Name: isolation, characterization, and total dna extraction to identify endophytic fungi in mycoheterotrophic plants
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我们感谢 FAPESP （2015/26479-6） 和 CNPq （447453/2014-9） 的资助。JLSM 感谢 CNPq 提供的生产力资助 （303664/2020-7）。MPP 感谢 Capes（硕士学位奖学金，流程 88887.600591/2021-00）和 CNPq。

1. 植物样本采集
<ol><li>培养依赖性方法的样品采集<ol><li>小心挖掘地下器官;这些可以是要收集的植物的根、茎、根茎或储存器官。除了高度压实的土壤外，还要手工收集这些样品。 注意：在此步骤中使用抹子或勺子等工具是不明智的，因为它会破坏真菌异养植物的脆弱结构，并可能导致组织受到非内生真菌的污染。</li>
		<li>尽可能多地收集地下器官。将样品保存在冷却容器内的?...

<ol>
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植物样品的表面灭虫是所提出的协议中最关键的阶段之一。PDA培养皿中不会受到上次洗涤的液滴污染，这是非常可取的。细菌经常在隔离皿中被观察到为污染物，通常比空气中的孢子真菌更多，因为内生细菌在植物组织中也很常见3,11。因此，在安装器官碎片时，在培养基中添加抗生素是必不可少的。当组合不同的抗生素类型时，可以获得更好的效果，?...

An erratum was issued for:&#160;<a href="https://www.jove.com/t/65135">Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants</a>. The Authors section was updated from:
Juliana Lishcka Sampaio Mayer
to:
Juliana Lischka Sampaio Mayer

An erratum was issued for:&#160;<a href="https://www.jove.com/t/65135">Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants</a>. The Authors section was updated.

Erratum: Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants

根据定义，内生真菌是那些栖息在植物器官和组织内部的不明显感染（即不会对其宿主造成伤害）的真菌1,2。这些真菌可以与寄主植物进行中性或有益的相互作用，可能赋予对病原体和不利环境条件的抵抗力，并可能有助于植物有益化合物（例如生长因子和其他植物激素）的合成1,3。菌根内生菌是与植物建立菌根关联的真菌，参与养分转移4.在兰科中，与菌根内生菌的相互作用是绝大多数物种种子萌发和该科所有植物幼苗建立的基础5.在这种情况下，真菌异养兰花代表了对其菌根伙伴的完全依赖性，因为它们依赖于这些真菌在其整个生命周期中的矿物质营养物质和碳化合物转移6.因此，在研究真菌异养生命策略时，分离和鉴定关联真菌是基础。此外，人们对真菌内生菌在真菌异养植物中的作用知之甚少，甚至对这些真菌的真正多样性知之甚少 7,8.
内生真菌的研究 可以通过不同的技术 进行，传统上被描述为不依赖培养或依赖，例如：（a）直接观察，（b）真菌分离和形态学和/或分子鉴定，以及（c）植物组织的总DNA提取和分子鉴定9。在直接观察（a）中，内生真菌可以通过光学或电子显微镜9在植物细胞和组织内部进行研究，因为Pena-Passos等人10详细介绍了不同的显微镜方案。通过分离方法（b），可以根据真菌内生菌的菌落、菌丝以及生殖或抗性结构形态来表征真菌内生菌。此外， 通过 分离技术，可以通过DNA提取、分子识别序列（条形码或指纹图谱）的扩增和测序11来对分离株进行分子鉴定。后一种技术 （c） 能够在植物组织内部（元条形码）中通过 DNA 提取对内生真菌进行分子鉴定，然后进行文库制备和测序12。
此外，真菌分离株可以使用来自自养或真菌异养兰花的种子进行共生发芽试验。这种应用的一个例子是 Sisti 等人进行的研究13，描述了 Pogoniopsis schenckii（一种真菌异养兰）的原生球茎发育的萌发和初始阶段，与其一些分离株（包括非菌根内生真菌）相关。Pena-Passos等人10在视频中详细介绍了应用的共生萌发方案。分离与不同植物器官相关的真菌可以对植物-真菌相互作用的性质进行不同的研究（例如，理解这种关联的生态或生理方面，以及调查从真菌到植物的营养转移）9。
第 1 节中介绍的方法基于地下器官样本的收集，因为这些器官在收集方面存在最大的困难，并且由于菌根内生菌定植于它们，因此它们引起了人们的极大兴趣。然而，包含的方案（步骤1.1和1.2）都可以应用于其他真菌异养植物器官（例如，根茎，花茎和果实）。步骤1.1中描述的收集方法被指定用于分离内生真菌（第2节）进行形态学表征（第4节和第5节）和/或总DNA提取以进行分离物鉴定（第6节）。另一方面，步骤1.2中描述的收集方法专门用于植物组织的总DNA提取，用于元条形码技术（第7节）。在第 3 节中，介绍了四种丝状真菌储存和保存方法，其中两种用于短期储存（3-6 个月），另外两种适合长期储存（>1 年）。形态学特征（第 4 节和第 5 节）可能与分子鉴定相关联，以加强它并提供有关真菌宏观和微观形态学的重要信息。 图1 总结了随后描述的集体方法。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/65135/65135fig01.jpg"> 图1：所提出方法的示意图。 通过培养依赖性和独立性方法进行植物收集和真菌分离、保存和分子鉴定。 <a href="https://www.jove.com/files/ftp_upload/65135/65135fig01large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Adhesive tape</td>
			<td></td>
			<td></td>
			<td>(from any company, for adhesive tape mount in micromorphological analyses)</td>
		</tr>
		<tr>
			<td>Ampicillin</td>
			<td>Sigma-Aldrich</td>
			<td>A5354</td>
			<td>(for installation of plant fragments; other antibiotics may be used - check step 2.2.1)</td>
		</tr>
		<tr>
			<td>Autoclave</td>
			<td></td>
			<td></td>
			<td>(from any company, for materials sterilization in many steps)</td>
		</tr>
		<tr>
			<td>Bacteriological agar</td>
			<td>Sigma-Aldrich</td>
			<td>A1296</td>
			<td>(for many steps)</td>
		</tr>
		<tr>
			<td>C1, C2, C3, C4, C5, and C6 solutions</td>
			<td>Qiagen</td>
			<td>12888-50</td>
			<td>(purchased with DNeasy PowerSoil kit)</td>
		</tr>
		<tr>
			<td>Centrifuge&#160;&#160;</td>
			<td>Merck/Eppendorf</td>
			<td>5810 G</td>
			<td>(for total DNA extraction from fungal isolates)</td>
		</tr>
		<tr>
			<td>Centrifuge tubes</td>
			<td>Merck</td>
			<td>CLS430828</td>
			<td>(for samples collection)</td>
		</tr>
		<tr>
			<td>Chloroform</td>
			<td>Sigma-Aldrich</td>
			<td>C2432</td>
			<td>(for total DNA extraction from fungal isolates)</td>
		</tr>
		<tr>
			<td>Congo red</td>
			<td>Supelco</td>
			<td>75768</td>
			<td>(for hyphae staining)</td>
		</tr>
		<tr>
			<td>Cryotubes</td>
			<td>Merck</td>
			<td>BR114831</td>
			<td>(for many steps)</td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Supelco</td>
			<td>100983</td>
			<td>It will be necessary to carry out the appropriate dilutions (for many steps)</td>
		</tr>
		<tr>
			<td>Ethylenediaminetetraacetic acid (EDTA)</td>
			<td>Sigma-Aldrich</td>
			<td>3609</td>
			<td>(for total DNA extraction from fungal isolates)</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>Merck</td>
			<td>WHA10010155</td>
			<td>(for many steps)</td>
		</tr>
		<tr>
			<td>Glass test tubes</td>
			<td>Merck</td>
			<td>CLS7082516</td>
			<td>(for cryopreservation in unhulled rice grains)</td>
		</tr>
		<tr>
			<td>Glass wool</td>
			<td>Supelco</td>
			<td>20411</td>
			<td>(for cryopreservation in unhulled rice grains)</td>
		</tr>
		<tr>
			<td>Glucose</td>
			<td>Sigma-Aldrich</td>
			<td>G8270</td>
			<td>Or dextrose (for cryopreservation in vermiculite)</td>
		</tr>
		<tr>
			<td>Glycerol</td>
			<td>Sigma-Aldrich</td>
			<td>G5516</td>
			<td>Or glycerin (for cryopreservation in vermiculite, for preparing LPCB)</td>
		</tr>
		<tr>
			<td>Isopropanol</td>
			<td>Sigma-Aldrich</td>
			<td>563935</td>
			<td>(for total DNA extraction from fungal isolates)</td>
		</tr>
		<tr>
			<td>Lactic acid</td>
			<td>Sigma-Aldrich</td>
			<td>252476</td>
			<td>(for preparing LPCB - hyphae staining)</td>
		</tr>
		<tr>
			<td>Lactophenol blue solution (LPCB)</td>
			<td>Sigma-Aldrich</td>
			<td>61335</td>
			<td>(for hyphae staining)</td>
		</tr>
		<tr>
			<td>Laminar flow hood</td>
			<td></td>
			<td></td>
			<td>(class I, from any company, for many steps)</td>
		</tr>
		<tr>
			<td>Light microscope</td>
			<td></td>
			<td></td>
			<td>(from any company, for hyphae observation)</td>
		</tr>
		<tr>
			<td>MB Spin Columns</td>
			<td>Qiagen</td>
			<td>12888-50</td>
			<td>(purchased with DNeasy PowerSoil kit)</td>
		</tr>
		<tr>
			<td>Methyl blue (cotton blue)</td>
			<td>Sigma-Aldrich</td>
			<td>M5528</td>
			<td>(for preparing LPCB - hyphae staining)</td>
		</tr>
		<tr>
			<td>Microcentrifuge tube (1.5 mL)</td>
			<td>Merck</td>
			<td>HS4323</td>
			<td>(for total DNA extraction from fungal isolates)</td>
		</tr>
		<tr>
			<td>Microcentrifuge tube (2 mL)</td>
			<td>Merck</td>
			<td>BR780546</td>
			<td>(for many steps)</td>
		</tr>
		<tr>
			<td>Mineral oil</td>
			<td></td>
			<td></td>
			<td>(for preservation of fungal isolates)</td>
		</tr>
		<tr>
			<td>Paper bags</td>
			<td></td>
			<td></td>
			<td>Average size 150 mm x 200 mm (for samples collection)</td>
		</tr>
		<tr>
			<td>Petri dish (Glass, 120 mm x 20 mm)</td>
			<td>Merck/Pyrex</td>
			<td>SLW1480/10D</td>
			<td>(autoclavable, for fungi slide culture, prefer higher ones)</td>
		</tr>
		<tr>
			<td>Petri dish (Glass, 50 mm x 17 mm)</td>
			<td>Merck/Aldrich</td>
			<td>Z740618</td>
			<td>(for purification of fungal isolates); alternatively: polystyrene petri dishes (sterile, &#947;-irradiated, non-autoclavable)</td>
		</tr>
		<tr>
			<td>Petri dish (Glass, 80 mm x 15 mm)</td>
			<td>Merck/Brand</td>
			<td>BR455732</td>
			<td>(for installation of plant fragments); alternatively: polystyrene petri dishes (sterile, &#947;-irradiated, non-autoclavable)</td>
		</tr>
		<tr>
			<td>Phenol</td>
			<td>Sigma-Aldrich</td>
			<td>P1037</td>
			<td>(for total DNA extraction from fungal isolates, for preparing LPCB)</td>
		</tr>
		<tr>
			<td>Porcelain mortar</td>
			<td>Sigma-Aldrich</td>
			<td>Z247464</td>
			<td>(for total DNA extraction from fungal isolates)</td>
		</tr>
		<tr>
			<td>Porcelain pestle</td>
			<td>Sigma-Aldrich</td>
			<td>Z247502</td>
			<td>(for total DNA extraction from fungal isolates)</td>
		</tr>
		<tr>
			<td>Potato dextrose agar (PDA)</td>
			<td>Millipore</td>
			<td>P2182</td>
			<td>(for many steps)</td>
		</tr>
		<tr>
			<td>PowerBead tubes</td>
			<td>Qiagen</td>
			<td>12888-50</td>
			<td>(purchased with DNeasy PowerSoil kit)</td>
		</tr>
		<tr>
			<td>Rapid mounting medium (Entellan)</td>
			<td>Sigma-Aldrich</td>
			<td>1.0796</td>
			<td>(for fungi slide culture)</td>
		</tr>
		<tr>
			<td>Silica gel</td>
			<td>Supelco</td>
			<td>717185</td>
			<td>(for cryopreservation in unhulled rice grains)</td>
		</tr>
		<tr>
			<td>Sodium chloride (NaCl)</td>
			<td>Sigma-Aldrich</td>
			<td>S9888</td>
			<td>(for total DNA extraction from fungal isolates)</td>
		</tr>
		<tr>
			<td>Sodium dodecyl sulfate (SDS)</td>
			<td>Sigma-Aldrich</td>
			<td>L3771</td>
			<td>Lauryl sulfate sodium salt (for total DNA extraction from fungal isolates)</td>
		</tr>
		<tr>
			<td>Sodium hypochlorite (w/ 2% active chlorine)</td>
			<td></td>
			<td></td>
			<td>(commercial product, for superficial desinfestation)</td>
		</tr>
		<tr>
			<td>Soil DNA extraction kit (DNeasy PowerSoil kit)</td>
			<td>Qiagen</td>
			<td>12888-50</td>
			<td>(for total DNA extraction from plant organs)</td>
		</tr>
		<tr>
			<td>Spectrophotometer - Nanodrop 2000/2000c</td>
			<td>ThermoFisher Scientific</td>
			<td>ND2000CLAPTOP</td>
			<td>(for total DNA extraction from plant organs)</td>
		</tr>
		<tr>
			<td>Stereomicroscope</td>
			<td></td>
			<td></td>
			<td>(=dissecting microscope, from any company, for macromorphological analyses)</td>
		</tr>
		<tr>
			<td>Tetracycline</td>
			<td>Sigma-Aldrich</td>
			<td>T7660</td>
			<td>(for installation of plant fragments)</td>
		</tr>
		<tr>
			<td>Thermoblock</td>
			<td>Merck/Eppendorf</td>
			<td>EP5362000035</td>
			<td>(or from other companies)</td>
		</tr>
		<tr>
			<td>Tissue homogenizer and cell lyzer</td>
			<td>SPEX SamplePrep</td>
			<td></td>
			<td>2010 Geno/Grinder - Automated Tissue Homogenizer and Cell Lyzer (for total DNA extraction from plant organs)</td>
		</tr>
		<tr>
			<td>Toluidine blue O</td>
			<td>Sigma-Aldrich/Harleco</td>
			<td>364-M</td>
			<td>(for hyphae staining)</td>
		</tr>
		<tr>
			<td>Trehalose</td>
			<td>Sigma-Aldrich</td>
			<td>T9531</td>
			<td>(for cryopreservation in vermiculite)</td>
		</tr>
		<tr>
			<td>Tris Base Solution (Tris)</td>
			<td>Sigma-Aldrich</td>
			<td>T1699</td>
			<td>(for total DNA extraction from fungal isolates)</td>
		</tr>
		<tr>
			<td>Unhulled rice grains</td>
			<td></td>
			<td></td>
			<td>(for cryopreservation)</td>
		</tr>
		<tr>
			<td>U-shaped glass rod</td>
			<td></td>
			<td></td>
			<td>(or an adaptation - check step 5.4.1, for fungi slide culture)</td>
		</tr>
		<tr>
			<td>Vermiculite</td>
			<td></td>
			<td></td>
			<td>Fine granulometry (for cryopreservation in vermiculite)</td>
		</tr>
		<tr>
			<td>Vortexer</td>
			<td>Sigma-Aldrich/BenchMixer</td>
			<td>BMSBV1000</td>
			<td>(for total DNA extraction from fungal isolates)</td>
		</tr>
		<tr>
			<td>Yeast extract</td>
			<td>Sigma-Aldrich</td>
			<td>Y1625</td>
			<td>(for cryopreservation in vermiculite)</td>
		</tr>
	</tbody>
</table>

isolation, characterization, and total dna extraction to identify endophytic fungi in mycoheterotrophic plants

霉菌异养植物是菌根依赖性最极端的形式之一，完全丧失了自养能力。与任何其他重要资源一样重要，与这些植物密切相关的真菌对它们来说是必不可少的。因此，研究真菌异养物种的一些最相关的技术是能够研究相关真菌的技术，尤其是那些栖息在根部和地下器官中的真菌。在这种情况下，通常应用鉴定培养依赖性和培养依赖性内生真菌的技术。分离真菌内生菌为形态学鉴定它们、分析它们的多样性和维持接种物提供了一种用于兰花种子共生萌发的方法。然而，众所周知，有大量不可培养的真菌栖息在植物组织中。因此，不依赖文化的分子鉴定技术提供了更广泛的物种多样性和丰度。本文旨在为启动两个调查程序提供必要的方法支持：一个是依赖于文化的调查程序，另一个是独立的调查程序。关于培养依赖性方案，详细介绍了从收集地点到实验室设施收集和维护植物样品的过程，以及从真菌异养植物的地下和地上器官中分离丝状真菌，保留分离物的集合，通过载玻片培养方法对菌丝进行形态学表征，以及通过总DNA提取对真菌进行分子鉴定。包括不依赖培养的方法，详细程序包括收集植物样本进行宏基因组分析和使用商业试剂盒从叶绿素植物器官中提取总DNA。最后，还建议使用连续性方案（例如，聚合酶链反应 [PCR]、测序）进行分析，并在此处介绍技术。

在分离方案中，考虑到最后一次洗涤时使用的水存在污染，并且在带有接种片段的培养皿中也检测到污染，根据污染物的类型，可以采取不同的措施（表1）。如果高度孢子化真菌污染物（也呈现加速生长）和强烈繁殖的细菌，对所选抗生素具有抗药性，则必须从一开始就重复此过程。相反，如果真菌污染物生长缓慢或没有孢子，则可以简单地避免在方案的后?...

Watch this Scientific Journal Video about Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants at JoVE.com

Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants

本文旨在为植物相关内生真菌的分离、分离株的长期保存、形态学表征和总 DNA 提取提供详细和充分的方案，以便随后进行分子鉴定和宏基因组分析。

分离、表征和总 DNA 提取以鉴定真菌异养植物中的内生真菌

Mycoheterotrophic plants present one of the most extreme forms of mycorrhizal dependency, having totally lost their autotrophic capacity. As essential as ...

这里介绍的技术对于研究与不同植物器官相关的内生真菌群落很有用。内生真菌分离可以识别它们，并且在共生萌发等其他技术中很有价值。首先，准备 8 至 10 厘米的 PDA 培养基培养皿，每升补充 3 毫升抗生素。使用前将培养基培养基培养皿在36摄氏度下孵育24小时，检查污染。第二天，使用标准程序对健康的真菌异养植物片段进行表面消毒，并将它们放入无菌培养皿中。为了评估样品表面除虫的功效，在PDA上接种几滴在样品最后一次洗涤期间使用的蒸馏水。接下来，使用火焰手术刀和镊子，将样品切成 0.2 厘米厚的碎片。对样品进行切片以放大与介质接触的表面积。将五个地下器官碎片放在补充抗生素的PDA板中，尽可能远离，不要接触培养皿边缘。用保鲜膜密封培养皿，并将它们存放在 25 至 27 摄氏度的黑暗中五天。传代培养前一天，使用每升 AA 培养基 5 至 7 克制备 5 厘米培养皿，并将平板在 36 摄氏度下孵育 24 小时。为了纯化真菌分离株，通过颜色、生长模式、质地和边缘形式区分 PDA 平板菌落。然后在培养皿的底部划定菌落边缘。还要用代码识别菌落。使用高压灭菌的木牙签的尖端，从已识别的真菌菌落的边缘回收少量菌丝体，并在AA板上划出条纹，在彼此和培养皿边缘相距一厘米处产生三个条纹。使用不干纸和铅笔在 AA 板上贴上适当的代码。密封板，然后在 25 至 27 摄氏度的黑暗中孵育三天。孵育后，仔细观察平板以识别形成单个菌落的细小菌丝。使用永久性标记划定单个菌落的区域。使用高压灭菌的牙签切割含有菌落的培养基的一部分，并将切割的体积转移到不含抗生素的新PDA培养皿的中心。在培养皿上贴上分离物的代码后，用保鲜膜密封它们，并在 25 至 27 摄氏度的黑暗中保持 7 至 14 天。要开始用Castellani的方法保存真菌分离物，将0.5毫升蒸馏水加入高压灭菌的2毫升微量离心管中。使用高压灭菌的牙签，从已经生长的纯化分离株的菌丝体边缘切下0.5×0.5厘米的长方体培养基。将四到六个长方体放入装有蒸馏水的微量离心管中。根据需要将试管存放在 25 摄氏度的黑暗中。需要时，回收一个长方体并将其放置在新PDA培养皿的中心，以培养储存的分离物。通过在干净的载玻片上滴一滴甲苯胺蓝-O染色剂来开始挑逗安装方法。在这种技术中可以使用不同的染色剂。使用高压灭菌的牙签，小心地从生长的分离物中去除一些菌丝，并将它们放入污渍滴中。放置盖玻片并在光学显微镜下进行分析。对于胶带安装方法，将一滴乳酚棉蓝色污渍放在干净的载玻片上。剪下一条透明胶带，其尺寸适合载玻片和中央污渍滴。将胶条的粘性表面引导到菌丝体表面，并尝试收集一些菌丝，不要按压或收集太多菌丝。然后将胶带粘在载玻片上，确保污渍与收集的菌丝接触。在胶带上方滴一滴水并放置盖玻片，然后在光学显微镜下分析载玻片。在PDA培养基上接种菌源异养植物器官片段五天后，观察到从碎片内部出现的小组丝状真菌菌丝体的生长。在接种纯化分离物的7至14天期间，在PDA上观察到纯真菌分离物菌落的离心生长，形成唯一的环状菌丝体。可能的污染很容易识别，损害了菌落在培养基中生长形式、颜色和色素产生的均匀性。从菌性异养兰花（Wullschlaegelia aphylla）的梭形根中分离出的菌落是不透明的，培养基中没有可扩散的色素或渗出物。菌落上侧为白色至灰色，下侧为褐色。菌丝体气生而丰富，边缘不规则且气生。该殖民地具有天鹅绒般的质地和没有宏观结构的皱纹地形。用乳酚棉蓝和甲苯胺蓝-O染色真菌菌丝体后，表现出在未染色菌丝体中不易看到的结构，便于鉴定。这里介绍的真菌分离方法也可以应用于绿色植物，以识别真菌内生菌。分离出的真菌可用于共生萌发试验。

Research

JoVE Journal

Biology

RNA Extraction from Neuroprecursor Cells Using the Bio-Rad Total RNA Kit

使用Bio - Rad公司的总RNA试剂盒从Neuroprecursor细胞的RNA提取

科研

生物学

Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA

Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA

核糖体束缚新生多肽的分离<em&gt;在体外</em&gt;确定沿mRNA的翻译暂停网站

The rate of translational elongation is non-uniform. mRNA secondary structure, codon usage and mRNA associated proteins may alter ribosome movement on the ...

Filtration Isolation of Nucleic Acids:  A Simple and Rapid DNA Extraction Method

核酸的分离过滤：一种简单快速的DNA提取方法

FINA, filtration isolation of nucleic acids, is a novel extraction method which utilizes vertical filtration via a separation membrane and absorbent pad ...

Modified Methods for Loading of High-Throughput DNA Extraction Plates Reduce Potential for Contamination

改进的高通量DNA提取板加载方法，降低污染的可能性

High-throughput DNA sequencing techniques have contributed substantially to advances in our understanding of relationships among microbial communities, ...

Systematic Approach to Identify Novel Antimicrobial and Antibiofilm Molecules from Plants' Extracts and Fractions to Prevent Dental Caries

从植物提取物和分数中识别新型抗菌和抗生物膜分子的系统方法，以防止牙科蛀牙

Natural products provide structurally different substances, with a myriad of biological activities. However, the identification and isolation of active ...

Profiling of H3K4me3 Modification in Plants using Cleavage under Targets and Tagmentation

在靶标和标记下使用裂解分析植物中的H3K4me3修饰

Epigenomic regulation at the chromatic level, including DNA and histone modifications, behaviors of transcription factors, and non-coding RNAs with their ...

Isolation and Characterization of Intact Phycobilisome in Cyanobacteria

蓝藻中完整藻胆体的分离与表征

In cyanobacteria, phycobilisome is a vital antenna protein complex that harvests light and transfers energy to photosystem I and II for photochemistry. ...

Isolation and Screening from Soil Biodiversity for Fungi Involved in the Degradation of Recalcitrant Materials

土壤生物多样性中参与顽固性物质降解的真菌的分离与筛选

Environmental pollution is an increasing problem, and identifying fungi involved in the bioremediation process is an essential task. Soil hosts an ...

Microscopy Techniques for Interpreting Fungal Colonization in Mycoheterotrophic Plants Tissues and Symbiotic Germination of Seeds

用于解释霉菌异养植物组织中真菌定植和种子共生萌发的显微镜技术

Structural botany is an indispensable perspective to fully understand the ecology, physiology, development, and evolution of plants. When researching ...