This work was supported by Biotech Risk Assessment Grant Program competitive grand no. 2006-39454-17438 to A. Snow, K. Mercer, and H. Alexander from the United States Department of Agriculture, National Institute of Food and Agriculture. Experiments using this method were conducted at and supported by the University of Kansas Field Station, a research unit of the Kansas Biological Survey and the University of Kansas. The authors would like to thank P. Jourdan and E. Regnier for helpful reviews on earlier versions of this manuscript. Additionally, this work was aided by the contributions of the staff at the University of Kansas Field Station, Waterman Farm at the Ohio State University (OSU), the USDA Ornamental Plant Germplasm Center at OSU, and the Seed Biology Lab in the Department of Horticulture and Crop Science at OSU, especially E. Renze, S. Stieve, A. Evans, and E. Grassbaugh, for technical support.

1.从多个物种或品种单一控制杂交种子收集注意:使用野性，混合动力，和作物类型作为母体（制种）母种向日葵 （向日葵）内15个十字型本例使用的种子。 <ol><li>在生长季节结束后，收集袋标有成熟的种子头。从谷壳和地点种子标有标准格式（ 即产妇点¯x父）亲十字型信封干净的种子。 </li><li>散装种子一起在大信封用的种子等量从同等数量的同一父母跨类型。例如，从10跨种100粒种子用相同的母系和父系父母。与识别（交叉型）标签上的信封。这些大信封大师的种子在第2节灌墓葬条使用。 </li></ol><table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody&gt; <tr><td></td><td colspan="3"> 母本 </td></tr><tr><td> 父本 </td><td>野:WXW </td><td> ˚F1:WXC </td><td>作物:CXC </td></tr><tr><td>野:WXW </td><td> 0％WXW </td><td> 25％F 1 xW，它们† </td><td> 50％CXW† </td></tr><tr><td> BC:WxF1或F1xW </td><td> 12.5％WxBC </td><td> 37.5％F 1 XBC </td><td> 62.5％CxBC </td></tr><tr><td> ˚F1:WXC </td><td> 25％WXF 1† </td><td> 50％F 1 xF的1 </td><td> 75％的CxF 1† </td></tr><tr><td> F 2:F 1 xF的1 </td><td> 25％WXF 2 </td><td> 50％F 1 xF的2 </td><td> 75％的CxF 2 </td></tr><tr><td>作物:CXC </td><td> 50％WXC† </td><td> 75％F 1 XC† </td><td> 100％CXC </td></tr></tbody></table> 表1.家长交Ť从人工授粉向日葵作物野杂交种生产ypes制作以手工授粉种子埋藏实验中使用。对于所有类型的交叉，母本列第一和父本第二上市。标有†跨类型与同一作物％等位基因，但不同的母本相互十字型对的一部分。表先前已发表于:吴佩慈，BA 等 。 （2015年）15。 2.创建自定义种子埋藏条注:在这个例子中，我们有三个去除日期的治疗和15重复，所以我们所需总45条。本例使用每带15车厢的话，我们需要足够的细孔涤纶织物或蚊帐容纳15，7×10cm的车厢。 见图1。 <ol><li>通过削减织物20×105厘米条每个种子带开始。 </li><li>测量和标记车厢，然后对折织物。使用HIGH-高温胶枪，封口一短（10厘米）结束。一旦胶水冷却略有下降，但不完全，按面料双方共同打造一个密封。 注意:高温胶和胶枪技巧可引起灼伤，如果与皮肤接触的延长。 </li><li>胶线从折叠边缘在每个隔间标记开短端。按两种织物双方共同沿着胶线，确保密封件不室之间的孔。种子车厢之间迁移造成实验误差。 注:现在，车厢已创建，展开的边缘应保持开放并准备接收种子和他们的身份信息（交叉型名称）。 </li><li>随机分配的种子交叉类型车厢每个带。 <ol><li>填充种子隔间具有均匀数目的种子（20为每个隔室在本实施例中使用）。对于标签，使用上切到合适的塑料飞镖园工业记号笔，使他们能够承受埋葬。 注:可使用的替代标记方法。 </li><li>每个隔室填充有种子和其标签后，立即密封隔室与高温胶枪。继续进行，直到整个种子带被完全密封。在用复制，嵌段和/或治疗信息的第一隔室标签整体带。 注:对于我们的研究，因为我们有时间清除，块和块中的复制条，我们包括所有这些信息。 </li><li>检查车厢和现货胶之间的缝隙上移动到下一个条之前需要。 </li></ol></li></ol><img alt="图1" src="/files/ftp_upload/54663/54663fig1.jpg" /> 图1.埋葬种子带图。显示了各个隔间跨类型识别埋藏带的例子。母本列第一与第二上市父本。颜色表示不同的母系血统，黄色的野生，蓝色杂种F 1，红色作物。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54663/54663fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 3.种子埋在条领域<ol><li>挖地块到指定深度的房子带。分配条，其中条带被随机地布置，以允许移除日期各个块以用作如在分析的副区的主要情节和隔室的身份（在本例中，十字型）。 </li><li>覆盖用土带。切割金属网金属布比超过埋条情节和地方大10厘米。使用U-引脚与地获得边缘和盖布边。 注意:这是覆盖排除穴居种天敌。作为另一种选择，美世等 13构造埋包围由于激烈的gopher压力种子带在自己的领域LOCA硬件布盒化）。 </li><li>将彩色验船师的标志来区分条的位置和拆除日期，以补充明确的书面记录。 </li></ol> 4.挖掘和条种子评估<ol><li>挖分配给给定移除日期种子条。保持在阴凉潮湿的环境条，直到他们到达实验室，种子的评估将发生。这可通过在湿报纸包裹片和放置在运输或过夜航运冷却器来实现。 </li><li>从冲洗淤泥带和土壤中水，直到条是不够干净外部碎片不会晦涩种子。 </li><li>用剪刀在一次划开一个隔间。计数发芽的种子，并删除它们。显然伯爵死种子（ 如腐烂，或腐烂的种子）。广场上标记，湿Petri网规模的吸墨纸未萌发的种子进一步区分未萌发，休眠和死亡的种子。 <ol><li>将培养皿萌发的种子成标准的发芽条件下生长室集:25℃/ 10℃，12小时昼/夜。允许1周种子发芽。任何做的，计为可行，但萌发的，因为他们没有在野外条件下发芽。 </li></ol></li><li>生长室处理后，测试采用四氮唑（TZ）13剩余未萌发的种子活力。要做到这一点，弥补了TZ适当的使用AOSA四氮唑测试手册14本研究物种％的溶液。对于这个例子，使用了1％的溶液。 <ol><li>切未萌发的种子的一半，并把一半是在胚和胚乳在含有足够的TZ解决方案覆盖种子培养皿中可见（种子可能是浮动的，这是正常的）。 </li><li>放置TZ的培养皿入设定在30℃的培养箱中，并等待3小时。注:较高的温度将导致更快的反应，如果需要的话，但overstaining是有风险的。 </li></ol></li> &lt;li&gt;从孵化器中取出培养皿和评估。在他们的胚胎已经染成红色的种子可萌发，同时未染色的种子都死了。关于区分边际案件和特定物种的详细咨询AOSA四氮唑测试手册。 </li></ol>

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The authors have nothing to disclose.

在这里，我们目前使用的种子埋藏带在现场预选的时间段来观察种子发芽，休眠和多样化种子储备的死亡方法。用条而不是单个数据包在于（1）条和隔间建设创造了单个数据包的速度的优势;和（2）的难易程度和在一个运动中除去多个隔间没有遗漏的分组或无意地去除一个的危险的速度。作为两个这里介绍的实施例中除去的日期是在冬季当土壤硬和现场条件是冷，这种方法降低了劳动和人为错误的可能性。种子带也通过允许研究人员随机在实验室隔室的位置在不能由人的错误的字段被改变的方式帮助实验物流。通过去除次选择，涵盖的预测自然萌发的时候我们是广度能够做出推论约种子库，在我们的例子中潜在的杂交种子在种子库依然存在，有助于未来的人口。这种方法的一个限制是，隔室之间的种子的一些迁移可能是不可避免的，这取决于种子的大小。出于这个原因，（和在协议中规定），该调查仔细检查室之间的高温胶密封件，以减少种子迁移是重要的。 种子埋藏实验的一些问题可以通过精明地思考时间来避免。选择在时间相关的研究物种的自然循环和学习目标，埋葬和去除日期。最好是尽快之后自然脱落的种子尽可能厂。去除日期应根据高峰出苗时间的正常萌发行为观察评估选择;增加较早的日期还使研究人员能够捕捉提早发芽。 BEC澳洲英语我们以前的工作曾表示，一些不适应的萌发产生的，由于完全缺乏一些交叉类型的休眠，我们选择了一个非常早期的拆除日期之后仅几个星期葬（去除1，秋末）15，16。 在各个分组条带的优点是，一打或更多个隔室可埋，除去，并评价为一体，从而允许用于涉及一切从多个删除日期，以灌溉或营养疗法复杂的实验设计。调查人员应注意构建块，让他们最关心的理解是主要影响治疗。我们的例子强调种子的行为随着时间的推移，但其他治疗方法可以应用，将需要考虑领域的布局，并在实验设计阶段复制块内安排。通过随机分配的块内带埋葬位置，拆除时间可以分析为主要功效。 &lt;P类="jove_content"&gt;除去种子条带后，制备的Petri尺寸吸墨纸的预标记栈评估开始前整条减少人为的错误。同样，开放和评估在一个时间内单个舱有助于保持精度。当把未萌发的种子放入生长室，可以肯定的是，昼夜温差匹配研究物种的最佳种子发芽的条件。而标准发芽条件下（25℃/ 10°C 12小时天/夜）可能是适当的多种物种的，这并不是对所有物种的情况。 考虑到种子染色模式的TZ治疗中的应用及评价强烈影响的研究成果。当生长室治疗后评估未萌发的种子，参考文献（或AOSA四唑测试手册14）TZ的一个给定的物种的正确浓度。虽然判断个人TZ染色小号电火工品，要记住的种子活力和种子的发芽，弱染色的种子可以指示呼吸的组织，这可能是无法从土壤出现并产生一幼苗能力之间的差别是很重要的。这说明，真正的休眠的种子是那些保持最佳发芽条件下萌发的，但测试时仍然强烈可行的。这些种子都可能有助于持久种子库，并在未来几年发芽的唯一种子。 许多这些方法的应用是可能超出这里概述。种子埋藏条可以定制几乎所有的实验装置和实验设计。它们可应用于比较不同物种，种子大小，或条件/处理。建立在这些方法都有加深许多物种的种子基本生物学知识的生态环境有关的潜力。

该方法的总的目标是可靠地评估种子生存随时间在野外条件下。 土壤种子库的分布无论是在土壤表面，地面枯枝落叶层内，或在土壤剖面分散的，可行的但萌发的种子，可以瞬时或多年坚持1,2的储备。当类似的这里提出的种子埋藏方法应用于使用几种十余种了17年的研究中，可行的种子在许多测试3种被发现。种子休眠是种子发芽直到对幼苗存活的条件适当的组合出现4块。剩下的休眠可以让种子恶劣条件下生存，如冬季气温低，营养限制，或季节性干旱，直到休眠释放外部触发允许发芽。触发器休眠释放可以从曝光而异用f左延长冷，化合物愤怒，或通过摩擦或动物胃酸5接触的种皮物理攻击。作为发芽线索可以是属或种特异性，通常从过去自然选择造成的，适应不良种子萌发是发生在不适当的时间，并可能导致种子或秧苗死亡率或幼苗生长不良。而休眠已被分类成若干基于休眠解除（ 例如 ，物理休眠，生理休眠）的机制的类型，6种子休眠保持在植物生物学的至少一个明白的主题。因此，现场研究，允许对个人的种子或相关的生态条件下的种子组的状态评估比那些单纯依靠在实验室标准发芽试验更高的解释力。 已知的种子特性研究开发能够洞察休眠机制。种子休眠的控制是合mplex，包括生理和形态遗传因素控制。虽然休眠机制的广度有充分的认识还有待阐明，一般的模式已经出现，涉及到两个植物激素赤霉素（GA）和脱落酸（ABA）7之间的反馈关系。在用于与生理组件到其休眠种子这个广义模型，GA用作信号为休眠解除，而ABA用于保持休眠状态。母体遗传效应以及母体的生长环境可以影响休眠等种子性状，如大小，通过母体产生的组织和发育信号8。母体产生的外部结构（或种子覆盖物）可以保持休眠状态，在生理线索相结合倍。由于母源种子覆盖物是由母体植物的基因控制的，他们可能不能反映种子的实际核基因组成。我们有我们ED农作物野生杂交的数组向日葵瘦果跨越梳理出种子的特性，这些孕妇与胎儿遗传效应9,10。因此，研究设计，包括品种多样，交叉型，或基因型可以收集有关信息生态与种子休眠，发芽和生存的基因。 种子萌发和生存的表型如何影响人口动态的一个重要例子可以在作物野杂区可以看出。栽培植物的驯化期间选择消除了大部分休眠并减少种子对生长期的外生存的能力。然而，在作物野杂区基因流动，或杂交，耕地和野生类型之间可以重新提出作物的等位基因（或基因变异）到一个野生种群，对种子库的潜在影响。培育和野生近缘种之间的杂交种作物野杂区发现的潜在可能拥有各种中间休眠表型，预计将种植（ 如冬季）11以外的生存条件，只有少数的表型。 这份手稿的目的是展示如何使用种子埋藏带的方法，我们可以评估发芽，休眠，以及一系列的种子种生存在不同的时间段进行调查野外条件下的自然变化。在我们的例子中，我们采用葵花籽从15作物野杂交类型，因为我们感兴趣的种子特性的孕产妇和胎儿遗传效应。

<table border="0" cellpadding="0" cellspacing="0" width="440">
	<colgroup>
		<col />
		<col />
		<col span="2" />
	</colgroup>
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:240px;">Small coin envelopes</td>
			<td style="width:72px;">Any</td>
			<td style="width:64px;"></td>
			<td style="width:64px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Large coin envelopes</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">fine meshed polyester mosquito netting</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">high-temperature glue gun</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">high-temperature glue stick refills</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Industrial permenant markers</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">plastic garden labels</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">scissors</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Shovel</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Metal mesh hardward cloth</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Surveyor&#39;s flags, multiple colors</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Wet newspaper</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">cooler</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">blotter paper</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">petri dishes</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Temp. controlled growth chamber</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">razor blades</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">petri dishes</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tetrazolium chloride</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">water</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">heat incubator</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

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.

交叉类型具有不同母体亲子和作物等位基因百分比（ 表1）在整个去除日期不同百分比萌发，萌发的和死的种子（图2和3）。使用未萌发的种子TZ测试，我们发现一些真正的休眠的种子在第二去除（早春）（ 表2），而由第三去除（春季）未萌发的种子都被认为是真正的休眠状态。 一般情况下，降低作物等位基因比例秋末（ 图2）减少适应不良的发芽。在早春除去，所有的交叉类型具有高发芽，但在他们的比例死和未萌发（ 图3）不同。尤其是跨类型的作物产妇的父母子女关系有较高的死亡率，而产妇野生跨类型的人更可能留未萌发的。在春天，发芽拒绝对一些交叉的类型。而反直觉，这可能是由于掩埋条较早发芽交叉类型中种子的死亡率。因此，死亡率为更作物状横类型更高（ 即 ，那些具有更高作物等位基因百分比）由作物母本产生（ 图3） <img alt="图2" src="/files/ftp_upload/54663/54663fig2.jpg" /> 图2.发芽在秋末先切除。在第一去除日期向日葵作物野生跨类型的萌发，在秋末。交叉类型由作为标记增加作物等位基因的百分比在y轴上组织。母本首先列出每个十字型。发芽最小二乘装置（以本身条），然后用相同的字母不显著不同使用图基 - 克莱默调整多重比较。图先前已发表于:小精灵E， 等人 （2015年）15。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54663/54663fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/54663/54663fig3.jpg" /> 图3.百分比发芽，萌发的，和死亡率第二去除（早春）和第三去除（春季）。发芽百分比早春（A），弹簧（B），百分比萌发的早春（C），弹簧（ D），和百分比死了早春（E）和春季（F）去除日期。向日葵作物野生混合交叉类型由增加作物等位基因百分比组织，用最作物状在y轴的顶部和最状野在y轴的底部。十字型母本列在第一位。最小二乘意味着随后在同一封信中萌发，萌发的和死种子（与SE条）不显著不同的使用杜克 - 克莱默调整多重比较。图先前已发表于:佩斯等人（2015）15。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54663/54663fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> 十字型 </td><td> TZ休眠 </td><td> 标准错误 </td></tr><tr><td> WXW </td><td> 0.071 </td><td> 0.0131 </td></tr><tr><td> WxBC </td><td> 0.041 </td><td> 0.0126 </td></tr><tr><td> WxF1 </td><td> 0.089 </td><td> 0.0136 </td></tr><tr><td> WxF2 </td><td> 0.054 </td><td> 0.0126 </td></tr><tr><td> F1xW </td><td> 0 </td><td> 0.0126 </td></tr><tr><td> F1xBC </td><td> 0.013 </td><td> 0.0126 </td></tr><tr><td> WXC </td><td> 0.075 </td><td> 0.0131 </td></tr><tr><td> F1xF1 </td><td> 0.007 </td><td> 0.0126 </td></tr><tr><td> F1xF2 </td><td> 0.013 </td><td> 0.0126 </td></tr><tr><td> CXW </td><td> 0 </td><td> 0.0126 </td></tr><tr><td> CxBC </td><td> 0 </td><td> 0.0126 </td></tr><tr><td> F1xC </td><td> 0 </td><td> 0.0126 </td></tr><tr><td> CxF1 </td><td> 0 </td><td> 0.0126 </td></tr><tr><td> CxF2 </td><td> 0 </td><td> 0.0126 </td></tr><tr><td> CXC </td><td> 0 </td><td> 0.0126 </td></tr></tbody></table> 表2.种子可能有助于持久种子库。最小二乘装置，用于在第二（早春）除去未萌发的种子的百分比休眠分数。种子确定采用四唑氯碱是可行的在有利的条件下催芽孵育后IDE。表先前已发表在:佩斯， 等人，（2015）15。

Watch this Scientific Journal Video about Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions at JoVE.com

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

在这里，我们提出了利用掩埋，标记种子带和四氮唑（TZ）的可行性试验在野外条件下评估种子存活，萌发和休眠的协议。

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 ...

reliable-method-for-assessing-seed-germination-dormancy-mortality

Nanyang Technological University

Research

JoVE Journal

Environment

10.4K 次观看.  Ohio State University. 在这里，我们提出了利用掩埋，标记种子带和四氮唑（TZ）的可行性试验在野外条件下评估种子存活，萌发和休眠的协议。 

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

Surface Renewal: An Advanced Micrometeorological Method for Measuring and Processing Field-Scale Energy Flux Density Data

Advanced micrometeorological methods have become increasingly important in soil, crop, and environmental sciences. For many scientists without formal training in atmospheric science, these techniques are relatively inaccessible. Surface renewal and other flux measurement methods require an understanding of boundary layer meteorology and extensive training in instrumentation and multiple data management programs. To improve accessibility of these techniques, we describe the underlying theory of surface renewal measurements, demonstrate how to set up a field station for surface renewal with eddy covariance calibration, and utilize our open-source turnkey data logger program to perform flux data acquisition and processing. The new turnkey program returns to the user a simple data table with the corrected fluxes and quality control parameters, and eliminates the need for researchers to shuttle between multiple processing programs to obtain the final flux data. An example of data generated from these measurements demonstrates how crop water use is measured with this technique. The output information is useful to growers for making irrigation decisions in a variety of agricultural ecosystems. These stations are currently deployed in numerous field experiments by researchers in our group and the California Department of Water Resources in the following crops: rice, wine and raisin grape vineyards, alfalfa, almond, walnut, peach, lemon, avocado, and corn.

Surface renewal is a micrometeorological method that is being used increasingly to determine energy fluxes, but its technical complexity makes it inaccessible to a broad audience. We describe the steps needed to set up and calibrate a surface renewal field station, to acquire and process data, and to correctly interpret results.

表面重建:一种先进的微气象法测量和处理油田规模能流密度数据

Advanced micrometeorological methods have become  increasingly important in soil, crop, and environmental sciences.  For many scientists without formal ...

科研

环境科学

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

Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is a biotrophic fungal pathogen and the causal agent of corn smut on maize. This disease is responsible for significant yield losses of approximately $1.0 billion annually in the U.S.1 Several methods including crop rotation, fungicide application and seed treatments are currently used to control corn smut2. However, host resistance is the only practical method for managing corn smut. Identification of crop plants including maize, wheat, and rice that are resistant to various biotrophic pathogens has significantly decreased yield losses annually3-5. Therefore, the use of a pathogen inoculation method that efficiently and reproducibly delivers the pathogen in between the plant leaves, would facilitate the rapid identification of maize lines that are resistant to U. maydis. As, a first step toward indentifying maize lines that are resistant to U. maydis, a needle injection inoculation method and a resistance reaction screening method was utilized to inoculate maize, teosinte, and maize x teosinte introgression lines with a U. maydis strain and to select resistant plants.
Maize, teosinte and maize x teosinte introgression lines, consisting of about 700 plants, were planted, inoculated with a strain of U. maydis, and screened for resistance. The inoculation and screening methods successfully identified three teosinte lines resistant to U. maydis. Here a detailed needle injection inoculation and resistance reaction screening protocol for maize, teosinte, and maize x teosinte introgression lines is presented. This study demonstrates that needle injection inoculation is an invaluable tool in agriculture that can efficiently deliver U. maydis in between the plant leaves and has provided plant lines that are resistant to U. maydis that can now be combined and tested in breeding programs for improved disease resistance.

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

The use of a needle injection method to inoculate maize and teosinte plants with the biotrophic pathogen Ustilago maydis is described. The needle injection inoculation method facilitates the controlled delivery of the fungal pathogen in between the plant leaves where the pathogen enters the plant through the formation of appresoria. This method is highly efficient, enabling reproducible inoculations with U. maydis.

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 ...

Preparation and Testing of Plant Seed Meal-based Wood Adhesives

Recently, the interest in plant seed meal-based products as wood adhesives has steadily increased, as these plant raw materials are considered renewable and environment-friendly. These natural products may serve as alternatives to petroleum-based adhesives to ease environmental and sustainability concerns. This work demonstrates the preparation and testing of the plant seed-based wood adhesives using cottonseed and soy meal as raw materials. In addition to untreated meals, water washed meals and protein isolates are prepared and tested. Adhesive slurries are prepared by mixing a freeze-dried meal product with deionized water (3:25 w/w) for 2 hr. Each adhesive preparation is applied to one end of 2 wood veneer strips using a brush. The tacky adhesive coated areas of the wood veneer strips are lapped and glued by hot-pressing. Adhesive strength is reported as the shear strength of the bonded wood specimen at break. Water resistance of the adhesives is measured by the change in shear strength of the bonded wood specimens at break after water soaking. This protocol allows one to assess plant seed-based agricultural products as suitable candidates for substitution of synthetic-based wood adhesives. Adjustments to the adhesive formulation with or without additives and bonding conditions could optimize their adhesive properties for various practical applications.

To facilitate the effort in seeking more economic and environment-friendly formulations of natural product-based wood adhesives, this work demonstrates the preparation and testing of plant seed-based wood adhesives. This protocol allows one to assess plant seed-based agricultural products as suitable candidates for the substitution of synthetic-based wood adhesives.

准备和植物种子膳食为基础的木材胶粘剂的测试

Recently, the interest in plant seed meal-based products as wood adhesives has steadily increased, as these plant raw materials are considered renewable ...

Calibrated Passive Sampling - Multi-plot Field Measurements of NH3 Emissions with a Combination of Dynamic Tube Method and Passive Samplers

Agricultural ammonia (NH3) emissions (90% of total EU emissions) are responsible for about 45% airborne eutrophication, 31% soil acidification and 12% fine dust formation within the EU15. But NH3 emissions also mean a considerable loss of nutrients. Many studies on NH3 emission from organic and mineral fertilizer application have been performed in recent decades. Nevertheless, research related to NH3 emissions after application fertilizers is still limited in particular with respect to relationships to emissions, fertilizer type, site conditions and crop growth. Due to the variable response of crops to treatments, effects can only be validated in experimental designs including field replication for statistical testing. The dominating ammonia loss methods yielding quantitative emissions require large field areas, expensive equipment or current supply, which restricts their application in replicated field trials. This protocol describes a new methodology for the measurement of NH3 emissions on many plots linking a simple semi-quantitative measuring method used in all plots, with a quantitative method by simultaneous measurements using both methods on selected plots. As a semi-quantitative measurement method passive samplers are used. The second method is a dynamic chamber method (Dynamic Tube Method) to obtain a transfer quotient, which converts the semi-quantitative losses of the passive sampler to quantitative losses (kg nitrogen ha-1). The principle underlying this approach is that passive samplers placed in a homogeneous experimental field have the same NH3 absorption behavior under identical environmental conditions. Therefore, a transfer co-efficient obtained from single passive samplers can be used to scale the values of all passive samplers used in the same field trial. The method proved valid under a wide range of experimental conditions and is recommended to be used under conditions with bare soil or small canopies (&#60;0.3 m). Results obtained from experiments with taller plants should be treated more carefully.

Calibrated Passive Sampling - Multi-plot Field Measurements of NH3 Emissions with a Combination of Dynamic Tube Method and Passive Samplers

Ammonia emissions are a major threat to the environment by eutrophication, soil acidification and fine particle formation and stem mainly from agricultural sources. This method allows ammonia loss measurements in replicated field trials enabling statistical analysis of emissions and of relationships between crop development and emissions.

校准被动采样 - NH多情节场测量<sub&gt; 3</sub&gt;与动态管法和被动采样的组合排放

Agricultural ammonia (NH3) emissions (90% of total EU emissions) are responsible for about 45% airborne eutrophication, 31% soil acidification and 12% ...

Sediment Core Sectioning and Extraction of Pore Waters under Anoxic Conditions

We demonstrate a method for sectioning sediment cores and extracting pore waters while maintaining oxygen-free conditions. A simple, inexpensive system is built and can be transported to a temporary work space close to field sampling site(s) to facilitate rapid analysis. Cores are extruded into a portable glove bag, where they are sectioned and each 1-3 cm thick section (depending on core diameter) is sealed into 50 ml centrifuge tubes. Pore waters are separated with centrifugation outside of the glove bag and then returned to the glove bag for separation from the sediment. These extracted pore water samples can be analyzed immediately. Immediate analyses of redox sensitive species, such as sulfide, iron speciation, and arsenic speciation indicate that oxidation of pore waters is minimal; some samples show approximately 100% of the reduced species, e.g. 100% Fe(II) with no detectable Fe(III). Both sediment and pore water samples can be preserved to maintain chemical species for further analysis upon return to the laboratory.

A protocol for sectioning sediment cores and extracting pore waters under anoxic conditions in order to permit analysis of redox sensitive species in both solids and fluids is presented.

沉积物核心切片和孔隙水萃取缺氧条件下

We demonstrate a method for sectioning sediment cores and extracting pore waters while maintaining oxygen-free conditions. A simple, inexpensive system is ...

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 ...

Measuring and Mapping Patterns of Soil Erosion and Deposition Related to Soil Carbonate Concentrations Under Agricultural Management

Spatial patterns of soil erosion and deposition can be inferred from differences in ground elevation mapped at appropriate time increments. Such changes in elevation are related to changes in near-surface soil carbonate (CaCO3) profiles. The objective is to describe a simple conceptual model and detailed protocol for repeatable field and laboratory measurements of these quantities. Here, accurate elevation is measured using a ground-based differential global positioning system (GPS); other data acquisition methods could be applied to the same basic method. Soil samples are collected from prescribed depth intervals and analyzed in the lab using an efficient and precise modified pressure-calcimeter method for quantitative analysis of inorganic carbon concentration. Standard statistical methods are applied to point data, and representative results show significant correlations between changes in soil surface layer CaCO3 and changes in elevation consistent with the conceptual model; CaCO3 generally decreased in depositional areas and increased in erosional areas. Maps are derived from point measurements of elevation and soil CaCO3 to aid analyses. A map of erosional and depositional patterns at the study site, a rain-fed winter wheat field cropped in alternating wheat-fallow strips, shows the interacting effects of water and wind erosion affected by management and topography. Alternative sampling methods and depth intervals are discussed and recommended for future work relating soil erosion and deposition to soil CaCO3.

Spatial patterns of soil erosion and deposition can be inferred from differences in ground elevation mapped at appropriate time increments. Such changes in elevation are related to changes in near-surface soil carbonates. Repeatable methods for field and laboratory measurements of these quantities and data analysis methods are described here.

农业经营条件下土壤侵蚀和沉积与土壤碳酸盐浓度关系的测量与制图

Spatial patterns of soil erosion and deposition can be inferred from differences in ground elevation mapped at appropriate time increments. Such changes ...

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 ...

Laboratory and Field Protocol for Estimating Sheet Erosion Rates from Dendrogeomorphology

Sheet erosion is among the crucial drivers of soil degradation. Erosion is controlled by environmental factors and human activities, which often lead to severe environmental impacts. The understanding of sheet erosion is, consequently, a worldwide issue with implications for both environment and economies. However, the knowledge on how erosion evolves in space and time is still limited, as well as its effects on the environment. Below, we explain a new dendrogeomorphological protocol for deriving eroded soil thickness (Ex) by acquiring accurate microtopographic data using both terrestrial laser scanning (TLS) and microtopographic profile gauges. Additionally, standard dendrogeomorphic procedures, dependent on anatomical variations in root rings, are utilized to establish the timing of exposure. Both TLS and microtopographic profile gauges are used to obtain ground surface pro&#64257;les, from which Ex is estimated after the threshold distance (TD) is determined, i.e., the distance between the root and the sediment knickpoint, which allows de&#64257;ning the lowering of the ground surface caused by sheet erosion. For each profile, we measured the height between the topside of the root and a virtual plane tangential to the ground surface. In this way, we intended to avoid small-scale impacts of soil deformation, which may be due to pressures exerted by the root system, or by the arrangement of exposed roots. This may provoke small amounts of soil sedimentation or erosion depending on how they physically affect the surface runoff. We demonstrate that an adequate microtopographic characterization of exposed roots and their associated ground surface is very valuable to obtain accurate erosion rates. This finding could be utilized to develop the best management practices designed to eventually halt or perhaps, at least, lessen soil erosion, so that more sustainable management policies can be put into practice.

Characterizing erosion from dendrogeomorphology has usually focused on accurately finding the starting time of root exposure, by examining macroscopic or cell level changes caused by exposure. Here, we offer a detailed description of different novel techniques to obtain more precise erosion rates from highly accurate microtopographic data.

利用树形地貌估算板材侵蚀速率的实验室和现场协议

Sheet erosion is among the crucial drivers of soil degradation. Erosion is controlled by environmental factors and human activities, which often lead to ...

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 ...