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Raphanusanin-mediated resistance to pathogens is light dependent in radish and Arabidopsis thaliana

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Abstract

Raphanusanin (Ra) is a light-induced inhibitor of hypocotyl growth that responds to unilateral blue light illumination in radish seedlings. We have previously shown that Ra regulates genes that are involved in common defense mechanisms. Many genes that are induced by Ra are also positively regulated by early blue light. To extend the understanding of the role of Ra in pathogen defense, we evaluated the effects of Ra on radish and Arabidopsis thaliana (A. thaliana) infected with the necrotrophic pathogen Botrytis cinerea (B. cinerea) and biotrophic pathogen Pseudomonas syringae (P. syringae). Radish and A. thaliana were found to be resistant to both pathogens when treated with Ra, depending on the concentration used. Interestingly, Ra-mediated resistance to P. syringae is dependent on light because Ra-treated seedlings exhibited enhanced susceptibility to P. syringae infection when grown in the dark. In addition to regulating the biotic defense response, Ra inhibited seed germination and root elongation and enhanced the growth of root hairs in the presence of light in radish and A. thaliana. Our data suggest that Ra regulates the expression of a set of genes involved in defense signaling pathways and plays a role in pathogen defense and plant development. Our results show that light may be generally required not only for the accumulation of Ra but also for its activation during the pathogen defense response.

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Abbreviations

Ra:

Raphanusanin

BL:

Blue light

4-MTBI:

4-Methylthio-3-butenyl isothiocyanate

4-MTBG:

4-Methylthio-3-butenyl glucosinolate

CSN3:

Cop-9 signalosome subunit 3

ERF:

Ethylene responsive factor

RMB2:

Radish myrosinase B 2

RsCSN3:

Raphanus sativus CSN3

DD-RT-PCR:

Differential display RT-PCR

PCD:

Programmed cell death

EST:

Expressed sequence tag

LD:

Long day

RH:

Relative humidity

HR:

Hypersensitive response

TCV:

Turnip Crinkle virus

MAMP:

Microbe-associated molecular patterns

CYP81F2:

Cytochrome P450, family 81, subfamily F, polypeptide 2

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Acknowledgments

This work was supported by grants from Japanese Ministry of Education, Science, and Culture and Sports (Monbukagakusho).

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  1. Moehninsi

    Present address: USDA/ARS, Vegetable and Forage Crops Production Research, Prosser, WA, 99350, USA

Authors and Affiliations

  1. Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki, 305-8572, Japan

    Moehninsi, Kenji Miura, Kosumi Yamada & Hideyuki Shigemori

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425_2014_2103_MOESM1_ESM.pptx

Supplemental Fig. S1 Ra-induced resistance to B. cinerea is independent of light in A. thaliana. a and b Representative images of intact Arabidopsis plants with the B. cinerea infection assay. a Leaves from 5-week-old plants treated with the indicated concentration of Ra on the abaxial side of the leaves at 3 days after inoculation with a 4-µl droplet of B. cinerea spores (105 spores ml−1) under dark (left panel) and under continuous BL (0.45 µmol m-2 s-1) (right panel) conditions. The Botrytis assay was performed by drop inoculation (see Methods). Toothpicks show the inoculation sites with different concentrations of Ra on intact plants. b Symptoms of B. cinerea infection on the leaves of intact plants. The symptoms of infection were identical to those on detached leaves, but the levels of infection were variable between intact leaves. BL = blue light. (PPTX 2479 kb)

425_2014_2103_MOESM2_ESM.pptx

Supplemental Fig. S2 Functional coupling of BL and Ra induces enhanced resistance to P. syringae in A. thaliana. a and b Representative images of intact Arabidopsis plants with the P. syringae infection assay. a Leaves from five-week-old plants treated with the indicated concentration of Ra on the abaxial side of the leaves at 5 days after inoculation with 5 x 107 cfu ml-1 of virulent strain of P. syringae pv. tomato Pst DC3000 under dark (left panel) and under BL (0.45 µmol m-2 s-1) (right panel) conditions. The Pseudomonas infection assay was performed by the pressure infiltration method (see "Materials and methods"), toothpicks show the inoculation sites with different concentrations of Ra on intact plants. b Symptoms of Pseudomonas infection at 5 days post-inoculation in A. thaliana leaves on intact plants. The symptoms of infection were identical to those on detached leaves, but the levels of infection were variable between intact leaves. BL = blue light; UNI = un-infected. (PPTX 5271 kb)

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Moehninsi, Miura, K., Yamada, K. et al. Raphanusanin-mediated resistance to pathogens is light dependent in radish and Arabidopsis thaliana . Planta 240, 513–524 (2014). https://doi.org/10.1007/s00425-014-2103-5

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