| 15N-nitrate Uptake Activity and Root-to-shoot Transport Assay in Rice
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Author:
Date:
2016-08-20
[Abstract] 15N is a nonradioactive heavy isotope of nitrogen, widely used for biochemical and physiological research in plants. For instance, 15N-KNO3 was used as the nitrogen source in plants in order to investigate nitrate uptake activity and transport from roots to shoots (Lin et al., 2008). Here, we describe a detailed pipeline used for labeling living rice (Oryza sativa) plants with 15N-KNO3 and determination of net nitrate uptake and transport activity, and this protocol was proved to be valid in Arabidopsis and rice (Lin et al., 2008; Hu et al., 2015).
[摘要] N是氮的非放射性重同位素，广泛用于植物的生化和生理研究。 例如，使用N-KNO 3作为植物中的氮源，以研究硝酸盐吸收活性和从根到芽的转运（Lin等人 。， 2008）。 在这里，我们描述了用于标记具有 N-KNO 3的活水稻（>稻）植物和测定硝酸盐硝酸盐吸收的详细管道 和转运活性，并且该方案在拟南芥（Arabidopsis）和水稻中被证明是有效的（Lin等人，2008; Hu等人， 2015）。
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| Isolation of Chloroplast Inner and Outer Envelope Membranes
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Author:
Date:
2015-02-20
[Abstract] The chloroplast is an important organelle found in plant cells that conduct photosynthesis. It is enclosed by a pair of closely spaced membranes, the double-membrane envelope, consisting of the inner membrane bounding the matrix or stroma and the outer membrane in contact with the cytoplasm. Like many bio-membranes, the chloroplast envelope plays an important role in mediating the complex interactions between the chloroplast and the cytoplasm. The envelope is also the site of various biosynthetic reactions, including the formation of the galactolipids, which are the major components of both envelope and the thylakoid membranes. The inner and outer envelope membranes have differences in both structure and function. For example, the outer membrane exhibits lower density of intramembranous ...
[摘要] 叶绿体是在进行光合作用的植物细胞中发现的重要细胞器。它被一对紧密间隔的膜包围，双膜包膜由包围基质或基质的内膜和与细胞质接触的外膜组成。像许多生物膜一样，叶绿体包膜在介导叶绿体和细胞质之间的复杂相互作用中起重要作用。包膜也是各种生物合成反应的位点，包括半乳糖脂的形成，半乳糖脂是包膜和类囊体膜的主要成分。内封套膜和外封套膜在结构和功能上都有差异。例如，外膜表现出比内膜剂量更低的膜内颗粒密度，表明外膜的蛋白质含量低。此外，外膜对于低分子量化合物是非特异性可渗透的，而内部膜对于这些化合物是不可渗透的，并且包含用于运输代谢物的几种易位体系统。
为了制备包膜，必须首先分离完整的叶绿体。然后通过以下方式分离内外膜：1）蛋白酶处理方法和2）基于外壳更轻且内膜更重的事实的离心方法。两种方法都需要首先分离完整的叶绿体。然而，离心分离可以得到纯的内外封套制剂，因此适合于随后的分析。此外，离心法可以避免在蛋白酶处理期间内皮多肽的破坏，因为一些蛋白酶可以进入内膜。此外，离心方法容易操作并且获得包含较少外部和内部信封膜的粘合区域的完全包封。在这里，我们描述了一种可靠的方法，用于从烟草中分离叶绿体的内和外包膜，这是相对不容易用于包膜分离的植物。
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