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Assessment of genetic and epigenetic changes following cryopreservation in papaya

  • Genetics and Genomics
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Abstract

A vitrification based cryopreservation technique for storage of in vitro shoot tips of papaya has been tested to ensure applicability across a range of genotypes and to assess the stability of both genotype and phenotype of such clonal material following cryopreservation. Shoot tips of 12 genotypes were cryopreserved, recovery rates were determined and resultant plants were screened for genetic and epigenetic changes. Genomic DNA structure was explored using polymerase chain reaction (PCR) based randomly amplified DNA fingerprinting (RAF), and methylation patterns were monitored using the amplified DNA methylation polymorphism (AMP) PCR technique. Plantlets were recovered following cryopreservation in all but one genotype and recovery rates of 61–73% were obtained from six genotypes. The regenerated plantlets showed varying levels of genomic DNA modifications (0–10.07%), and methylation modifications (0.52–6.62%) of detected markers. These findings have not been reported previously for papaya, and indicate some genotype dependent variability in DNA modifications occur following cryopreservation which may result in somaclonal variation.

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Abbreviations

RAF:

Randomly amplified DNA fingerprinting

AMP:

Amplified DNA methylation polymorphism

PVS2:

Plant vitrification solution 2

LN:

Liquid nitrogen

RFLP:

Restriction fragment length polymorphism

RAPD:

Randomly amplified polymorphic DNA

AFLP:

Amplified fragment length polymorphism

SSR:

Simple sequence repeats

MSAP:

Methylation-sensitive amplified polymorphism

BA:

6-Benzylaminopurine

IAA:

Indole-3-acetic acid

GA:

Gibberellic acid

DMSO:

Dimethylsulfoxide

CAPS:

Cleavage amplified polymorphic sequence

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Acknowledgments

I would like to thank Mahmoud Azimi, Shannon Dillon and Carl Ramage for valuable input and the International Plant Genetic Resources Institute (IPGRI) for project funding.

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Authors and Affiliations

  1. School of Biomolecular and Physical Sciences, Griffith University, Brisbane, Australia

    A. Kaity, S. E. Ashmore & R. A. Drew

  2. Biodiversity International, via Dei Tre Denari 472/a, 00057, Maccarese, Rome, Italy

    M. E. Dulloo

Authors
  1. A. Kaity
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  2. S. E. Ashmore
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  3. R. A. Drew
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  4. M. E. Dulloo
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Correspondence to A. Kaity.

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Communicated by S.A. Merkle.

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Kaity, A., Ashmore, S.E., Drew, R.A. et al. Assessment of genetic and epigenetic changes following cryopreservation in papaya. Plant Cell Rep 27, 1529–1539 (2008). https://doi.org/10.1007/s00299-008-0558-1

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  • DOI: https://doi.org/10.1007/s00299-008-0558-1

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