Antioxidant and anti-stress compounds improve regrowth of cryopreserved Rubus shoot tips

  • Esther E. Uchendu
  • Magfrat Muminova
  • Sandhya Gupta
  • Barbara M. Reed


Regrowth of plants after cryopreservation varies, and resulting regrowth ranges from poor to excellent. Oxidative stress is a potential cause of damage in plant tissues. Antioxidants and anti-stress compounds may improve regrowth by preventing or repairing the damage. Lipoic acid (LA), glutathione (GSH), glycine betaine (GB), and polyvinylpyrrolidone (PVP) were tested during cryopreservation of shoot tips using the plant vitrification solution 2 (PVS2) protocol. Two in vitro-grown blackberry cultivars were cold acclimated and then cryopreserved in liquid nitrogen (LN). The antioxidant and anti-stress compounds were added at four critical steps of the protocol: pretreatment, loading, rinsing, and regrowth. Three out of the four compounds significantly improved regrowth of cryopreserved shoot tips. Regrowth ranged from 40% to 50% for controls to >80% for treated shoot tips. LA (4-8 mM) produced high regrowth at pretreatment, loading, and rinsing for ‘Chehalem’ and at all steps for ‘Hull Thornless’. Recovery improved at all steps with GSH (0.16 mM) and GB (10 mM). PVP had a neutral or negative impact on regrowth. Overall addition of LA, GSH, and GB improved regrowth by ∼25% over the shoot tips cryopreserved using the regular PVS2 protocol (control). This study shows that adding non-vitamin antioxidants and anti-stress compounds during the PVS2-vitrification protocol improves regrowth of shoot cultures following cryopreservation. We recommend inclusion of antioxidants as part of standard cryopreservation protocols.


Blackberry Lipoic acid Germplasm Glutathione Glycine betaine Long-term storage Oxidative stress Polyvinylpyrrolidone 



This study was funded by CRIS project 5358-21000-03300D USDA-ARS. Thanks to Drs. Tony Chen and Ralph Reed for very useful advice and to the Ford Foundation International Fellowship Program and the University Club of Portland Inc. for awards to Esther Uchendu. Dr. Magfrat Muminova acknowledges the International Union of Biochemistry and Molecular Biology (IUBMB) for awarding a Wood-Whelan Fellowship.


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Copyright information

© The Society for In Vitro Biology 2010

Authors and Affiliations

  • Esther E. Uchendu
    • 1
    • 2
  • Magfrat Muminova
    • 3
  • Sandhya Gupta
    • 4
  • Barbara M. Reed
    • 5
  1. 1.Department of HorticultureOregon State UniversityCorvallisUSA
  2. 2.Department of Plant Agriculture, Ontario Agriculture CollegeUniversity of GuelphGuelphCanada
  3. 3.Department of BiotechnologyTashkent Chemical-Technology InstituteTashkentUzbekistan
  4. 4.Tissue Culture and Cryopreservation UnitNational Bureau of Plant Genetic ResourcesNew DelhiIndia
  5. 5.United States Department of Agriculture-Agricultural Research ServiceNational Clonal Germplasm RepositoryCorvallisUSA

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