We appreciate the assistance of Chris Parker who daily scanned the plants and Curtis Hubbard who maintained the plants in the greenhouse.

1.斑豆建立TSSM殖民地<ol><li>植物斑豆， 菜豆 ，在包含在温室的盆栽土的塑料托盘（56×28×5厘米3）， 如图1。 </li><li>与根据处理和复制标记棍棒标签的托盘。 </li><li>设置和维持温室温度至90°F和70％相对湿度。 </li><li> 如图2生长豆1-2枳叶阶段10。 </li><li>通过去除感染的叶子收集与螨虫滋生自然植物棉叶螨。 </li><li>地方红蜘蛛出没棉叶斑豆视需要，直到托盘所有的植物都与众多TSSMs出没。 </li></ol><img alt="图1" src="/files/ftp_upload/54314/54314fig1.jpg" /> 图1:在塑料种植斑豆托盘。斑豆种子种植在温室中的塑料托盘（56×28×5厘米3）和每天浇水。 <a href="http://ecsource.jove.com/files/ftp_upload/54314/54314fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/54314/54314fig2.jpg" /> 图2:平托豆三出复叶。后子叶从土壤中出现所形成的第一片真叶是简单的单叶或叶。随后的叶子是枳叶细齿提示。 <a href="http://ecsource.jove.com/files/ftp_upload/54314/54314fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 2.将TSSM到棉株<ol><li>生长非转Bt（未转基因）的棉花植物，以在塑料托盘4-5真叶阶段（56×28×5厘米），在温室中，如步骤1.3中指定并在图3中示出。 </li><li>在4-5真叶期转乘斑豆红蜘蛛殖民地到年轻的棉花植株。 <ol><li>转移群众3叶螨为轻度侵染的植物。注:在非常高的水平为患，红蜘蛛形成肿块或棉桃11和被发现挂在叶尖， 如图4所示。 <ol><li>将含下TSSM群众斑豆尖叶一盘。 </li><li>切的斑豆叶尖用剪刀，允许TSSM群众落入盘作为示于图5和6。 </li><li>转动盘倒置在棉花植物和抽头TSSM群众到塑料托盘中生长的棉花植物， 如图7。注:每个托盘包含〜100个棉花植株。 </li><li>随机分布3名TSSM群众的到棉花植株。 </li></ol></li><li>转移群众20对我来说二烯丙基侵染的植物。 <ol><li>将含下TSSM群众斑豆尖叶一盘。 </li><li>用剪刀剪去斑豆叶的提示，让他们掉进锅里。 </li><li>收集在底盘20名群众。 </li><li>打开锅倒在棉花植株并且挖掘出TSSM群众在温室募集到约100棉花植株。 </li></ol></li><li>转移群众40为严重侵染的植物。 <ol><li>将含下TSSM群众斑豆尖叶一盘。 </li><li>用剪刀剪去斑豆叶的提示，让他们掉进锅里。 </li><li>收集在盘40块。 </li><li>打开锅倒在棉厂，挖掘出TSSM群众到〜100棉花植株并且随机传播他们。 </li></ol></li></ol></li></ol><img alt="图3" src="/files/ftp_upload/54314/54314fig3.jpg" /> 图3.用4-5片真叶阶段的棉花植株。钍Ë子叶从土壤中出现如叶状结构面向彼此相反对幼苗茎。顶端分生组织出现过子叶和形成第一片真叶。 <a href="http://ecsource.jove.com/files/ftp_upload/54314/54314fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/54314/54314fig4.jpg" /> 图4. TSSM群众挂在枳豆叶。 TSSM生活在殖民地，当人口达到高密度，它们形成肿块或铃状结构，在为分散叶尖聚集。 <a href="http://ecsource.jove.com/files/ftp_upload/54314/54314fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/54314/54314fig5.jpg" /> 含有用剪刀TSSM质量图5.切割斑豆叶的提示。含TSSM群众枳豆叶技巧与剪刀为害棉花植株被拆除。 <a href="http://ecsource.jove.com/files/ftp_upload/54314/54314fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图6" src="/files/ftp_upload/54314/54314fig6.jpg" /> 图6.豆叶提示TSSM群众放在盘内。当试验植物中发现的与TSSM三叶豆叶提示足够多，他们被拆除，并放入油锅内。这些样品用于侵染处理的类别:轻，中，其中接收到的3,20和40质量TSSM的，分别重。 <a href="http://ecsource.jove.com/files/ftp_upload/54314/54314fig6large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> </a&gt; <img alt="图7" src="/files/ftp_upload/54314/54314fig7.jpg" /> 图7.关于泛倒挂。含枳豆叶技巧与TSSM将盘子天翻地覆棉花树冠侵扰测试工厂。 <a href="http://ecsource.jove.com/files/ftp_upload/54314/54314fig7large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 3.扫描TSSM被感染的棉株与多光谱光学传感器<ol><li>水平安装光学传感器到温室帧地板大约7英尺的如图8。设置扫描仪，并在36&quot; 植物冠层之间的距离，使用一个木匠的水平，以确保传感器器水平。 </li><li>将棉花植株不受侵染托盘在轮式推车。 </li><li>激活传感器开关和慢慢推CA直到托盘完全通过传感器头， 如图8中的传感器下室温。关闭开关。退车。 </li><li>重复步骤3.3三次，总共3次重复。 </li><li>重复此过程棉花所有托盘。 </li><li>在第1天重复扫描，第5天，第6天，第7天，第9天，第10天，第12天，第13天以及第14天治疗后（DAT）。提供的NDVI（归一化植被指数）的扫描值12。发送NDVI值和存储口袋PC，然后在文本格式下载到计算机上。 注:NDVI由下列公式计算:NDVI =（NIR - RED）/（NIR + RED），其中红色和NIR是在红色和近红外在660和770的频谱的光谱反射率值（0-255）纳米，分别。 </li></ol><img alt="图8" src="/files/ftp_upload/54314/54314fig8.jpg" /> 图8.亩用于定量测量与TSSM的变化的密度水平侵染的棉花植物的健康ltispectral光学传感器。与试验植物轮式手推车传感器头下缓慢遍历来获得光谱反射率值。 一个是传感器头; b为袖珍PC; c是电池舱和输入/输出端口; d为RS-232串行数据线和e是牛皮纸，以提供均匀的背景。 <a href="http://ecsource.jove.com/files/ftp_upload/54314/54314fig8large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 4.数据分析<ol><li>获得使用PROC手段程序13的最大NDVI值。计算值NDVI减少百分数使用0天作为参考值的观察的每一天。使用重复测量的PROC GLM程序分析数据&lt;SUP类= "外部参照"&gt; 13。 注:手段是利用邓肯的多范围检验在P = 0.05隔开。用相同的小写字母表示没有显著不同。 </li><li> 如图9执行数据14的图形说明。 </li></ol><img alt="图9" src="/files/ftp_upload/54314/54314fig9.jpg" /> 图9.减少百分数或治疗后NDVI相对变化到数天。 JMP软件用于图形地示出的变化百分比之间在NDVI相对于采样的天（DAT）的功能关系。 <a href="http://ecsource.jove.com/files/ftp_upload/54314/54314fig9large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

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E., L&#243;pez, J. D., Lan, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+evaluation+of+the+GreenSeeker+handheld+optical+sensor+to+variations+in+orientation+and+height+above+canopy.">Laboratory evaluation of the GreenSeeker handheld optical sensor to variations in orientation and height above canopy.</a> International Journal of Agricultural and Biological Engineering. 5 (1), 43-47 (2012).</li></ol>

The authors have nothing to disclose.

在该领域中进行以往，杀虫剂效力测试包括以不同速率施加，并与未经处理的支票的化学的几种治疗方法。用针对TSSM的龄和成虫阶段变化的毒性特征杀螨剂评估，以确定是否由它们引起的损害可以通过化学处理来减少。 TSSM收集样品并带到其中它们在显微镜下检查和TSSM的不同阶段进行计数并记录的实验室。采取宿主植物的足够样本以确定在每个治疗的损伤和具有统计学上可接受的精度区分它们是非常重要的。所需要的彼此区分处理的样本的数目取决于生物体的分布轮廓。在TSSM的高度片状分布导致了相当数量的样本地区之间的差异，许多工厂必须进行采样为了确保人口估计的重现性。然而，预算，人力，时间和统计精度是在采样技术，冲击的重要因素。它理应研究者以最佳地分配可用资源，用最少的成本，但与大多数的精度进行采样。 代替计数TSSM阶段昆虫学家视觉评分基于损坏的不同水平的无损伤的分的损坏。例如，一些研究人员提出二项抽样，其中感染的叶子只有比例进行评分，而不是每个叶片9，19叶螨的数量。其他基于叶子变红索引标尺，其从点画和变红到植物冠层19的广泛变化变红棉花由TSSM估计的损害。这些方法是任意的，轶事和损伤的程度的个体感知偏置。造成TSSM损害的更强大和定量评估，需要评估和处理与统计精度分开。 基于地面的多光谱光学传感器似乎是一种改进的采样工具以定量测定由TSSM造成的损害和对治疗不是由许多研究人员采用的视觉损害评分系统更精确地分离。然而，研究人员已经报道了数据密集型光谱遥感提供了许多光谱特征来识别和比较多光谱遥感这是较少的数据密集的带有两个波长度20，21检测的作物的应力和冠层的特性。使用高光谱仪，Reisig和戈弗雷发现NIR反射波长≈850纳米，在区分一个信息从频谱未侵染的棉花22节肢动物侵染的。在这项研究中，我们已经表明，多光谱反射率值（NIR值为≈770纳米）仅使用两个光谱带能够鉴定和表征与TSSM的不同密度侵染的棉花植物。此外，我们先前报道，多光谱光学传感器不仅能有效分离出没TSSM的广泛不同密度类别棉花植物，但也显示，螺甲螨是比一半在温室早季棉控制TSSM阿维菌素更有效最低的标示率23的速度。 多谱光传感器可以被安装在移动的研究平台和反射率值可以从植被檐处理区域没有人的主观性而获得。杀螨效力数据因此可以被没有太多的人力劳动获得。该NDVI数据可以轻松上传到计算机，并使用商业统计进行分析软件。 GPS接收器也可安装到所述移动平台来收集GPS为了产生场的损坏地图坐标。从植物冠层使用多光谱反射特征，所述多光谱光学传感器提供了鉴定和定量植物胁迫的快速和成本有效的手段。此外，该字段的一个大得多的区域可以在更短的时间内覆盖有植物冠层的空间分辨率更高的相对于传统的字段侦察。重要的是要记住的门槛损害水平TSSM棉花是从区域到美国区域变量是很重要的。例如，TSSM损伤将是干旱环境下更高如在加州相比中南部区域降雨量和高湿度经常为准24。因此，为了TSSM损坏由于产量损失将是可变的，所以是损伤阈值。然而，来自密西西比州，阿肯色州和田纳西州的报告显示，阈值水平对棉花TSSM似乎是当植物的30〜50％侵染和人口正在积极增加5。此外，田间生长的植物暴露于多种胁迫，包括水应力和节肢动物食草动物喂养和这些活动之间的相互作用可以显着减少工厂生产力和可能影响损伤阈值。该TSSM可降低气孔导度，光合作用和蒸腾速率棉花25。在温室中生长的植物通过UV光照射的影响，它不影响显著气孔功能，光合作用和篷形态26,27和很可能对植物胁迫的添加剂的效果。然而，TSSM能够避免UV辐射由具有访问栖植物冠层28，29 &lt;的下表面上保护免受太阳辐射/ SUP&gt;，30，在那里它驻留。 目标篷和相对于目标的传感器的方位上述光学传感器的高度是显著受多光谱光学传感器31所获得的反射率值的重要因素。例如，当移动的研究平台通过一排农作物字段遍历诸如当棉篷打开时，传感器可能产生，这取决于传感器的取向不同的结果，或者平行于或者垂直于该行。它也可能是土壤和其它背景材料可极大地影响传感器读数，特别是当所述传感器被垂直于行方向。为了获得从传感器最大响应，传感器头应线被定向以与直接的行之上。虽然垂直定向的光束将行更容易拿起回地面土壤反射，这可以，是可以接受的，然而，当棉花篷与茂密的植被关闭。此外，运营商应当遵循的81-122厘米的操作高度范围制造商的建议和定向在线传感器头与所述目标，以获得最大信号的响应。重要的是要在使用前的传感器电池充电是很重要的，或者它应保持插入到避免故障排除。低电池电量可能会产生错误的读数。

二斑叶螨， 二斑叶螨 （科赫）是许多领域和园艺植物1,2的多食性和国际化的有害。它生活在植物3，4的底部表面上的菌落里面织带。它已经从一个季节晚期到赛季初的害虫中期，美国南部在过去十年5进化而来的。 TSSM是棉的第 5 个最具破坏性的害虫和在2011年6，7引起的棉花和减少0.167％在美国产量57441包损失估计。它的生命周期短，高繁殖力和单倍体二倍体性别决定与消化和解毒外源性化学物质的能力相结合加剧了耐药性的发展，农药8。目前，杀螨剂保持为邻NLY可靠的控制机制T.螨的抑制。因此，现场昆虫学家不断评估疗效现有和新开发的杀螨剂。 通过叶螨损害的估计通常是通过计分在因为在手动计数螨遇到的困难的主观尺度的损伤进行。一些进行二项式采样，其中感染的叶子的唯一的比例进行评分而不是每叶9叶螨的数目。叶变红索引标尺，其从点画和发红变化以植被的广泛发红，被用来作为在估计损害的标准。对棉花T.螨的空间分布图案符合一个集群分布图案9。螨分布在棉花叶子由疏到大量聚集和现场条件下，依然如此。这种分布格局腠以其体积小，移动性和多产再现PLED使TSSM的枚举困难。为了定量地评价针对TSSM杀螨剂的效力需要用于螨密度的评估可靠的替代技术。 本研究的目的是通过使用光学传感器的多光谱由TSSM的不同密度损坏棉花植物中分离出来。我们的目的是确定地面光学传感器可以进行分类，并从那些由红蜘蛛损坏分开健康棉花植株。

<table><tbody><tr><td>GreenSeeker</td><td>Trimble Ag. Division&lt;br /&gt;Westminster, CO</td><td>Model 505</td><td>Red NDVI sensor</td></tr><tr><td>Pinto beans</td><td>Producer&#039;s Co-op., Bryan, TX</td><td>Not applicable</td><td>Free choice item</td></tr><tr><td>Deltapine cotton seeds</td><td>Brazos Bottom Crop Care, Caldwell, TX77836</td><td>Not applicable</td><td>436 RR; NonBt &amp;amp; RoundUp Ready</td></tr><tr><td>Plastic trays</td><td>BWI, Schulenberg, TX</td><td>FG1020NL7</td><td>56 cm x 28 cm</td></tr><tr><td>Label sticks</td><td>Gempler&#039;s, Janesville, WI 53547</td><td>Item # 151276</td><td>Durable spike-style pot markers</td></tr><tr><td>4-wheel Garden push cart</td><td>Farm Tek, Dyersville, IA 52040</td><td>Item # 108676</td><td>61 cm x 122 cm</td></tr></tbody></table>

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.

该传感器发射红光和红外光该光反过来获取反射从植物冠层背面。该反射光作为可见光和电磁光谱的近红外波段的定量测量，并且数字记录为NDVI（归一化植被指数）读数。的NDVI值从0变化到0.99。在该NDVI读健康的植物冠层较高。健康植被吸收可见光和反射近红外光，不健康的植被反射更多的可见光和更少的近红外光。 NDVI用作代用品光合作用活性和该光谱性质是高度相关于光合有效辐射15，16，17，18。 NDVI的非常低的值（0.1或以下）对应于岩石的贫瘠区域或砂，而适中的值（0.2〜0.3）表示灌木，草地和高值（0.6〜0.8）表示温带和热带植物。在光谱的红色和近红外区域下白天照明获得的反射率的测量。 NDVI从以下方程计算:NDVI =（NIR - RED）/（NIR + RED），其中红色和NIR是在660和770处的光谱反射率值（0-255）在红和近红外光谱，分别。传感器记录的反射率值，每100毫秒。从由传感器每次采取NDVI值的阵列剔除最大NDVI读数在分析来最小化背景的反射率，并提供一致的可重复的数值。 的数据的方差分析表明，轻轻地之间观察到在NDVI降低百分比显著差异，米相比于未处理的对照edially和重度侵染的棉花植物（F = 436.4; P &lt;0.0001; DF = 3，32）。这在测试期间观察天之间变化显著描述的植物活力的NDVI值（DF = 8，256 F = 1398.2; P &lt;0.0001）。另外，在NDVI值减少百分数呈负相关，以在观察期间（DAT）的治疗方法，但很少倾向于从该图案偏离，并与DAT显著相互作用（F = 201.5; P &lt;0.0001; DF = 24，256） 。对于没有DAT效果MANOVA测试标准是显著以及（威尔克的λ= 0.00913; F = 339.0; P &lt;0.0001; DF = 8，25）。类似地，DAT和治疗之间的相互作用是显著（威尔克的λ= 0.00101; F = 29.8; P &lt;0.0001; DF = 24，73）。 图9示出了如在观察期间通过NDVI值证明在植物活力百分比变化。正每在NDVI值百分比变化表示健康生长的植物，而负值表明植物的活力有所减少由于第一测量（ 即第0天）制成。在整个研究过程中的非受侵染的对照植物表现出增加的营养生长，而TSSM侵染的植物卫生随着时间的推移呈现下降。在表1所示的处理意味着分离显示，在治疗类别（轻，中，重）和对照之间在NDVI降低百分比没有可定义差异观察到直到第5天当侵染类从控制显著偏离并且此后这样压倒性保持。该数据证明，所述光学传感器可以代替劳动密集型人工取样的被有效地用于评估针对棉花杀螨剂治疗功效。 <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td colspan = "10"&gt;天处理（DAT）后</td></tr><tr><td>侵扰范畴</td><td> 1 </td><td>五</td><td> 6 </td><td> 7 </td><td> 9 </td><td> 10 </td><td> 12 </td><td> 13 </td><td> 14 </td></tr><tr><td>控制</td><td> 1.18±0.33A </td><td> 2.70±0.40A </td><td> 4.0±0.36A </td><td> 3.94±0.37a </td><td> 3.68±0.53A </td><td> 2.57±0.42A </td><td> 2.96±0.47A </td><td> 3.48±0.38A </td><td> 3.08±0.22A </td></tr><tr><td>光</td><td> -0.13±0.13b </td><td> -0.71±0.29b </td><td> -0.65±0.28b </td><td> -2.02±0.47b </td><td> -5.68±0.72b </td><td> -11.17±0.94b </td><td> -15.73±1.76b </td><td> -19.54±1.68b </td><td> -24.9±1.90b </td></tr><tr><td>中</td><td> -1.83±0.42c </td><td> -7.06±0.63c </td><td> -9.61±0.53c </td><td> -10.39±0.57c </td><td> -17.06±0.80℃ </td><td> -26.92±0.72c </td><td> -33.84±0.96c </td><td> -37.05±1.14c </td><td> -41.74±0.73c </td></tr><tr><td>重</td><td> -0.97±0.58bc </td><td> -11.76±0.29d </td><td> -13.83±0.86d </td><td> -15.20±0.63D </td><td> -25.0±1.0D </td><td> -34.63±0.54d </td><td> -39.07±0.94d </td><td> -42.68±0.62d </td><td> -46.71±0.63 </td></tr></tbody></table> 表1:棉花植物后在最大NDVI减少百分数与不同集群或TSSM的质量的数目侵染。在温室中的塑料托盘中生长的棉花植物与三类叶螨密度的侵染。类光接收3块或簇TSSM的每盘，类别介质接收每盘20层的质量和类重型接收40名群众每托盘。手段是从控制根据邓肯的MULT分离IPLE范围测试（P = 0.05）。装置随后通过相同的小写字母不是在概率的5％的水平显著不同。

Watch this Scientific Journal Video about Remote Sensing Evaluation of Two-spotted Spider Mite Damage on Greenhouse Cotton at JoVE.com

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

这手稿描述了有效地检测到早季棉与二斑叶螨种群的不同密度人工侵染损伤多光谱光学传感器。

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 ...

remote-sensing-evaluation-two-spotted-spider-mite-damage-on

Research

JoVE Journal

Environment

8.5K 次观看.  USDA-ARS, SPARC-AATRU. 这手稿描述了有效地检测到早季棉与二斑叶螨种群的不同密度人工侵染损伤多光谱光学传感器。 

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

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

The two-spotted spider mite, Tetranychus urticae, is a ubiquitous polyphagous arthropod herbivore that feeds on a remarkably broad array of species, with more than 150 of economic value. It is a major pest of greenhouse crops, especially in Solanaceae and Cucurbitaceae (e.g., tomatoes, eggplants, peppers, cucumbers, zucchini) and greenhouse ornamentals (e.g., roses, chrysanthemum, carnations), annual field crops (such as maize, cotton, soybean, and sugar beet), and in perennial cultures (alfalfa, strawberries, grapes, citruses, and plums)1,2. In addition to the extreme polyphagy that makes it an important agricultural pest, T. urticae has a tendency to develop resistance to a wide array of insecticides and acaricides that are used for its control3-7.
T. urticae is an excellent experimental organism, as it has a rapid life cycle (7 days at 27 &#176;C) and can be easily maintained at high density in the laboratory. Methods to assay gene expression (including in situ hybridization and antibody staining) and to inactivate expression of spider mite endogenous genes using RNA interference have been developed8-10. Recently, the whole genome sequence of T. urticae has been reported, creating an opportunity to develop this pest herbivore as a model organism with equivalent genomic resources that already exist in some of its host plants (Arabidopsis thaliana and the&#160;tomato Solanum lycopersicum)11. Together, these&#160;model organisms could provide insights into molecular bases of plant-pest interactions.
Here, an efficient method for quick and easy collection of a large number of adult female mites, their application on an experimental plant host, and the assessment of the plant damage due to spider mite feeding are described. The presented protocol enables fast and efficient collection of hundreds of individuals at any developmental stage (eggs, larvae, nymphs, adult males, and females) that can be used for subsequent experimental application.

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

Protocols for efficient preparation of homogenous samples of spider mites, infestation of experimental plants, and assessment of plant damage, as required for studies of plant-pest interaction were developed.

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 ...

科研

环境科学

Development of Metarhizium anisopliae as a Mycoinsecticide: From Isolation to Field Performance

A major concern when developing commercial mycoinsecticides is the kill speed compared to that of chemical insecticides. Therefore, isolation and screening for the selection of a fast-acting, highly virulent entomopathogenic fungus are important steps. Entomopathogenic fungi, such as Metarhizium, Beauveria, and Nomurea, which act by contact, are better suited than Bacillus thuringiensis or nucleopolyhedrosis virus (NPV), which must be ingested by the insect pest. In the present work, we isolated 68 Metarhizium strains from infected insects using a soil dilution and bait method. The isolates were identified by the amplification and sequencing of the ITS1-5.8S-ITS2 and 26S rDNA region. The most virulent strain of Metarhizium anisopliae was selected based on the median lethal concentration (LC50) and time (LT50) obtained in insect bioassays against III-instar larvae of Helicoverpa armigera. The mass production of spores by the selected strain was carried out with solid-state fermentation (SSF) using rice as a substrate for 14 days. Spores were extracted from the sporulated biomass using 0.1% tween-80, and different formulations of the spores were prepared. Field trials of the formulations for the control of an H. armigera infestation in pigeon peas were carried out by randomized block design. The infestation control levels obtained with oil and aqueous formulations (78.0% and 70.9%, respectively) were better than the 63.4% obtained with chemical pesticide.

Development of Metarhizium anisopliae as a Mycoinsecticide: From Isolation to Field Performance

Here, we report the different stages involved in the knowledge-based development of an effective mycoinsecticide, including the isolation, identification, screening, and selection of the "best-fit" entomopathogenic fungus, Metarhizium anisopliae, for the control of insect pests in agriculture.

的发展<em&gt; Metarhizium anisopliae</em&gt;作为防腐剂:从隔离到现场表现

A major concern when developing commercial mycoinsecticides is the kill speed compared to that of chemical insecticides. Therefore, isolation and ...

Methodology for Developing Life Tables for Sessile Insects in the Field Using the Whitefly, Bemisia tabaci, in Cotton As a Model System

Life tables provide a means of measuring the schedules of birth and death from populations over time. They also can be used to quantify the sources and rates of mortality in populations, which has a variety of applications in ecology, including agricultural ecosystems. Horizontal, or cohort-based, life tables provide for the most direct and accurate method of quantifying vital population rates because they follow a group of individuals in a population from birth to death. Here, protocols are presented for conducting and analyzing cohort-based life tables in the field that takes advantage of the sessile nature of the immature life stages of a global insect pest, Bemisia tabaci. Individual insects are located on the underside of cotton leaves and are marked by drawing a small circle around the insect with a non-toxic pen. This insect can then be observed repeatedly over time with the aid of hand lenses to measure development from one stage to the next and to identify stage-specific causes of death associated with natural and introduced mortality forces. Analyses explain how to correctly measure multiple mortality forces that act contemporaneously within each stage and how to use such data to provide meaningful population dynamic metrics. The method does not directly account for adult survival and reproduction, which limits inference to dynamics of immature stages. An example is presented that focused on measuring the impact of bottom-up (plant quality) and top-down (natural enemies) effects on the mortality dynamics of B. tabaci in the cotton system.

Methodology for Developing Life Tables for Sessile Insects in the Field Using the Whitefly, Bemisia tabaci, in Cotton As a Model System

Life tables allow quantification of the sources and rates of mortality in insect populations and contribute to understanding, predicting and manipulating population dynamics in agroecosystems. Methods for conducting and analyzing cohort-based life tables in the field for an insect with sessile immature life stages are presented.

用虱、烟粉虱在棉花中开发无梗昆虫生命表的方法作为模型系统

Life tables provide a means of measuring the schedules of birth and death from populations over time. They also can be used to quantify the sources and ...

Screening Cotton Genotypes for Reniform Nematode Resistance

A rapid non-destructive reniform nematode (Rotylenchulus reniformis) screening protocol is needed for the development of resistant cotton (Gossypium hirsutum) varieties to improve nematode management. Most protocols involve extracting vermiform nematodes or eggs from the cotton root system or potting soil to determine population density or reproduction rate. These approaches are generally time-consuming with a small number of genotypes evaluated. An alternative approach is described here in which the root system is visually examined for nematode infection. The protocol involves inoculating cotton seedling 7 days after planting with vermiform nematodes and determining the number of females attached to the root system 28 days after inoculation. Data are expressed as the number of females per gram of fresh root weight to adjust for variation in root growth. The protocol provides an excellent method for evaluating host-plant resistance associated with the ability of the nematode to establish an infection site; however, resistance that hinders nematode reproduction is not assessed. As with other screening protocols, variation is commonly observed in nematode infection among individual genotypes within and between experiments. Data are presented to illustrate the range of variation observed using the protocol. To adjust for this variation, control genotypes are included in experiments. Nonetheless, the protocol provides a simple and rapid method to evaluate host-plant resistance. The protocol has been successfully used to identify resistant accessions from the G. arboreum germplasm collection and evaluate segregating populations of more than 300 individuals to determine the genetics of resistance. A vegetative propagation method for recovering plants for resistance breeding was also developed. After removal of the root system for nematode evaluation, the vegetative shoot is replanted to allow the development of a new root system. More than 95% of the shoots typically develop a new root system with plants reaching maturity.

Here, a protocol is presented for the rapid non-destructive screening of cotton genotypes for reniform nematode resistance. The protocol involves visually examining the roots of nematode-infected cotton seedlings to determine infection response. The vegetative shoot from each plant is then propagated to recover plants for seed production.

筛选抗内虫抗性棉基因型

A rapid non-destructive reniform nematode (Rotylenchulus reniformis) screening protocol is needed for the development of resistant cotton (Gossypium ...

遗传学

Evaluating Dryocosmus Kuriphilus-induced Damage on Castanea Sativa

Dryocosmus kuriphilus Yasumatsu has become a major pest for Castaneasativa since its arrival in Europe. Its galling activity results in the formation of different gall types and prevents the development of normal shoots. Repeated and uncontrolled attacks cause, besides the production of galls and the attendant gall-related reduction in leaf area, progressive corruption of the branch architecture, including the death of branch parts, and an increase in dormant bud activation. Thus far, there have been few attempts to quantify branch architecture damage. Further, the different methods for assessing infestation degree (MAID) that have been developed focus only on the galls' presence and abundance.
Using the leaf area to sapwood area relationship as a green biomass indicator, we developed in a previous study a damage composite index (DCI) that takes into account the most important branch architectural features, allowing for realistic damage assessment during the entire epidemic process. 
The aim of this study is to present this novel method and highlight differences in the damage description with respect to other broadly used indices. Results show how the DCI depicts branch damage better, especially during the epidemic peak, compared to MAID, which tend to underestimate it. We conclude by suggesting how to properly evaluate the overall impact of the pest by means of our composite damage index, the infestation degree using classic methods, and crown transparency evaluations.

Evaluating Dryocosmus Kuriphilus-induced Damage on Castanea Sativa

It is common practice to assess the damage caused by Dryocosmus kuriphilus by considering the abundance of galls alone rather than by also taking related branch corruption into consideration. We propose a composite damage index that takes into account the most important branch features, thus enabling more realistic damage assessment.

栗瘿栗瘿致锥害的评价

Dryocosmus kuriphilus Yasumatsu has become a major pest for Castaneasativa since its arrival in Europe. Its galling activity results in the formation of ...

Methodology to Test Control Agents and Insecticides Against the Coffee Berry Borer Hypothenemus hampei

Prior to recommending insecticides to treat the coffee berry borer (CBB) Hypothenemus hampei, it is valuable to know the mortality and repellency of these insecticides against adult insects or their impact on reproductive output. However, currently available methods assess adult mortality only, limiting the selection of novel insecticides with a different mode of action. In this work, different experimental methods were examined to identify the diverse effects on the CBB under laboratory conditions. For this, green coffee fruits (GFs) were collected and disinfected by immersion in sodium hypochlorite solution followed by UV light irradiation. In parallel, CBB adults from a colony were disinfected by immersion in sodium hypochlorite solution. To assess fruit protection (preinfestation), the fruits were placed in plastic boxes, and the insecticides were applied. Then, the CBB adults were released at a rate of two CBBs per GF. The GFs were left under controlled conditions to evaluate CBB infestation and survival after 1, 7, 15, and 21 days. To evaluate insecticide efficacy after CBB infestation (postinfestation), CBB adults were released to the GFs in a 2:1 ratio for 3 h at 21 &#176;C. Infested fruits showing CBB adults with their abdomens partially exposed were selected and placed in 96-well racks, and the CBBs boring into the fruits were treated directly. After 20 days, the fruits were dissected, and the CBB biological stages inside each fruit were recorded. The GFs served as substrates that mimic natural conditions to evaluate toxic, chemical, and biological insecticides against the CBB.

Methodology to Test Control Agents and Insecticides Against the Coffee Berry Borer Hypothenemus hampei

A method using green coffee fruits (GFs) was developed to test the toxicity of insecticides against the coffee berry borer (CBB). Insecticides or toxic substances were applied to disinfected GFs before or after CBB infestation. Insect mortality, repellency, and reproductive capacity, in addition to other parameters, were evaluated.

针对咖啡浆果螟子下皮测试控制剂和杀虫剂 的方法

Prior to recommending insecticides to treat the coffee berry borer (CBB) Hypothenemus hampei, it is valuable to know the mortality and repellency of these ...

Evaluating the Effect of Pesticides on the Larvae of the Solitary Bees

Current ecological risk assessments of pesticides on pollinators have primarily considered only laboratory conditions. For the larvae of solitary bees, ingestion of provisions contaminated with pesticides may increase the mortality rate of the larvae, decrease the collection rate and the population of adult solitary bees in the next year from a demographic perspective. But there are limited studies on the effects of pesticides on the larvae of solitary bees. Therefore, understanding how pesticides influence the larvae of solitary bees should be considered an integral part of pesticide ecological risk assessment. This study presents a method to expose the larvae of solitary bee, Osmia excavata, to lethal or sublethal doses of pesticide, tracking larval weight gain, developmental duration, eclosion ability, and food consumption efficiency conversion of ingested food. To demonstrate the effectiveness of this method, the larvae of O. excavata were fed with provisions containing acute lethal and sublethal doses of chlorpyrifos. Then, the above indexes of the treated larvae were investigated. This technique helps to predict and mitigate the risk of pesticides to pollinators.

The present protocol explains a method to feed pesticide-contaminated provisions to the larvae of the solitary bees, Osmia excavata. The procedure examines the ecotoxicity of the pesticide to the larvae of the solitary bees.

评估农药对孤独蜜蜂幼虫的影响

Current ecological risk assessments of pesticides on pollinators have primarily considered only laboratory conditions. For the larvae of solitary bees, ...

生物化学

Bioassays for Monitoring Insecticide Resistance

Pest resistance to pesticides is an increasing problem because pesticides are an integral part of high-yielding production agriculture. When few products are labeled for an individual pest within a particular crop system, chemical control options are limited. Therefore, the same product(s) are used repeatedly and continual selection pressure is placed on the target pest. There are both financial and environmental costs associated with the development of resistant populations. The cost of pesticide resistance has been estimated at approximately $ 1.5 billion annually in the United States. This paper will describe protocols, currently used to monitor arthropod (specifically insects) populations for the development of resistance. The adult vial test is used to measure the toxicity to contact insecticides and a modification of this test is used for plant-systemic insecticides. In these bioassays, insects are exposed to technical grade insecticide and responses (mortality) recorded at a specific post-exposure interval. The mortality data are subjected to Log Dose probit analysis to generate estimates of a lethal concentration that provides mortality to 50% (LC50) of the target populations and a series of confidence limits (CL's) as estimates of data variability. When these data are collected for a range of insecticide-susceptible populations, the LC50 can be used as baseline data for future monitoring purposes. After populations have been exposed to products, the results can be compared to a previously determined LC50 using the same methodology.

This manuscript demonstrates and discusses techniques used to survey pesticide susceptibility and detect resistance to contact and systemic pesticides in arthropod pests.

生物测定对杀虫剂的抗药性监测

Pest resistance to pesticides is an increasing problem because pesticides are an integral part of high-yielding production agriculture. When few products ...

生物学

Protocols for Investigating the Host-tissue Distribution, Transmission-mode, and Effect on the Host Fitness of a Densovirus in the Cotton Bollworm

Many novel viruses have been discovered in animal hosts using next-generation sequencing technologies. Previously, we reported a mutualistic virus, Helicoverpa armigera densovirus (HaDV2), in a lepidopteran species, the cotton bollworm, Helicoverpa armigera (Hubner). Here, we describe the protocols that are currently used to study the effect of HaDV2 on its host. First, we establish a HaDV2-free cotton bollworm colony from a single breeding pair. Then, we orally inoculate some neonate larval offspring with HaDV2-containing filtered liquid to produce two colonies with the same genetic background: one HaDV2-infected, the other uninfected. A protocol to compare life table parameters (e.g., larval, pupal, and adult periods and fecundity) between the HaDV2-infected and -uninfected individuals is also presented, as are the protocols for determining the host-tissue distribution and transmission efficiency of HaDV2. These protocols would also be suitable for investigating the effects of other orally transmitted viruses on their insect hosts, lepidopteran hosts in particular.

Here, we present a protocol to investigate the host-tissue distribution, transmission mode, and effect on the host fitness of a densovirus within a lepidopteran species, the cotton bollworm. This protocol can also be used for studying the interaction between other orally-transmitted viruses and their insect hosts.

协议侦办主机组织分布，传播模式和效果上浓核病毒的主机健身的棉铃虫

Many novel viruses have been discovered in animal hosts using next-generation sequencing technologies. Previously, we reported a mutualistic virus, ...

免疫与感染

Agrobacterium-Mediated Virus-Induced Gene Silencing Assay In Cotton

Cotton (Gossypium hirsutum) is one of the most important crops worldwide. Considerable efforts have been made on molecular breeding of new varieties. The large-scale gene functional analysis in cotton has been lagged behind most of the modern plant species, likely due to its large size of genome, gene duplication and polyploidy, long growth cycle and recalcitrance to genetic transformation1. To facilitate high throughput functional genetic/genomic study in cotton, we attempt to develop rapid and efficient transient assays to assess cotton gene functions.
Virus-Induced Gene Silencing (VIGS) is a powerful technique that was developed based on the host Post-Transcriptional Gene Silencing (PTGS) to repress viral proliferation2,3. Agrobacterium-mediated VIGS has been successfully applied in a wide range of dicots species such as Solanaceae, Arabidopsis and legume species, and monocots species including barley, wheat and maize, for various functional genomic studies3,4. As this rapid and efficient approach avoids plant transformation and overcomes functional redundancy, it is particularly attractive and suitable for functional genomic study in crop species like cotton not amenable for transformation.
In this study, we report the detailed protocol of Agrobacterium-mediated VIGS system in cotton. Among the several viral VIGS vectors, the tobacco rattle virus (TRV) invades a wide range of hosts and is able to spread vigorously throughout the entire plant yet produce mild symptoms on the hosts5. To monitor the silencing efficiency, GrCLA1, a homolog gene of Arabidopsis Cloroplastos alterados 1 gene (AtCLA1) in cotton, has been cloned and inserted into the VIGS binary vector pYL156. CLA1 gene is involved in chloroplast development6, and previous studies have shown that loss-of-function of AtCLA1 resulted in an albino phenotype on true leaves7, providing an excellent visual marker for silencing efficiency. At approximately two weeks post Agrobacterium infiltration, the albino phenotype started to appear on the true leaves, with 100% silencing efficiency in all replicated experiments. The silencing of endogenous gene expression was also confirmed by RT-PCR analysis. Significantly, silencing could potently occur in all the cultivars we tested, including various commercially grown varieties in Texas. This rapid and efficient Agrobacterium-mediated VIGS assay provides a very powerful tool for rapid large-scale analysis of gene functions at genome-wide level in cotton.

We present the detailed protocol for Agrobacterium-mediated virus-induced gene silencing (VIGS) assay in cotton. The tobacco rattle virus (TRV)-derived VIGS vectors were deployed to induce RNA silencing of cotton GrCLA1, Cloroplastos alterados 1 gene. The albino phenotype caused by silencing GrCLA1 was observed at the seedling stage within 2 weeks after inoculation.

农杆菌介导的病毒诱导的基因沉默含量在棉花

Cotton (Gossypium hirsutum) is one of the most important crops worldwide. Considerable efforts have been made on molecular breeding of new varieties.  The ...

温室棉二斑叶螨危害的遥感评价

摘要

探索更多视频

此视频中的章节

二斑叶螨的应用

的发展

用虱、烟粉虱在棉花中开发无梗昆虫生命表的方法作为模型系统

筛选抗内虫抗性棉基因型

栗瘿栗瘿致锥害的评价

针对咖啡浆果螟子下皮测试控制剂和杀虫剂的方法

评估农药对孤独蜜蜂幼虫的影响

生物测定对杀虫剂的抗药性监测

协议侦办主机组织分布，传播模式和效果上浓核病毒的主机健身的棉铃虫

农杆菌介导的病毒诱导的基因沉默含量在棉花

温室棉二斑叶螨危害的遥感评价

摘要

探索更多视频

此视频中的章节

二斑叶螨的应用

的发展

用虱、烟粉虱在棉花中开发无梗昆虫生命表的方法作为模型系统

筛选抗内虫抗性棉基因型

栗瘿栗瘿致锥害的评价

针对咖啡浆果螟子下皮测试控制剂和杀虫剂 的方法

评估农药对孤独蜜蜂幼虫的影响

生物测定对杀虫剂的抗药性监测

协议侦办主机组织分布，传播模式和效果上浓核病毒的主机健身的棉铃虫

农杆菌介导的病毒诱导的基因沉默含量在棉花

针对咖啡浆果螟子下皮测试控制剂和杀虫剂的方法