Cryopreservation of Garcinia cowa shoot tips by vitrification

The effects of sucrose preculture and loading treatment on ultrastructural changes in meristematic cells

Lip Vun Yap, Normah Mohd. Noor, Mahani Mansor Clyde, Hoong Fong Chin

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The effects of sucrose preculture duration and loading treatment on tolerance of Garcinia cowa shoot tips to cryopreservation using the PVS2 vitrification solution were investigated. Ultrastructural changes in meristematic cells at the end of the preculture and loading steps were followed in an attempt to understand the effects of these treatments on structural changes in cell membranes and organelles. Increasing preculture duration on 0.3 M sucrose medium from 0 to 3 days enhanced tolerance to PVS2 solution from 5.6% (no preculture) to 49.2% (3-day preculture). However, no survival was observed after cryopreservation. Examination of meristematic cells by transmission electron microscopy revealed the progressive accumulation of an electron-dense substance in line with increasing exposure durations to 0.3 M sucrose preculture. Treatment with a loading solution (2 M glycerol + 0.4 M sucrose) decreased tolerance of shoot tips to PVS2 vitrification solution and had a deleterious effect on the ultrastructure of G. cowa meristematic cells. This study suggests that G. cowa meristematic cells may lose their structural integrity due to exposure to glycerol present in the loading solution at a 2 M concentration, either due to its high osmotic potential, or due to its cytotoxicity.

Original languageEnglish
Pages (from-to)188-196
Number of pages9
JournalCryo-Letters
Volume32
Issue number3
Publication statusPublished - May 2011

Fingerprint

Garcinia
Vitrification
Cryopreservation
vitrification
cryopreservation
Sucrose
sucrose
Glycerol
cells
glycerol
Therapeutics
duration
Transmission Electron Microscopy
osmotic pressure
Organelles
exposure duration
organelles
transmission electron microscopy
cell membranes
cytotoxicity

Keywords

  • Dehydration tolerance
  • Garcinia cowa shoot tips
  • Loading solution
  • Sucrose preculture
  • Transmission electron microscope (TEM)
  • Ultrastructure

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Anatomy

Cite this

Cryopreservation of Garcinia cowa shoot tips by vitrification : The effects of sucrose preculture and loading treatment on ultrastructural changes in meristematic cells. / Yap, Lip Vun; Mohd. Noor, Normah; Clyde, Mahani Mansor; Chin, Hoong Fong.

In: Cryo-Letters, Vol. 32, No. 3, 05.2011, p. 188-196.

Research output: Contribution to journalArticle

Yap, Lip Vun ; Mohd. Noor, Normah ; Clyde, Mahani Mansor ; Chin, Hoong Fong. / Cryopreservation of Garcinia cowa shoot tips by vitrification : The effects of sucrose preculture and loading treatment on ultrastructural changes in meristematic cells. In: Cryo-Letters. 2011 ; Vol. 32, No. 3. pp. 188-196.
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