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The bHLH transcription factor ILR3 modulates multiple stress responses in Arabidopsis

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ILR3 and PYE function in a regulatory network that modulates GLS accumulation under iron deficiency.

Abstract

The molecular processes involved in the cross talk between iron (Fe) homeostasis and other metabolic processes in plants are poorly understood. In Arabidopsis thaliana the transcription factor IAA-LEUCINE RESISTANT3 (ILR3) regulates iron deficiency response, aliphatic glucosinolate (GLS) biosynthesis and pathogen response. ILR3 is also known to interact with its homolog, POPEYE (PYE), which also plays a role in Fe response. However, little is known about how ILR3 regulates such diverse processes, particularly, via its interaction with PYE. Since GLS are produced as part of a defense mechanism against wounding pathogens, we examined pILR3::β-GLUCURONIDASE expression and found that Fe deficiency enhances the wound-induced expression of ILR3 in roots and that ILR3 is induced in response to the wounding pathogen, sugarbeet root cyst nematode (Heterodera schachtii). We also examined the expression pattern of genes involved in Fe homeostasis and aliphatic GLS biosynthesis in pye-1, ilr3-2 and pye-1xilr3-2 (pxi) mutants and found that ILR3 and PYE differentially regulate the expression of genes involved these processes under Fe deficiency. We measured GLS levels and sugarbeet root cyst nematode infection rates under varying Fe conditions, and found that long-chain GLS levels are elevated in ilr3-2 and pxi mutants. This increase in long-chain GLS accumulation is correlated with elevated nematode resistance in ilr3-2 and pxi mutants in the absence of Fe. Our findings suggest that ILR3 and PYE function in a regulatory network that controls wounding pathogen response in plant roots by modulating GLS accumulation under iron deficiency.

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Abbreviations

Fe:

Iron

bHLH TF:

Basic helix-loop-helix transcription factor

ILR3:

IAA-LEUCINE RESISTANT3

PYE:

POPEYE

GLS:

Glucosinolates

BAT5:

Bile acid transporter 5

MAM1:

Methylthioalkylmalate synthase1

CYP81A:

Cytochrome P450 81A

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Acknowledgements

The authors thank Dr. Bonnie Bartel (Rice University) for the distribution of pILR3:GUS seeds. We also thank Drs. Marcela Rojas-Pierce and Bob Franks (North Carolina State University) for critical reading of the manuscript. This work was supported by the US National Science Foundation (Grant No. #1120937), and the USDA National Institute of Food and Agriculture, Hatch Project (Accession No. 101090).

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Authors and Affiliations

  1. Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, 27695, USA

    Jeffrey W. Gillikin & Terri A. Long

  2. Department of Plant Sciences, University of California, Davis, CA, 95616, USA

    Baohua Li & Daniel Kliebenstein

  3. DynaMo Center, Copenhagen Plant Science Centre, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark

    Daniel Kliebenstein

  4. Department of Entomology and Plant Pathology, North Carolina State University, Raleigh, NC, 27695, USA

    Rozalynne Samira, Chunying Li & Eric Davis

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  1. Rozalynne Samira
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  2. Baohua Li
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  3. Daniel Kliebenstein
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  4. Chunying Li
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  6. Jeffrey W. Gillikin
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  7. Terri A. Long
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Contributions

TAL, RS, CL, and DK planned and designed the research; RS, BL and JWG performed experiments; CL and ED provided reagents, tools and guidance for experiments with Heterodera schachtii; RS, BL and CL analyzed data. RS and TAL wrote the manuscript with the input of DK, JWG and all other authors.

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Correspondence to Terri A. Long.

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Samira, R., Li, B., Kliebenstein, D. et al. The bHLH transcription factor ILR3 modulates multiple stress responses in Arabidopsis. Plant Mol Biol 97, 297–309 (2018). https://doi.org/10.1007/s11103-018-0735-8

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