Cryopreservation of dendrobium cruentum Rchb. f.

Name: Keiko Kagawa

LCSH: Dendrobium cruentum Rchb. f.

LCSH: Cryopreservation of organs, tissues, etc.

MeSH: Cryopreservation

MeSH: Dendrobium

MeSH: Dehydration

Abstract: Dendrobium cruentum Rchb. f. is a wild Thai orchid species that faces extinction. Due to the loss of natural habitats and the difficulty of its cultivation, practical conservation methods should be established. Cryopreservation, an important method for long-term storage of biological germplasm under ultra low temperature without deterioration, was studied for Dendrobium cruentum Rchb. f. The seeds of Dendrobium cruentum Rchb.f. at varying periods of 2, 3 and 4 months after pollination were cryopreserved by vitrification. The seeds were exposed to plant vitrification solution 2 (PVS2) for 0-90 min at 25±2℃ before being plunged into liquid nitrogen (LN). The water content of the seeds decreased from 81.6%FW to 13.1%FW as the seed pods matured. After cryopreservation, the germination rates of the seeds preserved at 2, 3 and 4 months after pollination were 0%, 13% and 32%, respectively. Maturity of seeds was an important factor for successful cryopreservation. After 6 months culturing, plantlets were transferred to the saranhouse. There was no significant difference in growth between control and cryopreserved plantlets. Protocorms of Dendrobium cruentum Rchb. f. were cryopreserved. With the vitrification method, protocorms were precultured on modified Vacin and Went (1949) medium supplemented with 0.06 (control), 0.3, 0.5, and 0.7 M sucrose for 1-5 d and then exposed to Loading Solution, and PVS2 for 0-420 min at 0±2℃ before being plunged into LN. In encapsulation-dehydration method, protocorms were encapsulated in calcium alginate before preculturing in sucrose supplemented medium and dehydration for 0-10h by sterile air from a laminar air-flow cabinet and then being plunged into LN. After cryopreservation, the survival rate of protocorms at water content of 65%FW, diameter 0.1-0.2mm was 33% (vitrification), 27% (encapsulation-dehydration). However, protocorms at water content of 87%FW, diameter 0.2-0.5mm did not survive. After 6 months culturing, plantlets were transferred to the saranhouse. There was no significant difference in growth between control and cryopreserved plantlets. In vitro protocorm-like bodies (PLBs) and shoot tips of Dendrobium cruentum Rchb. f. cryopreserved by vitrification, encapsulation-dehydration and encapsulation-vitrification had no survival after preculturing with high sucrose concentration medium. PLBs and shoot tips had high water content. Preconditioning by cold acclimation, ABA and glycerol before preculturing had no effect on acquiring dehydration tolerance.

Mahidol University

Address: NAKHON PATHOM

Email: liwww@mahidol.ac.th

Name: Sompop Prathanturarug

Role: Thesis Advisors

Created: 2006

Modified: 2553-06-24

Issued: 2010-06-05

วิทยานิพนธ์/Thesis

application/pdf

ISBN: 9740476872

CallNumber: TH K27c 2006

eng

DegreeName: Master of Science

Level: Master's Degree

Descipline: Plant Science

Grantor: Mahidol University

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