Open AccessA high‐throughput method of analyzing multiple plant defensive compounds in minimized sample mass
Author(s) -
Jack Chandra N.,
Rowe Shawna L.,
Porter Stephanie S.,
Friesen Maren L.
Publication year - 2019
Publication title -
applications in plant sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 23
ISSN - 2168-0450
DOI - 10.1002/aps3.1210
Subject(s) - biology , protocol (science) , solanum , population , herbivore , plant defense against herbivory , microbiology and biotechnology , metabolomics , computational biology , botany , biochemistry , bioinformatics , gene , medicine , alternative medicine , demography , pathology , sociology
Premise of the Study 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. Methods and Results Using Solanum lycopersicum and Medicago polymorpha 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. Conclusions 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.