Jack Chandra N, Rowe Shawna L, Porter Stephanie S, Friesen Maren L
Department of Plant Pathology Washington State University Pullman Washington 99164 USA.
Department of Plant Biology Michigan State University East Lansing Michigan 48824 USA.
Appl Plant Sci. 2019 Jan 8;7(1):e01210. doi: 10.1002/aps3.1210. eCollection 2019 Jan.
Current methods for quantifying herbivore-induced alterations in plant biochemistry are often unusable by researchers due to practical constraints. We present a cost-effective, high-throughput protocol to quantify multiple biochemical responses from small plant tissue samples using spectrophotometric techniques.
Using and leaves pre- and post-herbivory, we demonstrate that our protocol quantifies common plant defense responses: peroxidase production, polyphenol oxidase production, reactive oxygen species production, total protein production, and trypsin-like protease inhibition activity.
Current protocols can require 500 mg of tissue, but our assays detect activity in less than 10 mg. Our protocol takes two people 6 h to run any of the assays on 300 samples in triplicate, or all of the assays on 20 samples. Our protocol enables researchers to plan complex experiments that compare local versus systemic plant responses, quantify environmental and genetic variation, and measure population-level variation.
由于实际限制，研究人员通常无法使用当前量化食草动物诱导的植物生化变化的方法。我们提出了一种经济高效的高通量方案，使用分光光度技术从小植物组织样本中量化多种生化反应。
利用食草前后的[具体植物]叶片，我们证明我们的方案能够量化常见的植物防御反应：过氧化物酶产生、多酚氧化酶产生、活性氧产生、总蛋白产生以及类胰蛋白酶抑制活性。
当前方案可能需要500毫克组织，但我们的检测方法能在不到10毫克的组织中检测到活性。我们的方案让两个人在6小时内就能对300个样本进行任何一项一式三份的检测，或者对20个样本进行所有检测。我们的方案使研究人员能够规划复杂的实验，比较植物的局部与系统反应，量化环境和遗传变异，并测量种群水平的变异。
