| Extraction and Measurement of Abscisic Acid in a Unicellular Red Alga Cyanidioschyzon merolae
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Author:
Date:
2016-12-05
[Abstract] Abscisic acid (ABA) has been known as a phytohormone of land plants, which is synthesized in response to abiotic stresses and induces various physiological responses, but is also found from eukaryotic algae. Recently, we reported that a unicellular red alga Cyanidioschyzon merolae produced ABA, which prevented cell growth and enhanced salt stress tolerance (Kobayashi et al., 2016). This report describes the detailed method for the extraction and quantification of ABA in the model red alga C. merolae.
[摘要] 脱落酸（ABA）已知为陆生植物的植物激素，其响应于非生物胁迫合成并诱导各种生理反应，但也可从真核藻类中发现。最近，我们报道了单细胞红藻（Cyanidioschyzon merolae）产生ABA，其阻止细胞生长和增强盐胁迫耐受性（Kobayashi等人，2016）。该报告描述了在红海藻模型中提取和定量ABA的详细方法。 。
关键字：脱落酸，藻类， Cyanidioschyzon merolae ，HPLC，植物激素
] 植物激素ABA已在发散光合真核生物中发现，但单细胞藻类的功能仍不清楚。在最近的研究中，我们显示单细胞红藻。 melorae 通过本方案累积ABA以应答盐胁迫。这是用于从C中提取和定量ABA的第一已公布方案的细节。 merolae 。此协议针对 C进行了优化。基于陆地植物协议的。
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| PCR-based Assay for Genome Integrity after Methyl Methanesulfonate Damage in Physcomitrella patens
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Author:
Date:
2016-10-05
[Abstract] In plant cells, genomic DNA exists in three organelles: the nucleus, chloroplast, and mitochondrion. Genomic DNA can be damaged by endogenous and exogenous factors, but the damaged DNA can be repaired by DNA repair systems. To quantify the extent of their repair activity of on individual genomic DNA, a PCR-based assay utilizing long amplicons is valuable for evaluable. This assay is based on the inhibitory effects of methyl methanesulfonate (MMS)-induced DNA damage on the amplicons. This assay is useful for assessing DNA double-strand repair pathways, such as homologous recombination repair, as it detects DNA double-strand breaks produced by MMS in vivo.
[摘要] 在植物细胞中，基因组DNA存在于三个细胞器中：核，叶绿体和线粒体。基因组DNA可以被内源和外源因子损伤，但损伤的DNA可以通过DNA修复系统修复。为了量化其对单个基因组DNA的修复活性的程度，使用长扩增子的基于PCR的测定对于可评价是有价值的。该测定基于甲磺酸甲酯（MMS）诱导的DNA损伤对扩增子的抑制作用。该测定可用于评估DNA双链修复途径，例如同源重组修复，因为其检测由MMS在体内产生的DNA双链断裂。
[背景] 基因组DNA损伤的定量可用于分析DNA修复机制。该测定利用实时PCR定量核，叶绿体和线粒体DNA拷贝数以用于长PCR产物的标准化，与由Hunter等人先前的方案相比提供更准确的定量。 （2010）。
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| Purification of Flagellin from Acidovorax avenae and Analysis of Plant Immune Responses Induced by the Purified Flagellin
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Author:
Date:
2016-08-20
[Abstract] Plants sense potential pathogens by recognizing conserved pathogen-associated molecular patterns (PAMPs) that cause PAMP-triggered immunity (PTI) including the generation of reactive oxygen species, callose deposition, and expression of several PTI-related genes. Acidovorax avenae is a Gam-negative bacterium that causes a seedling disease characterized by the deposition of brown stripes on the sheaths of infected plants. We previously reported that flagellin isolated from the rice avirulent A. avenae N1141 strain induces PTI, while flagellin isolated from the rice virulent A. avenae K1 strain does not induce PTI. To examine the molecular mechanism of specific PTI induction by N1141 flagellin, highly purified flagellin from N1141 or K1 strains is required. Here, ...
[摘要] 植物通过识别引起PAMP触发的免疫（PTI）的保守的病原体相关分子模式（PAMP）来感测潜在的病原体，包括活性氧的产生，胼lose质沉积和几个PTI相关基因的表达。 酸性弧菌是一种革兰氏阴性细菌，其导致幼苗疾病，其特征在于在感染的植物的鞘上沉积棕色条纹。 我们以前报告了从无毒力的大米中分离的鞭毛蛋白。 avenae N1141菌株诱导PTI，而从水稻毒力分离的鞭毛蛋白。 avenae K1株不诱导PTI。 为了检查N1141鞭毛蛋白的特异性PTI诱导的分子机制，需要来自N1141或K1菌株的高度纯化的鞭毛蛋白。 在这里，我们描述了一种高质量的纯化方法。 avenae鞭毛蛋白，并用于PTI诱导研究。
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