ELISA-based quantitative detection of peptides in plant tissue v1

Le résumé fourni par la source

Phytocytokines play a critical role in the fine tuning of plant immunity and development. The final bioactive peptide form can vary from 1-5 kDa and is usually expressed in low amounts, making them difficult to detect with conventional methods. Here, we present an ELISA-based assay that allows rapid and cost-effective detection of naturally released peptides in plant tissues. This protocol is optimized for Zip1, a 17-amino-acid phytocytokine derived from Zea mays that elicits salicylic acid signaling in maize leaves. Using a custom peptide antibody, we designed an experimental pipeline to achieve a specific, selective and sensitive recognition of Zip1 allowing its detection in complex biological samples. Using this protocol, we are able to quantify in leaf material native Zip1-containing peptides with a detection limit in the nanogram range. Our methodology has a three-day turn-over and allows the use of raw extract from plant tissue. This method can be adapted for the detection and quantification of a variety of plant signaling peptides.

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