我们想感谢卡塔琳娜Cosker和斯蒂芬妮Courchesne有益的讨论。

试剂的制备<ol><li> 胶原涂层 ：胶原蛋白外衣P35组织培养板，并放置在烤箱在37 ° C时2天前润滑的分隔。最后胶原蛋白的浓度应在0.71毫克/毫升0.001 ñ盐酸稀释。然后，添加1毫升每盘的混合物。 </li><li> 油脂装载机 ：为了填补油脂装载机，一个60ml的注射器必须先与康宁真空润滑脂填充。使用注射器填补油脂装载机，包装箔，然后高压灭菌45分钟。 </li><li> 聚四氟乙烯分隔的分频器，可每次实验后重新使用，但首先必须正确地清洗。从板卸下分配器，擦去所有剩余的油脂和硫酸为2天。从酸中取出后，冲洗3倍的水，煮沸20分钟，晾干，在一个玻璃P100培养皿和20分钟高压灭菌的地方。 </li><li> N2 -甲基纤维素 ：称取甲基纤维素1.5克，放置在500毫升的瓶子。添加搅拌棒，高压灭菌20分钟就干（这一点所有的工作必须是无菌的）。接下来，添加无血清媒体（N 2）500毫升，在寒冷的房间中，搅拌直至溶解。分装成50毫升conicals和冻结在-20 ° C。对于股票的工作，分装50毫升conicals的一成1ML管和冻结在-20 ° C。 </li><li> DRG的媒体 ：DMEM培养液，5％热灭活马血清，1％青霉素链霉素。 </li><li> 100ng/mL DRGN媒体的神经生长因子（NGF）和脑源性神经营养因子（BDNF）的股票浓度为1mg/ml。稀释成每个人的背根神经节媒体神经营养因子1:10,000。文化可以仅在NGF的增长，这改变了神经元在文化生存的补充。 注意 ：当需要时，浓度（阿糖胞苷）ARAC 0.3uM最终浓度在1微米和使用。这将抑制雪​​旺氏细胞和其他神经胶质细胞的生长。 </li><li> 在10ng/ml DRGN媒体 ：100ng/mL DRGN媒体（1:10）稀释与DRG的媒体。 <img src="/files/ftp_upload/old/951_2008_10_27_6/951_Figure-01.png" alt="图1" width="392" height="294" /> 设置所需的工具图1 。 </li></ol>隔间商会成立（启动前清扫这个过程1-2天） <ol><li>请在涂胶原蛋白P35盘向外运动中的划伤。 </li><li> N2 -甲基广场30ul从头中间。设置菜放在一旁，直到分频器是润滑。 </li><li>将一个23号鲁尔存根适配器油脂装载机。抓住一双90 °角止血特氟隆分压器和奠定它在显微镜下所面临的分压器持平。分压器跟踪与油脂，确保每次适配器适配器是从上一步中的油脂插入一个新的起点上，以便有连续线的油脂（见图表）。一旦油脂被应用到整个分，依次准备P35菜肴之一倒挂，并把它使N2 -甲基中间的车厢。按底部的菜用的镊子。请务必按分压器内部四角（左上角，右下角，右上角，左下角，在图所示的“X”）。 <img src="/files/ftp_upload/old/951_2008_10_9_2/951_Figure-02.png" alt="图2" width="171" height="183" /> 图2：润滑分配器的步骤 注 ：重要的是用力按压足够的油脂，使一个完全密封的菜，但如果添加太多的压力，轴突不会跨到侧面车厢。 拿起止血，翻过来，松开分压器。最后，坚决附加分压器中间车厢底部聚焦显微镜下的菜。随着油脂加载器，使一个小的障碍（0.25厘米），因此，一旦细胞被放置在中间舱，他们不能泄露出去。 </li><li>后成立了文化，在每个侧面车厢DRG的媒体和地点在将保持在该细胞的孵化器。让文化坐了几个小时，然后检查有无泄漏。如果媒体已泄露到中间的车厢，然后文化是不可用的。 注 ：当第一次学习这种技术，它是重要的，成立超过一个实验需要的文化，几个将漏水。 <img src="/files/ftp_upload/old/951_2008_10_9_3/951_Figure-03.png" alt="图3" width="254" height="191" /> 图3：“好VS漏”文化 </li></ol>在隔间文化保持背根神经节神经元<ol><li> 第1天 ：100ng/mL DRGN媒体+ ARAC一边车厢，更换DRG的媒体。进行清扫，并添加细胞中心室（100000细胞）。 </li><li> 第2天 ：添加在10ng/ml媒体+ ARAC聚四氟乙烯分频器外，直到媒体中心舱的油脂阻隔与交流流体流动。 </li><li> 第3天 ：替换在一旁车厢媒体100ng/mL DRGN省略在10ng/ml DRGN省略ARAC ARAC和环绕声。 </li><li> 第6天 ：1ng/mL + ARAC和DRG的媒体+ ARAC环绕在一旁车厢替换的媒体。 </li><li> 第九天 ：进行试验使用。 <img src="/files/ftp_upload/old/951_2008_10_9_5/951_Figure-04.png" alt="图4" width="267" height="148" /> 图4：胞体和远端轴突的免疫组化图像 注意：改变媒体时，重要的是从每一方舱顶部的吸液。此外，从中间的车厢本身永远不会改变媒体，只有从环绕，并让它流在进入中心的油脂阻隔。 </li></ol>

<ol>
	<li>Campenot, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Independent+Control+of+the+Local+Environment+of+Somas+and+Neurites.">Independent Control of the Local Environment of Somas and Neurites.</a> Methods in Enzymology. 58, 302-307 (1979).</li><li>Watson, F. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurotrophins+use+the+Erk5+pathway+to+mediate+a+retrograde+survival+response.">Neurotrophins use the Erk5 pathway to mediate a retrograde survival response.</a> Nature Neuroscience. 4, 981-988 (2001).</li><li>Heerssen, H. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynein+motors+transport+activated+Trks+to+promote+survival+of+target-dependent+neurons.">Dynein motors transport activated Trks to promote survival of target-dependent neurons.</a> Nature Neuroscience. 7, 596-603 (2004).</li><li>Taylor, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+microfluidic+culture+platform+for+CNS+axonal+injury,+regeneration+and+transport.">A microfluidic culture platform for CNS axonal injury, regeneration and transport.</a> Nature Methods. 2, 599-605 (2006).</li><li>Park, J. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidic+culture+platform+for+neuroscience+research.">Microfluidic culture platform for neuroscience research.</a> Nat Protoc. 4, 2128-2136 (2006).</li><li>Ure, D. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retrograde+transport+and+steady-state+distribution+of+125I-nerve+growth+factor+in+rat+sympathetic+neurons+in+compartmented+cultures.">Retrograde transport and steady-state distribution of 125I-nerve growth factor in rat sympathetic neurons in compartmented cultures.</a> J Neuroscience. 4, 1282-1290 (1997).</li></ol>

在这段视频中，我们已经演示了如何编写和维护用于培养DRG神经元的隔间商会。处理得当，这个系统可以从轴突细胞体内分离，以研究神经营养因子，其中跨长轴突信号机制。由于车厢之间的隔离流体，它允许没有受到影响的其他车厢选择性刺激或一室的治疗。隔间腔文化可以支持其他类型的细胞，包括从颈上神经节，视网膜神经节神经元和皮层神经元交感神经元。空间理解神经营养因子信号转导的神经退行性?...

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		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
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<tr>
<td>collagen</td>
<td>Reagent</td>
<td>BD Biosciences</td>
<td>354249</td>
<td>&nbsp;</td>
<tr>
<td>N2-methylcellulose 400CPS</td>
<td>Reagent</td>
<td>Sel-Win Chemicals</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Teflon divider</td>
<td>Other</td>
<td>Tyler Research</td>
<td>CAMP10</td>
<td>many other types of dividers are available</td>
</tr>
<tr>
<td>Pin rake</td>
<td>Tool</td>
<td>Tyler Research</td>
<td>Camp-PR</td>
<td>&nbsp;</td>
<tr>
<td>Grease loader</td>
<td>Tool</td>
<td>Tyler Research</td>
<td>Camp-GLSS</td>
<td>&nbsp;</td>
<tr>
<td>DMEM</td>
<td>Reagent</td>
<td>Fisher</td>
<td>MT10017CV</td>
<td>&nbsp;</td>
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<td>NGF</td>
<td>Reagent</td>
<td>Peprotech</td>
<td>450-01</td>
<td>&nbsp;</td>
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<td>BDNF</td>
<td>Reagent</td>
<td>Peprotech</td>
<td>450-02</td>
<td>&nbsp;</td>
<tr>
<td>High vacuum grease</td>
<td>Reagent</td>
<td>Fisher</td>
<td>14-635-5D</td>
<td>&nbsp;</td>
<tr>
<td>AraC</td>
<td>Reagent</td>
<td>Sigma</td>
<td>C-1768</td>
<td>&nbsp;</td>
<tr>
<td>23 gauge luer stub adapter</td>
<td>Tool</td>
<td>Fisher</td>
<td>427565</td>
<td>&nbsp;</td>
<tr>
<td>90&deg; angle hemostats</td>
<td>Tool</td>
<td>Roboz</td>
<td>RS-7035</td>
<td>&nbsp;</td>		</tbody>
		</table>
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preparation and maintenance of dorsal root ganglia neurons in compartmented cultures

神经元的延伸，从细胞体拆下来支配靶组织，靶源性生长因子神经元的存活和功能的要求的轴突过程。神经营养因子具体要求，以维持生存和支配的感觉神经元的分化，但如何将这些目标源性神经营养因子的支配神经元细胞体沟通的问题已经超过30年的一个活跃的研究领域。神经营养因子的信号如何到达细胞体的最普遍接受的模型的建议，信令内涵体进行这个信号沿着轴突逆行。为了研究逆行运输，文化系统的最初设计由罗伯特Campenot，其中胞体从它们的轴突隔离。技术准备培养的感觉神经元概括选择性刺激神经元的终端发生以下靶源性神经营养因子在体内释放这些隔间商会。逆行信号的事件，需要长距离微管依赖逆行运输的神经退行性疾病的治疗具有重要意义。

Watch this Scientific Journal Video about Preparation and Maintenance of Dorsal Root Ganglia Neurons in Compartmented Cultures at JoVE.com

Preparation and Maintenance of Dorsal Root Ganglia Neurons in Compartmented Cultures

在这里，我们描述培养的背根神经节感觉神经元的隔间钱伯斯编写和维护的技术。

隔间文化背根神经节神经元的制备及维修

Neurons extend axonal processes that are far removed from the cell body to innervate target tissues, where target-derived growth factors are required for ...

preparation-maintenance-dorsal-root-ganglia-neurons-compartmented

Research

JoVE Journal

Biology

22.3K 次观看.  Dana Farber Cancer Institute. 在这里，我们描述培养的背根神经节感觉神经元的隔间钱伯斯编写和维护的技术。

视频: Preparation and Maintenance of Dorsal Root Ganglia Neurons in Compartmented Cultures

Preparation of Neuronal Cultures from Midgastrula Stage Drosophila Embryos

This video illustrates the procedure for making primary neuronal cultures from midgastrula stage Drosophila embryos. The methods for collecting embryos and their dechorionation using bleach are demonstrated. Using a glass pipet attached to a mouth suction tube, we illustrate the removal of all cells from single embryos. The method for dispersing cells from each embyro into a small (5 l) drop of medium on an uncoated glass coverslip is demonstrated. A view through the microscope at 1 hour after plating illustrates the preferred cell density. Most of the cells that survive when grown in defined medium are neuroblasts that divide one or more times in culture before extending neuritic processes by 12-24 hours. A view through the microscope illustrates the level of neurite outgrowth and branching expected in a healthy culture at 2 days in vitro. The cultures are grown in a simple bicarbonate based defined medium, in a 5% CO2 incubator at 22-24&deg;C. Neuritic processes continue to elaborate over the first week in culture and when they make contact with neurites from neighboring cells they often form functional synaptic connections. Neurons in these cultures express voltage-gated sodium, calcium, and potassium channels and are electrically excitable. This culture system is useful for studying molecular genetic and environmental factors that regulate neuronal differentiation, excitability, and synapse formation/function.

This video demonstrates the preparation of primary neuronal cultures from midgastrula stage Drosophila embryos. Views of live cultures show cells 1 hour after plating and differentiated neurons after 2 days of growth in a bicarbonate-based defined medium. The neurons are electrically excitable and form synaptic connections.

从Midgastrula舞台果蝇胚胎神经文化的制备

This video illustrates the procedure for making primary neuronal cultures from midgastrula stage Drosophila embryos. The methods for collecting embryos ...

科研

生物学

Preparation of Dissociated Mouse Cortical Neuron Cultures

This video will guide you through the process for generating cortical neuronal cultures from late embryo and early postnatal mouse brain. These cultures can be used for a variety of applications including immunocytochemistry, biochemistry, electrophysiology, calcium and sodium imaging, protein and/or RNA isolation. These cultures also provide a platform to study the neuronal development of transgenic animals that carry a late embryonic or postnatal lethal gene mutation. The procedure is relatively straight forward, requires some experience in tissue culture technique and should not take longer than two to three hours if you are properly prepared. Careful separation of the cortical rind from the thalamo-cortical fiber tract will reduce the number of unwanted non-neuronal cells. To increase yields of neuronal cells triturate the pieces of the cortical tissue gently after the enzyme incubation step. This is imperative as it prevents unnecessary injury to cells and premature neuronal cell death. Since these cultures are maintained in the absence of glia feeder cells, they also offer an added advantage of growing cultures enriched in neurons.

This video shows a procedure for generating neuronal cultures from late embryo and early postnatal mouse cortex. These cultures can be used for immunocytochemistry, biochemistry, electrophysiology, calcium and sodium imaging and provide a platform to study the neuronal development of transgenic animals that carry a postnatal lethal gene mutation.

游离的小鼠皮层神经元文化的制备

This video will guide you through the process for generating cortical neuronal cultures from late embryo and early postnatal mouse brain. These cultures ...

Preparation of Gene Gun Bullets and Biolistic Transfection of Neurons in Slice Culture

Biolistic transfection is a physical means of transfecting cells by bombarding tissue with high velocity DNA coated particles. We provide a detailed protocol for biolistic transfection of rat hippocampal slices, from the initial preparation of DNA coated bullets to the final shooting of the organotypic slice cultures using a gene gun. Gene gun transfection is an efficient and easy means of transfecting neurons and is especially useful for fluorescently labeling a small subset of cells in tissue slice. In this video, we first outline the steps required to coat gold particles with DNA. We next demonstrate how to line the inside of plastic tubing with the gold/DNA bullets, and how to cut this tubing to obtain the plastic cartridges for loading into the gene gun. Finally, we perform biolistic transfection of rat hippocampal slice cultures, demonstrating handling of the Bio-Rad Helios gene gun, and offering trouble shooting advice to obtain healthy and optimally transfected tissue slices.  

We describe a method for preparing DNA coated gold bullets and demonstrate the use of such bullets to biolistically transfect neurons in cultured hippocampal slices.

基因枪子弹的制备和神经元的基因枪转染切片文化

Biolistic transfection is a physical means of transfecting cells by bombarding tissue with high velocity DNA coated particles. We provide a detailed ...

Preparation of 2-dGuo-Treated Thymus Organ Cultures

In the thymus, interactions between developing T-cell precursors and stromal cells that include cortical and medullary epithelial cells are known to play a key role in the development of a functionally competent T-cell pool. However, the complexity of T-cell development in the thymus in vivo can limit analysis of individual cellular components and particular stages of development. In vitro culture systems provide a readily accessible means to study multiple complex cellular processes. Thymus organ culture systems represent a widely used approach to study intrathymic development of T-cells under defined conditions in vitro. Here we describe a system in which mouse embryonic thymus lobes can be depleted of endogenous haemopoeitic elements by prior organ culture in 2-deoxyguanosine, a compound that is selectively toxic to haemopoeitic cells. As well as providing a readily accessible source of thymic stromal cells to investigate the role of thymic microenvironments in the development and selection of T-cells, this technique also underpins further experimental approaches that include the reconstitution of alymphoid thymus lobes in vitro with defined haemopoietic elements, the transplantation of alymphoid thymuses into recipient mice, and the formation of reaggregate thymus organ cultures. (This article is based on work first reported Methods in Molecular Biology 2007, Vol. 380 pages 185-196).

This video demonstrates the dissection and removal of the fetal thymus as well the preparation of ex vivo cultures of 2-dGuo-treated thymus.

2 dGuo治疗胸腺器官培养的制备

In the thymus, interactions between developing T-cell precursors and stromal cells that include cortical and medullary epithelial cells are known to play ...

Protocol for Culturing Sympathetic Neurons from Rat Superior Cervical Ganglia (SCG)

The superior cervical ganglia (SCG) in rats are small, glossy, almond-shaped structures that contain sympathetic neurons.  These neurons provide sympathetic innervations for the head and neck regions and they constitute a well-characterized and relatively homogeneous population (4).  Sympathetic neurons are dependent on nerve growth factor (NGF) for survival, differentiation and axonal growth and the wide-spread availability of NGF facilitates their culture and experimental manipulation (2, 3, 6).  For these reasons, cultured sympathetic neurons have been used in a wide variety of studies including neuronal development and differentiation, mechanisms of programmed and pathological cell death, and signal transduction (1, 2, 5, and 6).  Dissecting out the SCG from newborn rats and culturing sympathetic neurons is not very complicated and can be mastered fairly quickly.  In this article, we will describe in detail how to dissect out the SCG from newborn rat pups and to use them to establish cultures of sympathetic neurons.  The article will also describe the preparatory steps and the various reagents and equipment that are needed to achieve this.

Protocol for Culturing Sympathetic Neurons from Rat Superior Cervical Ganglia SCG

This is a protocol describing how to isolate and culture primary sympathetic neurons from superior cervical ganglia (SCG) of newborn rat pups.

培养大鼠颈上神经节（SCG）的交感神经元的议定书

The superior cervical ganglia (SCG) in rats are small, glossy, almond-shaped structures that contain sympathetic neurons.  These neurons provide ...

Preparation of Aplysia Sensory-motor Neuronal Cell Cultures

The nervous system of the marine mollusk Aplysia californica is relatively simple, consisting of approximately 20,000 neurons. The neurons are large (up to 1 mm in diameter) and identifiable, with distinct sizes, shapes, positions and pigmentations, and the cell bodies are externally exposed in five paired ganglia distributed throughout the body of the animal. These properties have allowed investigators to delineate the circuitry underlying specific behaviors in the animal1. The monosynaptic connection between sensory and motor neurons is a central component of the gill-withdrawal reflex in the animal, a simple defensive reflex in which the animal withdraws its gill in response to tactile stimulation of the siphon. This reflex undergoes forms of non-associative and associative learning, including sensitization, habituation and classical conditioning. Of particular benefit to the study of synaptic plasticity, the sensory-motor synapse can be reconstituted in culture, where well-characterized stimuli elicit forms of plasticity that have direct correlates in the behavior of the animal2,3. Specifically, application of serotonin produces a synaptic strengthening that, depending on the application protocol, lasts for minutes (short-term facilitation), hours (intermediate-term facilitation) or days (long-term facilitation). In contrast, application of the peptide transmitter FMRFamide produces a synaptic weakening or depression that, depending on the application protocol, can last from minutes to days (long-term depression). The large size of the neurons allows for repeated sharp electrode recording of synaptic strength over periods of days together with microinjection of expression vectors, siRNAs and other compounds to target specific signaling cascades and molecules and thereby identify the molecular and cell biological steps that underlie the changes in synaptic efficacy.
An additional advantage of the Aplysia culture system comes from the fact that the neurons demonstrate synapse-specificity in culture4,5. Thus, sensory neurons do not form synapses with themselves (autapses) or with other sensory neurons, nor do they form synapses with non-target identified motor neurons in culture. The varicosities, sites of synaptic contact between sensory and motor neurons, are large enough (2-7 microns in diameter) to allow synapse formation (as well as changes in synaptic morphology) with target motor neurons to be studied at the light microscopic level.
In this video, we demonstrate each step of preparing sensory-motor neuron cultures, including anesthetizing adult and juvenile Aplysia, dissecting their ganglia, protease digestion of the ganglia, removal of the connective tissue by microdissection, identification of both sensory and motor neurons and removal of each cell type by microdissection, plating of the motor neuron, addition of the sensory neuron and manipulation of the sensory neurite to form contact with the cultured motor neuron.

Preparation of Aplysia Sensory-motor Neuronal Cell Cultures

Primary cultures of Aplysia sensory-motor neurons provide a model preparation for studying synapse formation and synaptic plasticity in vitro. This video demonstrates the identification and microdissection of sensory and motor neurons from Aplysia ganglia as well as the methods for establishing and maintaining sensory-motor neurons in culture.

制备海兔感觉运动神经元细胞培养

The nervous system of the marine mollusk Aplysia californica is relatively simple, consisting of approximately 20,000 neurons.  The neurons are large (up ...

Alternative Cultures for Human Pluripotent Stem Cell Production, Maintenance, and Genetic Analysis

Human pluripotent stem cells (hPSCs) hold great promise for regenerative medicine and biopharmaceutical applications. Currently, optimal culture and efficient expansion of large amounts of clinical-grade hPSCs are critical issues in hPSC-based therapies. Conventionally, hPSCs are propagated as colonies on both feeder and feeder-free culture systems. However, these methods have several major limitations, including low cell yields and generation of heterogeneously differentiated cells. To improve current hPSC culture methods, we have recently developed a new method, which is based on non-colony type monolayer (NCM) culture of dissociated single cells. Here, we present detailed NCM protocols based on the Rho-associated kinase (ROCK) inhibitor Y-27632. We also provide new information regarding NCM culture with different small molecules such as Y-39983 (ROCK I inhibitor), phenylbenzodioxane (ROCK II inhibitor), and thiazovivin (a novel ROCK inhibitor). We further extend our basic protocol to cultivate hPSCs on defined extracellular proteins such as the laminin isoform 521 (LN-521) without the use of ROCK inhibitors. Moreover, based on NCM, we have demonstrated efficient transfection or transduction of plasmid DNAs, lentiviral particles, and oligonucleotide-based microRNAs into hPSCs in order to genetically modify these cells for molecular analyses and drug discovery. The NCM-based methods overcome the major shortcomings of colony-type culture, and thus may be suitable for producing large amounts of homogeneous hPSCs for future clinical therapies, stem cell research, and drug discovery.

Here, we present human pluripotent stem cell (hPSC) culture protocols, based on non-colony type monolayer (NCM) growth of dissociated single cells. This new method, utilizing Rho-associated kinase inhibitors or the laminin isoform 521 (LN-521), is suitable for producing large amounts of homogeneous hPSCs, genetic manipulation, and drug discovery.

另类文化的人多能干细胞的生产，维护和遗传分析

Human pluripotent stem cells (hPSCs) hold great promise for regenerative medicine and biopharmaceutical applications. Currently, optimal culture and ...

Lateral Root Inducible System in Arabidopsis and Maize

Lateral root development contributes significantly to the root system, and hence is crucial for plant growth. The study of lateral root initiation is however tedious, because it occurs only in a few cells inside the root and in an unpredictable manner. To circumvent this problem, a Lateral Root Inducible System (LRIS) has been developed. By treating seedlings consecutively with an auxin transport inhibitor and a synthetic auxin, highly controlled lateral root initiation occurs synchronously in the primary root, allowing abundant sampling of a desired developmental stage. The LRIS has first been developed for Arabidopsis thaliana, but can be applied to other plants as well. Accordingly, it has been adapted for use in maize (Zea mays). A detailed overview of the different steps of the LRIS in both plants is given. The combination of this system with comparative transcriptomics made it possible to identify functional homologs of Arabidopsis lateral root initiation genes in other species as illustrated here for the CYCLIN B1;1 (CYCB1;1) cell cycle gene in maize. Finally, the principles that need to be taken into account when an LRIS is developed for other plant species are discussed.

Lateral Root Inducible System in Arabidopsis and Maize

The Lateral Root Inducible System (LRIS) allows for synchronous induction of lateral roots and is presented for Arabidopsis thaliana and maize.

侧根诱导系统<em&gt;拟南芥</em&gt;和玉米

Lateral root development contributes significantly to the root system, and hence is crucial for plant growth. The study of lateral root initiation is ...

Divergence of Root Microbiota in Different Habitats based on Weighted Correlation Networks

The root microbiome plays an important role in plant growth and environmental adaptation. Network analysis is an important tool for studying communities, which can effectively explore the interaction relationship or co-occurrence model of different microbial species in different environments. The purpose of this manuscript is to provide details on how to use the weighted correlation network algorithm to analyze different co-occurrence networks that may occur in microbial communities due to different ecological environments. All analysis of the experiment is performed in the WGCNA package. WGCNA is an R package for weighted correlation network analysis. The experimental data used to demonstrate these methods were microbial community data from the NCBI (National Center for Biotechnology Information) database for three niches of the rice (Oryza sativa) root system. We used the weighted correlation network algorithm to construct co-abundance networks of microbial community in each of the three niches. Then, differential co-abundance networks among endosphere, rhizoplane and rhizosphere soil were identified. In addition, the core genera in network were obtained by the "WGCNA" package, which plays an important regulated role in network functions. These methods enable researchers to analyze the response of microbial network to environmental disturbance and verify different microbial ecological response theories. The results of these methods show that the significant differential microbial networks identified in the endosphere, rhizoplane and rhizosphere soil of rice.

Network analysis was applied to evaluate the association of various ecological microbial communities, such as soil, water and rhizosphere. Presented here is a protocol on how to use the WGCNA algorithm to analyze different co-occurrence networks that may occur in the microbial communities due to different ecological environments.

基于加权相关网络的不同栖息地根微生物群的差异

The root microbiome plays an important role in plant growth and environmental adaptation. Network analysis is an important tool for studying communities, ...

An Improved Chemotaxis Assay for the Rapid Identification of Rhizobacterial Chemoattractants in Root Exudates

Chemotaxis identification is very important for the research and application of rhizosphere growth-promoting bacteria. We established a straightforward method to quickly identify the chemoattractants that could induce the chemotactic movement of rhizosphere growth-promoting bacteria on sterile glass slides via simple steps. Bacteria solution (OD600 = 0.5) and sterile chemoattractant aqueous solution were added dropwise on the glass slide at an interval of 1 cm. An inoculating loop was used to connect the chemoattractant aqueous solution to the bacterial solution. The slide was kept at room temperature for 20 min on the clean bench. Finally, the chemoattractant aqueous solution was collected for bacterial counting and microscopic observation. In this study, through multiple comparisons of experimental results, the method overcame multiple shortcomings of traditional bacterial chemotaxis methods. The method reduced the error of plate counting and shortened the experimental cycle. For the identification of chemoattractant substances, this new method can save 2-3 days compared to the traditional method. Additionally, this method allows any researcher to systematically complete a bacterial chemotaxis experiment within 1-2 days. The protocol can be considered a valuable resource for understanding plant-microbe interactions.

Here, we present an improved chemotaxis assay protocol. The goal of this protocol is to reduce the steps and costs of traditional bacterial chemotaxis methods and to serve as a valuable resource for understanding plant-microbe interactions.

一种用于快速鉴定根系分泌物中根细菌化学引诱剂的改进趋化性测定

Chemotaxis identification is very important for the research and application of rhizosphere growth-promoting bacteria. We established a straightforward ...