Cryopreservation of Rubber (Hevea Brasiliensis Muell.-Arg.) Zygotic Embryos Using Vitrification Technique
The study was carried out to develop a vitrification procedure for the cryopreservation of Hevea zygotic embryos. Vitrification generally involves three main steps, namely, loading, freezing in vitrification solution and unloading. This study evaluated the optimum loading and vitrification treatm...
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| Format: | Thesis |
| Language: | English English |
| Published: |
1999
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| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/10434/1/FP_1999_12_A.pdf |
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| Summary: | The study was carried out to develop a vitrification procedure for the
cryopreservation of Hevea zygotic embryos. Vitrification generally involves three
main steps, namely, loading, freezing in vitrification solution and unloading. This
study evaluated the optimum loading and vitrification treatments for Hevea embryos
for liquid nitrogen exposure and also to compare the effectiveness of vitrification
with other cryopreservation techniques.
The loading solutions tested were relatively toxic as the loaded embryos
showed lower viability and survival compared to the fresh embryos. However, the
effects of the four loading solutions up to 60 minutes exposure were similar. On
exposure to liquid nitrogen, none of the embryos survived.
The vitrification solution PVS2 was more effective than PVS and L solution
in vitrifying Hevea tissue in liquid nitrogen because only embryos treated with
PVS2 could survive after eight weeks culture. Longer time of exposure to
vitrification solution significantly increased the viability suggesting that more
tissues were vitrified with longer exposure. The optimum time of exposure to PVS2
was 80 minutes with 28.0% viability and 13.4% survival. The moisture content of
embryos after PVS2 exposure stabilised around 43.3-46.6%. Survival of frozen
Hevea embryos at such relatively high moisture confirmed the potential ofPVS2 for
vitrifying the tissue in liquid nitrogen.
Desiccation using fast and slow drying methods to reduce tissue moisture
did not improve survival of frozen embryos. Viability and survival of desiccated
embryos following vitrification treatment declined dramatically compared to the
control. For Hevea embryos, desiccation following vitrification treatment caused
more injuries to the tissues.
Comparison of the vitrification technique with other methods of
cryopreservation showed vitrification technique to be more effective with 81. 1%
viability and 51.2% survival after liquid nitrogen exposure. Naked desiccation,
desiccation following sucrose preculture and encapsulation dehydration, were
relatively ineffective. |
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