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Arabidopsis thaliana defense response to the ochratoxin A-producing strain (Aspergillus ochraceus 3.4412)

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Abstract

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OTA-producing strain Aspergillus ochraceus induc ed necrotic lesions, ROS accumulation and defense responses in Arabidopsis . Primary metabolic and defense-related proteins changed in proteomics. Ascorbate–glutathione cycle and voltage-dependent anion-selective channel proteins fluctuated.

Abstract

Mycotoxigenic fungi, as widespread contaminants by synthesizing mycotoxins in pre-/post-harvest infected plants and even stored commercial cereals, could usually induce plant–fungi defense responses. Notably, ochratoxin A (OTA) is a nephrotoxic, hepatotoxic, teratogenic, immunotoxic and phytotoxic mycotoxin. Herein, defense responses of model system Arabidopsis thaliana detached leaves to infection of Aspergillus ochraceus 3.4412, an OTA high-producing strain, were studied from physiological, proteomic and transcriptional perspectives. During the first 72 h after inoculation (hai), the newly formed hypersensitive responses-like lesions, decreased chlorophyll content, accumulated reactive oxygen species and upregulated defense genes expressions indicated the defense response was induced in the leaves with the possible earlier motivated jasmonic acid/ethylene signaling pathways and the later salicylic acid-related pathway. Moreover, proteomics using two-dimensional gel electrophoresis 72 hai showed 16 spots with significantly changed abundance and 13 spots corresponding to 12 unique proteins were successfully identified by MALDI-TOF/TOF MS/MS. Of these, six proteins were involved in basic metabolism and four in defense-related processes, which included glutathione-S-transferase F7, voltage-dependent anion-selective channel protein 3 (VDAC-3), osmotin-like protein OSM34 and blue copper-binding protein. Verified from proteomic and/or transcriptional perspectives, it is concluded that the primary metabolic pathways were suppressed with the ascorbate–glutathione cycle fluctuated in response to A. ochraceus and the modulation of VDACs suggested the possibility of structural damage and dysfunction of mitochondria in the process. Taken together, these findings exhibited a dynamic overview of the defense responses of A. thaliana to A. ochraceus and provided a better insight into the pathogen-resistance mechanisms in plants.

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Abbreviations

APX:

Ascorbate peroxidases

ASC:

Ascorbate

BCB protein:

Blue copper-binding protein

CSP41-a:

Chloroplast stem-loop binding protein of 41 kDa a

DAB:

3,3-Diaminobenzidine

DHAR:

Dehydroascorbate reductase

EIN:

Ethylene insensitive

ERF:

Ethylene-response factor

ET:

Ethylene

GADPH:

Glyceraldehydes-3-phosphate dehydrogenase

GDH:

Glutamate dehydrogenase

GR:

Glutathione reductase

GSH:

Reduced glutathione

GST:

Glutathione-S-transferase

Hai:

Hours after inoculation

HR:

Hypersensitive responses

H2DCFDA:

Dichlorodihydrofluorescein diacetate

H2O2 :

Hydrogen peroxide

IDH-I:

Isocitrate dehydrogenase [NAD] regulatory subunit 1

IEF:

Isoelectric focusing

JA:

Jasmonic acid

MDHAR:

Monodehydroascorbate reductase

NBT:

Nitroblue tetrazolium

OTA:

Ochratoxin A

OSM:

Osmotin-like protein

O2 :

Superoxide anion radical

PDF:

Plant defensin

ROS:

Reactive oxygen species

PR:

Pathogenesis-related

RT-PCR:

Real-time polymerase chain reaction

SA:

Salicylic acid

SGAT:

Serine-glyoxylate aminotransferase

TCA cycle:

Tricarboxylic acid cycle

VDAC-3:

Voltage-dependent anion-selective channel protein 3

2-DE:

Two-dimensional electrophoresis

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Acknowledgments

The authors thank all of the members of Wentao Xu’s laboratory who contributed to the project, the mass spectrometry support from the Beijing Proteome Research Center and the support projects from National Key Technology R&D Program (2012BAK08B04-01).

Conflict of interest

There are no conflicts of interest to declare.

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Authors and Affiliations

  1. Laboratory of Food Safety and Molecular Biology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, People’s Republic of China

    Junran Hao, Weihong Wu, Yan Wang, Zhuojun Yang, Yangjun Lv, Yanan Zhai, Jing Yang, Zhihong Liang, Kunlun Huang & Wentao Xu

  2. The Supervision Inspection and Testing Center of Genetically Modified Organisms, Ministry of Agriculture, Beijing, 100083, People’s Republic of China

    Kunlun Huang & Wentao Xu

  3. Beijing Key Laboratory of Nutrition, Health and Food Safety, Beijing, 100083, People’s Republic of China

    Junran Hao, Yang Liu, Yangjun Lv & Yanan Zhai

  4. Nutrition and Health Research Institute, COFCO Corporation, Beijing, 100083, People’s Republic of China

    Yang Liu

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  1. Junran Hao
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  2. Weihong Wu
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  3. Yan Wang
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  9. Zhihong Liang
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  10. Kunlun Huang
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  11. Wentao Xu
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Correspondence to Zhihong Liang or Wentao Xu.

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Communicated by Jim Register.

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Hao, J., Wu, W., Wang, Y. et al. Arabidopsis thaliana defense response to the ochratoxin A-producing strain (Aspergillus ochraceus 3.4412). Plant Cell Rep 34, 705–719 (2015). https://doi.org/10.1007/s00299-014-1731-3

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  • DOI: https://doi.org/10.1007/s00299-014-1731-3

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