Antioxidant and anti-stress compounds improve regrowth of cryopreserved <Emphasis Type="Italic">Rubus</Emphasis> shoot tips |
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Authors: | Esther E Uchendu Magfrat Muminova Sandhya Gupta Barbara M Reed |
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Institution: | (1) Department of Horticulture, Oregon State University, 4017 Ag Life Sciences Bldg., Corvallis, OR 97331-7304, USA;(2) Present address: Department of Plant Agriculture, Ontario Agriculture College, University of Guelph, E.C. Bovey Bldg. RM 4227, Guelph, ON, N1G 2W1, Canada;(3) Department of Biotechnology, Tashkent Chemical-Technology Institute, 23 Navoyi str., Tashkent, 100011, Uzbekistan;(4) Tissue Culture and Cryopreservation Unit, National Bureau of Plant Genetic Resources, Pusa Campus, New Delhi, 110 012, India;(5) United States Department of Agriculture-Agricultural Research Service, National Clonal Germplasm Repository, 33447 Peoria Rd, Corvallis, OR 97333-2521, USA; |
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Abstract: | 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. |
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