The authors thank Davy Opdenacker for technical assistance and photography. We greatly thank Dr. Annick Bleys for helpful suggestions to improve the manuscript. This work was financed by the Interuniversity Attraction Poles Programme IUAP P7/29 'MARS' from the Belgian Federal Science Policy Office, by the FWO grant G027313N and by the Agency for Innovation by Science and Technology, IWT (IR).

1. 拟南芥 LRIS协议注:该文是指"小"或"大"规模的实验。小规模的实验，如标记线的分析和组织学染色6，14，只需要少数样品。大规模的实验，如定量实时定量RT-PCR，微阵列9-11或RNA测序，需要样品的量越大。这样，的〜1000苗每个样品，使用由Vanneste 等人 11的量到根段的解剖后执行微阵列实验。 第1天 <ol><li> 拟南芥种子消毒 注意:选择的种子消毒以下步骤之一。它们中的任何适合的下一步骤的协议。气体灭菌（1.1.2）给出了消毒液（1.1.1）两个主要优点:它处理大量麻木的时候花费较少的时间种子呃，因为没有移液必须发生对单个样品;和灭菌的种子可以贮存更长的时间。但是，它需要一个干燥器和小心处理。 <ol><li> 液体消毒 <ol><li>倾种子在微离心管的期望数量。在2 ml微离心管使用多达20毫克（〜800种子）在1.5ml管或40毫克（〜1600种子）。 </li><li>加入1毫升70％的乙醇。反转或轻轻摇动2分钟。 </li><li>更换70％的乙醇用1ml灭菌溶液15分钟。 （准备新鲜消毒液:3.85毫升次氯酸钠（次氯酸钠）股票（12％）（注意:使用通风橱），5微升吐温20，高达10毫升水倒置管几次，以确保所有的种子来。在与溶液接触。 </li><li>在层流气流柜，替换用1ml无菌蒸馏水灭菌溶液和冲洗种子5次，每次5分钟，通过反复LY替换用1ml新鲜无菌蒸馏水。 </li></ol></li><li> 气体消毒 <ol><li>倒在2 ml微离心管的种子的所需数目。有几个独立的线时，请使用单独的管子，并安排他们在一个塑料微型离心管中或支架打开。如果种子在一个管的数目超过500（12.5毫克），细分的种子在管的适当数量，以确保成功的灭菌。确保相邻的管帽不相互覆盖。配合在一起的盒子的盖子的底部，因为它需要在干燥器以及用于杀菌。 </li><li>放置箱在碗干燥器的，以及一个玻璃烧杯（小心:使用通风柜）。 </li><li> （:使用通风柜小心）用100毫升12％的NaOCl填充该烧杯。放置在干燥器的盖子，但留下一个小孔，其中烧杯放置。 </li><li>加入3毫升37％盐酸（发烟）（CAUTION:使用通风柜）的烧杯中通过小光圈和迅速关闭干燥器。该解决方案将泡了一段时间，由于氯2气的形成。离开系统封闭O / N（或至少8小时）。 </li><li>除去干燥器的盖子。取出盒子的盖子盖上，避免在运输过程中受到污染。 </li><li>把盒子中一个空气层流柜，取下盖子，并留下的种子进行约1小时，在室温，以允许气体释放。 注意:种子可以安全地储存在干燥，密闭微离心管长达2周4℃。 </li></ol></li></ol></li><li> 侧根诱导拟南芥 <ol><li> 侧根抑制（NPA处理） <ol><li>准备在体外生长培养基。培养基含有半强度Murashige和Skoog盐混合物15，0.1 g / L的肌醇，10克/升蔗糖，和缓冲用0.5克/升2-（N-吗啉代）乙磺酸（MES）。 </li><li>调节pH值至5.7使用1M KOH。添加植物组织琼脂8克/升和高压灭菌的培养基20分钟，在121℃。 </li><li>准备新人民军的25毫米原液在二甲基亚砜（DMSO）（注意:使用手套）。在一个空气层流柜，冷却高压灭菌介质下降到约65℃（这是在该瓶子可以用手保持的温度），并添加25mM的NPA原液，得到10μM的NPA的终浓度。 </li><li>轻轻摇动瓶子，持续10秒均质一旦加入。倾50毫升每平方培养皿介质（12厘米×12厘米）。 注:如遇大规模的实验，应用尼龙网（20微米），以确保轻松传输。准备一个目（9厘米×9厘米）高压灭菌（图1A）。当高压灭菌，应用网格，以使用无菌镊子（图1B）的生长培养基中。用STE网格轻轻地推向生长培养基rilized drigalsky，以确保它是在与生长培养基（图1B）的良好接触（注意:在无菌条件下工作）。 </li><li>母猪在NPA-板的种子。 注:如遇大规模的实验计划，母猪2致密，排列排50颗种子，以方便取样（见1.2.2.3）。 <ol><li>如果液体消毒，播种在用200微升吸管NPA-板的种子。切断3 - 为了4毫米的在无菌条件下（或尖端的灭菌前）的吸头端部，以便能够吸管种子。吸管上下几次重新悬浮的种子，然后吸管大约150微升无菌水，用10 - 20种子和等到种子沉积物在尖端的底部。 （注意:在无菌条件下工作） 注意:当轻轻与尖端接触琼脂或网格，一滴水与种子将被从它（图1C）释放。 </li> &lt;li&gt;在案件气体消毒，用高压灭菌toothpicks.Pinch牙签放入琼脂采取的种子，以保证表面发粘前播下种子。拿起使用牙签将种子逐个和分发的种子在平板上（图1D）。或者，无菌水添加到种子，和母猪中1.2.1.5.1中描述（注意:工作在无菌条件下）。 </li></ol></li><li>密封与透气胶带将板在黑暗中存储它们板在4℃下至少2天，最大一周。这是必要的分层，并确保同步，并更有效的发芽。 注意:可替换地，种子可以在播种前进行分层，浸渍在蒸馏水在微离心管中，其中需要较少的冰箱的空间。在这种情况下，储存管至少一周，在4℃，播种（见1.2.1.7）后，立即将板到生长室。 第3天 </LI&gt; <li>移动板向生长室（连续光（110μE米-2 -1），21℃）5天（2天的发芽和生长的3天）在一个几乎垂直的位置（80到90度）以确保根生长上，而不是在介质。 第八天 </li></ol></li><li> 侧根感应（NAA处理） <ol><li>在1.2.1.1和1.2.1.2所述准备相同的生长培养基。制备NAA的DMSO的50mM储备溶液（小心:使用手套）。冷却该高压灭菌介质下降到65℃，并添加NAA溶液在培养基中，以获得10微米的NAA的终浓度（注意:工作在无菌条件下）。 <ol><li>轻轻摇动瓶子，持续10秒均质一旦加入。倾50毫升每平方培养皿介质（12厘米×12厘米）。 </li></ol></li><li>从含有补充了10μM的NPA的那些SUP生长培养基平板转移苗执行完成与10μMNAA。 <ol><li>钩弯镊子的手臂下的幼苗子叶，轻轻地从板中删除。只有转移苗其根部增长完全与NPA-生长培养基，最好向下接触。 </li><li>脱脂根在含有NAA的生长介质表面上一个小的距离。确保植物根与生长介质接触良好。如果需要的话，轻轻按下根部到生长培养基表面与镊子。 注:如遇大规模的实验，删除不符合网和传输通过解除在网用镊子上部的两个角接触全苗。确保网格是在与含有NAA的培养基良好接触由掠过网格在琼脂表面上一个小的距离（图1E）。 </li></ol></li><li>密封新板的透气胶带，并将其放置在生长室（CON连续的光（110μE米-2 -1），21℃）中进行诱导直到采样后的希望的时间一垂直位置。 注意:如果一个大规模的实验中，使用解剖刀一次全部直接切根段上的网格和轻轻刮网的表面进行采样它们。 </li></ol></li></ol></li></ol> 2. LRIS玉米协议<ol><li> 玉米籽粒分层 <ol><li>在黑暗中温育的玉米籽粒在4℃下至少1周。 注:此协议一直使用B73玉米自交系发展。 </li></ol></li><li> 玉米籽粒灭菌 第1天 注意:虽然玉米幼苗不必在无菌条件下将要生长，建议来灭菌的玉米籽粒并用无菌材料的工作，以防止在纸卷真菌生长系统。 <ol><li>放玉米核的必要数量在无菌玻璃烧杯中。 </li><li>加入100毫升消毒液（6％次氯酸钠水溶液）（注意:使用通风橱）。 </li><li>添加磁性搅拌棒并将该烧杯放在磁力搅拌器上以低搅拌速度（约250转）在RT 5分钟。 </li><li>通过用100ml新鲜无菌水代替溶液冲洗五次5分钟。 </li></ol></li><li> 播种玉米籽粒 <ol><li>戴上手套，以减少污染的危险性，并取纸巾卷。 </li><li>撕下的手巾纸2延伸到两片（约92厘米×24厘米）的总长度，并把他们放在彼此顶部在清洁的表面（ 图2A）。 </li><li>折叠片材翻一番的长度（大约92厘米×12厘米）（ 图2A）。 </li><li>用镊子来自t相关的前10分发在内核约两厘米见方他的纸张整个长度，同时保持8厘米每个内核与8厘米之间的内空间自由两端（图2B）。确保内核的胚根朝下且朝向纸（ 图2B）。 </li><li>轻轻摇动纸张上的长度，同时保持种子到位（ 图2B）。为了便于滚动，它是可选的，以第一喷纸用无菌水。 </li><li>把纸卷在大约6厘米直径和14厘米高度 （例如，加入250毫升离心管）灭菌玻璃管并将其放置在机架（图2C）。 </li></ol></li><li> 侧根诱导玉米 <ol><li>准备一个50毫米NPA储备液（溶于DMSO）（注意:使用手套）。 </li><li>对于每个管，使50μM的NPA溶液中加入从50mM的NPA原液125微升至125毫升无菌水在无菌玻璃烧杯中。 </li><li>搅拌均匀，倒入125个毫升50μM的NPA溶液在纸卷在管。纸卷将吸收的解决方案，并成为完全湿透。 注意:当使用纸卷和管与其他尺寸，相应地调整音量。液体应达到中途的管，以确保有足够的摄取。 </li><li>将纸卷系统在一个生长室三天（27℃，持续光照，相对湿度70％）。确保纸卷仍然通过增加额外的50微米的不良资产解决方案浸泡，由于溶液蒸发随着时间的推移（ 图2D）。 第4天 </li><li>准备一个50毫米NAA储备液（溶于DMSO）（注意:使用手套）。 </li><li>对于每个管中，加入从50毫萘乙酸库存125微升至125毫升无菌水在一灭菌的玻璃烧杯中制备50μM的萘乙酸溶液。 </li><li>轻轻挤压了大部分剩余的不良资产解决方案从纸卷，然后将米在无菌水5分钟，以洗涤出来（小心:使用手套）。轻轻挤压出大部分的水从纸辊。重复此洗涤步骤三次。 </li><li>搅拌均匀，倒在纸卷在管的50μMNAA的解决方案。确保它们完全浸透。 </li><li>将纸卷系统中的生长室（27℃，持续光照，相对湿度70％）为诱导后所需的持续时间。 </li></ol></li><li> 侧根诱导玉米不定冠根 注:LRIS上述诱导侧根在主根。然而，在种植玉米等单子叶植物中，初生根和胚胎种子根，以及它们的侧根，是幼苗发育过程中主要是很重要的。在植物生长的稍后阶段，后胚胎不定根出现在杆和还开发侧根形成一个新的根系统 16。该LRIS也可使用D由下面的胚胎主根一些小的调整，LRIS促使这些胚胎后不定冠根侧根。 <ol><li>为了使纸卷，创建论文与两个分别在其外侧层的手巾纸，并萌发纸内的两个层的夹层。放置灭菌内核（见步骤2.1至2.2）在两片发芽纸之间。发芽纸相比，手巾纸，将是不容易通过根毛和侧根，这将使得纸张的开幕推出后更容易在穿透。另一方面，手毛巾纸的两个外层将有利于液体吸收。 </li><li>将700毫升的玻璃试管辊和对他们倒无菌水，无不良资产。确保它们完全浸透。 </li><li>在发芽生长柜中消毒内核（见步骤2.4.9）。 </li><li>成长的幼苗，直到不定冠根开始出现（6天为B73）。确保纸辊由必要时加入无菌水保持湿润，在任何时候。 </li><li>转移到4天（见步骤2.4.1至2.4.4）的25微米的NPA溶液，然后由一个类似的感应侧根开始的通过更换用清洗步骤之后的50微米的NAA溶液的NPA溶液（参见步骤2.4 0.5〜2.4.9）。 </li></ol></li></ol>

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Biol. 1, 212-225 (2007).</li><li>Parizot, B., Beeckman, T., Crespi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genomics+of+root+development.">Genomics of root development.</a> Root Genomics and Soil Interactions. , 3-28 (2012).</li><li>Bargmann, B. O., Birnbaum, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescence+activated+cell+sorting+of+plant+protoplasts.">Fluorescence activated cell sorting of plant protoplasts.</a> Journal of visualized experiments : JoVE. , (2010).</li><li>Nakazono, M., Qiu, F., Borsuk, L. A., Schnable, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser-capture+microdissection,+a+tool+for+the+global+analysis+of+gene+expression+in+specific+plant+cell+types:+identification+of+genes+expressed+differentially+in+epidermal+cells+or+vascular+tissues+of+maize.">Laser-capture microdissection, a tool for the global analysis of gene expression in specific plant cell types: identification of genes expressed differentially in epidermal cells or vascular tissues of maize.</a> Plant Cell. 15, 583-596 (2003).</li><li>Ludwig, Y., Hochholdinger, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser+microdissection+of+plant+cells.">Laser microdissection of plant cells.</a> Methods Mol Biol. 1080, 249-258 (2014).</li><li>Beeckman, T., Engler, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+easy+technique+for+the+clearing+of+histochemically+stained+plant+tissue.">An easy technique for the clearing of histochemically stained plant tissue.</a> Plant Mol. Biol. Rep. 12, 37-42 (1994).</li><li>Ditengou, F. A., 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=Mechanical+induction+of+lateral+root+initiation+in+Arabidopsis+thaliana.">Mechanical induction of lateral root initiation in Arabidopsis thaliana.</a> Proc. Natl. Acad. Sci. USA. 105, 18818-18823 (2008).</li><li>Lucas, M., Godin, C., Jay-Allemand, C., Laplaze, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxin+fluxes+in+the+root+apex+co-regulate+gravitropism+and+lateral+root+initiation.">Auxin fluxes in the root apex co-regulate gravitropism and lateral root initiation.</a> J. Exp. Bot. 59, 55-66 (2008).</li><li>Wang, S., Taketa, S., Ichii, M., Xu, L., Xia, K., Zhou, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lateral+root+formation+in+rice+(Oryza+Sativa.+L.):+differential+effects+of+indole-3-acetic+acid+and+indole-3-butyric+acid.">Lateral root formation in rice (Oryza Sativa. L.): differential effects of indole-3-acetic acid and indole-3-butyric acid.</a> Plant Growth Regul. 41, 41-47 (2003).</li><li>Martinez-de la Cruz, E., Garcia-Ramirez, E., Vazquez-Ramos, J. M., Reyes de la Cruz, H., Lopez-Bucio, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxins+differentially+regulate+root+system+architecture+and+cell+cycle+protein+levels+in+maize+seedlings.">Auxins differentially regulate root system architecture and cell cycle protein levels in maize seedlings.</a> J Plant Physiol. 176, 147-156 (2015).</li><li>Beyer, E. M., Morgan, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+ethylene+on+uptake,+distribution,+and+metabolism+of+indoleacetic+acid-1-14C+and+-2-14C+and+naphthaleneacetic+acid-1-14C.">Effect of ethylene on uptake, distribution, and metabolism of indoleacetic acid-1-14C and -2-14C and naphthaleneacetic acid-1-14C.</a> Plant Physiol. 46, 157-162 (1970).</li><li>Delbarre, A., Muller, P., Imhoff, V., Guern, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+mechanisms+controlling+uptake+and+accumulation+of+2,4-dichlorophenoxy+acetic+acid,+naphthalene-1-acetic+acid,+and+indole-3-acetic+acid+in+suspension-cultured+tobacco+cells.">Comparison of mechanisms controlling uptake and accumulation of 2,4-dichlorophenoxy acetic acid, naphthalene-1-acetic acid, and indole-3-acetic acid in suspension-cultured tobacco cells.</a> Planta. 198, 532-541 (1996).</li><li>Bailey-Serres, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microgenomics:+genome-scale,+cell-specific+monitoring+of+multiple+gene+regulation+tiers.">Microgenomics: genome-scale, cell-specific monitoring of multiple gene regulation tiers.</a> Annu Rev Plant Biol. 64, 293-325 (2013).</li><li>Perez-Torres, C. A., 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=Phosphate+availability+alters+lateral+root+development+in+Arabidopsis+by+modulating+auxin+sensitivity+via+a+mechanism+involving+the+TIR1+auxin+receptor.">Phosphate availability alters lateral root development in Arabidopsis by modulating auxin sensitivity via a mechanism involving the TIR1 auxin receptor.</a> Plant Cell. 20, 3258-3272 (2008).</li></ol>

作者什么都没有透露。

在拟南芥 LRIS协议，它只能传输已经变得完全在与含NPA生长培养基接触的幼苗是重要的。这确保侧根开始被阻止在整个根长度。为了防止伤人传输过程中的小植株，弯曲钳子的臂能下的幼苗的子叶钩住。转让时，请确保幼苗根在与含NAA琼脂培养基充分接触。这可以通过掠过根在琼脂表面上一个小的距离来实现。这将确保一个有效的同步感应侧根萌生过的总根长。的根源可以种植没有他们的?...

根系统是植物生长的关键，因为它确保锚固的水分吸收和养分从​​土壤。因为根系统的扩展主要是依赖于生产侧根，其起始和形成已被广泛研究。侧根开始在柱鞘细胞的特定子集，称为创始人电池 1。在大多数双子叶植物，如拟南芥 ，这些细胞都位于原生木质部极2，而在单子叶植物，如玉米，它们被发现在韧皮部磁极3。创办人细胞通过增加生长素应答4标记，接着通过特定的细胞周期基因表达（例如， 细胞周期蛋白B1; 1 / CYCB1; 1），之后，它们经历了第一轮的不对称背斜司5。经过一系列的协调背斜与平周分裂，一侧根原基形成的最终将成为一个一utonomous侧根。的位置和侧根起始定时是但是不可预见的，因为这些事件既不丰富也不同步。这妨碍了使用分子的方法，例如转录研究这一过程。 为了解决这个问题，一个侧根诱导系统（LRIS）已经开发6,7。在这个系统中，幼苗首先用N - 1-naphthylphthalamic酸（NPA），其抑制生长素运输和积累，从而阻断侧根开始8。通过随后转移至苗含有合成植物生长素-1-萘乙酸（NAA）培养基中，整个柱鞘层响应该升高的生长素水平从而大规模诱导侧根启动细胞分裂6。因此，这一制度导致了快速，同步和广泛的侧根灌顶，方便收集富含晚期的特定阶段根样本拉尔根系发育。随后，这些样品可以被用来确定在侧根形成的全基因组表达谱。该LRIS取得了约侧根开始在拟南芥和玉米9-13，但需要已经显著的知识，这个系统应用到其他植物物种变得更基因组被测序更加明显，并且有越来越多的兴趣将知识传授给经济的重要种类。 这里， 拟南芥和玉米LRISs的详细方案给出。接着，使用该系统的一个例子是提供，通过说明如何从玉米LRIS获得转录数据可用于识别具有在不同的植物物种侧根发起期间的保守功能功能同系物。最后，指导方针，以优化LRIS其他植物物种提出了建议。

<table><tbody><tr><td>&lt;strong&gt;ARABIDOPSIS LRIS&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;Seeds&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Arabidopsis seeds</td><td></td><td></td><td>Col-0 ecotype</td></tr><tr><td>&lt;strong&gt;Gas sterilization of seeds&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>micro-centrifuge tubes 1.5 ml</td><td>SIGMA-ALDRICH</td><td>0030 125.215</td><td>Eppendorf microtubes 3810X, PCR clean</td></tr><tr><td>micro-centrifuge tubes 2 ml</td><td>SIGMA-ALDRICH</td><td>0030 120.094</td><td>Eppendorf Safe-Lock microcentrifuge tubes</td></tr><tr><td>hydrochloric acid</td><td>Merck KGaA</td><td>1,003,171,000</td><td>37% (fuming) for analysis EMSURE ACS,ISO,Reag. Ph Eu</td></tr><tr><td>glass desiccator</td><td>SIGMA-ALDRICH</td><td></td><td>Pyrex</td></tr><tr><td>glass beaker</td><td></td><td></td><td></td></tr><tr><td>plastic micro-centrifuge tubes box or holder</td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;Bleach sterilization of seeds&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>ethanol</td><td>Chem-Lab nv</td><td>CL00.0505.1000</td><td>Ethanol, abs. 100% a.r. dilute to 70%</td></tr><tr><td>sodium hypochlorite (NaOCl)</td><td>Carl Roth</td><td>9062.3</td><td>12%</td></tr><tr><td>Tween 20</td><td>SIGMA-ALDRICH</td><td>P1379</td><td></td></tr><tr><td>sterile water</td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;Growth medium&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Murashige and Skoog salt mixture</td><td>DUCHEFA Biochemie B.V.</td><td>M0221-0050</td><td></td></tr><tr><td>myo-inositol</td><td>SIGMA-ALDRICH</td><td>I5125-100G</td><td></td></tr><tr><td>2-(N-morpholino)ethanesulfonic acid (MES)</td><td>DUCHEFA Biochemie B.V.</td><td>M1503.0100</td><td></td></tr><tr><td>sucrose</td><td>VWR, Internation LLC</td><td>27483.294</td><td>D(+)-Sucrose Ph. Eur.</td></tr><tr><td>KOH</td><td>Merck KGaA</td><td>1050211000</td><td>pellets for analysis (max. 0.002% Na) EMSURE ACS,ISO,Reag. Ph Eur</td></tr><tr><td>Plant Tissue Culture Agar</td><td>LabM Limited</td><td>MC029</td><td></td></tr><tr><td>&lt;strong&gt;Lateral root induction chemicals&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>N-1-naphthylphthalamic acid (NPA)</td><td>DUCHEFA Biochemie B.V.</td><td>No. N0926.0250</td><td>10 &amp;micro;M (Arabidopsis)</td></tr><tr><td>1-naphthalene acetic acid (NAA)</td><td>DUCHEFA Biochemie B.V.</td><td>No. N0903.0050</td><td>10 &amp;micro;M (Arabidopsis)</td></tr><tr><td>dimethylsulfoxide (DMSO)</td><td>SIGMA-ALDRICH</td><td>494429-1L</td><td></td></tr><tr><td>&lt;strong&gt;Making a mesh for transfer&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>nylon mesh</td><td>Prosep byba</td><td></td><td>Synthetic nylon mesh 20 &amp;micro;m</td></tr><tr><td>&lt;strong&gt;Sowing and seedling handling&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>square petri dish plates</td><td>GOSSELIN</td><td>BP124-05</td><td>12 x 12 cm</td></tr><tr><td>50 ml DURAN tubes</td><td>SIGMA-ALDRICH</td><td>CLS430304</td><td>Corning&amp;nbsp;50 ml centrifuge tubes</td></tr><tr><td>drigalski</td><td>Carl Roth</td><td>K732.1</td><td></td></tr><tr><td>pipette</td><td></td><td></td><td></td></tr><tr><td>cut pipette tips</td><td>Daslab</td><td>162001X</td><td>Universal 200, cut off 5 mm of tip before autoclaving</td></tr><tr><td>breathable tape&amp;nbsp;</td><td>3M Deutschland GmbH</td><td>cat. no. 1530-1</td><td></td></tr><tr><td>tweezers</td><td>Fiers nv/sa</td><td>K342.1; K344.1</td><td>Dumont tweezers type a nr 5; Dumont tweezers type e nr 7</td></tr><tr><td>&lt;strong&gt;Growth conditions&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>growth room</td><td></td><td></td><td>21 &amp;deg;C, continuous light</td></tr><tr><td>&lt;strong&gt;Materials&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;MAIZE LRIS&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;Seeds&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Maize kernels</td><td></td><td></td><td>B-73</td></tr><tr><td>&lt;strong&gt;Bleach sterilization of kernels&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>glass beaker</td><td></td><td></td><td></td></tr><tr><td>magnetic stirrer&amp;nbsp;</td><td>Fiers nv/sa</td><td>C267.1</td><td></td></tr><tr><td>sodium hypochlorite (NaOCl)</td><td>Carl Roth</td><td>9062.3</td><td>12%</td></tr><tr><td>sterile water</td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;Lateral root induction chemicals&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>N-1-naphthylphthalamic acid (NPA)</td><td>DUCHEFA Biochemie B.V.</td><td>No. N0926.0250</td><td>50 &amp;micro;M (maize primary root), 25 &amp;micro;M (maize adventitious root)</td></tr><tr><td>1-naphthalene acetic acid (NAA)</td><td>DUCHEFA Biochemie B.V.</td><td>No. N0903.0050</td><td>50 &amp;micro;M (maize)</td></tr><tr><td>dimethylsulfoxide (DMSO)</td><td>SIGMA-ALDRICH</td><td>494429-1L</td><td></td></tr><tr><td>&lt;strong&gt;Sowing and seedling handling&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>paper hand towels</td><td>Kimberly-Clark Professional*</td><td>6681</td><td>SCOTT Hand Towels - Roll / White; sheet size (24 x 46 cm)</td></tr><tr><td>seed germination paper</td><td>Anchor Paper Company</td><td></td><td>10 X 15 38# seed germination paper</td></tr><tr><td>tweezers</td><td>Fiers nv/sa</td><td>K342.1; K344.1</td><td>Dumont tweezers type a nr 5; Dumont tweezers type e nr 7</td></tr><tr><td>250 ml (centrifuge) tubes</td><td>SCHOTT DURAN</td><td>2160136</td><td>approx. 5.6 cm diameter and 14.7 cm height&amp;nbsp;</td></tr><tr><td>700 ml tubes</td><td>DURAN GROUP</td><td>213994609</td><td>cylinders, round foot tube, D 60&amp;nbsp; x 250</td></tr><tr><td>rack</td><td></td><td></td><td>for maize tubes, home made</td></tr><tr><td>sterile water</td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;Growth conditions&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>growth cabinet</td><td></td><td></td><td>27 &amp;deg;C, continuous light, 70% relative humidity</td></tr></tbody></table>

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.

执行侧根启动过程的LRIS中的应用比较转录 在LRIS的一个应用是在侧根形成不同物种的比较和基因表达谱的关系。比较转录方法创建查明参与了不同种类的侧根发育过程中的同源基因的可能性。侧根开始，它由从包含在一个已形成的根轴的细胞的子集形成一个新的器官，是由被子植物共享，以控制它们的根构造的主...

Watch this Scientific Journal Video about Lateral Root Inducible System in Arabidopsis and Maize at JoVE.com

拟南芥和玉米侧根诱导系统生物学 |视频

Lateral Root Inducible System in Arabidopsis and Maize

侧根诱导系统<em&gt;拟南芥</em&gt;和玉米

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

lateral-root-inducible-system-in-arabidopsis-and-maize

Universidad de Cartagena UDEC

Research

Education

JoVE Journal

JoVE Core

Biology

Lateral Root Inducible System in Arabidopsis and Maize

Plant Biology

Plant Structure, Growth, and Nutrition

SCIENTISTS IN ACTION

13.7K 次观看.  Ghent. The Lateral Root Inducible System (LRIS) allows for synchronous induction of lateral roots and is presented for Arabidopsis thaliana and maize.

视频: Lateral Root Inducible System in Arabidopsis and Maize

Experimental Design for Laser Microdissection RNA-Seq: Lessons from an Analysis of Maize Leaf Development

Genes with important roles in development frequently have spatially and/or temporally restricted expression patterns. Often these gene transcripts are not detected or are not identified as differentially expressed (DE) in transcriptomic analyses of whole plant organs. Laser Microdissection RNA-Seq (LM RNA-Seq) is a powerful tool to identify genes that are DE in specific developmental domains. However, the choice of cellular domains to microdissect and compare, and the accuracy of the microdissections are crucial to the success of the experiments. Here, two examples illustrate design considerations for transcriptomics experiments; a LM RNA-seq analysis to identify genes that are DE along the maize leaf proximal-distal axis, and a second experiment to identify genes that are DE in liguleless1-R (lg1-R) mutants compared to wild-type. Key elements that contributed to the success of these experiments were detailed histological and in situ hybridization analyses of the region to be analyzed, selection of leaf primordia at equivalent developmental stages, the use of morphological landmarks to select regions for microdissection, and microdissection of precisely measured domains. This paper provides a detailed protocol for the analysis of developmental domains by LM RNA-Seq. The data presented here illustrate how the region selected for microdissection will affect the results obtained.

Many developmentally important genes have cell- or tissue-specific expression patterns. This paper describes LM RNA-seq experiments to identify genes that are differentially expressed at the maize leaf blade-sheath boundary and in lg1-R mutants compared to wild-type. The experimental considerations discussed here apply to transcriptomic analyses of other developmental phenomena.

激光显微切割RNA测序实验设计:从玉米叶片发展的经验分析

Genes with important roles in development frequently have spatially and/or temporally restricted expression patterns. Often these gene transcripts are not ...

科研

生物化学

Measuring Gene Expression in Bombarded Barley Aleurone Layers with Increased Throughput

The aleurone layer of barley grains is an important model system for hormone-regulated gene expression in plants. In aleurone cells, genes required for germination or early seedling development are activated by gibberellin (GA), while genes associated with stress responses are activated by abscisic acid (ABA). The mechanisms of GA and ABA signaling can be interrogated by introducing reporter gene constructs into aleurone cells via particle bombardment, with the resulting transient expression measured using enzyme assays. An improved protocol is reported that partially automates and streamlines the grain homogenization step and the enzyme assays, allowing significantly more throughput than existing methods. Homogenization of the grain samples is carried out using an automated tissue homogenizer, and GUS (&#946;-glucuronidase) assays are carried out using a 96-well plate system. Representative results using the protocol suggest that phospholipase D activity may play an important role in the activation of HVA1 gene expression by ABA, through the transcription factor TaABF1.

An improved protocol is presented for the measurement of transient gene expression from reporter constructs in barley aleurone cells after particle bombardment. The combination of automated grain grinding with 96-well plate enzyme assays provides high throughput for the procedure.

提高吞吐量的轰击大麦糊粉层层中基因表达的测定

The aleurone layer of barley grains is an important model system for hormone-regulated gene expression in plants. In aleurone cells, genes required for ...

遗传学

Inducible, Cell Type-Specific Expression in Arabidopsis thaliana Through LhGR-Mediated Trans-Activation

Inducible, tissue-specific expression is an important and powerful tool to study the spatio-temporal dynamics of genetic perturbation. Combining the flexible and efficient GreenGate cloning system with the proven and benchmarked LhGR system (here termed GR-LhG4) for the inducible expression, we have generated a set of transgenic Arabidopsis lines that can drive the expression of an effector cassette in a range of specific cell types in the three main plant meristems. To this end, we chose the previously developed GR-LhG4 system based on a chimeric transcription factor and a cognate pOp-type promoter ensuring tight control over a wide range of expression levels. In addition, to visualize the expression domain where the synthetic transcription factor is active, an ER-localized mTurquoise2 fluorescent reporter under control of the pOp4 or pOp6 promoter is encoded in driver lines. Here, we describe the steps necessary to generate a driver or effector line and demonstrate how cell type specific expression can be induced and followed in the shoot apical meristem, the root apical meristem and the cambium of Arabidopsis. By using several or all driver lines, the context specific effect of expressing one or multiple factors (effectors) under control of the synthetic pOp promoter can be assessed rapidly, for example in F1 plants of a cross between one effector and multiple driver lines. This approach is exemplified by the ectopic expression of VND7, a NAC transcription factor capable of inducing ectopic secondary cell wall deposition in a cell autonomous manner.

Inducible, Cell Type-Specific Expression in Arabidopsis thaliana Through LhGR-Mediated Trans-Activation

Here, we describe the generation and application of a set of transgenic Arabidopsis thaliana lines enabling inducible, tissue-specific expression in the three main meristems, the shoot apical meristem, the root apical meristem, and the cambium.

Lhgra 介导的反激活在拟南芥诱导、细胞类型特异性表达中的表达

Inducible, tissue-specific expression is an important and powerful tool to study the spatio-temporal dynamics of genetic perturbation. Combining the ...

Lignin Down-regulation of Zea mays via dsRNAi and Klason Lignin Analysis

To facilitate the use of lignocellulosic biomass as an alternative bioenergy resource, during biological conversion processes, a pretreatment step is needed to open up the structure of the plant cell wall, increasing the accessibility of the cell wall carbohydrates. Lignin, a polyphenolic material present in many cell wall types, is known to be a significant hindrance to enzyme access. Reduction in lignin content to a level that does not interfere with the structural integrity and defense system of the plant might be a valuable step to reduce the costs of bioethanol production. In this study, we have genetically down-regulated one of the lignin biosynthesis-related genes, cinnamoyl-CoA reductase (ZmCCR1) via a double stranded RNA interference technique. The ZmCCR1_RNAi construct was integrated into the maize genome using the particle bombardment method. Transgenic maize plants grew normally as compared to the wild-type control plants without interfering with biomass growth or defense mechanisms, with the exception of displaying of brown-coloration in transgenic plants leaf mid-ribs, husks, and stems. The microscopic analyses, in conjunction with the histological assay, revealed that the leaf sclerenchyma fibers were thinned but the structure and size of other major vascular system components was not altered. The lignin content in the transgenic maize was reduced by 7-8.7%, the crystalline cellulose content was increased in response to lignin reduction, and hemicelluloses remained unchanged. The analyses may indicate that carbon flow might have been shifted from lignin biosynthesis to cellulose biosynthesis. This article delineates the procedures used to down-regulate the lignin content in maize via RNAi technology, and the cell wall compositional analyses used to verify the effect of the modifications on the cell wall structure.

Lignin Down-regulation of Zea mays via dsRNAi and Klason Lignin Analysis

A double stranded RNA interference (dsRNAi) technique is employed to down-regulate the maize cinnamoyl coenzyme A reductase (ZmCCR1) gene to lower plant lignin content. Lignin down-regulation from the cell wall is visualized by microscopic analyses and quantified by the Klason method. Compositional changes in hemicellulose and crystalline cellulose are analyzed.

木质素下调<em&gt;玉米</em&gt;通过dsRNAi和Klason木质素分析

To facilitate the use of lignocellulosic biomass as an alternative bioenergy resource, during biological conversion processes, a pretreatment step is ...

生物工程

Inoculation Strategies to Infect Plant Roots with Soil-Borne Microorganisms

The rhizosphere harbors a highly complex microbial community in which plant roots are constantly challenged. Roots are in close contact with a wide variety of microorganisms, but studies on soil-borne interactions are still behind those performed on aboveground organs. Although some inoculation strategies for infecting model plants with model root pathogens are described in the literature, it remains difficult to get a comprehensive methodological overview. To address this problem, three different root inoculation systems are precisely described that can be applied to gain insights into the biology of root-microbe interactions. For illustration, Verticillium species (namely, V. longisporum and V. dahliae) were employed as root invading model pathogens. However, the methods can be easily adapted to other root colonizing microbes - both pathogenic and beneficial. By colonizing the plant xylem, vascular soil-borne fungi such as Verticillium spp. exhibit a unique lifestyle. After root invasion, they spread via the xylem vessels acropetally, reach the shoot, and elicit disease symptoms. Three representative plant species were chosen as model hosts: Arabidopsis thaliana, economically important oilseed rape (Brassica napus), and tomato (Solanum lycopersicum). Step-by-step protocols are given. Representative results of pathogenicity assays, transcriptional analyses of marker genes, and independent confirmations by reporter constructs are shown. Furthermore, the advantages and disadvantages of each inoculation system are thoroughly discussed. These proven protocols can assist in providing approaches for research questions on root-microbe interactions. Knowing how plants cope with microbes in the soil is crucial for developing new strategies to improve agriculture.

This protocol presents a detailed summary of strategies to inoculate plant roots with soil-borne microbes. Exemplified for the fungi Verticillium longisporum and Verticillium dahliae, three different root infection systems are described. Potential applications and possible downstream analyses are highlighted, and advantages or disadvantages are discussed for each system.

用土壤微生物感染植物根系的接种策略

The rhizosphere harbors a highly complex microbial community in which plant roots are constantly challenged. Roots are in close contact with a wide ...

环境科学

Translating Ribosome Affinity Purification (TRAP) to Investigate Arabidopsis thaliana Root Development at a Cell Type-Specific Scale

In this article, we give hands-on instructions to obtain translatome data from different Arabidopsis thaliana root cell types via the translating ribosome affinity purification (TRAP) method and consecutive optimized low-input library preparation.
As starting material, we employ plant lines that express GFP-tagged ribosomal protein RPL18 in a cell type-specific manner by use of adequate promoters. Prior to immunopurification and RNA extraction, the tissue is snap frozen, which preserves tissue integrity and simultaneously allows execution of time series studies with high temporal resolution. Notably, cell wall structures remain intact, which is a major drawback in alternative procedures such as fluorescence-activated cell sorting-based approaches that rely on tissue protoplasting to isolate distinct cell populations. Additionally, no tissue fixation is necessary as in laser capture microdissection-based techniques, which allows high-quality RNA to be obtained.
However, sampling from subpopulations of cells and only isolating polysome-associated RNA severely limits RNA yields. It is, therefore, necessary to apply sufficiently sensitive library preparation methods for successful data acquisition by RNA-seq.
TRAP offers an ideal tool for plant research as many developmental processes involve cell wall-related and mechanical signaling pathways. The use of promoters to target specific cell populations is bridging the gap between organ and single-cell level that in turn suffer from little resolution or very high costs. Here, we apply TRAP to study cell-cell communication in lateral root formation.

Translating Ribosome Affinity Purification TRAP to Investigate Arabidopsis thaliana Root Development at a Cell Type-Specific Scale

Translating ribosome affinity purification (TRAP) offers the possibility to dissect developmental programs with minimal processing of organs and tissues. The protocol yields high-quality RNA from cells targeted with a green fluorescent protein (GFP)-labeled ribosomal subunit. Downstream analysis tools, such as qRT-PCR or RNA-seq, reveal tissue and cell type-specific expression profiles.

翻译核糖体亲和纯化（TRAP）以研究细胞类型特异性尺度下阿拉伯生物基塔利亚纳根发育

In this article, we give hands-on instructions to obtain translatome data from different Arabidopsis thaliana root cell types via the translating ribosome ...

发育生物学

Imaging Spatial Reorganization of a MAPK Signaling Pathway Using the Tobacco Transient Expression System

Visualization of dynamic signaling events in live cells has been a challenge. We expanded an established transient expression system, the biomolecular fluorescent complementation (BiFC) assay in tobacco epidermal cells, from testing protein-protein interaction to monitoring spatial distribution of signal transduction in plant cells. In this protocol, we used the BiFC assay to show that the interaction and the signaling between the Arabidopsis MAPKKK YODA and MAPK6 occur at the plasma membrane. When the scaffold protein BASL was co-expressed, the YODA-MAPK interaction redistributed and spatially co-polarized with CFP-BASL. This modified tobacco expression system allows for quick examination of signaling localization and dynamic changes (less than 4 days) and can accommodate multiple of fluorescent protein colors (at least three). We also presented detailed methods to quantify protein distribution (asymmetric spatial localization, or &#34;polarization&#34;) in tobacco cells. This advanced tobacco expression system has a potential to be used widely for quick testing of dynamic signaling events in live plant cells.

At the subcellular level, signaling events are dynamically modulated by developmental and environmental cues. Here we describe a protocol that employs the tobacco transient expression system to monitor dynamic protein-protein interaction and to disclose spatial organization of signal transduction in plant cells.

成像使用烟草瞬时表达系统MAPK信号通路的空间重组

Visualization of dynamic signaling events in live cells has been a challenge. We expanded an established transient expression system, the biomolecular ...

生物学

Light Sheet Fluorescence Microscopy of Plant Roots Growing on the Surface of a Gel

One of the key questions in understanding plant development is how single cells behave in a larger context of the tissue. Therefore, it requires the observation of the whole organ with a high spatial- as well as temporal resolution over prolonged periods of time, which may cause photo-toxic effects. This protocol shows a plant sample preparation method for light-sheet microscopy, which is characterized by mounting the plant vertically on the surface of a gel. The plant is mounted in such a way that the roots are submerged in a liquid medium while the leaves remain in the air. In order to ensure photosynthetic activity of the plant, a custom-made lighting system illuminates the leaves. To keep the roots in darkness the water surface is covered with sheets of black plastic foil. This method allows long-term imaging of plant organ development in standardized conditions.

This protocol shows a plant sample preparation method for light-sheet microscopy. The setup is characterized by mounting the plant vertically on the surface of a gel and letting it grow in controlled bright conditions. This allows long-term observation of plant organ development in standardized conditions.

植物根系的轻质板材荧光显微镜生长在一个凝胶表面

One of the key questions in understanding plant development is how single cells behave in a larger context of the tissue. Therefore, it requires the ...

Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes

Demonstrated here is a detailed protocol for Agrobacterium-mediated genetic transformation of maize inbred lines using morphogenic genes Baby boom (Bbm) and Wuschel2 (Wus2). Bbm is regulated by the maize phospholipid transferase gene (Pltp) promoter, and Wus2 is under the control of a maize auxin-inducible (Axig1) promoter. An Agrobacterium strain carrying these morphogenic genes on transfer DNA (T-DNA) and extra copies of Agrobacterium virulence (vir) genes are used to infect maize immature embryo explants. Somatic embryos form on the scutella of infected embryos and can be selected by herbicide resistance and germinated into plants. A heat-activated cre/loxP recombination system built into the DNA construct allows for removal of morphogenic genes from the maize genome during an early stage of the transformation process. Transformation frequencies of approximately 14%, 4%, and 4% (numbers of independent transgenic events per 100 infected embryos) can be achieved for W22, B73, and Mo17, respectively, using this protocol.

Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes

Plant morphogenic genes can be used to improve genetic transformation of recalcitrant genotypes. Described here is an Agrobacterium-mediated genetic transformation (QuickCorn) protocol for three important public maize inbred lines.

使用形态基因的农业细菌 -顽固玉米近亲繁殖系的介导未成熟胚胎转化

Demonstrated here is a detailed protocol for Agrobacterium-mediated genetic transformation of maize inbred lines using morphogenic genes Baby boom (Bbm) ...

农杆菌介导的植物毛状根诱导和再生

Hairy Root Transformation and Regeneration in Arabidopsis thaliana and Brassica napus

Hairy root transformation represents a versatile tool for plant biotechnology in various species. Infection by an Agrobacterium strain carrying a Root-inducing (Ri) plasmid induces the formation of hairy roots at the wounding site after the transfer of T-DNA from the Ri plasmid into the plant genome. The protocol describes in detail the procedure of the injection-based hairy root induction in Brassica napus DH12075 and Arabidopsis thaliana Col-0. The hairy roots may be used to analyze a transgene of interest or processed for the generation of transgenic plants. Regeneration medium containing cytokinin 6-benzylaminopurine (5 mg/L) and auxin 1-naphthaleneacetic acid (8 mg/L) successfully elicits shoot formation in both species. The protocol covers the genotyping and selection of regenerants and T1 plants to obtain plants carrying a transgene of interest and free of T-DNA from the Ri plasmid. An alternative process leading to the formation of a composite plant is also depicted. In this case, hairy roots are kept on the shoot (instead of the natural roots), which enables the study of a transgene in hairy root cultures in the context of the whole plant.

作者聚焦:优化基于毛状根的转化方案以提高十字花科植物的效率

The protocol describes hairy root induction using Arabidopsis primary inflorescence stems and Brassica napus hypocotyls. The hairy roots can be cultured and used as explants to regenerate transgenic plants.

侧根诱导系统

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