Articles
Protective effect of reducing cryotube volume on Lilium lancifolium and Euonymus fortunei shoot tips survival after cryopreservation by vitrification
Article number
1104_39
Pages
255 – 258
Language
English
Abstract
Rapid freezing and rewarming are essential to achieve vitrification of internal solutes and of the surrounding cryoprotective medium during freezing and to avoid potentially damaging devitrification and recrystallization events during rewarming.
In order to evaluate heat exchange rate, a comparative experiment was established using two different volume cryotubes (1 mL and 200 µL) in vitrification of Euonymus fortunei and Lilium lancifolium shoot tips with the vitrification protocols established by our group before.
TTC (2,3,5-triphenyltetrazolium chloride) staining was used to check shoot tips survival by microscopic examination.
The results show that: using 200-μL cryotubes significantly improved viability of E. fortunei shoot tips after liquid nitrogen, with 27.1±3.1% of shoot tips stained red, compared to control 11.2±1.8% (PKLEINERDAN0.05); it also had a protective effect on L. lancifolium shoot tips survival, with 95.8±2.5% compared to 75.1±3.9% (PKLEINERDAN0.05). In conclusion, reducing cryotube volume in vitrification is an effective way to improve shoot tips survival after cryopreservation, and it may be related to higher cooling/warming rates achieved in using smaller volume cryotubes.
In order to evaluate heat exchange rate, a comparative experiment was established using two different volume cryotubes (1 mL and 200 µL) in vitrification of Euonymus fortunei and Lilium lancifolium shoot tips with the vitrification protocols established by our group before.
TTC (2,3,5-triphenyltetrazolium chloride) staining was used to check shoot tips survival by microscopic examination.
The results show that: using 200-μL cryotubes significantly improved viability of E. fortunei shoot tips after liquid nitrogen, with 27.1±3.1% of shoot tips stained red, compared to control 11.2±1.8% (PKLEINERDAN0.05); it also had a protective effect on L. lancifolium shoot tips survival, with 95.8±2.5% compared to 75.1±3.9% (PKLEINERDAN0.05). In conclusion, reducing cryotube volume in vitrification is an effective way to improve shoot tips survival after cryopreservation, and it may be related to higher cooling/warming rates achieved in using smaller volume cryotubes.
Authors
Xu Jin, Liu Yan, Li Bingling, Liu Qian, Shi Yin, Jia Mengxue, Shen Chen
Keywords
cryopreservation, vitrification, shoot tips, heat exchange rate
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