Insights into stress responses in mandarins triggered by Bacillus subtilis cyclic lipopeptides and exogenous plant hormones upon Penicillium digitatum infection

  • Paiboon Tunsagool
  • Warangkana JutidamrongphanEmail author
  • Narumon Phaonakrop
  • Janthima Jaresitthikunchai
  • Sittiruk Roytrakul
  • Wichitra LeelasuphakulEmail author
Original Article


Key message

Bacillus subtilis CLP extract activates defense gene expression and increases the unique protein production involving in pathways of ISR, SAR, ubiquitin-proteasome system, and glycolysis for stress responses in flavedo tissues.


Cyclic lipopeptides (CLPs) of Bacillus subtilis ABS-S14 had ability to activate plant defensive pathways, increase resistance and control green mold rot caused by Penicillium digitatum in mandarin fruit. The current study investigated transcriptional and proteomic data to highlight the unique induction effect of CLPs produced by B. subtilis ABS-S14 on the defense mechanism of mandarins in response to P. digitatum attack, and their differences from those following the exogenous plant hormone application. The proteomic patterns of the flavedo tissues as affected by Bacillus CLP extract, salicylic acid (SA), methyl jasmonate (MeJA), and ethephon (Et) were explored. qPCR analysis revealed the great effects of CLP extract in enhancing the transcription of PAL, ACS1, GLU, POD, and PR1. Tryptic peptides by LC–MS analysis between treatments with and without fungal infection were compared. B. subtilis CLP extract empowered the plant’s immune response to wound stress by the significant production of calmodulin-binding receptor-like cytoplasmic kinase 2, molybdenum cofactor sulfurase, and NAD+-dependent glyceraldehyde-3-phosphate dehydrogenase. Ubiquitin carrier protein abundance was developed only in the treated flavedo with CLP extract coupled with P. digitatum infection. The gene expression and overall proteome findings involving pathways of ubiquitin proteasome system, ISR, SAR, and energy production provide a new insight into the molecular mechanisms of the antagonist B. subtilis ABS-S14 inducing resistance against green mold in mandarins.


Proteomics Defense-related gene Mandarin Bacillus subtilis Cyclic lipopeptides Penicillium digitatum 



Abscisic acid


1-Aminocyclopropane-1-carboxylic acid oxidase


1-Aminocyclopropane-1-carboxylic acid synthase




Cyclic lipopeptides


Calmodulin-binding receptor-like cytoplasmic kinase 2








Induced systemic resistance


Jasmonic acid




Methyl jasmonate

MoCo sulfurase

Molybdenum cofactor sulfurase


Phenylalanine ammonia lyase




Pathogenesis-related protein 1


Reactive oxygen species


Salicylic acid


Systemic acquired resistance


Ubiquitin carrier protein



This work was financially supported by the National Royal Research Council of Thailand (Grant no.SCI600214S), P. Tunsagool gratefully acknowledges the grant of a TRF Royal Golden Jubilee PhD. student scholarship, and the University Academic Excellence Strengthening Program in Biochemistry of Prince of Songkla University (PSU), the PSU Graduate Fund. The authors thank Mr. Michael G. Currie for his assistance with English.

Compliance with ethical standards

Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Supplementary material

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Copyright information

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

Authors and Affiliations

  • Paiboon Tunsagool
    • 1
  • Warangkana Jutidamrongphan
    • 2
    Email author
  • Narumon Phaonakrop
    • 3
  • Janthima Jaresitthikunchai
    • 3
  • Sittiruk Roytrakul
    • 3
  • Wichitra Leelasuphakul
    • 1
    Email author
  1. 1.Department of BiochemistryPrince of Songkla UniversitySongkhlaThailand
  2. 2.Faculty of Environmental ManagementPrince of Songkla UniversitySongkhlaThailand
  3. 3.Proteomics Research Laboratory, National Center for Genetic Engineering and BiotechnologyNational Science and Technology Development Agency, Thailand Science Park (TSP)Pathum ThaniThailand

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