Funding by the German Environmental Foundation (DBU) is acknowledged. The authors thank Ute Kuhlicke for technical help with CLSM analysis and Birgit W&#252;rz, Rita Remer, and Jana Reichenbach for skilled experimental help. The authors would particularly like to thank Prof. Jan Roelof van der Meer and Dr. Robin Tecon for fruitful discussion and providing the bioreporter strain. It contributes to the &#8216;Chemicals in the Environment&#8217; (CITE) research program of the Helmholtz Association.

1.准备菜肴，幻灯片和孵化商会<ol><li>准备以下材料每缩影：一个大的塑料培养皿底部（D = 10厘米），一个被修改的（见步骤1.2）的小塑料培养皿底部（D = 5厘米）与盖和三腔一个计数室幻灯片。 </li><li>取的培养皿底部零件所需的数字（D ​​= 5厘米）。用锯恰好适合幻灯片（26毫米边长）拆下边缘的一部分。消毒系统，浸泡培养皿底部和盖在70％乙醇的O / N和干燥它们至少2小时在流动柜在UV光下。存储培养皿在一个密封的塑料容器，这也被暴露在UV光2小时。 </li><li>擦滑动用70％乙醇和所需数量的空气干燥。包裹滑动铝箔，并把它们在马弗炉中进行5小时，在450℃以除去可能的污染物。 </LI&gt; <li>准备玻璃孵化室带盖。清洗，用70％的乙醇和暴露室开放于UV光至少2小时在流动柜。 注：大干燥器（直径约30厘米），用盖子可以用于这一目的;每个干燥器可容纳多达10缩影设置。 </li></ol> 2.准备媒体与文化<ol><li>运行20平行缩影，得到百毫升改性胰蛋白胨-酵母培养基（MTY）14，约150毫升的21℃的基本培养基16（MM），将5ml基本培养基琼脂甲酯（MMA; 0.5％，重量/体积），三马铃薯葡萄糖琼脂平板上（PDA; 1.5％，重量/体积）为P.霉 ，4空白板，其中之一PDA平板含有2mg L -1酮，四琼脂糖平板（3％，重量/体积）含0.3克·毫升-1活性炭和40mg感在2毫克的部分。 </li><li>制备含50毫克卡那霉素的L -1 MTY（卡那霉素是requirED保持EGFP报告质粒）和MM含10毫米醋酸细菌培养。使用MM的无乙酸以制备0.5％的MMA为细菌在微观上固定和倾1.5％的PDA平板（各20毫升）中进行栽培的终极腐霉和缩影设置。 </li><li>接种P.一些菌丝霉三个新鲜PDA平板孵育在黑暗中在25℃下进行约72小时。然后确保这些板块都长满完全新鲜和柔软的菌丝体。 </li><li> B.启动两种文化sartisoli RP037-MCHE在50ml MTY的从甘油贮存。孵育在30℃，230转旋转烧瓶运动O / N。 </li><li>测量培养的光密度（OD）在578纳米和离心机在1000×g离心10分钟（为B. sartisoli RP037-MCHE，预期的OD 578为约1.0和细胞处于指数期）。弃上清液并重新悬浮细胞在MM中的一个适当的量来达到的OD 0.4 578。接种20毫升的MM含有50mg L-1用400微升重新悬浮的细胞的卡那霉素。孵育在30℃，230转旋转烧瓶运动O / N。 </li><li>衡量MM文化的OD 578（为B. sartisoli RP037-MCHE，预计OD 578大约是0.2和细胞在早期指数期）。离心培养在2000×g离心10分钟，并重新悬浮细胞中的MM适量以达到0.4的OD。混合50微升用500μl温暖，液体的MMA的细胞悬浮在2ml管中并储存在50℃，直到使用。用150微升的细胞中的MMA的每个缩影。 </li><li>倾含有活性碳的琼脂糖凝胶片。融化琼脂糖（3％;重量/体积）在双蒸水。用活性炭调匀（0.3克毫升-1），将混合物倒入塑料培养皿（D = 10厘米）。 </li></ol> 3.缩影安装注意：完整的缩影设置在图1中示意性地描绘的，请参阅图1为所有以下各个步骤。下面的步骤被描述一个样品（SAM）与菌丝传输载体和三种不同的控制设置（CON AIR中 ，CON NEG，CON POS）的制备。所有不同的设置的摘要可以在表1中可以找到。 <ol><li>取的培养皿和幻灯片期望数量和它们下一个流柜暴露于UV光30分钟。 </li><li>放置小培养皿内的大的，并通过小培养皿的间隙配合的幻灯片。取暖细胞悬浮液中的MMA的一个样本，并添加150微升到中间的一个滑动的空腔。直到所有幻灯片的中间空腔重复此步骤，充满了MMA。等待5分钟的MMA凝固。 </li><li>使用1厘米的打孔器冲出来的圆形贴片从空白PDA板。放置一个贴剂以2mm旁边的MMA的小皮氏培养皿内的距离。添加该促进在设置菌丝生长（PDA 3中图1）。 </li><li>切弯PDA补丁的机械屏障。使用干净和无菌小培养皿底部，并将其按入一个PDA板。用刮刀切另一个较小环以0.5厘米的距离成其导致一个PDA的环琼脂。 <ol><li>切出一块，这恰好装配到小培养皿中的缩影设置的间隙，并将其放置在滑动的顶部的间隙内。确保MMA和圆形的PDA屏障之间的距离为约2毫米。关闭小培养皿轻轻按压入在PDA屏障的盖子。 </li><li>重复步骤3.4和3.4.1与含有2g L的PDA板-1对放线菌酮。 注意：这是控制设置为空中运输（CON AIR）对bioreporter细胞，由于菌丝体是由放线菌酮抑制，不长满MMA。 </li></ol></li><li>使用1厘米的打孔器冲出来的是空白PDA板的圆形贴片。使用另一个0.5厘米打孔器切出第一补丁的中间。放置所产生的小的PDA型环以1mm旁边的PDA屏障（PDA 1中图1）的距离。 </li><li>添加2毫克流感进入PDA 1.用孔1厘米的打孔器切出从PDA板长满P.圆形补丁霉和放置一个补丁（PDA 2在图1中）底部向下到PDA的环，使得菌丝垫朝向FLU晶体。 <ol><li>无需重复添加FLU晶体上一步。 注意：这是阴性对照设置（CON NEG）来确定背景荧光bioreporter细胞。 </li></ol></li><li>使用1厘米的打孔器冲出来的圆形贴片含有活性炭琼脂。将4个贴片在每一个缩影设置，以进一步降低气体浓度FLU（ 图1）。 </li><li>准备阳性对照（CON POS）。一些FLU晶体添加到200微升的MMA细胞悬液，并把它放在一个空的幻灯片的一个腔。 </li><li>放置一个培养皿（D = 10厘米），用活性炭对孵育室的底部的一些粉末，以尽量减少在腔室中的气态FLU浓度。此外，放置在底部四到五50毫升烧杯用无菌水以保持高湿度的室中。 <ol><li>转移缩影设置成孵化室和孵化在25℃96小时。将样品随机在不同的生长室，以排除位置的影响。 注：此孵化时间AP层数为卵菌P.霉 。它可能是用于其它生物体不同。 </li></ol></li></ol> 4.激光共聚焦显微镜<ol><li>理想的情况是使用激光共聚焦显微镜的设置与正置显微镜。 注意：它可以配备有常规的激光器提供例如用于激励或具有可调白光激光源488，561和633纳米的线条。 </li><li>使用以下设置收集的图像：激发：490纳米（EGFP）和585纳米（mCherry）在适当的激光强度（同时激励）;发射范围：500〜550纳米（EGFP）和605至650纳米（mCherry）;物镜：63X 0.9 NA水浸入（由于其工作距离长）;步长大小对于z轴堆栈：0.5微米。 </li><li>开放缩影设置和使用手术刀去除PDA 1，2，3和PDA的屏障。放置在MMA的顶部玻璃盖玻片并按下它轻轻地除去气泡。在显微镜阶段安装幻灯片。加的水的液滴在th之上Ë盖玻片并用mCherry设置找bioreporter细胞样品中。 </li><li>获取样品的快速浏览和使用电热环比下查找表的红色（mCherry）和绿色（EGFP）信道优化信噪比。选择在样本的随机位置来分析bioreporter荧光。 注：菌丝可能显示自发荧光在bioreporter发射（ 例如，绿色自发荧光）17的范围内。一定要选择无菌丝地区，如果是这样的话。 </li><li>定义和在绿色（EGFP）和红色（mCherry）信道的各位置记录的z栈。通过长时间曝光，以萤光光或激光避免样品漂白。 <ol><li>对10个随机，均匀的分布位置，重复前面的步骤。 </li></ol></li><li>重复使用相同的设置进行测试轨道（SAM），CON AIR，CON NEG和CON POS的所有样品的所有步骤。 </li></oL&gt; 5.图像分析注意：要分析红色（mCherry）和绿色（EGFP）的荧光在记录的z栈，其中包括选择的自由软件ImageJ的18（http://rsb.info.nih.gov/ij/download.html）可以是使用。 <ol><li>请确保安装logi_tool插件（http://downloads.openmicroscopy.org/bio-formats/4.4.10/）。 </li><li>在ImageJ的打开文件。选择“拆分通道”并关闭绿色通道。用宏“mCherry区”作为补充代码文件到Z堆栈量化红色荧光区。在宏观调整首选设置（以粗体显示）。 <ol><li>输出与所测量的像素在阈值以上（以上所定义的最小尺寸）的Z-堆栈的表。 注：所有值的总和是一个红色像素（区域）中的叠层的总数。 </li></ol></li><li>在ImageJ的打开文件。选择“拆分通道”，并接近红色通道。使用宏<html> 216;绿色荧光蛋白INT“作为补充代码文件，以量化的绿色荧光强度Z-堆栈。在宏观调整首选设置（以粗体显示）。 <ol><li>输出与平均强度在Z堆栈的所有绿色物体高于阈值（及以上定义的最小尺寸）和区域的表格。为了计算在堆栈绿色像素的总强度，乘以各意味着由相应的区域的强度。 </li></ol></li><li>计算相对的eGFP感应（eGFP的REL）： <img alt="式（1）" src="/files/ftp_upload/52334/52334eq1.jpg" width="300px" /> （1） 注意：由于mCherry表达组成型，eGFP的相对的结果值是的eGFP的一定量的细胞的表达量的相对量度。也请参考表3的绿色荧光蛋白相对计算的进一步信息。 </li></ol>TLE“&gt; 6。化学定量注：微观设置也可被用于执行的易位FLU量有或没有非生物污染物水槽化学定量。 <ol><li>包装15毫升玻璃小瓶，1毫升GC / MS小瓶和插入铝箔。填充用Na 2 SO 4蒸发器皿;将所有在马弗炉中进行5小时，在450℃以除去可能的污染物。存储活化干燥器的用Na 2 SO 4内。 </li><li>切割PDMS涂覆的玻璃纤维成0.5厘米长。通过摇动四倍2小时的新鲜甲醇洗纤维。重复洗涤步骤再四次用超纯水和存储清洁纤维在超纯水中，在密封的玻璃小瓶中。 </li><li>制备1.5升的甲苯/丙酮（3:1，V / V）含有10μgL -1的菲-D 10。 </li><li>执行步骤1，2和3的协议。准备微观无bioreporter细胞来研究FLU运输（SAM（ - ）和CON AIR（ - ））中，用bioreporter细胞（SAM）来验证FLU降解。 <ol><li> 可选（无bioreporter细胞）：将镀膜玻璃纤维3 PDMS（通常用于SPME实验）作为内SAM的MMA人工和可量化的污染物接收器（ - ）用细镊子。 </li><li>删除使用细镊子玻璃纤维和每一个与插入转移到GC / MS小瓶。 </li><li>加入200μl甲苯和垂直摇O / N，并通过GC / MS分析。使用表2中提供的设置。 </li></ol></li><li>从中间空腔转移的MMA入15ml玻璃小瓶中，并加入约用Na 2 SO 4 3铲。调匀用抹刀和加入10毫升溶剂。旋涡的混合物，并在黑暗中晃动水平O / N。 <ol><li>转移1001，L样品放入GC / MS小瓶插入。添加10微升苊-D 08（10毫克L-1），并使用如在6.4.3中描述的相同的程序，通过GC / MS法进行分析。 </li></ol></li></ol>

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The authors declare that they have no competing financial interest.

所提出的缩影设置证明适合学习的空间分离的化学品的生物利用度，以降低摄取和运输的菌丝体后的生物。防止部分挥发性化合物的潜在的气相输送与细菌bioreporter细胞可以可视化，而不精心样品制备并因此与敏感的系统的干扰最小。同时，样品的化学分析可以容易地进行，允许有一个良好的控制获得的结果的和的总运输定量。然而，一些点必须仔细之前和在执行实验审议。很重要的一点就是保?...

人口稠密由多种微生物1,2的土壤如细菌。然而，在此栖息条件具有挑战性，特别是在水的供应3项。菌永久需要寻找在异构环境4的最佳条件，但没有连续水膜是导致受限可移动性5妨碍他们自由传播。此外，溶质的扩散率（ 例如，营养素）的不饱和的条件6下降低。因此，细菌和营养物往往物理上分离和营养辅助限于3。其结果是，传输载体的化学化合物，它不需要连续水相可有助于克服这些限制。事实上，很多微生物如真菌和卵菌已经开发出一种丝状增长方式，使他们通过空气填充孔隙成长从而达到和MOBIlizing也物理长距离分离营养素7和8含碳物质。他们甚至可以作为其提供糖和其他能源，以细菌9生物的运输载体。摄取和运输中菌丝生物体也已显示对疏水性有机污染物如多环芳烃（PAH）在终极腐霉 10或在丛枝菌根真菌11。由于PAH是无处不在，难溶于水的污染物在土壤中12，菌丝体介导的运输可能有助于提高生物利用度污染的潜在细菌降解。装置10而污染物输送的总量，可以通过化学直接定量，污染物通过菌丝体降解细菌和其它生物体输送的生物利用度不能容易地进行评估。 以下协议呈现给评估myce的影响的方法LIA对污染物的生物利用度要以直接的方式细菌降解;它允许收集有关污染物的微生物生态系统的时空影响的信息。我们描述了如何建立一个复杂的不饱和缩影系统通过将一个物理上分开PAH点源通过菌丝体的运输载体的PAH降解菌bioreporter模仿气水界面土壤。因为空中交通被排除，菌丝基于交通工具上的PAH的生物利用度对于细菌的效果，可以研究在一个孤立的方式。更详细地说，三环多环芳烃芴，菌丝生物体终极腐霉和细菌bioreporter 伯克霍尔德sartisoli RP037-MCHE 13中所描述的缩影设置施加。细菌B.最初构建sartisoli RP037-MCHE研究菲通量到电池14，并表示增强型绿色荧光蛋白（EGFP）的PAH通量的结果细胞中，而红色荧光mCherry表示组成。详细信息，记者建造由TECON 等 13在初步测试中给出，细菌没有透露游泳，只有非常缓慢蜂拥能力。它能够在作为致密悬浮在菌丝顶端施加缓慢迁移对终极腐霉的菌丝。因为细菌嵌入琼脂糖在以下方案，上菌丝迁移并没有出现。 使用共聚焦激光扫描显微镜（CLSM）中，bioreporter细菌，可以直接在微观可视和EGFP的表达可以相对于被量化到细胞的量（正比于mCherry信号）与该软件ImageJ的的帮助。这允许定性在不同的情景进行比较生物利用度（ 即，更高或更低）。 FLU被发现是后由P.菌丝运输生物利用霉 （ 即，它较阴性对照更高）。此外，该协议描述了如何量化通过化学手段菌丝体介导的转运的总量，并使用硅被覆的玻璃纤维（固相微萃取纤维）中相同的缩影验证污染物的生物利用度。使用此设置一个缩影成果已发表 ​​和体育相结合的探讨霉 ，芴和B. sartisoli RP037-MCHE 15。在这里，重点在于一个详细的方法说明和协议的潜在缺陷的鉴定潜在的进一步应用提供了这方面的知识。进一步的应用可涉及各种真菌，细菌种类（ 例如，从污染的场所），以及其它污染物（ 例如，农药）或杂质的供应（ 例如，年龄土壤）。

<table border="0" cellpadding="0" cellspacing="0" width="1158">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Name of Reagent/ Equipment</td>
			<td style="width:195px;">Company</td>
			<td style="width:128px;">Catalog Number</td>
			<td style="width:523px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Confocal Microscope</td>
			<td style="width:195px;">Leica</td>
			<td style="width:128px;"></td>
			<td>TCS SP5X, LAS AF - Version 2.6.1; or equivalent CLSM</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:313px;">GC HP 7890 Series GC and Agilent 5975C MSD</td>
			<td>Agilent</td>
			<td style="width:128px;"></td>
			<td>an equivalent GC/MS may be used</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">GC capillary column J&#38;W 121-5522&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td>Agilent</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Cork borer</td>
			<td style="width:195px;">Fisher Scientific</td>
			<td align="right">12863952</td>
			<td>or any other</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Cover slips</td>
			<td style="width:195px;">Marienfeld</td>
			<td style="width:128px;">107222</td>
			<td>High performance, No.1.5H</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">GC/MS insterts</td>
			<td style="width:195px;">WICOM</td>
			<td style="width:128px;">WIC 47080</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">GC/MS vials 2 ml</td>
			<td style="width:195px;">WICOM</td>
			<td style="width:128px;">WIC 41150</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Lids / septa for screw cap vials</td>
			<td style="width:195px;">DIONEX</td>
			<td>49463 / 049464&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Lids for GC/MS vials</td>
			<td style="width:195px;">WICOM</td>
			<td style="width:128px;">WIC 43948/B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Objective Slides</td>
			<td style="width:195px;">Menzel</td>
			<td style="width:128px;"></td>
			<td>ordinary</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:313px;">PDMS coated glass fibers</td>
			<td style="width:195px;">Polymicro Technologies, Inc.</td>
			<td style="width:128px;"></td>
			<td>V (PDMS) = 13.55 &#177; 0.02 &#181;L m-1</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:313px;">Petri Dishes small / big</td>
			<td style="width:195px;">Greiner</td>
			<td style="width:128px;">633-102 / 628-102</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Screw cap vials 40 ml</td>
			<td style="width:195px;">DIONEX</td>
			<td>48783</td>
			<td>other glass vials may be used</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Screw cap vials 60 ml</td>
			<td style="width:195px;">DIONEX</td>
			<td>48784</td>
			<td>other glass vials may be used</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:313px;">Acenaphthylene d08</td>
			<td style="width:195px;">Dr. Ehrenstorfer</td>
			<td style="width:128px;">C 20510100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Acetone</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">9372.2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Activated carbon</td>
			<td style="width:195px;">Sigma-Aldrich</td>
			<td style="width:128px;">242276-1kg</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Agarose</td>
			<td style="width:195px;">Carl Roth</td>
			<td align="right">2267.4</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Fluorene</td>
			<td style="width:195px;">Fluka</td>
			<td style="width:128px;">46880</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:313px;">Kanamycin sulfate</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">T832.2</td>
			<td>50 mg L-1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Methanol</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">P7171</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Minimal Medium:</td>
			<td style="width:195px;"></td>
			<td style="width:128px;"></td>
			<td>100 mL solution 1 + 25 mL solution 2 + 5 mL solution 3 ad. 1000 mL aqua dest</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">&#160; Solution 1</td>
			<td style="width:195px;"></td>
			<td style="width:128px;"></td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:313px;">&#160;&#160;&#160; Ammonium sulfate&#160;</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">3746.1</td>
			<td>5 g L-1&#160;</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:313px;">&#160;&#160;&#160; Magnesium chloride x 6 H2O</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">2189.1</td>
			<td>1 g L-1</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:313px;">&#160;&#160;&#160; Calcium nitrate x 4 H2O</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">P740.1</td>
			<td>0.5 g L-1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">&#160; Solution 2</td>
			<td style="width:195px;"></td>
			<td style="width:128px;"></td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:313px;">&#160;&#160;&#160; Disodium phosphate</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">P030.1</td>
			<td>55.83 g L-1</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:313px;">&#160;&#160;&#160; Monopotassium phosphate</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">3904.1</td>
			<td>20 g L-1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">&#160; Solution 3</td>
			<td style="width:195px;"></td>
			<td style="width:128px;"></td>
			<td>pH 6.0</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">&#160;&#160;&#160; Disodium EDTA</td>
			<td style="width:195px;">MERCK</td>
			<td style="width:128px;">1084180250</td>
			<td>0.8 g L-1</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">&#160;&#160;&#160; Iron(II) chloride x 4 H2O</td>
			<td style="width:195px;">MERCK</td>
			<td style="width:128px;">1038610250</td>
			<td>0.3 g L-1</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">&#160;&#160;&#160; Cobalt(II) chloride x 6 H2O</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">T889.3</td>
			<td>4 mg L-1</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">&#160;&#160;&#160; Manganese(II) chloride x 1 H2O&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">&#160; 4320.2</td>
			<td>10 mg L-1</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">&#160;&#160;&#160; Copper(II) sulfate</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">&#160;P023.1</td>
			<td>1 mg L-1</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">&#160;&#160;&#160; Sodium molybdate x 2 H2O</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">&#160;0274.1</td>
			<td>3 mg L-1</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">&#160;&#160;&#160; Zinc chloride</td>
			<td style="width:195px;">MERCK</td>
			<td style="width:128px;">1088160250</td>
			<td>2 mg L-1</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">&#160;&#160;&#160; Lithium chloride</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">P007.1</td>
			<td>0.5 mg L-1</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">&#160;&#160;&#160; Tin(II) chloride x 2 H2O</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">4473.1</td>
			<td>0.5 mg L-1</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">&#160;&#160;&#160; Boric acid</td>
			<td style="width:195px;">Riedel-de-Haen&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:128px;">11606</td>
			<td>1 mg L-1</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">&#160;&#160;&#160; Potassium bromide</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">A137.1</td>
			<td>2 mg L-1</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">&#160;&#160;&#160; Potassium iodide</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">6750.1</td>
			<td>2 mg L-1</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">&#160;&#160;&#160; Barium chloride</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">4453.1</td>
			<td>0.5 mg L-1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">MMA</td>
			<td style="width:195px;"></td>
			<td style="width:128px;"></td>
			<td>Minimal medium + agarose 0.2 %</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:313px;">Phenanthrene d10</td>
			<td style="width:195px;">Dr. Ehrenstorfer</td>
			<td style="width:128px;">C 20920100</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:313px;">Potato Dextrose Agar:</td>
			<td style="width:195px;"></td>
			<td style="width:128px;"></td>
			<td>24 g L-1 broth + bacto-agar 1.5 %; pH 6.8</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">&#160;&#160;&#160; Potato Dextrose broth</td>
			<td>Difco/ Beckton Dickinson</td>
			<td style="width:128px;">254920</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">&#160;&#160;&#160; Bacto-agar</td>
			<td>Difco/ Beckton Dickinson</td>
			<td align="right">214040</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Sodium acetate x 3 hydr.</td>
			<td style="width:195px;">Carl Roth</td>
			<td style="width:128px;">6779.1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Sodium sulfate</td>
			<td style="width:195px;">MERCK</td>
			<td style="width:128px;">&#160;1066495000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Toluene</td>
			<td style="width:195px;">MERCK</td>
			<td style="width:128px;">1083252500</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:313px;">mTY medium:</td>
			<td style="width:195px;"></td>
			<td style="width:128px;"></td>
			<td>3 g L-1 yeast extract, 5 g L-1 bacto tryptone and 50 mM NaCl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">&#160;&#160;&#160; Yeast extract</td>
			<td style="width:195px;">Merck</td>
			<td>1037530500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">&#160;&#160;&#160; Tryptone</td>
			<td style="width:195px;">Serva</td>
			<td>4864702</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">&#160;&#160;&#160; Sodium chloride</td>
			<td style="width:195px;">Carl Roth</td>
			<td>3957.1</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:313px;">imageJ with logi tool plugin</td>
			<td style="width:195px;"></td>
			<td style="width:128px;"></td>
			<td style="width:523px;">http://rsb.info.nih.gov/ij/download.html and http://downloads.openmicroscopy.org/bio-formats/4.4.10</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:313px;">Pythium ultimum strain 67-1</td>
			<td style="width:195px;"></td>
			<td style="width:128px;"></td>
			<td style="width:523px;">Obtained from the lab of Dr. Christoph Keel; Department of Fundamental Microbiology, University of Lausanne, Switzerland</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:313px;">Burkholderia sartisoli RP037-mChe</td>
			<td style="width:195px;"></td>
			<td style="width:128px;"></td>
			<td style="width:523px;">Obtained from the lab of Prof. Jan Roelof van der Meer; Department of Fundamental Microbiology, University of Lausanne, Switzerland</td>
		</tr>
	</tbody>
</table>

a whole cell bioreporter approach to assess transport and bioavailability of organic contaminants in water unsaturated systems

Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is not an appropriate measure for its availability; bioavailability rather depends on the dynamic interplay of potential mass transfer (flux) of a compound to a microbial cell and the capacity of the latter to degrade the compound. In water-unsaturated parts of the soil, mycelia have been shown to overcome bioavailability limitations by actively transporting and mobilizing organic compounds over the range of centimeters. Whereas the extent of mycelia-based transport can be quantified easily by chemical means, verification of the contaminant-bioavailability to bacterial cells requires a biological method. Addressing this constraint, we chose the PAH fluorene (FLU) as a model compound and developed a water unsaturated model microcosm linking a spatially separated FLU point source and the FLU degrading bioreporter bacterium Burkholderia sartisoli RP037-mChe by a mycelial network of Pythium ultimum. Since the bioreporter expresses eGFP in response of the PAH flux to the cell, bacterial FLU exposure and degradation could be monitored directly in the microcosms via confocal laser scanning microscopy (CLSM). CLSM and image analyses revealed a significant increase of the eGFP expression in the presence of P. ultimum compared to controls without mycelia or FLU thus indicating FLU bioavailability to bacteria after mycelia-mediated transport. CLSM results were supported by chemical analyses in identical microcosms. The developed microcosm proved suitable to investigate contaminant bioavailability and to concomitantly visualize the involved bacteria-mycelial interactions.

这里介绍的结果已经较早公布的15。请参考文章进行详细的机械和环境的讨论。 通过CLSM图像记录后，最大强度投影可使用相应的显微镜软件或ImageJ的获得样品的一个第一视觉印象和对照（ 图2）进行。以后，该数据集可以以显示由特定的可视化软件有意义特征被投影不同。阳性对照（CON POS）显示明显的绿色荧光蛋白的诱导（ 图...

Watch this Scientific Journal Video about A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems at JoVE.com

A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems

与鼻疽sartisoli RP037-MCHE全细胞bioreporter法的开发是为了检测有机污染物（ 即芴）供细菌降解菌丝桥空气填充孔隙中水不饱和模型系统主动转运后的分数。

全细胞Bioreporter方法​​评估交通有机污染物的生物利用度和水饱和系统

Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is ...

a-whole-cell-bioreporter-approach-to-assess-transport-bioavailability

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Environment

10.3K 次观看.  Helmholtz Centre for Environmental Research - UFZ. 与鼻疽sartisoli RP037-MCHE全细胞bioreporter法的开发是为了检测有机污染物（ 即芴）供细菌降解菌丝桥空气填充孔隙中水不饱和模型系统主动转运后的分数。 

视频: A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. I. Collection of Virus Samples

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. The standardized sampling component of the method concentrates viruses that may be present in water by passage of a minimum specified volume of water through an electropositive cartridge filter. The minimum specified volumes for surface and finished/ground water are 300 L and 1,500 L, respectively. A major method limitation is the tendency for the filters to clog before meeting the sample volume requirement. Studies using two different, but equivalent, cartridge filter options showed that filter clogging was a problem with 10% of the samples with one of the filter types compared to 6% with the other filter type. Clogging tends to increase with turbidity, but cannot be predicted based on turbidity measurements only. From a cost standpoint one of the filter options is preferable over the other, but the water quality and experience with the water system to be sampled should be taken into consideration in making filter selections.

EPA Method 1615 uses an electropositive filter to concentrate enteroviruses and noroviruses in environmental and drinking waters. This manuscript describes the procedure for collecting samples for Method 1615 analyses.

EPA方法1615测量肠道病毒和诺瓦克病毒发生水由文化和RT-qPCR的了。病毒样本的收集I.

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. ...

科研

环境科学

A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated (WATER-PAM) Fluorometry

Conventional methods for experimental manipulation of microalgae have employed large volumes of culture (20 ml to 5 L), so that the culture can be subsampled throughout the experiment1&#8211;7. Subsampling of large volumes can be problematic for several reasons: 1) it causes variation in the total volume and the surface area:volume ratio of the culture during the experiment; 2) pseudo-replication (i.e., replicate samples from the same treatment flask8) is often employed rather than true replicates (i.e., sampling from replicate treatments); 3) the duration of the experiment is limited by the total volume; and 4) axenic cultures or the usual bacterial microbiota are difficult to maintain during long-term experiments as contamination commonly occurs during subsampling.
The use of microtiter plates enables 1 ml culture volumes to be used for each replicate, with up to 48 separate treatments within a 12.65 x 8.5 x 2.2 cm plate, thereby decreasing the experimental volume and allowing for extensive replication without subsampling any treatment. Additionally, this technique can be modified to fit a variety of experimental formats including: bacterial-algal co-cultures, algal physiology tests, and toxin screening9&#8211;11. Individual wells with an alga, bacterium and/or co-cultures can be sampled for numerous laboratory procedures including, but not limited to: WATER-Pulse-Amplitude-Modulated (WATER-PAM) fluorometry, microscopy, bacterial colony forming unit (cfu) counts and flow cytometry. The combination of the microtiter plate format and WATER-PAM fluorometry allows for multiple rapid measurements of photochemical yield and other photochemical parameters with low variability between samples, high reproducibility and avoids the many pitfalls of subsampling a carboy or conical flask over the course of an experiment.

A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated WATER-PAM Fluorometry

The goal of this procedure is to demonstrate the reproducibility and adaptability of using a microtiter plate format for microalgal screening. This rapid screen combines WATER-Pulse-Amplitude-Modulated (WATER-PAM) fluorometry to measure photosynthetic yield as an indicator of Photosystem II (PSII) health with small volume bacterial-algal co-cultures.

小体积的生物测定评估细菌/浮游植物共培养使用水脉冲幅度调制（WATER-PAM）荧光

Conventional methods for experimental manipulation of microalgae have employed large volumes of culture (20 ml to 5 L), so that the culture can be ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a standardized method for use in multiple analytical laboratories during monitoring period 3 of the Unregulated Contaminant Monitoring Rule. Herein we present the protocol for extraction of viral ribonucleic acid (RNA) from water sample concentrates and for quantitatively measuring enterovirus and norovirus concentrations using reverse transcription-quantitative PCR (RT-qPCR). Virus concentrations for the molecular assay are calculated in terms of genomic copies of viral RNA per liter based upon a standard curve. The method uses a number of quality controls to increase data quality and to reduce interlaboratory and intralaboratory variation. The method has been evaluated by examining virus recovery from ground and reagent grade waters seeded with poliovirus type 3 and murine norovirus as a surrogate for human noroviruses. Mean poliovirus recoveries were 20% in groundwaters and 44% in reagent grade water. Mean murine norovirus recoveries with the RT-qPCR assay were 30% in groundwaters and 4% in reagent grade water.

Here we present a procedure to quantify enterovirus and norovirus in environmental and drinking waters using reverse transcription-quantitative PCR. Mean virus recovery from groundwater with this standardized procedure from EPA Method 1615 was 20% for poliovirus and 30% for murine norovirus.

EPA方法1615测量肠道病毒和诺瓦克病毒发生水由文化和RT-qPCR的了。第三部分。病毒检测用RT-qPCR的

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a ...

Soil Lysimeter Excavation for Coupled Hydrological, Geochemical, and Microbiological Investigations

Studying co-evolution of hydrological and biogeochemical processes in the subsurface of natural landscapes can enhance the understanding of coupled Earth-system processes. Such knowledge is imperative in improving predictions of hydro-biogeochemical cycles, especially under climate change scenarios. We present an experimental method, designed to capture sub-surface heterogeneity of an initially homogeneous soil system. This method is based on destructive sampling of a soil lysimeter designed to simulate a small-scale hillslope. A weighing lysimeter of one cubic meter capacity was divided into sections (voxels) and was excavated layer-by-layer, with sub samples being collected from each voxel. The excavation procedure was aimed at detecting the incipient heterogeneity of the system by focusing on the spatial assessment of hydrological, geochemical, and microbiological properties of the soil. Representative results of a few physicochemical variables tested show the development of heterogeneity. Additional work to test interactions between hydrological, geochemical, and microbiological signatures is planned to interpret the observed patterns. Our study also demonstrates the possibility of carrying out similar excavations in order to observe and quantify different aspects of soil-development under varying environmental conditions and scale.

This study presents an excavation method for investigating subsurface hydrological, geochemical, and microbiological heterogeneity of a soil lysimeter. The lysimeter simulates an artificial hillslope which was initially under homogeneous condition and had been subjected to approximately 5,000 mm of water over eight cycles of irrigation in an 18-month period.

土壤蒸渗仪开挖加之水文，地球化学和微生物调查

Studying co-evolution of hydrological and biogeochemical processes in the subsurface of natural landscapes can enhance the understanding of coupled ...

Implementation of Portable Emissions Measurement Systems (PEMS) for the Real-driving Emissions (RDE) Regulation in Europe

Vehicles are tested in controlled and relatively narrow laboratory conditions to determine their official emission values and reference fuel consumption. However, on the road, ambient and driving conditions can vary over a wide range, sometimes causing emissions to be higher than those measured in the laboratory. For this reason, the European Commission has developed a complementary Real-Driving Emissions (RDE) test procedure using the Portable Emissions Measurement Systems (PEMS) to verify gaseous pollutant and particle number emissions during a wide range of normal operating conditions on the road. This paper presents the newly-adopted RDE test procedure, differentiating six steps: 1) vehicle selection, 2) vehicle preparation, 3) trip design, 4) trip execution, 5) trip verification, and 6) calculation of emissions. Of these steps, vehicle preparation and trip execution are described in greater detail. Examples of trip verification and the calculations of emissions are given.

Implementation of Portable Emissions Measurement Systems PEMS for the Real-driving Emissions RDE Regulation in Europe

The European Commission has developed a Real-Driving Emissions (RDE) test procedure to verify pollutant emissions during real-world vehicle operation using the Portable Emissions Measurement Systems (PEMS). This paper presents the experimental procedures required by the newly-adopted RDE test.

便携式排放测量系统（PEMS）在欧洲的真实驾驶排放（RDE）规例的执行情况

Vehicles are tested in controlled and relatively narrow laboratory conditions to determine their official emission values and reference fuel consumption. ...

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

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

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

用于去除与农业和城市径流中地表水毒性有关的污染物的植被处理系统

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

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 ...

Improving Infrared Spectroscopy Characterization of Soil Organic Matter with Spectral Subtractions

Soil organic matter (SOM) underlies numerous soil processes and functions. Fourier transform infrared (FTIR) spectroscopy detects infrared-active organic bonds that constitute the organic component of soils. However, the relatively low organic matter content of soils (commonly &#60; 5% by mass) and absorbance overlap of mineral and organic functional groups in the mid-infrared (MIR) region (4,000-400 cm-1) engenders substantial interference by dominant mineral absorbances, challenging or even preventing interpretation of spectra for SOM characterization. Spectral subtractions, a post-hoc mathematical treatment of spectra, can reduce mineral interference and enhance resolution of spectral regions corresponding to organic functional groups by mathematically removing mineral absorbances. This requires a mineral-enriched reference spectrum, which can be empirically obtained for a given soil sample by removing SOM. The mineral-enriched reference spectrum is subtracted from the original (untreated) spectrum of the soil sample to produce a spectrum representing SOM absorbances. Common SOM removal methods include high-temperature combustion (&#39;ashing&#39;) and chemical oxidation. Selection of the SOM removal method carries two considerations: (1) the amount of SOM removed, and (2) absorbance artifacts in the mineral reference spectrum and thus the resulting subtraction spectrum. These potential issues can, and should, be identified and quantified in order to avoid fallacious or biased interpretations of spectra for organic functional group composition of SOM. Following SOM removal, the resulting mineral-enriched sample is used to collect a mineral reference spectrum. Several strategies exist to perform subtractions depending on the experimental goals and sample characteristics, most notably the determination of the subtraction factor. The resulting subtraction spectrum requires careful interpretation based on the aforementioned methodology. For many soil and other environmental samples containing substantial mineral components, subtractions have strong potential to improve FTIR spectroscopic characterization of organic matter composition.

SOM underlies many soil functions and processes, but its characterization by FTIR spectroscopy is often challenged by mineral interferences. The described method can increase the utility of SOM analysis by FTIR spectroscopy by subtracting mineral interferences in soil spectra using empirically obtained mineral reference spectra.

利用光谱减法改进土壤有机质的红外光谱表征

Soil organic matter (SOM) underlies numerous soil processes and functions. Fourier transform infrared (FTIR) spectroscopy detects infrared-active organic ...

Single-throughput Complementary High-resolution Analytical Techniques for Characterizing Complex Natural Organic Matter Mixtures

Natural organic matter (NOM) is composed of a highly complex mixture of thousands of organic compounds which, historically, proved difficult to characterize. However, to understand the thermodynamic and kinetic controls on greenhouse gas (carbon dioxide [CO2] and methane [CH4]) production resulting from the decomposition of NOM, a molecular-level characterization coupled with microbial proteome analyses is necessary. Further, climate and environmental changes are expected to perturb natural ecosystems, potentially upsetting complex interactions that influence both the supply of organic matter substrates and the microorganisms performing the transformations. A detailed molecular characterization of the organic matter, microbial proteomics, and the pathways and transformations by which organic matter is decomposed will be necessary to predict the direction and magnitude of the effects of environmental changes. This article describes a methodological throughput for comprehensive metabolite characterization in a single sample by direct injection Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS), gas chromatography mass spectrometry (GC-MS), nuclear magnetic resonance (NMR) spectroscopy, liquid chromatography mass spectrometry (LC-MS), and proteomics analysis. This approach results in a fully-paired dataset which improves statistical confidence for inferring pathways of organic matter decomposition, the resulting CO2 and CH4 production rates, and their responses to environmental perturbation. Herein we present results of applying this method to NOM samples collected from peatlands; however, the protocol is applicable to any NOM sample (e.g., peat, forested soils, marine sediments, etc.).

This protocol describes a single throughput for complementary analytical and omics techniques culminating in a fully-paired characterization of natural organic matter and microbial proteomics in different ecosystems. This approach permits robust comparisons for identifying metabolic pathways and transformations important for describing greenhouse gas production and predicting responses to environmental change.

用于表征复杂天然有机物混合物的单吞吐量互补高分辨率分析技术

Natural organic matter (NOM) is composed of a highly complex mixture of thousands of organic compounds which, historically, proved difficult to ...

Microplot Design and Plant and Soil Sample Preparation for 15Nitrogen Analysis

Many&#160;nitrogen fertilizer studies evaluate the overall effect of a treatment on end-of-season measurements such as grain yield or cumulative N losses. A stable isotope approach is necessary to follow and quantify the fate of fertilizer derived N (FDN) through the soil-crop system. The purpose of this paper is to describe a small-plot research&#160;design utilizing non-confined 15N enriched microplots for multiple soil and plant sampling events over two growing seasons and provide sample collection, handling, and processing protocols for total 15N analysis. The methods were demonstrated using a replicated study from south-central Minnesota planted to corn (Zea mays L.). Each treatment consisted of six corn rows (76 cm row-spacing) 15.2 m long with a microplot (2.4 m by 3.8 m) embedded at one end. Fertilizer-grade urea was applied at 135 kg N&#8729;ha-1 at planting, while the microplot received urea enriched to 5 atom % 15N. Soil and plant samples were taken several times throughout the growing season, taking care to minimize cross-contamination by using separate tools and physically separating unenriched and enriched samples during all procedures. Soil and plant samples were dried, ground to pass through a 2 mm screen, and then ground to a flour-like consistency using a roller jar mill. Tracer studies require additional planning, sample processing time and manual labor, and incur higher costs for 15N enriched materials and sample analysis than traditional N studies. However, using the mass balance approach, tracer studies with multiple in-season sampling events allow the researcher to estimate FDN distribution through the soil-crop system and estimate unaccounted-for FDN from the system.

Microplot Design and Plant and Soil Sample Preparation for 15Nitrogen Analysis

A microplot design for 15N tracer research is described to accommodate multiple in-season plant and soil sampling events. Soil and plant sample collection and processing procedures, including grinding and weighing protocols, for 15N analysis are put forth.

微图设计及植物和土壤样品制备，进行15次氮气分析

Many&#160;nitrogen fertilizer studies evaluate the overall effect of a treatment on end-of-season measurements such as grain yield or cumulative N losses. ...

全细胞Bioreporter方法​​评估交通有机污染物的生物利用度和水饱和系统

全细胞Bioreporter方法评估交通有机污染物的生物利用度和水饱和系统