| 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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| Salinity and Drought Treatment Assays in Kenaf (Hibiscus cannabinus L.)
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Author:
Date:
2016-09-05
[Abstract] Salinity and drought are the two main factors that cause fiber yield and quality losses in kenaf. It is reported that salinity and drought can affect more than 10% of arable land and cause a global decline in the average yields of major crops by more than 50%. Therefore, understanding plant tolerance of drought and salinity is of fundamental importance and has become the focus of intensive research. This protocol describes a simple and reproducible protocol to imitate natural salinity and drought stress under soil conditions. Even though the water-culture method is most frequently used for salinity and drought treatments, the soil-culture method in this study was more applicable to studying natural stress conditions.
[摘要] 盐度和干旱是导致红麻产量和质量损失的两个主要因素。 据报道，盐度和干旱可影响10％以上的耕地，导致主要作物平均单产下降50％以上。 因此，了解植物对干旱和盐分的耐受性具有根本重要性，已成为深入研究的重点。 该协议描述了在土壤条件下模拟天然盐度和干旱胁迫的简单和可重复的方案。 即使水培法最常用于盐分和干旱处理，本研究中的土壤培养方法更适用于研究自然胁迫条件。
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| Transformation of the Cyanobacterium Leptolyngbya boryana by Electroporation
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Author:
Date:
2015-12-20
[Abstract] Leptolyngbya boryana (L. boryana) (formerly Plectonema boryanum) is a versatile, filamentous cyanobacterium that has the ability to fix nitrogen under microoxic conditions and to grow heterotrophically with glucose in the dark, providing an excellent system to investigate photosynthesis, nitrogen fixation, and their regulatory mechanisms. While L. boryana is not naturally transformable different from the unicellular cyanobacterium Synechocystis sp. PCC 6803, it can be transformed by electroporation. Here we describe the transformation of L. boryana by electroporation to isolate mutants in which a targeted gene is disrupted.
[摘要] Leptolyngbya boryana （ L。boryana ）（以前称为Plectonema boryanum）是一种多用途的丝状蓝细菌，具有在微氧条件下固氮的能力， 在黑暗中与葡萄糖异养生长，提供了研究光合作用，固氮和它们的调节机制的优良系统。 而 L。 boryana 不是天然可转化的，不同于单细胞蓝细菌集胞藻。 PCC 6803，它可以通过电穿孔转化。 这里我们描述了 L的变换。 boryana通过电穿孔分离其中靶基因被破坏的突变体。
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