Abstract

Alkaline soils comprise 30% of the earth and have low plant-available iron (Fe) concentration, and can cause iron deficiency chlorosis (IDC). IDC causes soybean yield losses of $260 million annually. However, it is not known whether molecular responses to IDC are equivalent to responses to low iron supply. IDC tolerant and sensitive soybean lines provide a contrast to identify specific factors associated with IDC. We used RNA-seq to compare gene expression under combinations of normal pH (5.7) or alkaline pH (7.7, imposed by 2.5 mM bicarbonate, or pH 8.2 imposed by 5 mM bicarbonate) and normal (25 μM) or low (1 μM) iron conditions from roots of these lines. Thus, we were able to treat pH and Fe supply as separate variables. We also noted differential gene expression between IDC sensitive and tolerant genotypes in each condition. Classical iron uptake genes, including ferric-chelate reductase (FCR) and ferrous transporters, were upregulated by both Fe deficiency and alkaline stress, however, their gene products did not function well at alkaline pH. In addition, genes in the phenylpropanoid synthesis pathway were upregulated in both alkaline and low Fe conditions. These genes lead to the production of fluorescent root exudate (FluRE) compounds, such as coumarins. Fluorescence of nutrient solution increased with alkaline treatment, and was higher in the IDC tolerant line. Some of these genes also localized to previously identified QTL regions associated with IDC. We hypothesize that FluRE become essential at alkaline pH where the classical iron uptake system does not function well. This work could result in new strategies to screen for IDC tolerance, and provide breeding targets to improve crop alkaline stress tolerance.

Highlights

  • Iron (Fe) is an essential micronutrient for plants

  • While Fe is usually abundant in soil, its solubility is low in alkaline soils, which can lead to iron deficiency chlorosis (IDC), a functional Fe deficiency where leaves are yellow instead of green (Mengel, 1994)

  • This study has indicated a correlation between fluorescent root exudate (FluRE) quantity and IDC tolerance, and has indicated specific genes that may be involved in the regulation, synthesis, and efflux of these compounds

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Summary

Introduction

Iron (Fe) is an essential micronutrient for plants. Iron deficient plants increase root Fe uptake capacity by one of two strategies (Kobayashi and Nishizawa, 2012); grasses use a chelationuptake strategy, while non-grass monocots and dicots employ what is classically considered a reduction-uptake strategy. Alkaline pH and Bic decrease solubility, and in turn availability of Fe, and may inhibit Fe uptake gene expression and/or function (Lucena et al, 2007; Hsieh and Waters, 2016). Plants that are Fe deficient from low Fe supply at normal pH have chlorotic leaves, but it is not clear that this chlorosis is equivalent to IDC under alkaline conditions. Proteomic, or metabolomic studies have included alkaline pH in combination with low Fe supply to induce IDC (Rellán-Álvarez et al, 2010; Rodríguez-Celma et al, 2011a; Rodríguez-Celma et al, 2013; Schmidt et al, 2014). Our first objective was to treat Fe supply and nutrient solution pH as separate variables to determine whether alkaline IDC conditions and low Fe supply affect soybean root gene expression in the same manner

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