Division of Mechatronics, Biostatistics and Sensors (MeBioS), Department of Biosystems (BIOSYST), Katholieke Universiteit Leuven, Willem de Croylaan 42, bus 2428, B-3001 Leuven, Belgium.
Plant Methods. 2011 Jun 23;7:17. doi: 10.1186/1746-4811-7-17.
The foundations for ethylene research were laid many years ago by researchers such as Lizada, Yang and Hoffman. Nowadays, most of the methods developed by them are still being used. Technological developments since then have led to small but significant improvements, contributing to a more efficient workflow. Despite this, many of these improvements have never been properly documented.
This article provides an updated, integrated set of protocols suitable for the assembly of a complete picture of ethylene biosynthesis, including the measurement of ethylene itself. The original protocols for the metabolites 1-aminocyclopropane-1-carboxylic acid and 1-(malonylamino)cyclopropane-1-carboxylic acid have been updated and downscaled, while protocols to determine in vitro activities of the key enzymes 1-aminocyclopropane-1-carboxylate synthase and 1-aminocyclopropane-1-carboxylate oxidase have been optimised for efficiency, repeatability and accuracy. All the protocols described were optimised for apple fruit, but have been proven to be suitable for the analysis of tomato fruit as well.
This work collates an integrated set of detailed protocols for the measurement of components of the ethylene biosynthetic pathway, starting from well-established methods. These protocols have been optimised for smaller sample volumes, increased efficiency, repeatability and accuracy. The detailed protocol allows other scientists to rapidly implement these methods in their own laboratories in a consistent and efficient way.
乙烯研究的基础是多年前由 Lizada、Yang 和 Hoffman 等研究人员奠定的。如今，他们开发的大多数方法仍在使用。自那时以来的技术发展带来了微小但显著的改进，有助于提高工作效率。尽管如此，这些改进中的许多从未得到妥善记录。
本文提供了一套经过更新的、集成的方案，适用于组装完整的乙烯生物合成图，包括乙烯本身的测量。代谢物 1-氨基环丙烷-1-羧酸和 1-（丙二酰氨基）环丙烷-1-羧酸的原始方案已经过更新和缩小规模，而用于确定关键酶 1-氨基环丙烷-1-羧酸合酶和 1-氨基环丙烷-1-羧酸氧化酶体外活性的方案已针对效率、可重复性和准确性进行了优化。所有描述的方案均针对苹果果实进行了优化，但已证明它们也适用于番茄果实的分析。
这项工作汇集了一套详细的测量乙烯生物合成途径成分的综合方案，从成熟的方法开始。这些方案已针对较小的样本量、提高效率、可重复性和准确性进行了优化。详细的方案允许其他科学家以一致和有效的方式在自己的实验室中快速实施这些方法。
