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Endogenous salicylic acid accumulation is required for chilling tolerance in cucumber (Cucumis sativus L.) seedlings

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Abstract

Salicylic acid (SA) is an important plant hormone, and its exogenous application can induce tolerance to multiple environmental stresses in plants. In this study, we examine the potential involvement of endogenous SA in response to chilling in cucumber (Cucumis sativus L.) seedlings. A low temperature of 8 °C induces a moderate increase in endogenous SA levels. Chilling stimulates the enzymatic activities and the expression of genes for phenylalanine ammonia-lyase (PAL) and benzoic acid-2-hydroxylase rather than isochorismate synthase. This indicates that the PAL enzymatic pathway contributes to chilling-induced SA production. Cucumber seedlings pretreated with SA biosynthesis inhibitors accumulate less endogenous SA and suffer more from chilling damage. The expression of cold-responsive genes is also repressed by SA inhibitors. The reduction in stress tolerance and in gene expression can be restored by the exogenous application of SA, confirming the critical roles of SA in chilling responses in cucumber seedlings. Furthermore, the inhibition of SA biosynthesis under chilling stress results in a prolonged and enhanced hydrogen peroxide (H2O2) accumulation. The application of exogenous SA and the chemical scavenger of H2O2 reduces the excess H2O2 and alleviates chilling injury. In contrast, the protective effects of SA are negated by foliar spraying with high concentrations of H2O2 and an inhibitor of the antioxidant enzyme. These results suggest that endogenous SA is required in response to chilling stress in cucumber seedlings, by modulating the expression of cold-responsive genes and the precise induction of cellular H2O2 levels.

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Abbreviations

3-AT:

3-Amino-1,2,4-triazole

AOPP:

l-α-Aminooxy-β-phenylpropionic acid

APX:

Ascorbate peroxidase

BA:

Benzoic acid

BA2H:

Benzoic acid 2-hydroxylase

CA:

Carbonic anhydrase

cAPX:

Cytosolic APX

CAT:

Catalase

CBF:

C-repeat binding factor

COR47 :

Cold-regulated 47

DMTU:

Dimethylthiourea

ETR:

Electron transport rate

GA:

Gibberellin

GA2ox:

GA 2-oxidase

GA20ox:

GA 20-oxidase

ICS:

Isochorismate synthase

MDA:

Malondialdehyde

P5CS:

Delta 1-pyrroline-5-carboxylate synthase

P5CR:

Pyrroline-5-carboxylate reductase

PAC:

Paclobutrazol

PAL:

Phenylalanine-ammonia-lyase

PR :

Pathogenesis-related

RAB18 :

Responsive to ABA 18

RbcS:

Ribulose-1,5-bisphosphate carboxylase oxygenase small subunit

SA:

Salicylic acid

SAG:

SA O-β-glucoside

SOD:

Superoxide dismutase

SuSy:

Sucrose synthase

UPLC:

Ultra-performance liquid chromatography

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Acknowledgments

We thank the editors and reviewers for their critical reading and constructive suggestions. This work is supported by the Young Scientists Fund of the National Natural Science Foundation of China (No. 31101548), the Special Fund for Agro-scientific Research in the Public Interest (No. 201303014), the International Science and Technology Cooperation Program of China (2010DFB30550) and the China Agriculture Research System (CARS-25). This work is also supported by the Key Laboratory of Horticultural Crop Biology and Germplasm Innovation, Ministry of Agriculture.

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  1. Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing, 100081, Peoples’ Republic of China

    Chun-Juan Dong, Liang Li, Qing-Mao Shang, Xin-Yan Liu & Zhi-Gang Zhang

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  1. Chun-Juan Dong
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  2. Liang Li
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  3. Qing-Mao Shang
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Correspondence to Chun-Juan Dong or Qing-Mao Shang.

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Dong, CJ., Li, L., Shang, QM. et al. Endogenous salicylic acid accumulation is required for chilling tolerance in cucumber (Cucumis sativus L.) seedlings. Planta 240, 687–700 (2014). https://doi.org/10.1007/s00425-014-2115-1

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