Hydrogen peroxide is involved in methane-induced tomato lateral root formation

  • Yingying Zhao
  • Yihua Zhang
  • Feijie Liu
  • Ren Wang
  • Liqin Huang
  • Wenbiao Shen
Original Article


Key message

Pharmacological and molecular evidence reveals a novel role of methane (CH4) gas in root organogenesis, the induction of lateral root (LR) formation, and this response might require hydrogen peroxide (H2O2) synthesis.


Although plants can produce CH4 and release this to atmosphere, the beneficial role(s) of CH4 are not fully elucidated. In this study, the fumigation with CH4 not only increased NADPH oxidase activity and H2O2 production, but also induced tomato lateral root primordial formation and thereafter LR development. However, exogenously applied argon and nitrogen failed to influence LR formation. Above responses triggered by CH4 were sensitive to the removal of endogenous H2O2 with dimethylthiourea (DMTU; a membrane-permeable scavenger of H2O2), suggesting the hypothesis that CH4’s effect on LR formation could be mediated by endogenous H2O2. Diphenylene iodonium (DPI) inhibition of the H2O2 generating enzyme NADPH oxidase attenuated H2O2 synthesis and impaired LR formation in response to CH4, confirming the requirement of NADPH oxidase-dependent H2O2. Meanwhile, the alterations of endogenous H2O2 concentrations failed to influence CH4 production in tomato seedlings. Molecular evidence revealed that CH4-induced SlCDKA1, SlCYCA2;1, and SlCYCA3;1 transcripts, and -decreased SlKRP2 mRNA were impaired by DMTU or DPI. Contrasting changes in LR formation-related miR390a and miR160 transcripts and their target genes, including SlARF4 and SlARF16, were observed. Together, our pharmacological and molecular evidence suggested the requirement of H2O2 synthesis in CH4-triggered tomato LR formation, partially via the regulation of cell cycle regulatory genes, miRNA-, and tasiRNA-modulated gene expression.


Hydrogen peroxide Lateral root formation Methane Tomato 





Auxin response factors






Diphenylene iodonium


2′,7′-Dichlorofluorescein diacetate


Hydrogen peroxide


Lateral root


Lateral root primordia


Laser scanning confocal microscopy






Primary root


Quantitative real-time RT-PCR


Reactive oxygen species


Transacting short-interfering RNAs



This work was supported by the Natural Science Foundation of Jiangsu Province (Grant no.: BK20181317) and the National Natural Science Foundation of China (31772292).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.

Supplementary material

299_2019_2372_MOESM1_ESM.doc (40 kb)
Supplementary material 1 (DOC 28 KB)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Yingying Zhao
    • 1
  • Yihua Zhang
    • 1
  • Feijie Liu
    • 1
  • Ren Wang
    • 2
  • Liqin Huang
    • 3
  • Wenbiao Shen
    • 1
  1. 1.College of Life Sciences, Laboratory Center of Life SciencesNanjing Agricultural UniversityNanjingChina
  2. 2.Institute of BotanyJiangsu Province and Chinese Academy of SciencesNanjingChina
  3. 3.College of SciencesNanjing Agricultural UniversityNanjingChina

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