我们感谢埃米尔·伊马诺维奇博士提供的实验室和温室援助。我们还感谢雪莉·弗林特-加西亚博士提供玉米x特奥辛特内向线。

1. 植物材料的增长
<ol><li>选择植物线进行接种和筛选。本作品使用了两条玉米线、五条特奥辛特线和四十条具有非特异性抗性玉米x特奥辛特线（表1）。</li>
	<li>植物种子用于实验（美国梅迪斯 注射）和控制（注水）针注射接种实验。为每个工厂生产线这样做。</li>
	<li>在小平底中为每株植物线种植四个种子（复制品），用手指将种子推入土壤约1/2英寸，并轻轻覆盖土壤（图1A和 1B）。不要把土壤包在种子上。将种子种植得更深或将土壤包装在种子上可能会导致苗木出现问题。</li>
	<li>把种子浇进土壤里。确保土壤浸泡，浇水后种子留在土壤下。</li>
	<li>浇水后，将植物放置在生长室中，白天和黑夜环境分别为 28/20 °C 温度和 14/10 小时的光周期，约 500 微米/m2秒   的光合作用活性辐射位于树冠顶部。白天和晚上的相对湿度分别保持在70%和90%左右。</li>
	<li>将所有植物保持在同一生长室中，以保持整个实验的生长环境一致。</li>
	<li>10天后，将植物从生长室中取出，并用针头注射接种方法为植物接种 美国梅迪斯 细胞悬浮培养。注：玉米植物可以在种植8-10天后7天接种。然而，7天后，特奥辛特植物太小。因此，在种植10天后给玉米和特奥辛特植物接种疫苗，以在实验中保持一致性（见第2.12步）。</li>
</ol>2. 针头注射接种
<ol><li>在层流罩中完成所有工作。从冰柜储存中取出 美国甘 油库存。使用无菌循环和条纹甘油库存 的美国梅迪斯 野生型菌株1/2（交配类型a1b1）和2/9（交配类型a2b2，近同源到1/2）土豆脱氧脱糖（PDA）板。单独维护菌株。</li>
	<li>将带有 美国梅迪斯 的 PDA 板在 30 °C 孵化器中划出两天。如果使用不同的生物营养病原体使用适当的菌株，介质和生长条件。监测病原体在两天内的生长，以确保 美国梅迪斯 菌株生长良好。</li>
	<li>两天后从孵化器中取出PDA板。板块应具有良好的病原体生长，并包含单一菌落（图2A）。获得单一殖民地很重要。如果单个菌落不存在，则以较低的浓度对板块进行恢复。</li>
	<li>在层流罩中完成所有工作。使用无菌牙签从 PDA 板中为每个菌株选择一个菌落。将含有单个菌落的牙签放入 3 毫升土豆脱糖汤 （PDB） 中。建议有2-3种文化。</li>
	<li>将 3 毫升 PDB 培养物放入 30 °C 孵化器/摇床中，以 200 rpm 的速度放置两天。监测文化在两天内的增长，以确保文化的增长。文化应该显得很阴。</li>
	<li>从孵化器/摇床中取出液体培养物，测量OD600 的浓度，以确保细胞生长到1.0（+1 x 107 细胞/毫升）17的OD。</li>
	<li>将 美国梅迪斯 细胞悬浮培养物最终浓度为 1 x 106 细胞/毫升，使用水在最后 30 毫升培养量中。这种浓度持续导致植物的良好感染与病原体细胞悬浮培养。17 
</li>
</ol>注意：使用不同的病原体菌株时，应测试各种细胞悬浮浓度，以确定接种18，19所需的适当细胞滴定器。细胞悬浮培养的最终浓度可以用作滴答声的起点。应通过对感染良好的植物表型进行可视化来验证病原细胞悬浮培养物的适当浓度（图3A-E）。
<ol start="8"><li>在接种前混合两种 美国可能菌 株的等量。如果使用一种病原菌株继续步骤 2.9。为每次接种实验准备新的 美国梅迪斯 细胞悬浮培养物，并在两天后丢弃细胞悬浮培养物。</li>
	<li>对于实验性针头注射接种，通过将细胞悬浮培养物引入注射器，用 美国梅迪斯 细胞悬浮培养物填充3毫升注射器。</li>
	<li>对于控制针注射接种，用水填充一个3毫升注射器17。使用相同的程序进行实验针注射接种。</li>
	<li>在每 3 毫升注射器的末端附加 0.457 mm x 1.3 厘米皮下针。选定的针头尺寸将提供植物叶之间的细胞悬浮培养，对植物组织造成的损害最小。</li>
	<li>种植10天后从生长室中取出实验和控制植物，准备注射针头接种（图2B）（见第1.7步）。</li>
	<li>小心地将含有美国梅迪斯细胞悬浮培养物的皮下针插入位于土壤线正上方90°角的实验植物的茎中。插入针头，直到针头在茎的中间。不要将针头穿过茎（图2C）。</li>
	<li>给实验植物注射约100微克 的美国梅迪斯 细胞悬浮培养18，19。这会略有不同，这取决于幼苗的高度。细胞悬浮培养将穿过茎，进入植物的旋转。细胞悬浮培养将在植物的旋转中可见。继续向每个工厂注入 100 μl 的细胞悬浮培养，直到 3 毫升注射器为空。</li>
	<li>注射后，小心地从植物茎中取出针头。从现在空的 3 毫升注射器中取出针头，并装满水。将针头重新连接到注射器上，将水推过针头，以去除可能夹在针尖的任何植物组织。</li>
	<li>每个实验工厂重复步骤 2.9-2.15。通过注水，遵循对照工厂的相同协议。</li>
	<li>将接种的实验和控制植物放回生长室。每天通过润湿土壤而不是植物组织来浇水。</li>
	<li>每天检查植物，以检测病原体发展和植物抗药性反应。</li>
</ol>3. 电阻反应筛查
<ol><li>使用 1 到 5 阻力反应额定等级对接种后 7 天、10 天、14 天和 21 天的每株植物的电阻反应进行评分和记录。疾病严重程度随着额定等级上的数字值增加而增加（表2）。1C（叶氯化）、1A（叶花青素生产）或2（小叶胆）耐药反应表示耐药性。A 3（茎胆）、4（基底胆）或5（植物死亡）耐药反应表示易感性（图3A-E和表2）18，19。</li>
	<li>对实验和控制工厂进行评分，并记录阻力反应评分。</li>
	<li>比较实验和控制装置的抗药性反应。选择具有 1C、1A 或 2 阻力反应等级的实验植物。这些植物被认为对 美的18，19具有抗药性。</li>
	<li>重复整个实验以验证植物表型。</li>
</ol>

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作者没有什么可透露的。

在这项研究中，用于将一株 美国麦地的 针头注射接种方法成功输送到700株玉米和青金植物的茎中。此外，还使用修订后的抗病等级表来筛选植物和检测病原体发展。由于使用这两种方法，在700种玉米和特奥辛特植物中发现了抗 美国梅迪斯 的植物线，这些植物现在可以在育种计划中结合和测试，以提高抗病性。
与大多数接种方法一样，从同一系植物中复制相同?...

植物真菌病是农业面临的最突出的威胁之一。由于世界人口的粮食需求，开发抗病能力提高的作物的需求正在增加。植物病原体自然感染田间作物，引起疾病，对作物产量产生负面影响。结果表明，识别和利用抗性植物可以提高抗性，减少产量损失。许多植物品种，包括玉米、小麦、水稻和高梁，都通过给植物接种植物病原体和选择抗药性线7来识别抗性品种。因此，开发和使用有效的接种方法将使许多植物能够接种疫苗和筛选耐药性。各种接种方法已经使用，包括浸接种，管道病原体细胞悬浮培养到植物的旋转，针注射接种8-11。每种方法，病原体必须可靠地引入植物叶之间，病原体进入植物通过形成的复原体，以确保病原体的发展和植物感染12，13。
浸渍接种方法包括将植物幼苗浸入病原体细胞悬浮培养中，而管状方法要求将病原体细胞悬浮培养进入植物幼苗的旋转。但是，这两种方法都存在问题。首先，这两种方法都取决于病原体从叶面自然运动到植物组织，这是高度可变的。大多数病原体通过植物叶表面的口腔开口或伤口自然进入植物。然而，病原体通过叶面上的气雾和/或伤口穿透植物叶表面的能力存在显著变化。因此，无法用任何一种接种方法控制病原体渗透，这可能导致数据不一致。其次，在筛选大量植物时，将幼苗浸入病原体细胞悬浮培养物中可能非常耗时，并可能限制可筛选的植物数量。相反，此处描述的针头注射接种协议在植物叶之间提供病原体细胞悬浮培养，促进14型阿普索的形成。然后，病原体利用新开发的阿普西里亚进入植物，消除病原体渗透问题。此外，针头注射接种协议为已经接种了 美国梅迪斯 的玉米和青金植物提供了一系列表型，并表现出良好的感染。表型可以用作标记，以确定病原体细胞悬浮培养的最佳浓度，从而在不同的实验中和之间产生一致的植物表型。
植物接种后，与病原体细胞悬浮培养物，植物通常被筛选，以检测耐药或易感表型8-11，15。虽然疾病评级尺度被广泛用于筛选和分类植物表型，但等级等级因所分析的病原体而异。因此，一个疾病等级表协议建立为美国可能迪斯和玉米相互作用可用于类似的真菌病原体16。
本系列协议详细介绍了针头注射接种与 美国可能迪斯 细胞悬浮培养和抗病反应筛查玉米，硫酸盐，玉米x特异丙辛特内引线。目前的协议不限于针注射接种 美国可能迪斯 到玉米植物，但可用于相对任何真菌病原体和植物物种。因此，将这两种方法的细节包括在同一协议中将使研究人员能够直接利用协议进行接种和筛选，或操纵原始协议，以更好地适应病原体和植物物种的利益。

<table border="1">
	<tbody>
		<tr>
			<td>Name of Reagent/Material</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Seed for plants</td>
			<td></td>
			<td></td>
			<td>Collected from original crosses</td>
		</tr>
		<tr>
			<td>Growth chamber</td>
			<td>Conviron</td>
			<td>PGR14 REACH-IN</td>
			<td></td>
		</tr>
		<tr>
			<td>Planting flats</td>
			<td>Hummert International</td>
			<td>14-3385-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Soil (3 parts pine bark; 1 part peat moss with perlite)</td>
			<td>Hummert International</td>
			<td>10-1059-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Laminar flow hood</td>
			<td>Lab Conoco</td>
			<td>70875372</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycerol stock of pathogen (U. maydis) or fungal pathogen of interest</td>
			<td></td>
			<td></td>
			<td>Stocks were grown from original culture</td>
		</tr>
		<tr>
			<td>Sterile loop</td>
			<td>Fisher Scientific</td>
			<td>S17356A</td>
			<td></td>
		</tr>
		<tr>
			<td>Potato dextrose agar (PDA) plates</td>
			<td>Fisher Scientific</td>
			<td>R454311</td>
			<td></td>
		</tr>
		<tr>
			<td>Incubator set to 30 &#176;C</td>
			<td>Fisher Scientific</td>
			<td>11-690-650F</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile toothpicks</td>
			<td>Walmart</td>
			<td></td>
			<td>Purchased from Walmart and sterilized by autoclave</td>
		</tr>
		<tr>
			<td>Potato dextrose broth (PDB)</td>
			<td>Fisher Scientific</td>
			<td>ICN1008617</td>
			<td></td>
		</tr>
		<tr>
			<td>Incubator-shaker set to 30 &#176;C</td>
			<td>New Brunswick</td>
			<td>14-278-179</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrophotometer</td>
			<td>Fisher Scientific</td>
			<td>4001000</td>
			<td></td>
		</tr>
		<tr>
			<td>U. maydis&#160;cell suspension culture (1 x 106 cells/ml)</td>
			<td></td>
			<td></td>
			<td>Grown from glycerol stock as described in the methods</td>
		</tr>
		<tr>
			<td>3 ml Syringes</td>
			<td>Becton Dickinson</td>
			<td>309606</td>
			<td></td>
		</tr>
		<tr>
			<td>.457 mm x 1.3 cm Hypodermic needles</td>
			<td>Kendall Brands</td>
			<td>8881250321</td>
			<td></td>
		</tr>
	</tbody>
</table>

a rapid and efficient method for assessing pathogenicity of ustilago maydis on maize and teosinte lines

玉米是全世界主要的谷物作物。然而，易感性生物营养病原体是提高生产力的主要制约因素。美国麦迪斯是一种生物营养真菌病原体，也是玉米上玉米的因果因子。这种疾病导致美国每年损失约10亿美元。 然而，宿主电阻是管理玉米污迹的唯一实用方法。对各种生物营养病原体具有抗药性的作物作物，包括玉米、小麦和水稻的鉴定，每年可显著减少3-5个产量损失。因此，使用病原体接种方法，有效和可重复地将病原体输送到植物叶之间，将有助于快速识别抗美国玉米的玉米线。作为，向确定抗U.maydis的玉米线迈出的第一步，采用针注射接种法和抗性反应筛选法，为玉米、青金石和玉米x特奥辛特引进线接种，并选择抗性植物。
种植了由大约700株植物组成的玉米、青金石和玉米x特奥辛特自给系，接种了 一株美国梅迪斯，并进行了抗药性筛查。接种和筛选方法成功地确定了三条抗 美国梅迪斯的青金石线。这里介绍了玉米、青金石和玉米x青金石引种线的详细针头注射接种和抗性反应筛查方案。这项研究表明，针头注射接种是农业中一种宝贵的工具，可以有效地在植物叶之间输送 美国梅迪斯 ，并提供了对 美的 抗药性的植物线，现在可以在育种计划中结合和测试，以提高抗病性。

成功的针头注射接种可以通过可视化用美国梅迪斯（实验）接种的植物的表型来确定。大多数实验植物都容易感染美国梅迪斯病毒。易感植物表现出非常严重的疾病发展，表现为茎和基底胆形成与黑色三叶草孢子（图3D和3E，表2）。由于疾病的严重性，几种植物在接种后死亡。确定了三条抗美国梅迪斯的玉米x特奥辛特自入系。对于抗美...

Watch this Scientific Journal Video about A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines at JoVE.com

A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

据介绍，使用针头注射法为玉米和青金植物注射生物营养病原体 Ustilago。 针头注射接种方法通过形成病原体，促进病原体进入植物的植物叶子之间的真菌病原体的可控输送。这种方法效率很高，使 美国梅迪斯的可重复接种成为可能。

评估 乌西拉戈 病原性的快速高效方法，可在玉米和特奥辛特线上使用

Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is ...

a-rapid-efficient-method-for-assessing-pathogenicity-ustilago-maydis

Research

JoVE Journal

Environment

A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

8.4K 次观看.  University of Georgia. 据介绍，使用针头注射法为玉米和青金植物注射生物营养病原体 Ustilago。 针头注射接种方法通过形成病原体，促进病原体进入植物的植物叶子之间的真菌病原体的可控输送。这种方法效率很高，使 美国梅迪斯的可重复接种成为可能。

视频: A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

Use of Chironomidae (Diptera) Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies

Rapid bioassessment protocols using benthic macroinvertebrate assemblages have been successfully used to assess human impacts on water quality. Unfortunately, traditional benthic larval sampling methods, such as the dip-net, can be time-consuming and expensive. An alternative protocol involves collection of Chironomidae surface-floating pupal exuviae (SFPE). Chironomidae is a species-rich family of flies (Diptera) whose immature stages typically occur in aquatic habitats. Adult chironomids emerge from the water, leaving their pupal skins, or exuviae, floating on the water&#8217;s surface. Exuviae often accumulate along banks or behind obstructions by action of the wind or water current, where they can be collected to assess chironomid diversity and richness. Chironomids can be used as important biological indicators, since some species are more tolerant to pollution than others. Therefore, the relative abundance and species composition of collected SFPE reflect changes in water quality. Here, methods associated with field collection, laboratory processing, slide mounting, and identification of chironomid SFPE are described in detail. Advantages of the SFPE method include minimal disturbance at a sampling area, efficient and economical sample collection and laboratory processing, ease of identification, applicability in nearly all aquatic environments, and a potentially more sensitive measure of ecosystem stress. Limitations include the inability to determine larval microhabitat use and inability to identify pupal exuviae to species if they have not been associated with adult males.

Use of Chironomidae Diptera Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies

Rapid bioassessment protocols using benthic macroinvertebrates are often used to monitor and assess water quality. An efficient protocol involves collections of Chironomidae surface-floating pupal exuviae (SFPE). Here, techniques for field collection, laboratory processing, slide mounting, and identification of Chironomidae SFPE are described.

使用摇蚊科（双翅目）的表面浮蛹蜕为快速生物议定书水体

Rapid bioassessment protocols using benthic macroinvertebrate assemblages have been successfully used to assess human impacts on water quality. ...

科研

环境科学

The Use of an Automated System (GreenFeed) to Monitor Enteric Methane and Carbon Dioxide Emissions from Ruminant Animals

Ruminant animals (domesticated or wild) emit methane (CH4) through enteric fermentation in their digestive tract and from decomposition of manure during storage. These processes are the major sources of greenhouse gas (GHG) emissions from animal production systems. Techniques for measuring enteric CH4 vary from direct measurements (respiration chambers, which are highly accurate, but with limited applicability) to various indirect methods (sniffers, laser technology, which are practical, but with variable accuracy). The sulfur hexafluoride (SF6)&#160;tracer gas method is commonly used to measure enteric CH4 production by animal scientists and more recently, application of an Automated Head-Chamber System (AHCS) (GreenFeed, C-Lock, Inc., Rapid City, SD), which is the focus of this experiment, has been growing. AHCS is an automated system to monitor CH4 and carbon dioxide (CO2) mass fluxes from the breath of ruminant animals. In a typical AHCS operation, small quantities of baiting feed are dispensed to individual animals to lure them to AHCS multiple times daily. As the animal visits AHCS, a fan system pulls air past the animal&#8217;s muzzle into an intake manifold, and through an air collection pipe where continuous airflow rates are measured. A sub-sample of air is pumped out of the pipe into non-dispersive infra-red sensors for continuous measurement of CH4 and CO2 concentrations. Field comparisons of AHCS to respiration chambers or SF6 have demonstrated that AHCS produces repeatable and accurate CH4 emission results, provided that animal visits to AHCS are sufficient so emission estimates are representative of the diurnal rhythm of rumen gas production. Here, we demonstrate the use of AHCS to measure CO2 and CH4 fluxes from dairy cows given a control diet or a diet supplemented with technical-grade cashew nut shell liquid.

The Use of an Automated System GreenFeed to Monitor Enteric Methane and Carbon Dioxide Emissions from Ruminant Animals

Accuracy and precision of the techniques used to measure methane emissions from ruminant animals are critically important for the success of greenhouse gas mitigation efforts. This manuscript describes the principles and operation of an automated system to monitor methane and carbon dioxide mass fluxes from the breath of ruminant animals.

自动系统（绿色饲料）的用于监视肠道的甲烷和二氧化碳从反刍动物排放

Ruminant animals (domesticated or wild) emit methane (CH4) through enteric fermentation in their digestive tract and from decomposition of manure during ...

Administering and Detecting Protein Marks on Arthropods for Dispersal Research

Monitoring arthropod movement is often required to better understand associated population dynamics, dispersal patterns, host plant preferences, and other ecological interactions. Arthropods are usually tracked in nature by tagging them with a unique mark and then re-collecting them over time and space to determine their dispersal capabilities. In addition to actual physical tags, such as colored dust or paint, various types of proteins have proven very effective for marking arthropods for ecological research. Proteins can be administered internally and/or externally. The proteins can then be detected on recaptured arthropods with a protein-specific enzyme-linked immunosorbent assay (ELISA). Here we describe protocols for externally and internally tagging arthropods with protein. Two simple experimental examples are demonstrated: (1) an internal protein mark introduced to an insect by providing a protein-enriched diet and (2) an external protein mark topically applied to an insect using a medical nebulizer. We then relate a step-by-step guide of the sandwich and indirect ELISA methods used to detect protein marks on the insects. In this demonstration, various aspects of the acquisition and detection of protein markers on arthropods for mark-release-recapture, mark-capture, and self-mark-capture types of research are discussed, along with the various ways that the immunomarking procedure has been adapted to suit a wide variety of research objectives.

Proteins are often used to mark arthropods for dispersal research. Methods for administering internal and external protein marks on arthropods are demonstrated. Protein mark detection techniques, either through indirect or sandwich enzyme-linked immunosorbent assays (ELISAs), are also illustrated.

管理和检测蛋白标记的节肢动物的扩散研究

Monitoring arthropod movement is often required to better understand associated population dynamics, dispersal patterns, host plant preferences, and other ...

Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester fabric and glue can be folded and glued to construct a strip of compartments that house seeds and identifying information, while allowing contact with soil leachate, water, microorganisms, and ambient temperature. Strips may be constructed with a wide range of compartment numbers and sizes and allow the researcher to house a variety of genotypes within a single species, different species, or seeds that have experienced different treatments. As opposed to individual seed packets, strips are more easily retrieved as a unit. While replicate packets can be included within a strip, different strips can act as blocks or can be retrieved at different times for observation of seed behavior over time. We used a high temperature glue gun to delineate compartments and sealed the strips once the seed and tags identifying block and removal times were inserted. The seed strips were then buried in the field at the desired depth, with the location marked for later removal. Burrowing animal predators were effectively excluded by use of a covering of metal mesh hardware cloth on the soil surface. After the selected time interval for burial, strips were dug up and seeds were assessed for germination, dormancy and mortality. While clearly dead seeds can often be distinguished from ungerminated living ones by eye, dormant seeds were conclusively identified using a standard Tetrazolium chloride colorimetric test for seed viability.

Here we present a protocol for assessing seed survivorship, germination and dormancy under field conditions using buried, labeled seed strips and tetrazolium chloride (TZ) viability testing.

对于在野外条件下评估种子萌发，休眠和死亡率可靠的方法

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester ...

Protocol for Assessing the Relative Effects of Environment and Genetics on Antler and Body Growth for a Long-lived Cervid

Cervid phenotype can be placed into one of two categories: efficiency, which promotes survival over extravagant morphometric growth, and luxury, which promotes growth of large weaponry and body size. Populations of the same species display each phenotype depending on environmental conditions. Although antler and body size of male white-tailed deer (Odocoileus virginianus) varies by physiographic region in Mississippi, USA and is strongly correlated with regional variation in nutritional quality, the effects of population-level genetics from native stocks and previous re-stocking efforts cannot be disregarded. This protocol describes how we designed a controlled study, where other factors that influence phenotype, such as age and nutrition, are controlled. We brought wild-caught pregnant females and six-month-old fawns from three distinct physiographic regions in Mississippi, USA to the Mississippi State University Rusty Dawkins Memorial Deer Unit. Deer from the same region were bred to produce a second generation of offspring, allowing us to assess generational responses and maternal effects. All deer ate the same high-quality (20% crude protein deer pellet) diet ad libitum. We uniquely marked each neonate and recorded body mass, hind foot, and total body length. Each subsequent fall, we sedated individuals via remote injection and sampled the same morphometrics plus antlers of adults. We found that all morphometrics increased in size from first to second generation, with full compensation of antler size (regional variation no longer present) and partial compensation of body mass (some evidence of regional variation) evident in the second generation. Second generation males that originated from our poorest quality soil region displayed about a 40% increase in antler size and about a 25% increase in body mass when compared to their wild harvested counterparts. Our results suggest phenotypic variation of wild male white-tailed deer in Mississippi are more related to differences in nutritional quality than population-level genetics.

Phenotypic differences among cervid populations may be related to population-level genetics or nutrition; discerning which is difficult in the wild. This protocol describes how we designed a controlled study where nutritional variation was eliminated. We found that phenotypic variation of male white-tailed deer was more limited by nutrition than genetics.

评估环境和遗传因素对鹿茸和长寿命的鹿的身体生长的相对影响的议定书 》

Cervid phenotype can be placed into one of two categories: efficiency, which promotes survival over extravagant morphometric growth, and luxury, which ...

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

Dimethylated thioarsenicals such as dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV), which are produced by the metabolic pathway of dimethylarsinic acid (DMAV) thiolation, have been recently found in the environment as well as human organs. DMMTAV and DMDTAV can be quantified to determine the ecological effects of dimethylated thioarsenicals and their stability in environmental media. The synthesis method for these compounds is unstandardized, making replicating previous studies challenging. Furthermore, there is a lack of information about storage techniques, including storage of compounds without species transformation. Moreover, because only limited information about synthesis methods is available, there may be experimental difficulties in synthesizing standard chemicals and performing quantitative analysis. The protocol presented herein provides a practically modified synthesis method for the dimethylated thioarsenicals, DMMTAV and DMDTAV, and will help in the quantification of species separation analysis using high performance liquid chromatography in conjunction with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). The experimental steps of this procedure were modified by focusing on the preparation of chemical reagents, filtration methods, and storage.

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

This article presents modified experimental protocols for dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV) synthesis, inducing dimethylarsinic acid (DMAV) thiolation through mixing of DMAV, Na2S, and H2SO4. The modified protocol provides an experimental guideline, thereby overcoming limitations of the synthesis steps that could have caused experimental failures in quantitative analysis.

使用 DMAv为环境应用 DMMTAv和 DMDTAv的准备: 合成、纯化和确认

Dimethylated thioarsenicals such as dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV), which are produced by the metabolic ...

The Hawaii Protocol for Scientific Monitoring of Coffee Berry Borer: a Model for Coffee Agroecosystems Worldwide

Coffee berry borer (CBB) is the most devastating insect pest for coffee crops worldwide. We developed a scientific monitoring protocol that is aimed at capturing and quantifying the dynamics and impact of this invasive insect pest as well as the development of its host plant across a heterogeneous landscape. The cornerstone of this comprehensive monitoring system is timely georeferenced data collection on CBB movement, coffee berry infestation, mortality by the fungus Beauveria bassiana, and coffee plant phenology via a mobile electronic data recording application. This electronic data collection system allows field records to be georeferenced through built-in global positioning systems, and is backed by a network of weather stations and records of farm management practices. Comprehensive monitoring of CBB and host plant dynamics is an essential part of an area-wide project in Hawaii to aggregate landscape-level data for research to improve management practices. Coffee agroecosystems in other parts of the world that experience highly variable environmental and socioeconomic factors will also benefit from implementing this protocol, in that it will drive the development of customized integrated pest management (IPM) to manage CBB populations.

Comprehensive monitoring of coffee berry borer and host plant dynamics is essential for aggregating landscape-level data to improve management of this invasive pest. Here, we present a protocol for scientific monitoring of coffee berry borer movement, infestation, mortality, coffee plant phenology, weather, and farm management via a mobile electronic data recording application.

夏威夷咖啡莓科学监测协议: 全球咖啡农田模型

Coffee berry borer (CBB) is the most devastating insect pest for coffee crops worldwide. We developed a scientific monitoring protocol that is aimed at ...

A Flexible Low Cost Hydroponic System for Assessing Plant Responses to Small Molecules in Sterile Conditions

A wide range of studies in plant biology are performed using hydroponic cultures. In this work, an in vitro hydroponic growth system designed for assessing plant responses to chemicals and other substances of interest is presented. This system is highly efficient in obtaining homogeneous and healthy seedlings of the C3 and C4 model species Arabidopsis thaliana and Setaria viridis, respectively. The sterile cultivation avoids algae and microorganism contamination, which are known limiting factors for plant normal growth and development in hydroponics. In addition, this system is scalable, enabling the harvest of plant material on a large scale with minor mechanical damage, as well as the harvest of individual parts of a plant if desired. A detailed protocol demonstrating that this system has an easy and low-cost assembly, as it uses pipette racks as the main platform for growing plants, is provided. The feasibility of this system was validated using Arabidopsis seedlings to assess the effect of the drug AZD-8055, a chemical inhibitor of the target of rapamycin (TOR) kinase. TOR inhibition was efficiently detected as early as 30 min after an AZD-8055 treatment in roots and shoots. Furthermore, AZD-8055-treated plants displayed the expected starch-excess phenotype. We proposed this hydroponic system as an ideal method for plant researchers aiming to monitor the action of plant inducers or inhibitors, as well as to assess metabolic fluxes using isotope-labeling compounds which, in general, requires the use of expensive reagents.

A simple, versatile, and low-cost in vitro hydroponic system was successfully optimized, enabling large-scale experiments under sterile conditions. This system facilitates the application of chemicals in a solution and their efficient absorption by roots for molecular, biochemical, and physiological studies.

一种灵活的低成本水培系统在无菌条件下评价植物对小分子的响应

A wide range of studies in plant biology are performed using hydroponic cultures. In this work, an in vitro hydroponic growth system designed for ...

High-throughput, Microscale Protocol for the Analysis of Processing Parameters and Nutritional Qualities in Maize (Zea mays L.)

Maize is an important grain crop in the United States and worldwide. However, maize grain must be processed prior to human consumption. Furthermore, whole grain composition and processing characteristics vary among maize hybrids and can impact the quality of the final processed product. Therefore, in order to produce healthier processed food products from maize, it is necessary to know how to optimize processing parameters for particular sets of germplasm to account for these differences in grain composition and processing characteristics. This includes a better understanding of how current processing techniques impact the nutritional quality of the final processed food product. Here, we describe a microscale protocol that both simulates the processing pipeline to produce cornflakes from large flaking grits and allows for the processing of multiple grain samples simultaneously. The flaking grits, the intermediate processed products, or final processed product, as well as the corn grain itself, can be analyzed for nutritional content as part of a high-throughput analytical pipeline. This procedure was developed specifically for incorporation into a maize breeding research program, and it can be modified for other grain crops. We provide an example of the analysis of insoluble-bound ferulic acid and p-coumaric acid content in maize. Samples were taken at five different processing stages. We demonstrate that sampling can take place at multiple stages during microscale processing, that the processing technique can be utilized in the context of a specialized maize breeding program, and that, in our example, most of the nutritional content was lost during food product processing.

High-throughput, Microscale Protocol for the Analysis of Processing Parameters and Nutritional Qualities in Maize Zea mays L.

Here, we present a microscale protocol for processing grain samples and for incorporating this microscale approach into a high-throughput analytical pipeline. This is a higher throughput adaptation of currently available protocols.

玉米加工参数和营养品质分析的高通量、微尺度协议 (玉蜀黍)

Maize is an important grain crop in the United States and worldwide. However, maize grain must be processed prior to human consumption. Furthermore, whole ...

Assessing the Particulate Matter Removal Abilities of Tree Leaves

Based on the conventional cleaning methods (water cleaning (WC) + brush cleaning (BC)), this study evaluated the influence of ultrasonic cleaning (UC) on collecting various sized particulate matter (PM) retained on leaf surfaces. We further characterized the retention efficiency of leaves to various sized PM, which will help to assess the abilities of urban trees to remove PM from ambient air quantitatively. 
Taking three broadleaf tree species (Ginkgo biloba, Sophora japonica, and Salix babylonica) and two needleleaf tree species (Pinus tabuliformis and Sabina chinensis) as the research objects, leaf samples were collected 4 days (short PM retention period) and 14 days (long PM retention period) after the latest rainfall. PM retained on the leaf surfaces was collected by means of WC, BC, and UC in sequence. Then, retention efficiencies of leaves (AEleaf) to three types of the various sized PM, including easily removable PM (ERP), difficult-to-remove PM (DRP), and totally removable PM (TRP), were calculated. Only around 23%-45% of the total PM retained on leaves could be cleaned off and collected by WC. When the leaves were cleaned through WC+BC, the underestimation of the PM retention capacity of different tree species was in the range of 29%-46% for various sized PM. Almost all PM retained on leaves could be removed if UC was supplemented to WC+BC. 
In conclusion, if the UC was complemented after the conventional cleaning methods, more PM on leaf surfaces could be eluted and collected. The procedure developed in this study can be used for assessing the PM removal abilities of different tree species.

The ultrasonic cleaning method was applied to elute the particulate matter (PM) retained on leaf surfaces after PM was eluted by the conventional cleaning methods (water cleaning only or water cleaning plus brush cleaning). The methodology can help to improve the estimation accuracy for PM retention capacity of leaves.

树叶颗粒物去除能力的评价

Based on the conventional cleaning methods (water cleaning (WC) + brush cleaning (BC)), this study evaluated the influence of ultrasonic cleaning (UC) on ...

评估 乌西拉戈 病原性的快速高效方法，可在玉米和特奥辛特线上使用

评估乌西拉戈病原性的快速高效方法，可在玉米和特奥辛特线上使用