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Abstract
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| The aim of this study has been to apply and to assess the capacity of the CRED-RA technique (¿Coupled restriction enzyme digestion and random amplification¿) (Cai et al., 1996) to obtain the methylation patterns of chrysanthemum cryopreserved shoots, and by comparing methylation patterns to analyse the epigenetic stability of the cryopreserved material in relation to control non-treated tissues. The methodology of this technique is based on the capacity of certain restriction enzymes (Hpa II and Msp I) to recognize the same cut sequence, but with different restriction capacity depending on the methylation status of the cytosine residues. Subsequently, PCR amplifications, using arbitrary primers, are produced with the digested DNA. These amplifications could show different bands patterns according to the DNA methylation status of the original sample. The results obtained in this work showed a high genetic and epigenetic stability of the chrysanthemum samples studied. Nevertheless, two out of six samples, derived from the encapsulation-dehydration treatment, presented different band-patterns compared to their controls when the CRED-RA technique was applied. These differences cannot be correlated to a possible somaclonal variation since the RAPD data did not show any genetic change. No variation was detected in the four samples derived from the vitrification method, although to conclude that this protocol implies a higher stability, compared to encapsulation-dehydration method, further analyses would be necessary. | |
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International
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Si |
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Congress
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COST 871, Working Group 2, Integration of cryopreservation in genebank strategies |
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960 |
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Place
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Gatersleben, Alemania |
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Reviewers
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Si |
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ISBN/ISSN
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0143-2044 |
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Start Date
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07/09/2009 |
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End Date
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11/09/2009 |
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From page
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85 |
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To page
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85 |
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CryoLetters 31(1) |