Name: detection of phytophthora capsici in irrigation water using loop-mediated isothermal amplification
Uploaded: 2020-06-25T13:00:00
Description: Watch this Scientific Journal Video about Detection of Phytophthora capsici in Irrigation Water using Loop-Mediated Isothermal Amplification at JoVE.com

这项工作得到了佐治亚州蔬菜商品委员会项目ID#FP00016659的财政支持。作者感谢佐治亚大学季平生博士和俄亥俄州立大学的安妮·多伦斯博士提供 纯正的菲托波拉spp文化。我们还感谢王丽和德洛里斯·维尼在整个研究中的技术援助。

1. 使用便携式循环 介质 等温扩增从灌溉水中对植物浮游水进行现场检测
<ol><li>设置泵和过滤器<ol>
<li>将滤瓶连接到连接到手泵的管子上，以便当泵激活时，空气会通过过滤瓶口拉进。</li>
		<li>将 Buchner 漏斗放入滤瓶口的橡胶塞中，并将适当大小的滤纸放入布赫纳漏斗中，以便空气通过滤纸输送。滤纸的保留尺寸应为 15 μm。 注：滤纸必须适合布赫纳漏斗的边缘，以便最少的水会流经滤?...

<ol>
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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plant+pathogens+in+recycled+irrigation+water+in+commercial+plant+nurseries+and+greenhouses:+their+detection+and+management.">Plant pathogens in recycled irrigation water in commercial plant nurseries and greenhouses: their detection and management.</a> Irrigation Science. 29 (4), 267-297 (2011).</li><li>Aragaki, M., Uchida, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphological+distinctions+between+Phytophthora+capsici+and+P.+tropicalis+sp.+nov.">Morphological distinctions between Phytophthora capsici and P. tropicalis sp. nov.</a> Mycologia. 93 (1), 137-145 (2001).</li><li>Tomlinson, J., Boonham, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+of+LAMP+for+detection+of+plant+pathogens.">Potential of LAMP for detection of plant pathogens.</a> CAB Reviews Perspectives in Agriculture Veterinary Science Nutrition and Natural Resources. 3 (066), 1-7 (2008).</li><li>Li, P., 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+PCR-based+assay+for+distinguishing+between+A1+and+A2+mating+types+of+Phytophthora+capsici.">A PCR-based assay for distinguishing between A1 and A2 mating types of Phytophthora capsici.</a> Journal of the American Society for Horticultural Science. 142 (4), 260-264 (2017).</li><li>Dong, Z., 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=Loop-mediated+isothermal+amplification+assay+for+sensitive+and+rapid+detection+of+Phytophthora+capsici.">Loop-mediated isothermal amplification assay for sensitive and rapid detection of Phytophthora capsici.</a> Canadian Journal of Plant Pathology. 37 (4), 485-494 (2015).</li><li>Khan, 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=Comparative+evaluation+of+the+LAMP+assay+and+PCR-based+assays+for+the+rapid+detection+of+Alternaria+solani.">Comparative evaluation of the LAMP assay and PCR-based assays for the rapid detection of Alternaria solani.</a> Frontiers in Microbiology. 9, 2089 (2018).</li><li>Sowmya, N., Thakur, M., Manonmani, H. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+and+simple+DNA+extraction+method+for+the+detection+of+enterotoxigenic+Staphylococcus+aureus+directly+from+food+samples:+comparison+of+PCR+and+LAMP+methods.">Rapid and simple DNA extraction method for the detection of enterotoxigenic Staphylococcus aureus directly from food samples: comparison of PCR and LAMP methods.</a> Journal of Applied Microbiology. 113 (1), 106-113 (2012).</li><li>Waliullah, S., 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=Comparative+analysis+of+different+molecular+and+serological+methods+for+detection+of+Xylella+fastidiosa+in+blueberry.">Comparative analysis of different molecular and serological methods for detection of Xylella fastidiosa in blueberry.</a> PLOS ONE. 14 (9), 0221903 (2019).</li><li>B&#246;hm, 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=Real-time+quantitative+PCR:+DNA+determination+in+isolated+spores+of+the+mycorrhizal+fungus+Glomus+mosseae+and+monitoring+of+Phytophthora+infestans+and+Phytophthora+citricola+in+their+respective+host+plants.">Real-time quantitative PCR: DNA determination in isolated spores of the mycorrhizal fungus Glomus mosseae and monitoring of Phytophthora infestans and Phytophthora citricola in their respective host plants.</a> Journal of Phytopathology. 147, 409-416 (1999).</li><li>Klimczak, L., Prell, H. 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P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=LAMP+detection+of+four+plant-pathogenic+oomycetes+and+its+application+in+lettuce+fields.">LAMP detection of four plant-pathogenic oomycetes and its application in lettuce fields.</a> Plant Disease. 103 (2), 298-307 (2019).</li><li>Aglietti, C., 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=Real-time+loop-mediated+isothermal+amplification:+an+early-warning+tool+for+quarantine+plant+pathogen+detection.">Real-time loop-mediated isothermal amplification: an early-warning tool for quarantine plant pathogen detection.</a> AMB Express. 9 (1), 50 (2019).</li><li>Almasi, M. 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对植物病菌的灌溉水进行测试，是种植者使用灌溉池塘和再生水的关键一步。灌溉池为一些植物病菌提供了一个水库和繁殖地，因为过量的灌溉水从田间流向池塘，携带任何可能存在16、27,的病原体。在大型水源中检测植物病原体的传统方法是使用悬浮在池塘中的易感宿主组织（如水果、叶子）为病原体设置诱饵，等待感染发生，然后...

由于水成本增加和用水背后的环境问题，农场和托儿所的回收用水正变得越来越受欢迎。为种植者开发了许多灌溉方法，以减少植物病的传播和发生。无论水源（灌溉或降水），径流都会产生，许多蔬菜和苗圃种植者都有一个池塘来收集和回收径流1。这创造了一个水库，为可能的病原体积累有利于病原体的传播，当再生水被用来灌溉作物2，3，4。,3,4Oomycete植物病原体特别受益于这种做法，因为动物孔会积聚在水中，主要分散孢子是自动的，但需要地表水5，6，7。,6,7植物性植物素是一种异种病原体，以不同的方式影响大量的多叶和库伯特作物。通常，症状是阻尼幼苗，根和冠腐烂;然而，在黄瓜，南瓜，甜瓜，南瓜，西瓜，茄子和胡椒等作物，整个收成可能会因为水果腐烂而损失。虽然已知有检测这种植物病原体的方法，但大多数都要求已经发生感染，这为时已晚，任何预防性杀菌剂都不能产生显著的影响10。
传统的灌溉水测试方法，用于检测和诊断目标微生物是一种过时的方法，当速度和敏感性是成功的关键和有利可图的作物生产11，12。,12易受靶向病原体影响的植物组织（例如，P. 辣椒茄子）附着在经过改良的陷阱上，该陷阱在被移除和检查感染之前被悬浮在灌溉池中较长时间。然后将植物组织的样品镀在半选择性介质（PARPH）上，并孵育成培养，然后使用复合显微镜13进行形态识别。其他植物病原体有其它类似的检测方法，使用选择性培养，并在分培养14、15之前，先将少量受污染的水电镀。这些方法需要2到6周，几轮亚培养来分离生物体，并体验植物预防诊断，以便能够识别每个物种的关键形态特征。由于水源中其他微生物的干扰等因素，这些传统方法对受P.辣椒污染的灌溉水检测不成功。一些快速生长的微生物，如Pythium spp.和水传播的细菌，可以在盘子里过度生长，使P. capsici无法检测到16，17。,17
本研究的目的是开发一种敏感和特定的分子方法，可用于田间和实验室设置，以检测灌溉水中的P.辣椒动物孔。该协议包括开发一个新的循环介质等温放大（LAMP）底像集，能够特别放大P.卡普西奇，基于1121基对（bp）片段的P.capsici18，19。,19与为这项研究开发20的测定相比，使用了由董等人（2015年）开发的LAMP底像。
LAMP检测是一种相对较新的分子检测形式，已被证明比常规聚合酶链反应（PCR）21更快速、更灵敏、更特异性。一般来说，传统的PCR测定不能检测低于500份（1.25皮克/μL）;相比之下，先前的研究表明，LAMP的灵敏度可以比传统PCR高10至1，000倍，并且能够轻松检测出1 fg/μL的基因组DNA22，23。,23此外，通过使用便携式加热块进行放大和色度染料改变阳性样品的颜色（无需电泳），可以快速（通常 30 分钟内）和现场（现场）进行测定。本研究中，我们利用滤波器提取方法比较了PCR和LAMP测定的灵敏度。拟议的检测方法使研究人员和推广剂能够在不到两个小时的时间轻松检测不同水源中P.辣椒孢子的存在。该测定方法证明比传统的PCR更敏感，通过检测种植者使用的灌溉水中的病原体存在，得到了原地验证。这种检测方法将使种植者能够估计用于灌溉的各种水源中病原体的存在和种群密度，防止破坏性爆发和经济损失。

<table>
	<tbody>
		<tr>
			<td>Agarose gel powder</td>
			<td>Thomas Scientific</td>
			<td>C997J85</td>
			<td></td>
		</tr>
		<tr>
			<td>Buchner funnel</td>
			<td>Southern Labware</td>
			<td>JBF003</td>
			<td></td>
		</tr>
		<tr>
			<td>Bullet Blender</td>
			<td>Next Advance</td>
			<td>BBX24</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge 5430</td>
			<td>Eppendorf</td>
			<td>22620509</td>
			<td></td>
		</tr>
		<tr>
			<td>Chloroform</td>
			<td>Fischer Scientific</td>
			<td>C298-500</td>
			<td></td>
		</tr>
		<tr>
			<td>CTAB solution</td>
			<td>Biosciences</td>
			<td>786-565</td>
			<td></td>
		</tr>
		<tr>
			<td>Dneasy Extraction Kit</td>
			<td>Qiagen</td>
			<td>69104</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter Flask</td>
			<td>United</td>
			<td>FHFL1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter Paper</td>
			<td>United Scientific Supplies</td>
			<td>FPR009</td>
			<td></td>
		</tr>
		<tr>
			<td>Gel Green 10000X</td>
			<td>Thomas Scientific</td>
			<td>B003B68 (1/EA)</td>
			<td></td>
		</tr>
		<tr>
			<td>Genie III</td>
			<td>OptiGene</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Hand pump</td>
			<td>Thomas Scientific</td>
			<td>1163B06</td>
			<td></td>
		</tr>
		<tr>
			<td>Iso-amyl Alcohol</td>
			<td>Fischer Scientific</td>
			<td>BP1150-500</td>
			<td></td>
		</tr>
		<tr>
			<td>LAVA LAMP master mix</td>
			<td>Lucigen</td>
			<td>30086-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnetic bead DNA extraction</td>
			<td>Genesig</td>
			<td>genesigEASY-EK</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnetic Separator</td>
			<td>Genesig</td>
			<td>genesigEASY-MR</td>
			<td></td>
		</tr>
		<tr>
			<td>polyvinylpyrrolidone</td>
			<td>Sigma Aldrich</td>
			<td>PVP40-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Primers</td>
			<td>Sigma Aldrich</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Prism Mini Centrifuge</td>
			<td>Labnet</td>
			<td>C1801</td>
			<td></td>
		</tr>
		<tr>
			<td>T100 Thermal Cycler</td>
			<td>Bio-Rad</td>
			<td>1861096</td>
			<td></td>
		</tr>
		<tr>
			<td>UV Gel Doc</td>
			<td>Analytik Jena</td>
			<td>849-00502-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Warmstart Colorimetric Dye</td>
			<td>Lucigen</td>
			<td>E1800m</td>
			<td></td>
		</tr>
		<tr>
			<td>Wide Mini ReadySub-Cell GT Cell</td>
			<td>Bio-Rad</td>
			<td>1704489EDU</td>
			<td></td>
		</tr>
		<tr>
			<td>70% isopropanol</td>
			<td>Fischer Scientific</td>
			<td>A451-1</td>
			<td></td>
		</tr>
	</tbody>
</table>

detection of phytophthora capsici in irrigation water using loop-mediated isothermal amplification

植物性浮肿是一 种毁灭性的奥米妥特病原体，它影响许多重要的冬虫夏菜和库伯特作物，每年给蔬菜生产造成重大经济损失。 植物浮游生物是 土壤传播的，由于其长期生存结构（卵源和衣原体）能够抵抗风化和降解，在菜地中是一个长期存在的问题。分散的主要方法是通过生产动物孔，这是单细胞的，旗状孢子，可以游过表面或充满水的土壤毛孔的薄膜，可以积累在水坑和池塘。因此，灌溉池可能是病原体和疾病爆发的最初点源。在 灌溉水中检测P. 辣椒很难使用基于传统文化的方法，因为环境中存在的其他微生物， 如Pythium spp.，通常过度生长 P.辣椒 ，使其检测不到。为了确定水源 （灌溉 水、径流等）中是否存在P.辣椒孢子，我们开发了一种基于手泵的滤纸（8-10μm）方法，该方法捕获病原体的孢子（zoospore），后来用于通过一种新型的循环调节等温（LAMP）分析来放大病原体的DNA，该测定法旨在对 P. 这种方法可以放大和检测低浓度为1.2×102 的DNA，其灵敏度是传统PCR的40倍。在测试密切相关的物种时，没有获得交叉放大。LAMP 还使用色度 LAMP 主混合染料进行，显示可用肉眼读取的结果，以便现场快速检测。该协议可适用于通过受污染的灌溉系统居住、积累或分散的其他病原体。

LAMP 方法的优化 在这项研究中，我们利用便携式 循环介质等温 扩解（LAMP）测定，检测了灌溉水中存在植物浮肿。首先，通过测试不同的LAMB底因浓度[F3，B3（每个0.1±0.5μM），对拟议的LAMB测定进行了优化;LF、LB（每磅0.5~1.0 μM）和FIP、BIP（每片0.8~2.4 μM）、持续时间（30~70分钟）和温度（55~70 °C）。本研究使用的最终 LAMP 底木组合是：每个 F3 和 B3 底注的 0.2 μM，每个环路-F 和?...

Watch this Scientific Journal Video about Detection of Phytophthora capsici in Irrigation Water using Loop-Mediated Isothermal Amplification at JoVE.com

Detection of Phytophthora capsici in Irrigation Water using Loop-Mediated Isothermal Amplification

我们开发了一种 利用 滤纸DNA提取方法检测水源中植物性浮肿的方法，并结合循环中等温扩增（LAMP）测定法，可现场或实验室进行分析。

使用 循环介质等温扩 增检测灌溉水中的植物浮肿

Phytophthora capsici is a devastating oomycete pathogen that affects many important solanaceous and cucurbit crops causing significant economic losses in ...

植物病原检测是植物病管理的关键一步。该协议为灌溉水污染提供了一种快速检测方法，该方法由一种常见的水性战斗机病原体引起。利用这种技术，特定的病原体，如植物性病原体，可以处理，并快速检测从我们大量的灌溉水，同时留在现场。演示这个程序的将是欧文哈德森，一个硕士学生从我的实验室，和苏米亚瓦利乌拉，博士后研究员从博士平西G的实验室，建立一个泵和过滤器现场植物托波拉卡皮奇检测灌溉水。将滤瓶连接到连接到手泵的管子上，并将布赫纳漏斗安装到过滤瓶口的橡胶塞中。然后将一块大小合适的滤纸放入漏斗中，其保留尺寸为 15 微米。获取水样，从目标水源收集灌溉水。水可能有少量的碎屑，但沉淀物或土壤并不明显。在使用手泵形成吸力以将水通过漏斗时，将 50 到 1000 毫升的试水缓慢地倒在漏斗内的过滤纸上。过滤所有水后，使用钳子从漏斗中取出滤纸。用无菌剪刀把纸剪成小块。将8至12张纸在400微升的萃取缓冲液中，在1.5毫升的管子中，在室温下孵育5分钟。漩涡或以其他方式每分钟搅拌纸张10秒。在孵育结束时，使用钳子用尽可能少的缓冲液取出纸张。然后用下一组滤纸重复许可证。在从滤纸样本中提取DNA时，将20微升蛋白酶K和10微升每微升RNA10纳米图添加到样品中，并在室温下孵育溶液15分钟，每三分钟进行涡流或摇动。在孵化结束时，在样品中加入500微升的磁珠，通过摇动混合。在室温下五分钟后，将管子和磁性分离器机架放置两分钟，然后取出并丢弃上经剂。从磁性分离器上拆下管，并在 500 微升洗涤缓冲液中大力重新悬浮磁珠。30 秒后，将管子返回磁铁，等待两分钟，然后丢弃上经剂。使用 500 微升的洗涤缓冲液和 500 微升的 80% 乙醇重复洗涤，正如刚刚证明的。空气在室温下驱动磁珠颗粒15分钟。在孵化结束时，将珠子重新悬浮在50微升洗脱缓冲液中，并移液1分钟，然后将管子放回磁铁上两分钟，以便收集上经剂。如表所示，在准备LAMP底流器混合后，在2.5微升的底材混合12.5微升的熔岩LAMP主混合一微升提取的DNA和9微升的双蒸馏水到PCR管，并设置一个正和一阴性对照。在64摄氏度的热博客中孵育所有样品45分钟，或在孵化结束时将精灵三个扩增仪器孵育，每个放大样品的5微升负载到1%的加糖凝胶上，并在紫外线成像机器上成像结果波段。如果使用热量测量染料，则查看颜色变化以确定结果为正或负。如果使用便携式放大仪器，请查看屏幕上的放大图以确定结果。在64度运行LAMS检测的光学时间是45分钟，因为检测结果为正的最低浓度直到40分钟才被放大，而高浓度在20分钟内被放大。扩增可以在标有核酸污渍的 1%agarose 凝胶上确认。如图所示，传统 PCR 的灵敏度比 LAMP 小 40 倍。检测4.8倍10至第三动物园孔每毫升浓度最低。在这项分析中，从佐治亚州蒂夫特县用于商业蔬菜生产的七个池塘中采集了水样。其中三个显示了积极的LAMLAM结果。然而，当用传统的PCR测试样品时，只在三个阳性样本中的一个中检测到动物孔隙。在表中，总结了使用灵敏度、时间、所需准备、材料和成本等变量的检测方法之间的差异。LAMP 是测试方法中成本较低的方法，也是最快的方法，与传统 PCR 扩增相比，只需 30 到 60 分钟的放大时间。使用该协议从密切相关的UMI种子病原体中提取的DNA导致任何样品的检测，确认底源的特异性。确保不要过快地倒出样品水，在将提取的DNA添加到LAMS管以限制污染时要小心。一旦其他水性病原体已经开发出其特定的底套，过滤和灯酸方法就可以适应其他病原体。

Research

JoVE Journal

Environment

Detection of Phytophthora capsici in Irrigation Water using Loop-Mediated Isothermal Amplification

Colorimetric Paper-based Detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from Large Volumes of Agricultural Water

Colorimetric Paper-based Detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from Large Volumes of Agricultural Water

比色纸为基础的检测<em&gt;大肠杆菌</em&gt;<em&gt;沙门氏菌</em&gt;属，和<em&gt;单核细胞增生李斯特氏菌</em&gt;从农业用水大容量

This protocol describes rapid colorimetric detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from large volumes (10 L) of ...

科研

环境科学

Laboratory-determined Phosphorus Flux from Lake Sediments as a Measure of Internal Phosphorus Loading

实验室确定的磷通量湖泊沉积物作为内部磷载入一个测量

Eutrophication is a water quality issue in lakes worldwide, and there is a critical need to identify and control nutrient sources. Internal phosphorus (P) ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. I. Collection of Virus Samples

EPA方法1615测量肠道病毒和诺瓦克病毒发生水由文化和RT-qPCR的了。病毒样本的收集I.

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. ...

A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated (WATER-PAM) Fluorometry

A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated WATER-PAM Fluorometry

小体积的生物测定评估细菌/浮游植物共培养使用水脉冲幅度调制（WATER-PAM）荧光

Conventional methods for experimental manipulation of microalgae have employed large volumes of culture (20 ml to 5 L), so that the culture can be ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR

EPA方法1615测量肠道病毒和诺瓦克病毒发生水由文化和RT-qPCR的了。第三部分。病毒检测用RT-qPCR的

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a ...

A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters

A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters

双工数字PCR方法的同时定量<em&gt;肠球菌。</em&gt;和人的粪便相关HF183标记的水域

This manuscript describes a duplex digital PCR assay (EntHF183 dPCR) for simultaneous quantification of Enterococcus spp. and the human fecal-associated ...

Screening for Endocrine Activity in Water Using Commercially-available In Vitro Transactivation Bioassays

Screening for Endocrine Activity in Water Using Commercially-available In Vitro Transactivation Bioassays

在用水筛查内分泌活动市售的<em&gt;体外</em&gt;式激活生物测定

In vitro transactivation bioassays have shown promise as water quality monitoring tools, however their adoption and widespread application has been ...

A Simple Planting Technique for Re-establishing Trees Where Frequent Inundation Occurs

常淹发生的重构树简易造林技术

Many of the Everglades tree islands have lost elevation over the past century and most of their trees have died such that they are now covered with ...

Bioindication Testing of Stream Environment Suitability for Young Freshwater Pearl Mussels Using In Situ Exposure Methods

Bioindication Testing of Stream Environment Suitability for Young Freshwater Pearl Mussels Using In Situ Exposure Methods

用原位曝光法测定淡水珍珠贻贝 Bioindication 的流环境适宜性

Knowledge of habitat suitability for freshwater mussels is an important step in the conservation of this endangered species group. We describe a protocol ...

Optimized Procedure for Determining the Adsorption of Phosphonates onto Granular Ferric Hydroxide using a Miniaturized Phosphorus Determination Method

用小型化磷测定法测定膦酸在粒状氢氧化铁上吸附的优化方法

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric ...

使用 循环介质等温扩 增检测灌溉水中的植物浮肿

使用循环介质等温扩增检测灌溉水中的植物浮肿