The infiltration-centrifugation technique for extraction of apoplastic fluid from plant leaves using Phaseolus vulgaris as an example.
The infiltration-centrifugation technique for extraction of apoplastic fluid from plant leaves using Phaseolus vulgaris as an example.
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DOI：
10.3791/52113
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发表时间：
2014-12-19
期刊：
影响因子：
--
通讯作者：
Preston GM
中科院分区：
文献类型：
--
作者：
O'Leary BM;Rico A;McCraw S;Fones HN;Preston GM
质外体是植物组织中位于质膜外并包括细胞壁的独特的细胞外区室。植物叶片的质外体区室是几个重要生物过程的场所，包括细胞壁形成、细胞营养和水的吸收和输出、植物-内生菌相互作用和对病原体的防御反应。渗透-离心法是分析各种植物可溶性质外体组成的可靠技术。通过该方法获得的流体通常称为质外体洗涤液（AWF）。以下方案描述了用于从菜豆（菜豆）cv.嫩绿的叶子。讨论了该方法的局限性和其他植物物种的协议的优化。AWF可以用于广泛的下游实验，这些实验旨在表征质外体的组成以及它如何响应植物物种和基因型，植物发育和环境条件而变化，或者确定微生物如何在质外体液体中生长并对其组成的变化做出反应。
The apoplast is a distinct extracellular compartment in plant tissues that lies outside the plasma membrane and includes the cell wall. The apoplastic compartment of plant leaves is the site of several important biological processes, including cell wall formation, cellular nutrient and water uptake and export, plant-endophyte interactions and defence responses to pathogens. The infiltration-centrifugation method is well established as a robust technique for the analysis of the soluble apoplast composition of various plant species. The fluid obtained by this method is commonly known as apoplast washing fluid (AWF). The following protocol describes an optimized vacuum infiltration and centrifugation method for AWF extraction from Phaseolus vulgaris (French bean) cv. Tendergreen leaves. The limitations of this method and the optimization of the protocol for other plant species are discussed. Recovered AWF can be used in a wide range of downstream experiments that seek to characterize the composition of the apoplast and how it varies in response to plant species and genotype, plant development and environmental conditions, or to determine how microorganisms grow in apoplast fluid and respond to changes in its composition.