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Nitric oxide production and its functional link with OIPK in tobacco defense response elicited by chitooligosaccharide

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Abstract

Chitooligosaccharide (COS) or oligochitosan has been shown to induce tobacco defense responses which are connected with nitric oxide (NO) and OIPK (oligochitosan-induced Ser/Thr protein kinase). The aim of this study was to reveal the relationship between NO production and OIPK pathway in the defense response of tobacco elicited by COS. NO generation was investigated by epidermal strip bioassay and fluorophore microscope using fluorophore diaminofluorescein diacetate (DAF-2DA). Tobacco epidermal cells treated with COS resulted in production of NO, which was first present in chloroplast, then in nucleus, finally in the whole cell; this NO production was sensitive to NO scavenger cPTIO and the mammalian NO synthase (NOS) inhibitor l-NAME, suggesting that NOS-like enzyme maybe involved in NO generation in tobacco epidermal cells. However, NOS and nitrate reductase (NR, EC 1.6.6.1) inhibitors reduced NO content in tobacco leaves by using NO Assay Kit, suggesting both NOS and NR were involved in NO production in tobacco leaves. Using a pharmacological approach and western blotting, we provide evidence that NO acts upstream of OIPK expression. NO scavenger, NOS inhibitor partly blocked the activation of OIPK and the activities of several defense-related enzymes induced by COS; treatment with NO donor sodium nitroprusside (SNP) induced the activation of OIPK and enhanced the defense systems. The results suggest that COS is able to induce NO generation, which results in up-regulation the activities of some defense-related enzymes through an OIPK-dependent or independent pathway.

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Acknowledgments

This work was financially supported by Hi-Tech Research and Development Program of China (Nos. 2006AA10A213 and 2007AA091601), the Knowledge Innovation Program of the Chinese Academy of Sciences (No. KSCX2-YW-G-041 and No. KSCX2-YW-N-007), Natural Science Fund of Liaoning province (No. 20082153), National Department Public Benefit Research Foundation of China (No. 200903052), Special Funds for Scientific Research on Public Causes in Agriculture (200903052), and Scientific and Technological Major Special Project “Significant Creation of New Drugs”(2009ZX9501-011).

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Authors and Affiliations

  1. Liaoning Provincial Key Laboratory of Carbohydrates, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Zhongshan Road 457, Dalian, 116023, People’s Republic of China

    Hongyan Zhang, Xiaoming Zhao, Jinli Yang, Heng Yin, Wenxia Wang, Hang Lu & Yuguang Du

  2. Graduate University of the Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Hongyan Zhang

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  1. Hongyan Zhang
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  2. Xiaoming Zhao
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  3. Jinli Yang
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  7. Yuguang Du
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Correspondence to Xiaoming Zhao or Yuguang Du.

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Communicated by J. Zou.

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Zhang, H., Zhao, X., Yang, J. et al. Nitric oxide production and its functional link with OIPK in tobacco defense response elicited by chitooligosaccharide. Plant Cell Rep 30, 1153–1162 (2011). https://doi.org/10.1007/s00299-011-1024-z

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