Name: choice and no-choice bioassays to study the pupation preference and emergence success of ectropis grisescens
Uploaded: 2018-10-30T13:00:00
Description: Watch this Scientific Journal Video about Choice and No-Choice Bioassays to Study the Pupation Preference and Emergence Success of Ectropis grisescens at JoVE.com

我们感谢玉珍、圣哲和燕骏 (华南农业大学林业与景观建筑学院) 对昆虫饲养和实验设置的帮助。这项工作由中国国家自然科学基金 (赠款号 31600516)、广东自然科学基金 (赠款号 2016A030310445) 和广东省科技规划项目 (批准号 2015A020208010) 资助。.

1. 水分选择生物鉴定, 以确定化蛹偏好的E. grisescens
<ol><li>
获得grisescens的成熟的末龄幼虫
	<ol>
<li>茶树 (山茶) 的鲜芽 (30-40 厘米长)。将 25-30 芽插入250毫升三角烧瓶中。用自来水灌满烧瓶。在塑料盆中放入 3-4 烧瓶 (带茶树) (上部: 直径51厘米; 底面: 直径40厘米; 高度:16 厘米)。</li>
		<li>释放 1000-2000 幼虫 (第二到第五龄) 的实验室殖民地的grisescens到茶叶芽在每?...

<ol>
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H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detecting+deep+divergence+in+seventeen+populations+of+tea+geometrid+(Ectropis+obliqua+Prout)+in+China+by+COI+mtDNA+and+cross-breeding.">Detecting deep divergence in seventeen populations of tea geometrid (Ectropis obliqua Prout) in China by COI mtDNA and cross-breeding.</a> PloS One. 9 (6), e99373 (2014).</li><li>Ma, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+tea+geometrid+(Ectropis+grisescens)+pheromone+gland+extracts+using+GC-EAD+and+GC&#215;+GC/TOFMS.">Analysis of tea geometrid (Ectropis grisescens) pheromone gland extracts using GC-EAD and GC&#215; GC/TOFMS.</a> Journal of Agricultural and Food Chemistry. 64 (16), 3161-3166 (2016).</li><li>Zhang, G. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Asymmetrical+reproductive+interference+between+two+sibling+species+of+tea+looper:+Ectropis+grisescens+and+Ectropis+obliqua.">Asymmetrical reproductive interference between two sibling species of tea looper: Ectropis grisescens and Ectropis obliqua.</a> Bulletin of Entomological Research. , (2016).</li><li>Luo, Z. X., Li, Z. Q., Cai, X. M., Bian, L., Chen, Z. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+of+premating+isolation+between+two+sibling+moths:+Ectropis+grisescens+and+Ectropis+obliqua+(Lepidoptera:+Geometridae).">Evidence of premating isolation between two sibling moths: Ectropis grisescens and Ectropis obliqua (Lepidoptera: Geometridae).</a> Journal of Economic Entomology. 110 (6), 2364-2370 (2017).</li><li>Li, Z. Q., 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=Chemosensory+gene+families+in+Ectropis+grisescens+and+candidates+for+detection+of+Type-II+sex+pheromones.">Chemosensory gene families in Ectropis grisescens and candidates for detection of Type-II sex pheromones.</a> Frontiers in Physiology. 8, (2017).</li><li>Chen, L. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Research+on+structure+of+soil+particle+by+hydrometer+method.">Research on structure of soil particle by hydrometer method.</a> Environmental Science Survey. 29 (4), 97-99 (2010).</li><li>Kucera, M., Malmgren, B. 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在一些害虫6、9、22、23中对不同土壤变量的化蛹偏好进行了研究。例如, 为了研究桔小实 tryoni (乔安妮弗罗加) (双翅目: 实蝇科) 成熟幼虫在不同土壤湿度条件下的偏好, Hulthen 和克拉克22设置 3 x 3 拉丁方形设计, 其中包含9个容器, 填充土壤的 0%, 75%, 或100% 场容量, 和25成熟的幼虫...

与昆虫幼虫和成虫阶段相比, 由于蛹的移动能力有限, 蛹阶段非常脆弱, 无法迅速逃离危险的环境。玉米螟下面是一个共同的战略使用的不同群体的昆虫 (例如, 在命令双翅目1,2,3,4, 鞘翅目5, 膜翅目6,缨翅目7, 和鳞翅目8,9,10,11,12) 保护他们免受地面捕食者和环境危害。其中许多是严重的农林害虫1、2、3、4、5、6、7、8 ,9,10,11,12。这些土壤玉米螟昆虫的成熟幼虫通常离开主人, 落在地上, 漫步寻找一个合适的地点, 挖洞进入土壤, 并为玉米螟8,10建造一个蛹室。
茶套,蛾 grisescens (鳞翅目: 尺蛾科), 是茶树山茶属植物中最重要的柱坑害虫之一.13。虽然这一物种在1894年首次被描述, 它已被错误地识别为蛾真菌Prout (鳞翅目: 尺蛾科) 在过去的几十年14,15。最近的一些研究14,15,16描述了两种同级物种之间的形态学、生物学和地理分布的差异。例如, 张等。15报告说, 该地区的倾斜主要发生在中国三个省份 (安徽、江苏和浙江) 的边界, 而grisescens的分布较斜的大得多.因此, grisescens造成的经济损失在很大程度上被忽视了, 这一害虫的知识需要被广泛修订和更新16,17,18,19.我们以前的研究表明, E. grisescens更喜欢在土壤中化蛹, 但也可以在土壤不可用时化蛹 (无化蛹基质条件)11,12。
本文提供了一个循序渐进的过程, 以 (1) 确定grisescens的化蛹偏好, 以响应的因素, 如基底类型和水分含量, 通过使用多选择生物鉴定, 和 (2) 确定非生物因素的影响grisescens生物鉴定的化蛹行为及出现的成功。所有这些生物鉴定都是在控制良好的实验室条件下进行的。此外, 这些生物鉴定也适用于评估其他因素对不同土壤玉米螟昆虫化蛹行为和存活的影响。

<table>
	<tbody>
		<tr>
			<td>Triangular flask</td>
			<td>Bomex Chemical (Shanghai) Co., LTD</td>
			<td>99</td>
			<td>250 mL</td>
		</tr>
		<tr>
			<td>Plastic basin</td>
			<td>Chahua, Fuzhou, China</td>
			<td>100</td>
			<td>upper side: 51 cm in diameter; bottom side: 40 cm in diameter; height: 16 cm</td>
		</tr>
		<tr>
			<td>Zip lock bags</td>
			<td>Glad, Guangzhou, China</td>
			<td>126/133</td>
			<td></td>
		</tr>
		<tr>
			<td>Polypropylene containers</td>
			<td>Youyou Plastic Factory, Taian, China</td>
			<td>139/155/160/161/190</td>
			<td>upper side: 20.0 cm [L] &#215; 13.5 cm [W], bottom side: 17.0 cm [L] &#215; 10.0 cm [W], height: 6.5 cm</td>
		</tr>
		<tr>
			<td>Waterproof polyviny chloride sheet</td>
			<td>Yidimei, Shanghai, China</td>
			<td>141</td>
			<td></td>
		</tr>
		<tr>
			<td>Tape</td>
			<td>V-tech, Guangzhou, China</td>
			<td>VT-710</td>
			<td></td>
		</tr>
		<tr>
			<td>Oven drier</td>
			<td>Kexi, Shanghai, China</td>
			<td>KXH-202-3A</td>
			<td></td>
		</tr>
		<tr>
			<td>Environmental chamber</td>
			<td>Life Apparatus, Ningbo, China</td>
			<td>PSX-280H</td>
			<td></td>
		</tr>
	</tbody>
</table>

choice and no-choice bioassays to study the pupation preference and emergence success of ectropis grisescens

许多昆虫生活在地面之上作为幼虫和成人, 并在地下化蛹。与他们生命周期的上述阶段相比, 在土壤中化蛹环境因素对这些昆虫的影响较少。蛾 grisescens (鳞翅目: 尺蛾科) 是茶树的一种严重害虫, 在华南造成了巨大的经济损失。本文所述的协议旨在通过多种选择生物鉴定研究, 即成熟的末龄grisescens幼虫是否能鉴别土壤变量, 如基底类型和含水量, 并通过无选择来确定生物鉴定, 基质类型和含水量对化蛹行为的影响以及 grisescens 的出现成功.研究结果将加深对grisescens化蛹生态学的认识, 并可为抑制grisescens种群的土壤管理策略带来深刻的见解。此外, 还可以对这些生物鉴定进行修改, 研究各种因素对土壤玉米螟害虫化蛹行为和存活的影响。

生物鉴定的水分选择表明, 与80% 水分砂相比, grisescens个体 pupated 在 5%-35%-水分砂中的含量明显增加 (图 2a)。然而, 有相当多的人更倾向于在土壤中或内化蛹 (沙质壤土1和2和淤泥壤土), 具有中间含水量 (图 2b - 2d)。
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/58126/58126fig2.jpg"><...

Watch this Scientific Journal Video about Choice and No-Choice Bioassays to Study the Pupation Preference and Emergence Success of Ectropis grisescens at JoVE.com

Choice and No-Choice Bioassays to Study the Pupation Preference and Emergence Success of Ectropis grisescens

选择与无选择生物鉴定研究蛾 grisescens的化蛹偏好及出现的成功

在这里, 我们提出了一个协议, 以研究蛾 grisescens成熟幼虫的化蛹偏好, 以响应土壤因子 (如基质类型和含水量) 使用选择生物鉴定。我们还提出了一种无选择生物鉴定的协议, 以确定影响 grisescens 化蛹行为和存活的因素.

Many insects live above the ground as larvae and adults and as pupate below the ground. Compared to the above-ground stages of their life cycles, less ...

该方法可针对不同环境因素，提供与茶叶毛毛虫、Ectropis毛虫的幼崽生态相关的重要信息，如它们的幼崽行为和幼崽生存。该技术的主要优点是，它允许您使用一种简单的方法，如测量的幼崽偏好和Ectropis格里森的产生成功。首先，从茶树中切出新鲜芽。然后用自来水填充 250 毫升三角烧瓶，然后将烧瓶放入烧瓶中。将三到四个烧瓶转移到塑料盆中。接下来，从E.grisescens的实验室群落中释放1000到2000只幼虫，放到茶笋的叶子上。在受控的实验室条件下保持幼虫。获得从芽的叶子上落下的成熟，最后一个星的幼虫，并沿着盆地底部积极漫步。在80摄氏度的烤箱烘干机中消毒土壤和沙子三天。然后在50摄氏度下将土壤和沙子干燥数周。当重量不随着时间的推移而变化时，基板样品将充分干燥。用木刺和砂浆研磨干燥的土壤。然后通过三毫米筛筛选沙子和地面土壤。将所需量的蒸馏水加入包含基材的密封袋中，并彻底混合。接下来，使用PVC板将聚丙烯容器分成六个相等的腔室。固定 PVC 板材，用热胶密封裂纹。接下来，用相同类型的基材填充不同水分含量的六个腔室。使用小块胶带将四到六片新鲜叶子粘贴到聚丙烯容器盖的内表面。接下来，释放30个成熟的最后一个星幼虫，粘贴到密室容器的盖子上的叶子上。小心地把盖子翻过来，盖住容器。在 26 摄氏度下，在 14 到 10 个光/暗光的室内保持生物测定竞技场。在第五天，计算每个腔室基板表面的幼崽数量。拆除生物测定并计算幼崽的数量。首先，使用PVC板材将聚丙烯容器分成四个腔室。然后，用热胶固定并密封PVC板材。用四种类型的基材填充容器的腔室，其水分含量相同，并随机分配订单。然后，在将新鲜茶叶翻到容器上之前，先用胶带将新鲜茶叶粘贴到盖子上。接下来，将幼虫转移到叶子上，记录和分析文本协议中概述的数据。准备 12 个基板水分含量处理后，将基板添加到塑料容器中。然后释放15个成熟的最后一星幼虫到每个竞技场的基质。在 26 摄氏度下，在 14 到 10 个光/暗光的室内维护每个生物测定竞技场。第三天，计算每个竞技场基底表面的幼虫数量和任何死幼虫的数量。最后，记录每天出现成人的数量，直到15天没有新成人出现。在该协议中，利用水分和基底选择生物测定来研究E.格里森的幼崽偏好。水分选择生物测定表明，与80%的水分砂相比，在5%和35%的水分砂上或体内的个体多。然而，明显有更多的个体喜欢在土壤中或土壤内进行中间水分含量的幼崽。底材选择生物测定表明，在20%的水分条件下和80%的水分条件下，沙子是E.grisescens的首选。然而，在50%的水分条件下，在四个基材中没有发现吸附偏好。经过开发，该技术为昆虫学领域的研究人员探索许多土壤虫的繁殖生态学铺平了道路。

Research

JoVE Journal

Environment

Choice and No-Choice Bioassays to Study the Pupation Preference and Emergence Success of Ectropis grisescens

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters (Homarus americanus)

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus

一种非侵入性的方法<em&gt;原位</em&gt;交配成功的女美国龙虾​​（测定<em&gt;美洲螯龙虾</em&gt;）

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American ...

科研

环境科学

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 ...

The Use of High-resolution Infrared Thermography (HRIT) for the Study of Ice Nucleation and Ice Propagation in Plants

The Use of High-resolution Infrared Thermography HRIT for the Study of Ice Nucleation and Ice Propagation in Plants

高分辨率红外热成像（HRIT）的冰核和冰传播的植物研究中的应用

Freezing events that occur when plants are actively growing can be a lethal event, particularly if the plant has no freezing tolerance. Such frost events ...

Empirical, Metagenomic, and Computational Techniques Illuminate the Mechanisms by which Fungicides Compromise Bee Health

经验、基因和计算技术阐明了杀菌剂危害蜜蜂健康的机制

Growers often use fungicide sprays during bloom to protect crops against disease, which exposes bees to fungicide residues. Although considered ...

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 ...

Using Pharmacological Manipulation and High-precision Radio Telemetry to Study the Spatial Cognition in Free-ranging Animals

使用药理操纵和高精度无线电遥测研究空间认知中自由放养的动物

An animal&#39;s ability to perceive and learn about its environment plays a key role in many behavioral processes, including navigation, migration, ...

Ecosystem Fabrication (EcoFAB) Protocols for The Construction of Laboratory Ecosystems Designed to Study Plant-microbe Interactions

Ecosystem Fabrication EcoFAB Protocols for The Construction of Laboratory Ecosystems Designed to Study Plant-microbe Interactions

用于研究植物-微生物相互作用的实验室生态系统制造 (EcoFAB) 协议

Beneficial plant-microbe interactions offer a sustainable biological solution with the potential to boost low-input food and bioenergy production. A ...

Experimental Study of the Relationship Between Particle Size and Methane Sorption Capacity in Shale

页岩中粒径与甲烷吸附能力关系的实验研究

The amount of adsorbed shale gas is a key parameter used in shale gas resource evaluation and target area selection, and it is also an important standard ...

Generating Homo- and Heterografts Between Watermelon and Bottle Gourd for the Study 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 ...