| Title: | A high-throughput method of analyzing multiple plant defensive compounds in minimized sample mass |
| Author(s): | Jack CN; Rowe SL; Porter SS; Friesen ML; |
| Address: | Department of Plant Pathology Washington State University Pullman Washington 99164 USA. Department of Plant Biology Michigan State University East Lansing Michigan 48824 USA. School of Biological Sciences Washington State University Vancouver Washington 98686 USA. Department of Crop and Soil Sciences Washington State University Pullman Washington 99164 USA |
| ISSN/ISBN: | 2168-0450 (Print) 2168-0450 (Electronic) 2168-0450 (Linking) |
| Abstract: | "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" |
| Keywords: | microplate peroxidase plant defense response polyphenol oxidase protease inhibitors protein quantification; |
| Notes: | "PubMed-not-MEDLINEJack, Chandra N Rowe, Shawna L Porter, Stephanie S Friesen, Maren L eng T32 GM110523/GM/NIGMS NIH HHS/ 2019/01/30 Appl Plant Sci. 2019 Jan 8; 7(1):e01210. doi: 10.1002/aps3.1210. eCollection 2019 Jan" |
