| A Low-volume Hydroponic Protocol for Maize and A Sensitive Bud-length Assay for Root-to-Shoot Impacts of The Strigolactone Analog, rac-GR24
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Author:
Date:
2020-08-20
[Abstract] Hydroponic approaches have been a boon for root research by facilitating root-feeding studies as well as secretion analysis. Results have aided our understanding of root-shoot signaling, transport, hormone function and more. However, existing approaches are often restricted to small plants and seedlings by prohibitive cost or availability of experimental substrates. In addition to this, research on the hormone, strigolactone (SL) has been constrained in species like maize by a lack of specific assays for quantifying responses. Here a low-volume hydroponic approach was developed for growing maize plants to the 3-leaf stage (about 2.5 weeks and 12 cm tall at the 2nd leaf collar) using 4 plants with 1 L of aerated media. This protocol also allowed development of an assay for ...
[摘要] [摘要 ] 通过促进根饲研究和分泌分析，水培方法已成为根研究的福音。结果有助于我们理解根射击信号，转运，激素功能等。然而，由于成本过高或实验底物的可获得性，现有方法通常仅限于小型植物和幼苗。除此之外，研究激素，独脚金（SL）已经在玉米等品种由于缺乏量化响应特定试验的限制。这里的低体积水培方法被用于玉米植物生长到3叶阶段（约2.5周和12厘米高的2开发第二LEA f领）使用4种植物和1升充气培养基。该协议还允许开发一种针对SL 类似物rac -GR24 的从根到芽影响的测定方法，该方法对玉米芽上侧芽的生长具有显着的，易于量化的影响。该协议的优势包括经济有效的简便性和精确度，可以量化从根到茎的SL 响应。此外，小体积水培方法可以很容易地用于玉米或其他高粱，C4草（如高粱，甘蔗和桔）的连续或脉冲型根饲研究，根标记实验和/或分泌物采样。
[背景 ] 的用于开发这里介绍的协议初始目的是量化的激素，根对枝条影响独脚金内酯（SL）在玉米中。推定松香内酯在该物种的驯化中起着核心作用，这是由于该激素对植物结构的贡献，以及因此对现代玉米的单茎形态造成的影响（关等人，2012 ）。为了解决这种可能性，需要一种系统，该系统将1 ）允许在这种大草种的初始营养生长期间进行根饲研究，2 ）最小化使用的SL 类似物的数量和成本（rac -GR24），以及3 ...
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| Manganese Superoxide Dismutase Activity Assay in the Yeast Saccharomyces cerevisiae
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Author:
Date:
2020-03-05
[Abstract] Superoxide dismutases (SODs) act as a primary defence against reactive oxygen species (ROS) by converting superoxide anion radicals (O2-) into molecular oxygen (O2) and hydrogen peroxide (H2O2). Members of this enzyme family include CuZnSODs, MnSODs, FeSODs, and NiSODs, depending on the nature of the cofactor that is required for proper activity. Most eukaryotes, including yeast, possess CuZnSOD and MnSOD. This protocol aims at assessing the activity of the yeast Saccharomyces cerevisiae MnSOD Sod2p from cellular extracts using nitroblue tetrazolium staining. This method can be used to estimate the cellular bioavailability of Mn2+ as well as to evaluate the redox state of the cell.
[摘要] [摘要 ] 超氧化物歧化酶（SOD能）充当主防御针对反应性氧物质（ROS）通过转换的超氧阴离子自由基（O 2 - ）为分子氧（O 2 ）和过氧化氢（H 2 ? 2 ）。这种酶的家庭成员包括CuZnSODs ，MnSODs ，FeSODs 和NiSODs ，这取决于是需要适当的活动辅助因子的性质。大多数真核生物，包括酵母，都具有CuZnSOD 和MnSOD 。该协议旨在评估酵母的活性 使用硝基蓝四唑染色法从细胞提取物中提取酿酒酵母MnSOD Sod2p 。该方法可用于估计Mn 2+ 的细胞生物利用度以及评估细胞的氧化还原状态。
[背景 ] 的SODs被定义为减少正常有氧代谢为氧气和过氧化氢期间形成的氧的有害自由基含金属的抗氧化剂酶。：这些酶是基于需要作为辅因子进行适当的酶活性的金属分类CuZnSODs ，MnSODs ，FeSODs ，和NiSODs 。在酿酒酵母中，有两个S OD ：CuZn-Sod1p和Mn-Sod2p（Abreu和Cabelli ...
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| Analysis of Metals in Whole Cells, Thylakoids and Photosynthetic Protein Complexes in Synechocystis sp. PCC6803
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Author:
Date:
2018-06-20
[Abstract] Metals are essential in many biological processes, including oxygenic photosynthesis. Here we described a method to measure the metal pool in whole cells and thylakoids, including the bioactive pool in intact photosynthetic protein complexes in the model oxygenic cyanobacterium Synechocystis PCC6803. In the first part of the protocol, whole cells and thylakoid membranes are carefully prepared, in which the total metal concentrations are measured by inductively coupled plasma triple-quadrupole mass spectrometry (ICP-QQQ-MS). In the second part of the protocol, isolated thylakoids are solubilized to release the integral membrane proteins and the metal binding protein complexes. These intact photosynthetic protein complexes are subjected to size exclusion chromatography (SEC) and ...
[摘要] 金属在许多生物过程中是必不可少的，包括含氧光合作用。 在这里我们描述了测量全细胞和类囊体中金属库的方法，包括模型含氧蓝藻PCH6803中完整光合蛋白复合体中的生物活性库。 在方案的第一部分中，仔细制备全细胞和类囊体膜，其中通过电感耦合等离子体三重四极杆质谱（ICP-MS / MS）测量总金属浓度。 在该方案的第二部分，分离的类囊体被溶解以释放完整的膜蛋白和金属结合蛋白复合物。 将这些完整的光合蛋白复合物进行尺寸排阻色谱（SEC），并通过ICP-MS / MS进行连接分析尺寸分离的复合物中的金属结合。
【背景】氧光合作用的过程需要金属，因为它们在光合电子传递链中作为辅因子和催化剂的基本功能。光合装置需要Fe-S簇，血红素桥Fe和非血红素Fe形式的铁（Fe），可溶性移动电子载体蛋白质质体蓝素中的铜（Cu），叶绿素中的镁（Mg）， ...
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