Measurement of Transcripts Associated with Photorespiration and Related Redox Signaling

Abstract

To study photorespiration and to characterize related components, gene expression analysis is a central approach. An overview of the experimental setup, protocols, and methods we use to investigate photorespiration-associated gene expression is presented. Within this chapter, we describe simple procedures to experimentally alter the photorespiratory flux and provide protocols for transcriptomic analysis with a focus on genes encoding photorespiratory proteins as well as those induced by photorespiratory hydrogen peroxide (HO). Examples of typical results are presented and their significance to understanding redox signaling is discussed.

Non‐targeted 13C metabolite analysis demonstrates broad re‐orchestration of leaf metabolism when gas exchange conditions vary

“…Second, since PAL is cytosolic (as well as other enzymes of caffeate synthesis), it eventually leads to a net export of redox power from the chloroplast, whereas tyrosine synthesis generates NADPH in the chloroplast. Third, phenylpropanoids can play the role of antioxidants and this may be beneficial at high photorespiration (Mhamdi, Kerchev, Willems, Noctor, & Van Breusegem, 2017; Noctor & Mhamdi, 2017). In particular, lignin precursors (hydroxycinnamate family members) can participate in quenching radical ROS via monodehydroascorbate reductase (Sakihama, Ji, Sano, Asada, & Yamasaki, 2000).…”

TheArabidopsismediator complex subunit 8 regulates oxidative stress responses

“…This analysis revealed that the med8 mutation did not further promote perturbation of the redox state of glutathione, as both oxidized and reduced glutathione were similar in cat2 and cat2 med8 (Figure 3D). Secondly, we quantified four oxidative signaling marker transcripts known to be induced by H 2 O 2 (Noctor et al, 2016;Mhamdi et al, 2017). Three of these transcripts were highly induced in cat2 med8 plants, hinting at a stronger activation of oxidative signaling in these conditions, above the cat2 level (Figure 3D).…”

Metabolite modification in oxidative stress responses: A case study of two defense hormones

“…RNA concentrations were determined using Nanodrop and 1 µg of RNA was used to perform reverse transcription using the qScript cDNA Synthesis Kit (Quantabio). Selected genes were chosen for qPCR analysis according to [32]. Two or three independent experiments were performed, each with three biological replicates and each with at least 12 plants per genotype.…”