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The Gossypium hirsutum WRKY gene GhWRKY39-1 promotes pathogen infection defense responses and mediates salt stress tolerance in transgenic Nicotiana benthamiana

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Abstract

Key message

Our results indicate that overexpression of the GhWRKY39 - 1 gene enhances resistance to pathogen infection and tolerance to high salt and oxidative stress in transgenic Nicotiana benthamiana.

Abstract

WRKY transcription factor genes play significant roles in the response to biotic and abiotic stresses. Cotton (Gossypium hirsutum) is an important fiber and oil crop worldwide. We isolated and characterized GhWRKY39-1, which is a group IId WRKY gene that is present as a single copy in the cotton genome. Quantitative PCR analyses indicated that GhWRKY39-1 was induced by pathogen infection, defense-related signaling molecules, and abiotic stresses, such as NaCl and methyl viologen. An analysis of the subcellular localization of the GhWRKY39-1 protein indicated that it localized to the nucleus. Furthermore, constitutive overexpression of GhWRKY39-1 in Nicotiana benthamiana conferred a greater resistance to infection by both the bacterial pathogen Ralstonia solanacearum and the fungal pathogen Rhizoctonia solani. The transgenic plants also exhibited elevated mRNA levels of several pathogen-related (PR) genes, including PR1c, PR2 and PR4. Moreover, transgenic plants displayed an enhanced tolerance to salt and oxidative stress and elevated expression of several oxidation-related genes, including APX, CAT, GST and SOD. Overall, these results indicate that GhWRKY39-1 functions as a positive regulator of plant defense against pathogen infection and responses to salt stress and reactive oxygen species.

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Abbreviations

APX:

Ascorbate peroxidase

At:

Arabidopsis thaliana

CAT:

Catalase

Ca:

Capsicum annuum

CTAB:

Cetyltrimethyl ammonium bromide

DAB:

3,3′-Diaminobenzidine

GFP:

Green fluorescence protein

Gh:

Gossypium hirsutum

Gm:

Glycine max

GST:

Glutathione-S-transferase

MeJA:

Methyl jasmonate

MS medium:

Murashige and Skoog medium

MV:

Methyl viologen

Nb:

Nicotiana benthamiana

NBT:

Nitro blue tetrazolium

NPR1:

Non-expression of PR1

OE:

Over expression

ORF:

Open reading frame

Os :

Oryza sativa

PR:

Pathogenesis-related

RACE:

Rapid amplification of cDNA ends

ROS:

Reactive oxygen species

Ralstonia solanacearum :

R. solanacearum

Rhizoctonia solani :

R. solani

RT-PCR:

Reverse transcription-PCR

Q-PCR:

Quantitative PCR

SA:

Salicylic acid

SOD:

Superoxide dismutase

UTR:

Untranslated region

WT:

Wild-type

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Acknowledgments

This work was financially supported by the Genetically Modified Organisms Breeding Major Projects of China [grant numbers 2009ZX08009-092B] and the National Natural Science Foundation of China [grant number 31171837].

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Authors and Affiliations

  1. Shandong Agricultural University, College of Life Sciences, Shandong, China

    Weina Shi, Lili Hao, Jing Li, Dongdong Liu, Xingqi Guo & Han Li

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  1. Weina Shi
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  2. Lili Hao
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  3. Jing Li
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  4. Dongdong Liu
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  6. Han Li
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Correspondence to Han Li.

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Communicated by Z.-Y. Wang.

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Shi, W., Hao, L., Li, J. et al. The Gossypium hirsutum WRKY gene GhWRKY39-1 promotes pathogen infection defense responses and mediates salt stress tolerance in transgenic Nicotiana benthamiana . Plant Cell Rep 33, 483–498 (2014). https://doi.org/10.1007/s00299-013-1548-5

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