Effects of osmotic pretreatments on oxidative stress, antioxidant profiles and cryopreservation of olive somatic embryos

Paul T. Lynch, Ayesha Siddika, Jason W. Johnston, Susan M. Trigwell, Aradhana Mehra, Carla Benelli, Maurizio Lambardi, Erica E. Benson

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Abstract

A three-day pretreatment of olive somatic embryos (SE) with 0.75 M sucrose, combined with cryoprotection (0.5 M DMSO, 1 M sucrose, 0.5 M glycerol and 0.009 M proline) and controlled rate cooling, supported regrowth (as 34.6% fresh weight gain) and resumption of embryo development after cryopreservation. Pretreatment with mannitol or sorbitol did not support regrowth. Profiles of sugars, proline, antioxidant enzymes, Reactive oxygen species (ROS), secondary oxidation products and ethylene were constructed for the most successful (0.75 M) pretreatment series. Sucrose was the optimal pretreatment for supporting recovery, it also elevated glutathione reductase (GR) activity compared to controls, whereas superoxide dismutase (SOD), catalase and guaiacol peroxidase activities remained relatively unchanged. Superoxide dismutase activity was higher in SE pretreated with sucrose, compared with those pretreated with polyols; H2O2 was enhanced in SE pretreated with sorbitol and sucrose compared to mannitol. The overall trend for ethylene and OH production revealed their levels were highest in SE pretreated with polyols albeit, for individual treatments this was not always the case. Generally, pretreatments did not significantly change embryo secondary oxidation profiles of ThioBarbituric Acid Reactive Substances (TBARS) and Schiff's bases. In combination these studies suggest oxidative processes may influence regrowth of cryopreserved olive SE and that optimal pretreatments could, in part, increase tolerance by an overall enhancement of endogenous antioxidants (particularly GR), proline and sugars. Highlights ► Biomarkers may be used to make evidence based decisions to optimize olive cryostorage. ► Osmotica can affect antioxidant, ROS, sugar and proline profiles in olive somatic embryos. ► Data suggests a putative involvement of ROS in cryopreservation-induced stress in olive cultures.
Original languageEnglish
Pages (from-to)47-56
Number of pages10
JournalPlant Science
Volume181
Issue number1
DOIs
StatePublished - Jul 2011
Externally publishedYes

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somatic embryos
Cryopreservation
Oxidative Stress
Embryonic Structures
Antioxidants
pretreatment
sucrose
Sucrose
proline
antioxidants
Proline
regrowth
cryopreservation
reactive oxygen species
sugars
Reactive Oxygen Species
Carbohydrates
polyols
sorbitol
glutathione-disulfide reductase

Cite this

Lynch, P. T., Siddika, A., Johnston, J. W., Trigwell, S. M., Mehra, A., Benelli, C., ... Benson, E. E. (2011). Effects of osmotic pretreatments on oxidative stress, antioxidant profiles and cryopreservation of olive somatic embryos. Plant Science, 181(1), 47-56. DOI: 10.1016/j.plantsci.2011.03.009

Lynch, Paul T.; Siddika, Ayesha; Johnston, Jason W.; Trigwell, Susan M.; Mehra, Aradhana; Benelli, Carla; Lambardi, Maurizio; Benson, Erica E. / Effects of osmotic pretreatments on oxidative stress, antioxidant profiles and cryopreservation of olive somatic embryos.

In: Plant Science, Vol. 181, No. 1, 07.2011, p. 47-56.

Research output: Contribution to journalArticle

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abstract = "A three-day pretreatment of olive somatic embryos (SE) with 0.75 M sucrose, combined with cryoprotection (0.5 M DMSO, 1 M sucrose, 0.5 M glycerol and 0.009 M proline) and controlled rate cooling, supported regrowth (as 34.6% fresh weight gain) and resumption of embryo development after cryopreservation. Pretreatment with mannitol or sorbitol did not support regrowth. Profiles of sugars, proline, antioxidant enzymes, Reactive oxygen species (ROS), secondary oxidation products and ethylene were constructed for the most successful (0.75 M) pretreatment series. Sucrose was the optimal pretreatment for supporting recovery, it also elevated glutathione reductase (GR) activity compared to controls, whereas superoxide dismutase (SOD), catalase and guaiacol peroxidase activities remained relatively unchanged. Superoxide dismutase activity was higher in SE pretreated with sucrose, compared with those pretreated with polyols; H2O2 was enhanced in SE pretreated with sorbitol and sucrose compared to mannitol. The overall trend for ethylene and OH production revealed their levels were highest in SE pretreated with polyols albeit, for individual treatments this was not always the case. Generally, pretreatments did not significantly change embryo secondary oxidation profiles of ThioBarbituric Acid Reactive Substances (TBARS) and Schiff's bases. In combination these studies suggest oxidative processes may influence regrowth of cryopreserved olive SE and that optimal pretreatments could, in part, increase tolerance by an overall enhancement of endogenous antioxidants (particularly GR), proline and sugars. Highlights ► Biomarkers may be used to make evidence based decisions to optimize olive cryostorage. ► Osmotica can affect antioxidant, ROS, sugar and proline profiles in olive somatic embryos. ► Data suggests a putative involvement of ROS in cryopreservation-induced stress in olive cultures.",
author = "Lynch, {Paul T.} and Ayesha Siddika and Johnston, {Jason W.} and Trigwell, {Susan M.} and Aradhana Mehra and Carla Benelli and Maurizio Lambardi and Benson, {Erica E.}",
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Lynch, PT, Siddika, A, Johnston, JW, Trigwell, SM, Mehra, A, Benelli, C, Lambardi, M & Benson, EE 2011, 'Effects of osmotic pretreatments on oxidative stress, antioxidant profiles and cryopreservation of olive somatic embryos' Plant Science, vol 181, no. 1, pp. 47-56. DOI: 10.1016/j.plantsci.2011.03.009

Effects of osmotic pretreatments on oxidative stress, antioxidant profiles and cryopreservation of olive somatic embryos. / Lynch, Paul T.; Siddika, Ayesha; Johnston, Jason W.; Trigwell, Susan M.; Mehra, Aradhana; Benelli, Carla; Lambardi, Maurizio; Benson, Erica E.

In: Plant Science, Vol. 181, No. 1, 07.2011, p. 47-56.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effects of osmotic pretreatments on oxidative stress, antioxidant profiles and cryopreservation of olive somatic embryos

AU - Lynch,Paul T.

AU - Siddika,Ayesha

AU - Johnston,Jason W.

AU - Trigwell,Susan M.

AU - Mehra,Aradhana

AU - Benelli,Carla

AU - Lambardi,Maurizio

AU - Benson,Erica E.

PY - 2011/7

Y1 - 2011/7

N2 - A three-day pretreatment of olive somatic embryos (SE) with 0.75 M sucrose, combined with cryoprotection (0.5 M DMSO, 1 M sucrose, 0.5 M glycerol and 0.009 M proline) and controlled rate cooling, supported regrowth (as 34.6% fresh weight gain) and resumption of embryo development after cryopreservation. Pretreatment with mannitol or sorbitol did not support regrowth. Profiles of sugars, proline, antioxidant enzymes, Reactive oxygen species (ROS), secondary oxidation products and ethylene were constructed for the most successful (0.75 M) pretreatment series. Sucrose was the optimal pretreatment for supporting recovery, it also elevated glutathione reductase (GR) activity compared to controls, whereas superoxide dismutase (SOD), catalase and guaiacol peroxidase activities remained relatively unchanged. Superoxide dismutase activity was higher in SE pretreated with sucrose, compared with those pretreated with polyols; H2O2 was enhanced in SE pretreated with sorbitol and sucrose compared to mannitol. The overall trend for ethylene and OH production revealed their levels were highest in SE pretreated with polyols albeit, for individual treatments this was not always the case. Generally, pretreatments did not significantly change embryo secondary oxidation profiles of ThioBarbituric Acid Reactive Substances (TBARS) and Schiff's bases. In combination these studies suggest oxidative processes may influence regrowth of cryopreserved olive SE and that optimal pretreatments could, in part, increase tolerance by an overall enhancement of endogenous antioxidants (particularly GR), proline and sugars. Highlights ► Biomarkers may be used to make evidence based decisions to optimize olive cryostorage. ► Osmotica can affect antioxidant, ROS, sugar and proline profiles in olive somatic embryos. ► Data suggests a putative involvement of ROS in cryopreservation-induced stress in olive cultures.

AB - A three-day pretreatment of olive somatic embryos (SE) with 0.75 M sucrose, combined with cryoprotection (0.5 M DMSO, 1 M sucrose, 0.5 M glycerol and 0.009 M proline) and controlled rate cooling, supported regrowth (as 34.6% fresh weight gain) and resumption of embryo development after cryopreservation. Pretreatment with mannitol or sorbitol did not support regrowth. Profiles of sugars, proline, antioxidant enzymes, Reactive oxygen species (ROS), secondary oxidation products and ethylene were constructed for the most successful (0.75 M) pretreatment series. Sucrose was the optimal pretreatment for supporting recovery, it also elevated glutathione reductase (GR) activity compared to controls, whereas superoxide dismutase (SOD), catalase and guaiacol peroxidase activities remained relatively unchanged. Superoxide dismutase activity was higher in SE pretreated with sucrose, compared with those pretreated with polyols; H2O2 was enhanced in SE pretreated with sorbitol and sucrose compared to mannitol. The overall trend for ethylene and OH production revealed their levels were highest in SE pretreated with polyols albeit, for individual treatments this was not always the case. Generally, pretreatments did not significantly change embryo secondary oxidation profiles of ThioBarbituric Acid Reactive Substances (TBARS) and Schiff's bases. In combination these studies suggest oxidative processes may influence regrowth of cryopreserved olive SE and that optimal pretreatments could, in part, increase tolerance by an overall enhancement of endogenous antioxidants (particularly GR), proline and sugars. Highlights ► Biomarkers may be used to make evidence based decisions to optimize olive cryostorage. ► Osmotica can affect antioxidant, ROS, sugar and proline profiles in olive somatic embryos. ► Data suggests a putative involvement of ROS in cryopreservation-induced stress in olive cultures.

U2 - 10.1016/j.plantsci.2011.03.009

DO - 10.1016/j.plantsci.2011.03.009

M3 - Article

VL - 181

SP - 47

EP - 56

JO - Plant Science

T2 - Plant Science

JF - Plant Science

SN - 0168-9452

IS - 1

ER -

Lynch PT, Siddika A, Johnston JW, Trigwell SM, Mehra A, Benelli C et al. Effects of osmotic pretreatments on oxidative stress, antioxidant profiles and cryopreservation of olive somatic embryos. Plant Science. 2011 Jul;181(1):47-56. Available from, DOI: 10.1016/j.plantsci.2011.03.009