Cryopreservation of zygotic embryos of a Japanese terrestrial orchid (Bletilla striata) by vitrification

The seeds of a Japanese terrestrial orchid (Bletilla striata Rchb.f.) were germinated and cultured on solidified new Dogashima (ND) medium for 10 days. These embryos were then precultured on ND medium supplemented with 0.3 M sucrose for 3 days at 25°C in continuous dark. The embryos were then overla...

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Published in:Plant cell reports 1997-09, Vol.16 (11), p.754-757
Main Authors: ISHIKAWA, K, HARATA, K, MII, M, SAKAI, A, YOSHIMATSU, K, SHIMOMURA, K
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Embryos
Eukaryotic cell cultures
Fundamental and applied biological sciences. Psychology
In vitro embryo culture
Methods. Procedures. Technologies
Plant cells and fungal cells
Sucrose
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creator ISHIKAWA, K
HARATA, K
MII, M
SAKAI, A
YOSHIMATSU, K
SHIMOMURA, K
description The seeds of a Japanese terrestrial orchid (Bletilla striata Rchb.f.) were germinated and cultured on solidified new Dogashima (ND) medium for 10 days. These embryos were then precultured on ND medium supplemented with 0.3 M sucrose for 3 days at 25°C in continuous dark. The embryos were then overlaid with a mixture of 2 M glycerol and 0.4 M sucrose for 15 min at 25°C and finally dehydrated with highly concentrated vitrification solution (PVS2) for 3 h at 0°C prior to immersion into liquid nitrogen for 30 min. After rapid warming, the embryos were washed with liquid ND medium supplemented with 1.2 M sucrose for 20 min and then plated on ND medium. Successfully vitrified and warmed embryos developed into normal plantlets. The rate of plant regeneration amounted to about 60%. This vitrification method appears to be a promising technique for cryopreservation of orchids.
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subjects Biological and medical sciences
Biotechnology
Embryos
Eukaryotic cell cultures
Fundamental and applied biological sciences. Psychology
In vitro embryo culture
Methods. Procedures. Technologies
Plant cells and fungal cells
Sucrose
title Cryopreservation of zygotic embryos of a Japanese terrestrial orchid (Bletilla striata) by vitrification
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