This project was funded by the Government of Canada through Genome Canada and the Ontario Genomics Institute (OGI-046), and Ontario Research Fund&#8211;Global Leadership in Genomics and Life Sciences GL2-01-035 (to M.G. and V.G.). T.V.L. is a postdoctoral fellow of the Fund for Scientific Research Flanders (FWO).

1，维护了叶螨种群的注：蜘蛛螨饲养加州红芸豆（ 菜豆 ）。 <ol><li>种植豆类植物的种子从2-3周前为患。 </li><li>与受侵染植物混用这些植物;成螨会迅速拓殖新鲜的植物材料。 </li><li>删除旧的侵染豆科植物每隔7-10天，用新鲜的植物取代。 </li></ol> 2，收集成年女性螨<ol><li>使用洗涤方法收集螨<ol><li>为了收集螨虫，放置一个新鲜的豆科植物与植物侵染接触实验前1〜2天。 </li><li>用20〜30鲜豆类植物，收集1000〜2000成年女性螨。 </li><li>用自来水在RT制备吐温20的溶液中于0.001％。 </li><li>在吐温20溶液清洗出没的豆类植物，使用2〜3个工厂的时间。 </li><li&gt;每个工厂洗涤2〜3倍，以收集所有叶螨。 </li><li>完成步骤2.1.4在10分钟内，以避免螨死亡率。 </li></ol></li><li>成年女性螨分离的螨过滤悬浮液通过定义网格大小筛<ol><li>准备以下网目尺寸的筛子：500微米（以去除碎片）和300微米（收集雌性成年螨）（ 图1A）。 </li><li>通过500微米的筛子过滤悬浮液。 </li><li>通过300微米的筛子重新筛选暂停。成年女性螨虫将被保留。 </li><li>沾了300微米筛成年女性螨在干净的自来水，除去吐温20。 </li><li>在上面的筛子的底部单层散布螨。 </li><li>用纸巾从筛的网眼和侧面除去过量的水。注意：这样做是因为螨虫需要，以恢复迅速干透即可。 </li><li>准备一个组件， 如图1所示乙;这将允许螨自由移动，但会阻止他们逃跑。 </li><li>将筛上，是用来作为支撑筛的顶螨。 </li><li>环绕声底筛与水，以防止螨虫从分散。注：螨将开始恢复后约5分钟。后约30分钟，移动螨虫的数量是足以启动集合（ 图1D）。螨应在1小时内被收集，因为它们产生的丝，将与集合中产生干扰。 </li></ol></li></ol> 3，植物侵染螨注意：一旦成年雌性螨已痊愈，它们可用于植物侵染。有两种方法用来侵扰实验工厂与成年女性螨：a）使用细刷（协议第3.1节）和b）用泵或真空线（协议第3.2节）。 <ol><li>螨虫为患使用Brush 注：红蜘蛛为害用刷子被用于应用程序的每株最高30螨。 <ol><li>使用大小00或更细的软毛圆艺术画笔。 </li><li>湿刷R​​T自来水。 </li><li>轻轻地捡起筛螨使用画笔笔尖，并将它们传送到工厂。 </li></ol></li><li>用泵螨感染 注意：植物侵染由泵/真空线螨收集时，使用超过30个螨虫被应用。对于这种方法，可以用一个真空泵用于微电子元件拾取或真空管线。它保持恒定气流具有足够的强度和（2-4磅）是重要的。 <ol><li>附1毫升移液管尖，以连接到泵或真空线用1.5ml的离心管中切的底部作为泵/真空管和所述顶端之间的适配器管中。 </li><li>将枪头和适配器（1.5毫升离心管）之间的一块纸巾。注：这样做的目的是为了里面吨螨虫陷阱他的枪头在抽吸过程，同时也减少了气流（ 图1C）。 </li><li>直接使用枪头筛收集螨。 </li><li>另外，从使用立体镜感染的叶子收集螨一个接一个。 </li><li>当已收集螨所需数量的，从管取出枪头，确保了一块纸巾在移液管尖的背面原状。 </li><li>通过点击枪头和结块的螨虫一起收集螨。然后，将它们放置在叶片上;几秒钟内，螨虫会开始驱散在叶片上。 </li></ol></li></ol> 4，记录和评估植物损伤<ol><li>录影厂损伤 注：对于A。拟南芥 ，厂房损坏，经过4天的螨虫喂养评估。损坏记录为褪绿斑的总表面积。对于损坏的测量，整个莲座被切割和扫描。描述扫描参数是一个爱普生V30扫描仪，但将作为一个很好的起点，任何类似的平板扫描仪。保留所有实验的扫描参数不变，允许个别实验运行之间的比较。 <ol><li>覆盖扫描仪床上，透明片，以防止污染扫描器表面的植物材料。 </li><li>切割整个莲座和地点上的扫描仪床上所以叶子的正面的（上）侧朝向扫描仪的光源和捕获元件。另外，如果花环过于密集捕获单个叶片不重叠，解剖莲座成单个叶片或不重叠的叶子用细剪刀组放置在扫描仪上床前。多个莲座可以同时被扫描。 </li><li>覆盖玫瑰花或树叶用一张白纸，以防止遵守和扫描器盖板的污染。 </li><li>关闭扫描仪盖板，并以下列参数进行扫描分辨率：1,200 DPI;色彩模式：Adobe RGB的;亮度：25;文件类型：最高质量JPEG格式。 </li><li>保存图像文件，用于后续的损害分析。 </li></ol></li><li>对植物的损伤定量 注：损伤的面积是使用Photoshop手工计算。由于叶片的形状和症状的颜色的强度作为螨馈送的结果，大的差异，自动的方法是不可靠的。因此，网格的方法是用来评估对植物的损伤： <ol><li>打开工厂用Photoshop的图像。 </li><li>创建一个新层，然后覆盖扫描图像为0.25 mm×0.25毫米区划网格（视图或显示 - 显示 - 网格，键盘快捷键Ctrl +&#39;）。 </li><li>在一个覆盖层，利用标记点（使用“铅笔工具”，快捷键B）损坏叶面积。用于网格下面这有损坏覆盖超过一半的网格单元中的每方一一个点。点的大小是在像素定义（ 即 </eM&gt;每点52像素）。 </li><li>当电网中所有的广场，展示植物的伤害，涵盖至少有一半的单元格中都标有一个圆点，使用直方图工具（窗口 - 直方图），以确定该层上的像素的总数。直方图工具允许一个测量的像素的数目。由于每一个点是由像素的恒定和已知的数来表示，直方图工具将测量的总像素数，代表点的总数，并且，通过扩展，标志着正方形的总数。 </li><li>标计算使用公式“点”的数量： 点阵数=每点像素的像素/数总数。 </li><li>通过1平方网格的区域在图像上标记相乘的点的数量，作为一个点对应于电网的1平方计算损害的总面积。 </li></ol></li></ol>

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The authors have nothing to disclose.

该视频演示了用于分离和侵扰的植物有大量的成年女性螨的协议。虽然我们使用甲。此协议芥 ，它可用于任何植物的叶螨相互作用系统，目前正在顺利也适用于番茄和葡萄（ 葡萄 ）的植物。该协议的产量可再现的结果，表明收集的螨虫是可比的生理状态（ 图3）。 虽然这些协议是简单的执行，有几个关键的步骤需要特别注意，因为它们会影?...

植物病虫害的相互作用具有重要的科学和经济重要性的话题。它使用两作物植物（如番茄）和模式植物，A.历史研究拟南芥 。在这两种情况下，植物虫害的易感性可以是食草动物后发作或间接地通过对害虫表现的评估测量直接通过植物的表型评估。 植物敏感性的直接测量以前用于使用各种方法的一些害虫种类。例如，鳞翅目幼虫取食被测量为所消耗的任小菜蛾 （小菜蛾）或粉纹夜蛾 （卷心菜尺蠖），用肉眼与电网12的帮助下植物组织的部分的估计。还有，也有利用的叶损害与随后的定量图像分析的数字成像方法。这种方法是在使用A.研究与西花蓟马 （西花蓟马）13，Scaptomyza黄花 （叶采果蝇）14和T拟南芥互动妮 15。 植物易感性间接测量被广泛应用于植物病虫害的互动研究。例如，易感A的拟南芥对桃蚜桃蚜取食通常是通过害虫产卵量和互动16,17后植物的大体形态的描述，分析评估。 A的另一个典型的间接指标拟南芥敏感性害虫是草食动物的干或湿重评估。这个参数是常用来表征取食lepidoterans，如菜粉蝶 （白色小），P.小菜蛾 ，或T。妮在其幼虫或蛹的阶段15,17。 红蜘蛛是细胞铁含量eders。螨虫引起的损伤是公认的褪绿斑点，颜色从白色到浅绿色的集合。植物宿主红蜘蛛虫食的易感性先前评估或者间接地通过对红蜘蛛性能日内进行分析后虫蛀18,19，或直接使用的植物星期后虫蛀18的大体形态，或使用数字树叶的成像曝光几天螨与随后的自动图像分析19。这些方法被开发并用于番茄植株和T之间的相互作用的研究螨，和通常使用少量的蜘蛛螨（5-15每处理）从混合螨种群收集和使用软毛刷被放置在叶片表面上。然而，这些方法不适合于研究，其中需要更大数目的螨虫被应用。此外，在叶片图像直接处理在图像分析软件如Adobe Photoshop（圣何塞，加利福尼亚州）的ImageJ或20可用于番茄损伤分析中，这些协议需要修改以适用于具有表面反射率大于或浅色的并且具有高度可见的毛状体（叶片例如 ， 拟南芥 ）上自动选择褪绿斑，标志着植物伤害干扰。此外，可与以前的方法很容易地利用蜘蛛螨的发育阶段被限制在最普遍的和易于识别的成年女性和排除利用其他发育阶段。 对植物叶螨相互作用的高通量分析的第一个关键步骤是建立可重复的，简单的和可靠的协议，挑战与植物叶螨和可靠的评估互动的结果。 在这段视频中，可快速方便地收集大量图片的有效方法成年女性螨的误码率，他们的实验植物宿主和植物造成的损害红蜘蛛喂养评估上的应用进行了描述。所提出的协议可实现快速和有效地收集数百人在任何发育阶段（卵，幼虫，若虫，成年男性和女性），可以用于后续的实验应用。此外，这些协议可以被应用到任何螨宿主植物，但在A的情况下是特别展示拟南芥 。

<table border="0" cellpadding="0" cellspacing="0" width="1157">
	<tbody>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:231px;">Plant material:</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:231px;">California red kidney bean</td>
			<td style="width:275px;">Stokes, Thorold, ON, Canada</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">Two week old, well infested with spider mites two or three days before use. Other cultivars of Phaseolus vulgaris can be used.</td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:231px;">Chemicals:</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Tween 20</td>
			<td>Sigma-Aldrich</td>
			<td style="width:125px;">P9416</td>
			<td style="width:527px;">1% stock solution is prepared to simplify aliquoting</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:231px;">Tap water</td>
			<td>Any supplier</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">At room temperature, heat- and cold-shock affect mite survival rate and peformance</td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:231px;">Other materials and equipment:</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Plastic tray</td>
			<td>Any supplier</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Set of scissors</td>
			<td>Any supplier</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">2 L beakers</td>
			<td>Any supplier</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Paper towels</td>
			<td>Any supplier</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Sets of sieves</td>
			<td style="width:275px;">Manufactured in house</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">Detailed instructions are available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Thin brush</td>
			<td>Any supplier</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Pipettes</td>
			<td>Any supplier</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Pipette tips</td>
			<td>Any supplier</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">0.2 and 1 mL</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:231px;">1.5 mL centrifuge tubes</td>
			<td>Any supplier</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">-</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:231px;">Air pump</td>
			<td>Any supplier</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">Aquarium type pump with inverted air flow. Vacuum line can be used. Required pressure drop is approx. 2-4 psi</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Stereoscope</td>
			<td>Any supplier</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">-</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:231px;">Scanner</td>
			<td>Epson</td>
			<td style="width:125px;">V30</td>
			<td style="width:527px;">Any flatbed scanner allowing necessary degree of control over scan quality. We use Epson V30 for our experiments.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:231px;">Computer</td>
			<td>Any supplier</td>
			<td style="width:125px;">NA</td>
			<td style="width:527px;">Windows or OS X PC which is compatible with scanner hardware and Adobe Photoshop software.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Adobe Photoshop software</td>
			<td style="width:275px;">Adobe Systems Inc., San Jose, CA, USA</td>
			<td style="width:125px;">various</td>
			<td style="width:527px;">Any version with Histogram tool included.</td>
		</tr>
	</tbody>
</table>

application of two-spotted spider mite tetranychus urticae for plant-pest interaction studies

二斑叶螨， 二斑叶螨 ，是一种普遍存在的多食性节肢动物食草动物，饲料是一个非常广泛的物种阵列，拥有超过150经济价值上。它是温室作物的主要害虫，特别是在茄科和葫芦科 （ 例如 ，西红柿，茄子，辣椒，黄瓜，西葫芦）和温室观赏植物（ 如玫瑰，菊花，康乃馨），年大田作物（如玉米，棉花，大豆，糖用甜菜），和多年生栽培（苜蓿，草莓，葡萄，柑橘，和李子）1,2。除了 ​​极端的polyphagy，使得它的一个重要农业害虫，T。螨有一种倾向，产生耐药性了用于其控制3-7杀虫剂和杀螨剂的广泛。 T.螨是一个很好的实验生物，因为它有一个快速的生命周期（7天，27°C）并且能够容易地维持在高密度在实验室。的方法来测定基因表达（包括原位杂交和抗体染色）和失活的利用RNA干扰红蜘蛛内源基因的表达已开发8-10。近日，T的全基因组序列螨有报道，创造了一个机会发展这种害虫草食动物作为模式生物与已经在一些寄主植物（ 拟南芥和番茄茄lycopersicum）11的存在相当于基因组资源。总之，这些模式生物可以提供深入了解植物害虫相互作用的分子基础。 在这里，可快速方便地收集了大量的成年女性螨，他们的实验厂主机上的应用程序，和植物造成的损害红蜘蛛喂养评估的有效方法进行了描述。所提出的协议连接埃布尔斯快速和有效地收集数百人在任何发育阶段（卵，幼虫，若虫，成年男性和女性），可以用于后续的实验应用。

用20〜30出没的豆科植物，可以用筛子收集约2,000成年女性螨。侵扰10个工厂，每个工厂20螨所需要的时间大约是15分钟，如果用刷子转移螨虫。的收集和应用方法的组合示于图2。 该协议产生的植物的损害可重复的结果，表明收集的螨虫是相似的生理状态，适合于植物叶螨相互作用研究（ 图3 A）。评估协议的再现性，使用的洗涤方法，其中，20螨置于3?...

Watch this Scientific Journal Video about Application of Two-spotted Spider Mite Tetranychus urticae for Plant-pest Interaction Studies at JoVE.com

Application of Two-spotted Spider Mite Tetranychus urticae for Plant-pest Interaction Studies

协议对红蜘蛛，实验植物的侵染和植物的损害评估的均匀样品的制备效率的要求，对植物病虫害的相互作用研究，开发。

二斑叶螨的应用<em&gt;二斑叶螨</em&gt;植物病虫害的相互作用研究

The two-spotted spider mite, Tetranychus urticae, is a ubiquitous polyphagous arthropod herbivore that feeds on a remarkably broad array of species, with ...

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Environment

Application of Two-spotted Spider Mite Tetranychus urticae for Plant-pest Interaction Studies

17.6K 次观看.  The University of Western Ontario. 协议对红蜘蛛，实验植物的侵染和植物的损害评估的均匀样品的制备效率的要求，对植物病虫害的相互作用研究，开发。 

视频: Application of Two-spotted Spider Mite Tetranychus urticae for Plant-pest Interaction Studies

Physical, Chemical and Biological Characterization of Six Biochars Produced for the Remediation of Contaminated Sites

The physical and chemical properties of biochar vary based on feedstock sources and production conditions, making it possible to engineer biochars with specific functions (e.g. carbon sequestration, soil quality improvements, or contaminant sorption). In 2013, the International Biochar Initiative (IBI) made publically available their Standardized Product Definition and Product Testing Guidelines (Version 1.1) which set standards for physical and chemical characteristics for biochar. Six biochars made from three different feedstocks and at two temperatures were analyzed for characteristics related to their use as a soil amendment. The protocol describes analyses of the feedstocks and biochars and includes: cation exchange capacity (CEC), specific surface area (SSA), organic carbon (OC) and moisture percentage, pH, particle size distribution, and proximate and ultimate analysis. Also described in the protocol are the analyses of the feedstocks and biochars for contaminants including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), metals and mercury as well as nutrients (phosphorous, nitrite and nitrate and ammonium as nitrogen). The protocol also includes the biological testing procedures, earthworm avoidance and germination assays. Based on the quality assurance / quality control (QA/QC) results of blanks, duplicates, standards and reference materials, all methods were determined adequate for use with biochar and feedstock materials. All biochars and feedstocks were well within the criterion set by the IBI and there were little differences among biochars, except in the case of the biochar produced from construction waste materials. This biochar (referred to as Old biochar) was determined to have elevated levels of arsenic, chromium, copper, and lead, and failed the earthworm avoidance and germination assays. Based on these results, Old biochar would not be appropriate for use as a soil amendment for carbon sequestration, substrate quality improvements or remediation.

Biochar is a carbon-rich material used as a soil amendment with the ability to sustainably sequester carbon, improve substrate quality and sorb contaminants. This protocol describes the 17 analytical methods used for the characterization of biochar, which is required prior to large scale implementation of these amendments in the environment.

物理，化学和六Biochars生产的生物表征污染场地修复的

The physical and chemical properties of biochar vary based on feedstock sources and production conditions, making it possible to engineer biochars with ...

科研

环境科学

Characterization and Application of Passive Samplers for Monitoring of Pesticides in Water

Five different water passive samplers were calibrated under laboratory conditions for measurement of 124 legacy and current used pesticides. This study provides a protocol for the passive sampler preparation, calibration, extraction method and instrumental analysis. Sampling rates (RS) and passive sampler-water partition coefficients (KPW) were calculated for silicone rubber, polar organic chemical integrative sampler POCIS-A, POCIS-B, SDB-RPS and C18 disk. The uptake of the selected compounds depended on their physicochemical properties, i.e., silicone rubber showed a better uptake for more hydrophobic compounds (log octanol-water partition coefficient (KOW) &#62; 5.3), whereas POCIS-A, POCIS-B and SDB-RPS disk were more suitable for hydrophilic compounds (log KOW &#60; 0.70).

A protocol about the characterization and application of five different passive sampling devices is presented.

表征和被动采样的应用在农药水监测

Five different water passive samplers were calibrated under laboratory conditions for measurement of 124 legacy and current used pesticides. This study ...

Remote Sensing Evaluation of Two-spotted Spider Mite Damage on Greenhouse Cotton

The objective of this study was to evaluate a ground-based multispectral optical sensor as a remote sensing tool to assess foliar damage caused by the two-spotted spider mite (TSSM), Tetranychus urticae Koch, on greenhouse grown cotton. TSSM is a polyphagous pest which occurs on a variety of field and horticultural crops. It often becomes an early season pest of cotton in damaging proportions as opposed to being a late season innocuous pest in the mid-southern United States. Evaluation of acaricides is important for maintaining the efficacy of and preventing resistance to the currently available arsenal of chemicals and newly developed control agents. Enumeration of spider mites for efficacy evaluations is laborious and time consuming. Therefore, subjective visual damage rating is commonly used to assess density of spider mites. The NDVI (Normalized Difference Vegetation Index) is the most widely used statistic to describe the spectral reflectance characteristics of vegetation canopy to assess plant stress and health consequent to spider mite infestations. Results demonstrated that a multispectral optical sensor is an effective tool in distinguishing varying levels of infestation caused by T. urticae on early season cotton. This remote sensing technique may be used in lieu of a visual rating to evaluate insecticide treatments.

This manuscript describes a multispectral optical sensor that effectively detected damage to early season cotton artificially infested with varying densities of the two-spotted spider mite populations.

温室棉二斑叶螨危害的遥感评价

The objective of this study was to evaluate a ground-based multispectral optical sensor as a remote sensing tool to assess foliar damage caused by the ...

Modification and Application of a Leaf Blower-vac for Field Sampling of Arthropods

Rice fields host a large diversity of arthropods, but investigating their population dynamics and interactions is challenging. Here we describe the modification and application of a leaf blower-vac for suction sampling of arthropod populations in rice. When used in combination with an enclosure, application of this sampling device provides absolute estimates of the populations of arthropods as numbers per standardized sampling area. The sampling efficiency depends critically on the sampling duration. In a mature rice crop, a two-minute sampling in an enclosure of 0.13 m2 yields more than 90% of the arthropod population. The device also allows sampling of arthropods dwelling on the water surface or the soil in rice paddies, but it is not suitable for sampling fast flying insects, such as predatory Odonata or larger hymenopterous parasitoids. The modified blower-vac is simple to construct, and cheaper and easier to handle than traditional suction sampling devices, such as D-vac. The low cost makes the modified blower-vac also accessible to researchers in developing countries.

Assessment of arthropod abundance in crops is critical for investigating population dynamics and species interactions. Here we describe the modification and application of a leaf blower-vac for suction sampling of arthropods in rice.

修改和叶鼓风机-VAC的节肢动物领域采样中的应用

Rice fields host a large diversity of arthropods, but investigating their population dynamics and interactions is challenging. Here we describe the ...

Use of a Filter Cartridge for Filtration of Water Samples and Extraction of Environmental DNA

Recent studies demonstrated the use of environmental DNA (eDNA) from fishes to be appropriate as a non-invasive monitoring tool. Most of these studies employed disk fiber filters to collect eDNA from water samples, although a number of microbial studies in aquatic environments have employed filter cartridges, because the cartridge has the advantage of accommodating large water volumes and of overall ease of use. Here we provide a protocol for filtration of water samples using the filter cartridge and extraction of eDNA from the filter without having to cut open the housing. The main portions of this protocol consists of 1) filtration of water samples (water volumes &#8804;4 L or &gt;4 L); (2) extraction of DNA on the filter using a roller shaker placed in a preheated incubator; and (3) purification of DNA using a commercial kit. With the use of this and previously-used protocols, we perform metabarcoding analysis of eDNA taken from a huge aquarium tank (7,500 m3) with known species composition, and show the number of detected species per library from the two protocols as the representative results. This protocol has been developed for metabarcoding eDNA from fishes, but is also applicable to eDNA from other organisms.

We describe a protocol for filtration of water samples with a filter cartridge and extraction of environmental DNA (eDNA) without having to cut open the housing to remove the filter. This protocol is developed for metabarcoding eDNA from fishes, but is also applicable to eDNA from other organisms.

滤芯水过滤样品和环境DNA的提取使用

Recent studies demonstrated the use of environmental DNA (eDNA) from fishes to be appropriate as a non-invasive monitoring tool. Most of these studies ...

Vegetated Treatment Systems for Removing Contaminants Associated with Surface Water Toxicity in Agriculture and Urban Runoff

Urban stormwater and agriculture irrigation runoff contain a complex mixture of contaminants that are often toxic to adjacent receiving waters. Runoff may be treated with simple systems designed to promote sorption of contaminants to vegetation and soils and promote infiltration. Two example systems are described: a bioswale treatment system for urban stormwater treatment, and a vegetated drainage ditch for treating agriculture irrigation runoff. Both have similar attributes that reduce contaminant loading in runoff: vegetation that results in sorption of the contaminants to the soil and plant surfaces, and water infiltration. These systems may also include the integration of granulated activated carbon as a polishing step to remove residual contaminants. Implementation of these systems in agriculture and urban watersheds requires system monitoring to verify treatment efficacy. This includes chemical monitoring for specific contaminants responsible for toxicity. The current paper emphasizes monitoring of current use pesticides since these are responsible for surface water toxicity to aquatic invertebrates.

This article summarizes the design attributes and the effectiveness of treatment systems that treat urban stormwater and agriculture irrigation runoff to remove pesticides and other contaminants associated with aquatic toxicity.

用于去除与农业和城市径流中地表水毒性有关的污染物的植被处理系统

Urban stormwater and agriculture irrigation runoff contain a complex mixture of contaminants that are often toxic to adjacent receiving waters. Runoff may ...

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current measures to combat such pathogens have proved too conservative and, thus, not sufficiently effective, and they can potentially pose environmental risks. Therefore, it is extremely necessary to identify host-resistance factors to assist in controlling plant diseases naturally through the identification of resistant germplasm, the isolation and characterization of resistance genes, and the molecular breeding of resistant cultivars. In this regard, there is need to establish an accurate, rapid, and large-scale inoculation method to breed and develop plant resistance genes. The rice blast fungal pathogen Magnaporthe grisea causes severe disease symptoms and yield losses. Recently, M. grisea has emerged as a model organism for studying the mechanisms of plant-fungal pathogen interactions. Hence, we report the development of a plant virulence test method that is specific for M. grisea. This method provides for both spray inoculation with a conidial suspension and wounding inoculation with mycelium cubes or droplets of conidial suspension. The key step of the wounding inoculation method for detached rice leaves is to make wounds on plant leaves, which avoids any interference caused by host penetration resistance. This spray/wounding protocol contributes to the rapid, accurate, and large-scale screening of the pathotypes of M. grisea isolates. This integrated and systematic plant infection method will serve as an excellent starting point for gaining a broad perspective of issues in plant pathology.

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

Here, we present a protocol to test plant virulence with the plant pathogen Magnaporthe grisea. This report will contribute to the large-scale screening of the pathotypes of fungi isolates and serve as an excellent starting point for understanding the resistant mechanisms of plants during molecular breeding.

植物感染试验: 以植物病原菌为载体的喷雾和伤口介导接种

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current ...

Rearing and Long-Term Maintenance of Eristalis tenax Hoverflies for Research Studies

With an estimated 6000 species worldwide, hoverflies are ecologically important as alternative pollinators to domesticated honeybees. However, they are also a useful scientific model to study motion vision and flight dynamics in a controlled laboratory setting. As the larvae develop in organically polluted water, they are useful models for investigating investment in microbial immunity. While large scale commercial breeding for agriculture already occurs, there are no standardized protocols for maintaining captive populations for scientific studies. This is important as commercial captive breeding programs focusing on mass output during peak pollination periods may fail to provide a population that is consistent, stable and robust throughout the year, as is often needed for other research purposes. Therefore, a method to establish, maintain and refresh a captive research population is required. Here, we describe the utilization of an artificial hibernation cycle, in addition to the nutritional and housing requirements, for long term maintenance of Eristalis tenax. Using these methods, we have significantly increased the health and longevity of captive populations of E. tenax compared to previous reports. We furthermore discuss small scale rearing methods and options for optimizing yields and manipulating population demographics.

Rearing and Long-Term Maintenance of Eristalis tenax Hoverflies for Research Studies

The overall goal of these procedures is to establish, maintain and refresh a captive population of Eristalis tenax in a research setting.

研究性学习的Eristalis 管Hoverflies 的饲养和长期维护

With an estimated 6000 species worldwide, hoverflies are ecologically important as alternative pollinators to domesticated honeybees. However, they are ...

Experimental Protocol for Examining Behavioral Response Profiles in Larval Fish: Application to the Neuro-stimulant Caffeine

Fish models and behaviors are increasingly used in the biomedical sciences; however, fish have long been the subject of ecological, physiological and toxicological studies. Using automated digital tracking platforms, recent efforts in neuropharmacology are leveraging larval fish locomotor behaviors to identify potential therapeutic targets for novel small molecules. Similar to these efforts, research in the environmental sciences and comparative pharmacology and toxicology is examining various behaviors of fish models as diagnostic tools in tiered evaluation of contaminants and real-time monitoring of surface waters for contaminant threats. Whereas the zebrafish is a popular larval fish model in the biomedical sciences, the fathead minnow is a common larval fish model in ecotoxicology. Unfortunately, fathead minnow larvae have received considerably less attention in behavioral studies. Here, we develop and demonstrate a behavioral profile protocol using caffeine as a model neurostimulant. Though photomotor responses of fathead minnows were occasionally affected by caffeine, zebrafish&#160;were markedly more sensitive for photomotor and locomotor endpoints, which responded at environmentally relevant levels. Future studies are needed to understand comparative behavioral sensitivity differences among fish with age and time of day, and to determine whether similar behavioral effects would occur in nature and be indicative of adverse outcomes at the individual or population levels of biological organization.

Here, we present a protocol to examine larval zebrafish and fathead minnow locomotor activities and photomotor responses (PMR) using an automated tracking software. When incorporated in common toxicology bioassays, analyses of these behaviors provide a diagnostic tool to examine chemical bioactivity. This protocol is described using caffeine, a model neurostimulant.

检测幼虫行为反应剖面的实验协议: 在神经刺激咖啡因中的应用

Fish models and behaviors are increasingly used in the biomedical sciences; however, fish have long been the subject of ecological, physiological and ...

Generating Homo- and Heterografts Between Watermelon and Bottle Gourd for the Study of Cold-responsive MicroRNAs

MicroRNAs (miRNAs) are endogenous small non-coding RNAs of about 20 - 24 nt, known to play important roles in plant development and adaptation. There is an accumulating evidence showing that the expressions of certain miRNAs are altered when grafting, an agricultural practice commonly used by farmers to improve crop tolerance to biotic and abiotic stresses. Bottle gourd is an inherently climate-resilient crop compared to many other major cucurbits, including watermelon, rendering it one of the most widely used rootstocks for the latter. The recent advancement of high-throughput sequencing technologies has provided great opportunities to investigate cold-responsive miRNAs and their contributions to heterograft advantages; yet, adequate experimental procedures are a prerequisite for this purpose. Here, we present a detailed protocol for efficiently generating homo- and heterografts between the cold-susceptible watermelon and the cold-tolerant bottle gourd, in addition to methods of tissue sampling, data generation, and data analysis. The presented methods are also useful for other plant-grafting systems, to interrogate miRNA regulations under various environmental stresses, such as heat, drought, and salinity.

Here we present a detailed protocol for efficiently making homo- and heterografts between watermelon and bottle gourd, in addition to methods of tissue sampling, data generation, and data analysis, for the investigation of cold-responsive microRNAs.

西瓜与瓶装葫芦之间的同源和异种移植在冷反应微 rna 研究中的应用

MicroRNAs (miRNAs) are endogenous small non-coding RNAs of about 20 - 24 nt, known to play important roles in plant development and adaptation. There is ...