, Volume 244, Issue 4, pp 831–841 | Cite as

Stomatal closure induced by phytosphingosine-1-phosphate and sphingosine-1-phosphate depends on nitric oxide and pH of guard cells in Pisum sativum

  • Mallikarjuna Rao Puli
  • Pidakala Rajsheel
  • Vetcha Aswani
  • Srinivas Agurla
  • Kazuyuki Kuchitsu
  • Agepati S. RaghavendraEmail author
Original Article


Main conclusion

Phyto-S1P and S1P induced stomatal closure in epidermis of pea ( Pisum sativum ) by raising the levels of NO and pH in guard cells.

Phosphosphingolipids, such as phytosphingosine-1-phosphate (phyto-S1P) and sphingosine-1-phosphate (S1P), are important signaling components during drought stress. The biosynthesis of phyto-S1P or S1P is mediated by sphingosine kinases (SPHKs). Although phyto-S1P and S1P are known to be signaling components in higher plants, their ability to induce stomatal closure has been ambiguous. We evaluated in detail the effects of phyto-S1P, S1P and SPHK inhibitors on signaling events leading to stomatal closure in the epidermis of Pisum sativum. Phyto-S1P or S1P induced stomatal closure, along with a marked rise in nitric oxide (NO) and cytoplasmic pH of guard cells, as in case of ABA. Two SPHK inhibitors, DL-threo dihydrosphingosine and N’,N’-dimethylsphingosine, restricted ABA-induced stomatal closure and prevented the increase of NO or pH by ABA. Modulators of NO or pH impaired both stomatal closure and increase in NO or pH by phyto-S1P/S1P. The stomatal closure by phyto-S1P/S1P was mediated by phospholipase D and phosphatidic acid (PA). When present, PA elevated the levels of pH, but not NO of guard cells. Our results demonstrate that stomatal closure induced by phyto-S1P and S1P depends on rise in pH as well as NO of guard cells. A scheme of signaling events initiated by phyto-S1P/S1P, and converging to cause stomatal closure, is proposed.


Abscisic acid LCB signaling Pea Phosphosphingolipids Sphingosine kinase Stomata 



Abscisic acid

DL-threo DHS

DL-threo dihydrosphingosine




Long chain base


Long chain base phosphate


Nitric oxide


Phosphatidic acid


Phospholipase D






Slow anion channel


Sphingosine kinase



This work was supported by grants (to ASR) from the Department of Biotechnology (No. BT/PR9227/PBD/16/748/2007), Council of Scientific and Industrial Research (CSIR, No. 38(1195)/08/EMR-II), JC Bose Fellow of Department of Science and Technology (No. SR/S2/JCB-06/2006), and Department of Science and Technology-Japanese Society for Promotion of Science (No. DST/INT/JSPS/P-121/10) project (to ASR & KK). MRP, PR, VA, and SA were all supported by Research Fellowships from CSIR/UGC, New Delhi, India. The facilities in our Department and School were supported by grants from DST-FIST, UGC-SAP-CAS and DBT-CREBB, all from New Delhi, India. We thank Ms Nalini, Technical Assistant, Central Instrumentation Laboratory, for her help in using the confocal microscope.

Compliance with ethical standards

Conflict of interest

The authors have no conflicts of interest to declare.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Mallikarjuna Rao Puli
    • 1
  • Pidakala Rajsheel
    • 1
  • Vetcha Aswani
    • 1
  • Srinivas Agurla
    • 1
  • Kazuyuki Kuchitsu
    • 2
  • Agepati S. Raghavendra
    • 1
    Email author
  1. 1.Department of Plant Sciences, School of Life SciencesUniversity of HyderabadHyderabadIndia
  2. 2.Department of Applied Biological SciencesTokyo University of ScienceNodaJapan

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