A Protocol for Flavonols, Kaempferol and Quercetin, Staining in Plant Root Tips
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Author:
Date:
2020-10-05
[Abstract] Flavonols are a subclass of flavonoids of the group of plant secondary metabolites. In planta, flavonols play various functions such as antioxidant and natural regulator of auxin polar transport. Many lines of evidence have shown that flavonols also contribute to human health in anti-oxidation, anti-inflammation, and even prevention some types of cancer. Several methods have been utilized to measure flavonols such as high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), and diphenylboric acid-2-aminoethyl ester (DPBA) staining. While HPLC or LC-MS can quantitatively determine the level of flavonols, DPBA staining can provide an in-situ view of flavonols accumulation in the plants. In this protocol, a detailed procedure for ...
[摘要] [摘要]黄酮醇是植物次生代谢物中黄酮类的一个亚类。在植物中,黄酮醇具有多种功能,例如抗氧化剂和生长素极性转运的天然调节剂。许多证据表明,黄酮醇还有助于抗氧化,抗炎症甚至预防某些类型的癌症,从而对人类健康做出贡献。几种方法已用于测量黄酮醇,例如高效液相色谱(HPLC),液相色谱-质谱(LC-MS)和二苯硼酸-2-氨基乙基酯(DPBA)染色。HPLC或LC-MS可以定量测定黄酮醇的含量 DPBA染色可以提供植物中黄酮醇积累的原位视图。在该协议中,描述了对拟南芥根尖中的黄酮醇进行染色的详细步骤。将五天大的拟南芥幼苗浸入含有DPBA的溶液中,然后在共聚焦显微镜下观察黄酮醇(山ka酚和槲皮素)。
[背景]在拟南芥,黄酮醇是从缩合反应对香豆酰基辅酶A的一个分子和丙二酰-CoA的三个分子之间生物合成。Lewis等。(2011年)发现槲皮素而不是山奈酚是根型根茎植物生长素转运的抑制剂。此外,黄酮醇也可作为植物和人类的有效抗氧化剂。先前的研究调查了DPBA与黄酮醇相互作用时可以发出荧光(Sheahan和Rechnitz,1992)。基于此,DPBA已被广泛应用于检测植物中黄酮醇的积累(Nguyen等,2013、2015和2016 ; ...
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Artificial Inoculation of Epichloë festucae into Lolium perenne, and Visualisation of Endophytic and Epiphyllous Fungal Growth
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Author:
Date:
2018-09-05
[Abstract] Natural hosts for the fungal endophyte Epichloë festucae include Festuca rubra (fine fescue) and Festuca trachyphylla (hard fescue). Some strains also form stable associations with Lolium perenne (perennial ryegrass). L. perenne is a suitable host to study fungal endophyte–grass interactions, such as endophytic fungal growth within the plant and epiphyllous growth on the plant surface. Here we provide a detailed protocol based on work by, for artificial inoculation of E. festucae into L. perenne, and newly developed staining and visualization techniques for observing endophytic and epiphyllous hyphae and the expressorium, an appressorium-like structure used by the fungus to exit the plant. The staining method uses a combination of ...
[摘要] 真菌内生菌Epichloëfestucae的天然宿主包括 Festuca rubra (细羊茅)和 Festuca trachyphylla (硬羊茅)。一些菌株也与黑麦草(多年生黑麦草)形成稳定的关联。 升。 perenne 是研究真菌内生菌 - 草相互作用的合适宿主,例如植物内生真菌生长和植物表面的叶生生长。在这里,我们提供了一个基于工作的详细协议,用于 E的人工接种。 festucae 进入 L. perenne ,以及新开发的染色和可视化技术,用于观察内生和附生的菌丝和表达菌,这是一种类似真菌的结构,用于离开植物。染色方法使用葡聚糖结合苯胺蓝二铵盐(AB)和几丁质结合小麦胚凝集素缀合的Alexa Fluor ® 488-(WGA-AF488)的组合。该协议将是研究Epichloë -grass相互作用的有用工具,特别是不同Epichloë - 草关联,各种内生菌 - 宿主发育阶段的比较,以及分析突变Epichloë菌株。
【背景】Latch和Christensen(1985)开发了一种用Epichloë内生菌人工感染草的方案。这项工作一直是广泛的 E的基础。 festucae Fl1- L. perenne 共生研究(Scott et al。,2012年综述)。在我们最近的工作中,我们描述了一种新发现的真菌结构,即能够实现 E的表达式。 ...
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Quantification of Thrips Damage Using Ilastik and ImageJ Fiji
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Author:
Date:
2018-04-20
[Abstract] Quantification of insect damage is an essential measurement for identifying resistance in plants. In screening for host plant resistance against thrips, the total damaged leaf area is used as a criterion to determine resistance levels. Here we present an objective novel method for analyzing thrips damage on leaf disc using the freely available software programs Ilastik and ImageJ. The protocol was developed in order to screen over 40 Capsicum lines for resistance against Frankliniella occidentalis (Western Flower Thrips) and Thrips tabaci (Onion thrips).
[摘要] 昆虫损伤的定量是鉴定植物抗性的重要指标。 在筛选宿主植物对蓟马的抗性时,将总的受损叶面积用作确定抗性水平的标准。 在这里我们提出了一种客观的新方法,用免费提供的软件程序Ilastik和ImageJ来分析叶盘上的蓟马损伤。 该协议的开发是为了筛选超过40个辣椒品系对西方花蓟马和西蓟马蓟马(洋葱蓟马)的抗性。
【背景】昆虫损伤的定量是鉴定植物抗性的重要指标。在筛选宿主植物对蓟马抗性的程序中,总损伤叶面积被用作确定抗性水平的标准。蓟马损害的特点是银色斑点与完整叶片区域显示出高对比度,但喂食点还包括从深绿色到棕色的深色区域。这些叶片的逐渐变色太细微,无法用诸如Winfolia等程序精确量化( http:// www .regentinstruments.com / assets / winfolia_software.html )或ImageJ(Rasband,2011)。结果,蓟马损害通常由评估样品的个体评分。将样本分为表示损害数量的类别(Mirnezhad等人,2010; Maharijaya等人,2011和2012),或损害估计为最接近的1 (Leiss等人,2009; ...
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