, Volume 247, Issue 1, pp 201–214 | Cite as

Characterization of CcSTOP1; a C2H2-type transcription factor regulates Al tolerance gene in pigeonpea

  • Abhijit Arun Daspute
  • Yuriko Kobayashi
  • Sanjib Kumar Panda
  • Bashasab Fakrudin
  • Yasufumi Kobayashi
  • Mutsutomo Tokizawa
  • Satoshi Iuchi
  • Arbind Kumar Choudhary
  • Yoshiharu Y. Yamamoto
  • Hiroyuki KoyamaEmail author
Original Article


Main conclusion

Al-responsive citrate-transporting CcMATE1 function and its regulation by CcSTOP1 were analyzed using NtSTOP1 -KD tobacco- and pigeonpea hairy roots, respectively, CcSTOP1 binding sequence of CcMATE1 showed similarity with AtALMT1 promoter.

The molecular mechanisms of Aluminum (Al) tolerance in pigeonpea (Cajanus cajan) were characterized to provide information for molecular breeding. Al-inducible citrate excretion was associated with the expression of MULTIDRUGS AND TOXIC COMPOUNDS EXCLUSION (CcMATE1), which encodes a citrate transporter. Ectopic expression of CcMATE1-conferred Al tolerance to hairy roots of transgenic tobacco with the STOP1 regulation system knocked down. This gain-of-function approach clearly showed CcMATE1 was involved in Al detoxification. The expression of CcMATE1 and another Al-tolerance gene, ALUMINUM SENSITIVE 3 (CcALS3), was regulated by SENSITIVE TO PROTON RHIZOTOXICITY1 (CcSTOP1) according to loss-of-function analysis of pigeonpea hairy roots in which CcSTOP1 was suppressed. An in vitro binding assay showed that the Al-responsive CcMATE1 promoter contained the GGNVS consensus bound by CcSTOP1. Mutation of GGNVS inactivated the Al-inducible expression of CcMATE1 in pigeonpea hairy roots. This indicated that CcSTOP1 binding to the promoter is critical for CcMATE1 expression. The STOP1 binding sites of both the CcMATE1 and AtALMT1 promoters contained GGNVS and a flanking 3′ sequence. The GGNVS region was identical in both CcMATE1 and AtALMT1. By contrast, the 3′ flanking sequence with binding affinity to STOP1 did not show similarity. Putative STOP1 binding sites with similar structures were also found in Al-inducible MATE and ALMT1 promoters in other plant species. The characterized Al-responsive CcSTOP1 and CcMATE1 genes will help in pigeonpea breeding in acid soil tolerance.


Aluminum Cis-elements Citrate Hairy roots MATE 



Aluminum-activated malate transporter 1


Aluminum-sensitive 3


Multidrug and toxic compound exclusion


Organic acid


Sensitive to proton rhizotoxicity 1



This work was funded by a scientific research Grant from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (15H04468).

Supplementary material

425_2017_2777_MOESM1_ESM.docx (5.2 mb)
Supplementary material 1 (DOCX 5371 kb)


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Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Abhijit Arun Daspute
    • 1
  • Yuriko Kobayashi
    • 1
  • Sanjib Kumar Panda
    • 2
  • Bashasab Fakrudin
    • 3
  • Yasufumi Kobayashi
    • 4
  • Mutsutomo Tokizawa
    • 1
  • Satoshi Iuchi
    • 5
  • Arbind Kumar Choudhary
    • 6
  • Yoshiharu Y. Yamamoto
    • 1
  • Hiroyuki Koyama
    • 1
    Email author
  1. 1.Laboratory of Plant Cell Technology, Faculty of Applied Biological SciencesGifu UniversityGifuJapan
  2. 2.Plant Molecular Biotechnology Laboratory, Department of Life Science and BioinformaticsAssam UniversitySilcharIndia
  3. 3.Department of Biotechnology and Crop Improvement, Post Graduate CentreUniversity of Horticultural SciencesBengaluruIndia
  4. 4.Department of Biosciences and BioengineeringIndian Institute of TechnologyGuwahatiIndia
  5. 5.RIKEN BioResource CenterIbarakiJapan
  6. 6.ICAR Research Complex for Eastern RegionPatnaIndia

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