| In situ Hybridization (ISH) in Preparasitic and Parasitic Stages of the Plant-parasitic Nematode Meloidogyne spp.
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Author:
Date:
2018-03-20
[Abstract] The spatio-temporal expression pattern of a gene provides important indications to better understand its biological function. In situ hybridization (ISH) uses a labeled complementary single-stranded RNA or DNA probe to localize gene transcripts in a whole organism, a whole organ or a section of tissue. We adapted the ISH technique to the plant parasite Meloidogyne spp. (root-knot nematode) to visualize RNAs both in free-living preparasitic juveniles and in parasitic stages settled in the plant tissues. We describe each step of the probe synthesis, digoxigenin (DIG) labeling, nematode extraction from plant tissue, and ISH procedure.
[摘要] 基因的时空表达模式为更好地理解其生物学功能提供了重要的指示。 原位杂交（ISH）使用标记的互补单链RNA或DNA探针来定位整个生物体，整个器官或一部分组织中的基因转录物。 我们将ISH技术应用于植物寄生虫
【背景】到目前为止，植物寄生性线虫的稳定转化尚未成功。 ISH能够在整个装载的Meloidogyne spp中分析体内时空基因表达。线虫。这些根结线虫在土壤中以微小蚓状幼虫（J2）形式孵化并感染宿主植物根部。 J2s穿透根部并迁移到根部维管柱状细胞。幼虫定居在根部，发育成J3和J4寄生幼鱼，诱导分化专化饲养细胞。线虫最终发育成梨形雌性，将在根表面释放数百个卵。在这里，我们报告了一个详细的协议来检测准备性整体安装J2s和寄生阶段中的单个RNA分子。寄生虫阶段的ISH需要在感染根部提取线虫前一天采取额外的程序。我们描述了在线虫整个组织中使用地高辛（DIG）标记的cDNA探针检测转录物。
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| Saccharification Protocol for Small-scale Lignocellulosic Biomass Samples to Test Processing of Cellulose into Glucose
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Author:
Date:
2016-01-05
[Abstract] Second generation biofuels are derived from inedible lignocellulosic biomass of food and non-food crops. Lignocellulosic biomass is mainly composed of cell walls that contain a large proportion of cellulosic and hemicellulosic polysaccharides. An interesting route to generate biofuels and bio-based materials is via enzymatic hydrolysis of cell wall polysaccharides into fermentable sugars, a process called saccharification. The released sugars can then be fermented to fuels, e.g., by use of yeast.
To test the saccharification efficiency of lignocellulosic biomass on a lab-scale, a manual saccharification protocol was established that uses only small amounts of biomass and a low concentration of enzyme. This protocol can be used for different plant species like Arabidopsis ...
[摘要] 第二代生物燃料衍生自食物和非食物作物的不可食用的木质纤维素生物质。木质纤维素生物质主要由含有大比例的纤维素和半纤维素多糖的细胞壁组成。生成生物燃料和生物基材料的有趣途径是通过将细胞壁多糖酶水解成可发酵的糖，这一过程称为糖化。释放的糖然后可以例如通过使用酵母发酵成燃料。
为了在实验室规模上测试木质纤维素生物质的糖化效率，建立了仅使用少量生物质和低浓度酶的手动糖化方案。该方案可以用于不同的植物物种，例如拟南芥（Arabidopsis thaliana），烟草，玉米和杨树。低的酶浓度使得可以检测糖化产量的微妙改进并分析水解的速度。虽然包括特定的酸和碱预处理，但是糖化步骤可以在任何其它预处理之后。由于不需要先进的设备，该方案可以在许多实验室进行以分析糖化产量。该方案最初在Van Acker等人（2013）中描述。
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