive and non-invasive quantification of root growth and soil water content in rhizotrons.</a> Journal of Plant Nutrition and Soil Science. 165 (5), 573-581 (2002).</li><li>Dresb&#248;ll, D. B., Thorup-Kristensen, K., McKenzie, B. M., Dupuy, L. X., Bengough, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Timelapse+scanning+reveals+spatial+variation+in+tomato+(Solanum+lycopersicum+L.)+root+elongation+rates+during+partial+waterlogging.">Timelapse scanning reveals spatial variation in tomato (Solanum lycopersicum L.) root elongation rates during partial waterlogging.</a> Plant and Soil. 369 (1-2), 467-477 (2013).</li><li>Wu, 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=RhizoChamber-Monitor:+a+robotic+platform+and+software+enabling+characterization+of+root+growth.">RhizoChamber-Monitor: a robotic platform and software enabling characterization of root growth.</a> Plant Methods. 14 (1), 44 (2018).</li><li>Rogers, S. W., Moorman, T. B., Ong, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescent+In+Situ+Hybridization+and+Micro-autoradiography+Applied+to+Ecophysiology+in+Soil.">Fluorescent In Situ Hybridization and Micro-autoradiography Applied to Ecophysiology in Soil.</a> Soil Science Society of America Journal. 71 (2), 620-631 (2007).</li><li>Eickhorst, T., Tippk&#246;tter, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+microorganisms+in+undisturbed+soil+by+combining+fluorescence+in+situ+hybridization+(FISH)+and+micropedological+methods.">Detection of microorganisms in undisturbed soil by combining fluorescence in situ hybridization (FISH) and micropedological methods.</a> Soil Biology and Biochemistry. 40 (6), 1284-1293 (2008).</li><li>Spohn, M., Kuzyakov, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distribution+of+microbial-+and+root-derived+phosphatase+activities+in+the+rhizosphere+depending+on+P+availability+and+C+allocation+-+Coupling+soil+zymography+with+14C+imaging.">Distribution of microbial- and root-derived phosphatase activities in the rhizosphere depending on P availability and C allocation - Coupling soil zymography with 14C imaging.</a> Soil Biology and Biochemistry. 67, 106-113 (2013).</li><li>Lv, G., Kang, Y., Li, L., Wan, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+irrigation+methods+on+root+development+and+profile+soil+water+uptake+in+winter+wheat.">Effect of irrigation methods on root development and profile soil water uptake in winter wheat.</a> Irrigation Science. 28 (5), 387-398 (2010).</li><li>Asseng, S., Ritchie, J. T., Smucker, A. J. M., Robertson, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Root+growth+and+water+uptake+during+water+deficit+and+recovering+in+wheat.">Root growth and water uptake during water deficit and recovering in wheat.</a> Plant and Soil. 201 (2), 265-273 (1998).</li><li>Hernandez-Ramirez, 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=Root+Responses+to+Alterations+in+Macroporosity+and+Penetrability+in+a+Silt+Loam+Soil.">Root Responses to Alterations in Macroporosity and Penetrability in a Silt Loam Soil.</a> Soil Science Society of America Journal. 78 (4), 1392-1403 (2014).</li><li>Zhang, Y. L., Wang, Y. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Soil+enzyme+activities+with+greenhouse+subsurface+irrigation.">Soil enzyme activities with greenhouse subsurface irrigation.</a> Pedosphere. 16 (4), 512-518 (2006).</li><li>Robinson, D., Hodge, A., Griffiths, B. S., Fitter, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plant+root+proliferation+in+nitrogen-rich+patches+confers+competitive+advantage.">Plant root proliferation in nitrogen-rich patches confers competitive advantage.</a> Proceedings of the Royal Society of London B: Biological Sciences. 266 (1418), 431-435 (1999).</li><li>Lobet, G., Draye, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+scanning+procedure+enabling+the+vectorization+of+entire+rhizotron-grown+root+systems.">Novel scanning procedure enabling the vectorization of entire rhizotron-grown root systems.</a> Plant Methods. 9, 1 (2013).</li><li>Swarup, R., Wells, D. M., Bennett, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Root+Gravitropism.">Root Gravitropism.</a> Plant Roots: The Hidden Half. , (2013).</li><li>Smit, A. L., Bengough, A. G., Engels, C., van Noordwijk, M., Pellerin, S., van de Geijn, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Root Methods: A Handbook. , (2000).</li><li>van Dusschoten, D., 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+3D+Analysis+of+Plant+Roots+Growing+in+Soil+Using+Magnetic+Resonance+Imaging1[OPEN].">Quantitative 3D Analysis of Plant Roots Growing in Soil Using Magnetic Resonance Imaging1[OPEN].</a> Plant Physiology. 170 (3), 1176-1188 (2016).</li><li>Metzner, 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=Direct+comparison+of+MRI+and+X-ray+CT+technologies+for+3D+imaging+of+root+systems+in+soil:+potential+and+challenges+for+root+trait+quantification.">Direct comparison of MRI and X-ray CT technologies for 3D imaging of root systems in soil: potential and challenges for root trait quantification.</a> Plant Methods. 11, 17 (2015).</li></ol>

本协议中描述的 rhizoboxes 可用于回答根和根际科学中的各种问题, 并在别处发现了不同用途10、20、21、22、23,24,25. 其他研究人员捕获了 rhizoboxes21、25、26</sup...

根系在植物养分采集中起着至关重要的作用, 但与地面上的同类相比往往被忽视。鉴于根系建造和维护的大量碳成本, 植物在觅食是值得投资的地方, 进化出了形成根系的机制。因此, 根系可以通过在热点中增殖、上调吸收率和快速转运养分向韧皮部进一步输送1, 从而高效、动态地挖掘资源斑块。可塑性反应在植物种类或基因型2、3和根据所涉及的养分的化学形式中有很大差异4,5。应进一步探讨根系可塑性的变化, 因为了解不同土壤资源的复杂根系反应, 可以告知育种和管理策略, 以提高农业的养分利用效率。
尽管它对了解植物系统的必要性和相关性, 但在相关的尺度上可视化和量化根塑性会带来技术挑战。从土壤中挖掘根冠 ("shovelomics"6) 是一种常见的方法, 但细根利用土壤骨料之间的小孔隙, 而开挖不可避免地导致这些脆弱的根的某种程度的损失。此外, 破坏性的收获使得不可能随着时间的推移在一个根系统中发生变化。原位成像方法, 如 X 射线计算机断层扫描允许直接可视化的根和土壤资源在高空间分辨率7, 但昂贵, 需要专门的设备。水培试验避免了从土壤中提取根的约束, 但与土壤的机械约束和生物物理复杂性相比, 在水性介质中根系形态和结构的差异8、9。最后, 在这些人工介质中, 根际过程和功能不能与发育可塑性相结合。
我们提出了一个用于构造和使用 rhizoboxes (窄的、透明的矩形容器) 的协议, 作为一种低成本、可定制的方法来表征土壤中的根系生长。特别设计的框架鼓励根在后面板上优先增长, 由于安歇, 提高根长度测量的准确性。Rhizoboxes 通常用于研究根系生长和根际相互作用10,11,12, 但这里提出的方法提供了简单的优势, 其单室设计和廉价材料, 旨在研究对局部营养素的根系反应。然而, 该方法也可用于研究一系列其他根和根际过程, 如内/种间竞争, 化合物的空间分布, 微生物, 或酶活性。在本文中, 我们研究了玉米杂交种对15个 n-标记豆科植物残留斑块的基因型差异, 并突出了代表性结果, 验证了 rhizobox 方法。

<table border="0" cellpadding="0" cellspacing="0" width="863">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">1.27 cm diameter PVC pipe</td>
			<td style="width:123px;">JM Eagle</td>
			<td style="width:119px;">530048</td>
			<td style="width:407px;">305 cm per box, cut into lengths as specified in the protocol</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">PVC side elbows</td>
			<td style="width:123px;">Lasco</td>
			<td>315498</td>
			<td>2 per box</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">PVC 90-degree elbows</td>
			<td style="width:123px;">Charlotte</td>
			<td style="width:119px;">PVC 02300 0600</td>
			<td>4 per box</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">PVC T joints</td>
			<td style="width:123px;">Charlotte</td>
			<td style="width:119px;">PVC 02402 0600</td>
			<td>4 per box</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Extruded acrylic panes</td>
			<td style="width:123px;">TAP Plastics</td>
			<td style="width:119px;">N/A</td>
			<td>2 per box, 0.64 cm thick x 40.5 cm wide x 61 cm long</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">HDPE spacers (sides)</td>
			<td style="width:123px;">TAP Plastics</td>
			<td style="width:119px;">N/A</td>
			<td>2 per box, 0.64 cm thick x 2.5 cm wide x 57 cm long</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">HDPE spacers (bottom)</td>
			<td style="width:123px;">TAP Plastics</td>
			<td style="width:119px;">N/A</td>
			<td>1 per box, 0.64 cm thick x 2.5 cm wide x 40.5 cm long</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">HDPE spacers (patch)</td>
			<td style="width:123px;">TAP Plastics</td>
			<td style="width:119px;">N/A</td>
			<td>2 per box, 0.64 cm thick x 3.8 cm wide x 28 cm long</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Polyester batting</td>
			<td style="width:123px;">Fairfield</td>
			<td>#A-X90</td>
			<td>2.5 cm x 40.5 cm strip per box</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">20-thread screws</td>
			<td style="width:123px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>3.2 cm long, 0.64 cm diameter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Washers</td>
			<td style="width:123px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>0.64 cm internal diameter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hex nuts</td>
			<td style="width:123px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>sized to fit the screws</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Light deprivation fabric</td>
			<td style="width:123px;">Americover, Inc.</td>
			<td>Bold 8WB26.5</td>
			<td>1 piece 95 cm wide and 69 cm long per box</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sand</td>
			<td style="width:123px;">Quikrete</td>
			<td>No. 1113</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Field soil</td>
			<td style="width:123px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Transparencies for tracing</td>
			<td style="width:123px;">FXN</td>
			<td>FXNT1319100S</td>
			<td>One per side of the box to be traced</td>
		</tr>
	</tbody>
</table>

an optimized rhizobox protocol to visualize root growth and responsiveness to localized nutrients

根是众所周知的难以研究。土壤既是一种视觉和机械屏障, 在没有破坏性收获或昂贵设备的情况下, 很难在原地跟踪根。我们提出了一种可自定义且经济实惠的 rhizobox 方法, 可在一段时间内实现根系生长的无损可视化, 特别适合研究本地化资源修补程序的根塑性。通过对含有15个 n-标记豆科植物残渣的斑块的塑性响应进行了评价, 验证了玉米基因型的变化。方法描述了随着时间的推移获得代表性的发育测量值, 测量含资源和控制斑块中的根长度密度, 计算根系生长速率, 并通过植物根和芽来确定15N 的回收率。本文还讨论了该方法的优点、注意事项和潜在的未来应用。尽管必须注意确保实验条件不偏向根增长数据, 但在这里提出的 rhizobox 协议如果进行得足够注意细节, 就会产生可靠的结果。

像预期的那样, 根在盒子后面的生长优先。盒背面的总追踪根长度介于400到 1956 cm 之间, 与盒前93-758 厘米相比。在扫描的根长度和追踪的根长度之间, 在盒子的前部、盒子的背面, 计算出成对皮尔逊相关系数, 并使用前后的总和来确定跟踪是否准确地反映了总根长度 (n = 23, 如在一个盒子里的植物在实验中死了)。扫描的总根长度与盒背面的追踪根长度显著相关 (<strong class="xfig...

Watch this Scientific Journal Video about An Optimized Rhizobox Protocol to Visualize Root Growth and Responsiveness to Localized Nutrients at JoVE.com

An Optimized Rhizobox Protocol to Visualize Root Growth and Responsiveness to Localized Nutrients

一种优化的 Rhizobox 协议, 用于可视化根系生长和对局部营养素的响应

在原位可视化和测量根系生长是极具挑战性的。我们提出了一种可自定义的 rhizobox 方法, 可根据营养丰富的时间跟踪根系发育和增殖。该方法用于分析玉米根系可塑性对有机氮源反应的基因型差异。

Roots are notoriously difficult to study. Soil is both a visual and mechanical barrier, making it difficult to track roots in situ without destructive ...

这种方法可以帮助回答根和Rhizsphere科学领域的关键问题，例如根如何回应局部的营养物质斑块。该技术的主要优点是，它允许重复的，非侵入性测量在土壤中的单个根系，但它不需要专门的设备，它相对便宜。收集 Rhizobox 的零件以进行装配。其中包括由透明丙烯酸制成、约40厘米、61厘米的正面和背面面板。此时，后面板顶部有侧面和底部的空格。空间器与面板中以前钻过的孔对齐。切割聚酯填充的长度，沿 Rhizobox 的底部。将衬垫放在底部垫垫正上方的后面板上，以帮助防止泄漏。将顶部面板放在垫片上，以将衬垫放置到位。现在获取硬件来组装 Rhizobox。每个钻孔都使用螺钉、两个垫圈和六角螺母。将垫圈放在前面板上，然后将螺钉拧紧到后面板上。另一个垫圈和十六进制螺母。为避免土壤流失，请确保所有螺钉都很紧。此外，创建两个空格来形成处理和控制补丁。用高密度聚乙烯板制作，并在顶部固定螺钉，仅允许部分插入。对于光剥夺和减少热量，准备一个保护箱，可以包围盒子。最后，构建一个框架，鼓励根在后面板上生长。此 PVC 框架以大约 55 度的角度将盒子保持。准备好田间土壤和沙子的均质混合物。从一个大袋子开始，里面装着每个 Rhizobox 的基底混合物。每个盒子标有两个小拉链袋，用于处理和控制补丁。使用秤，重达 30 克的土砂基材。将基板转移到控制袋中。同样，在治疗袋中加入30克基板。然后称一克氮15标签氮源，并将其添加到处理袋中。将基板和氮气源彻底混合。使用一个 Rhizobox 和它的补丁贴片器。将两个空格器插入 Rhizobox，直到螺钉防止进一步运动。标记 Rhizobox 侧面的空格基底边缘的深度。然后删除下一步的空格。使用漏斗将 Rhizobox 从大袋基材填充到标记的深度。缓慢均匀地来回移动漏斗，以均匀地填充 Rhizobox。当基板处于显著深度时，将空间器放入包装盒中。它们应该距离盒子的左右两侧五厘米。继续灌装盒子，直到基板距离盒子顶部约五厘米。接下来，用150毫升的水或用手，安排在包装盒中的基板缓慢灌溉。让它休息几个小时或过夜，使水扩散，每个空间周围的区域是彻底湿。此时，拆下空格，为贴片留空空腔。现在，磁带透明膜到盒子的外面。使用透明度将一个修补程序标记为处理，另一个修补程序标记为控件。然后使用漏斗和准备好的控制基板填充控制补丁。这是填充后标记的控制修补程序。继续用准备好的治疗基板以同样的方式填充处理补丁。完成后，使用永久标记跟踪透明度胶片上每个修补程序的边界。接下来，用大袋中剩余的基材均匀地填充 Rhizobox。通过跟踪透明度上的基板顶部完成。盒子准备好后，移植发芽的种子。首先，使用狭窄的铲子在 Rhizobox 的中心挖一个洞。这个洞应该深2.5厘米。将发芽的种子放在里面，确保基是直接向下。跟踪附加到框的透明度上种子的位置。盖上种子，然后用高达50毫升的去化水浇水。接下来，将框封闭在它的情况下，以防止光在生长过程中与根相互作用。将盒子放在准备的框架上。将 Rhizobox 放在框架上，以鼓励向后框架增长。每三到四天的植物生长后取回盒子。移除外壳后，每次使用不同的彩色笔跟踪植物可见的根。与本示例一样，此步骤要保持一致和系统化。将盖盒返回到其支架，并确保其位于其原始方向。在这里，24 个 Rhizobox 背面的跟踪根的总根长度根据使用扫描仪和计算机软件测量的根长度绘制。框前面跟踪根图案的类似图显示根在框的背面优先生长。此图中总根长度对数的一致斜率表示不同框之间的增长率相似。以下是一种玉米基因型的根长密度数据。控制片的密度小于氮处理片的密度。实验比较了六种独立玉米基因型的根长密度。在所有情况下，与控制贴片相比，氮 15 标记处理贴片中的根长密度更大。该协议可以与其他方法相结合，如植物学或浮游原位杂交，以可视化酶活性或微生物的分布。

Research

JoVE Journal

Environment

Optimize Flue Gas Settings to Promote Microalgae Growth in Photobioreactors via Computer Simulations

优化烟气设置通过电脑模拟来促进微藻生长的光生物反应器中

Flue gas from power plants can promote algal cultivation and reduce greenhouse gas emissions1. Microalgae not only capture solar energy more efficiently ...

科研

环境科学

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy

优化负染：通过电子显微镜检查小和不对称蛋白结构的高通量协议

Structural determination of proteins is rather challenging for proteins with molecular masses between 40 - 200 kDa. Considering that more than half of ...

A Protocol for Conducting Rainfall Simulation to Study Soil Runoff

一个协议，用于开展降雨模拟研究土壤径流

Rainfall is a driving force for the transport of environmental contaminants from agricultural soils to surficial water bodies via surface runoff. The ...

Measuring Fluxes of Mineral Nutrients and Toxicants in Plants with Radioactive Tracers

植物与放射性示踪剂测量矿质营养和毒物的通量

Unidirectional influx and efflux of nutrients and toxicants, and their resultant net fluxes, are central to the nutrition and toxicology of plants. ...

A Push-pull Protocol to Reduce Colonization of Bird Nest Boxes by Honey Bees

推挽议定书蜜蜂会影响家禽的殖民化巢箱

Introduction of the invasive Africanized honey bee (AHB) into the Neotropics is a serious problem for many cavity nesting birds, specifically parrots. ...

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

Ø的底栖交易所<sub&gt; 2</sub&gt;，N<sub&gt; 2</sub&gt;和营养盐使用小的核心孵化

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in ...

Laboratory Simulation of an Iron(II)-rich Precambrian Marine Upwelling System to Explore the Growth of Photosynthetic Bacteria

Laboratory Simulation of an IronII-rich Precambrian Marine Upwelling System to Explore the Growth of Photosynthetic Bacteria

一个铁（II）的实验室模拟的前寒武纪富海洋上升流系统探索光合细菌生长

A conventional concept for the deposition of some Precambrian Banded Iron Formations (BIF) proceeds on the assumption that ferrous iron [Fe(II)] upwelling ...

Soil Lysimeter Excavation for Coupled Hydrological, Geochemical, and Microbiological Investigations

土壤蒸渗仪开挖加之水文，地球化学和微生物调查

Studying co-evolution of hydrological and biogeochemical processes in the subsurface of natural landscapes can enhance the understanding of coupled ...

A Gusseted Thermogradient Table to Control Soil Temperatures for Evaluating Plant Growth and Monitoring Soil Processes

一个插角Thermogradient表来控制土壤温度为评估植物生长和监测土壤过程

Thermogradient tables were first developed in the 1950s primarily to test seed germination over a range of temperatures simultaneously without using a ...

Video Tracking Protocol to Screen Deterrent Chemistries for Honey Bees

用于蜂蜜蜜蜂筛选化学成分的视频跟踪协议

The European honey bee, Apis mellifera L., is an economically and agriculturally important pollinator that generates billions of dollars annually. Honey ...