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Recovering polyploid papaya in vitro regenerants as screened by flow cytometry

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Abstract

Several protocols have been proposed for in vitro propagation of papaya, either based on somatic embryogenesis or shoot organogenesis. It is well-known that tissue culture-based approaches are frequently associated with somaclonal variation. Whether on the one hand this phenomenon can preclude further stages of in vitro culture, on the other hand it can generate useful genetic variability for crop improvement. However, somaclonal variation analyses are limited in papaya tissue culture. The DNA ploidy level of 250 papaya somatic embryogenesis-derived plantlets from immature zygotic embryos was analyzed by flow cytometry. In vitro-grown and greenhouse seed-derived plantlets were used as diploid standards. Flow cytometry unambiguously evidenced euploid (diploid, mixoploid, triploid and tetraploid) and aneuploid papaya plantlets, indicating that in vitro culture conditions can lead the occurrence of somaclonal variation. Additionally, the two subsequent flow cytometry analyses showed that the DNA ploidy level remained stable in all cloned papaya plantlets during the successive subcultures in the multiplication medium.

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Abbreviations

ABA:

Abscisic acid

BAP:

6-Benzylaminopurine

2,4-D:

Dichlorophenoxyacetic acid

NAA:

α-Naphthaleneacetic acid

DAPI:

4′,6′-diamidino-2-phenylindole

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Acknowledgements

The authors are grateful to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brasília, DF, Brazil) and Caliman Agrícola SA (Linhares, ES, Brazil) for financial support, and Prof. Marcio G.C. Costa for critical reading this manuscript.

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Authors and Affiliations

  1. Laboratório de Citogenética e Citometria, Departamento de Biologia Geral, Universidade Federal de Viçosa, Vicosa, MG, 36570-000, Brazil

    Wellington Ronildo Clarindo, Carlos Roberto de Carvalho, Fernanda Santos Araújo & Isabella Santiago de Abreu

  2. Laboratório de Cultura de Tecidos, Departamento de Biologia Vegetal/BIOAGRO, Universidade Federal de Viçosa, Vicosa, MG, 36570-000, Brazil

    Wagner Campos Otoni

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  1. Wellington Ronildo Clarindo
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  2. Carlos Roberto de Carvalho
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  5. Wagner Campos Otoni
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Correspondence to Carlos Roberto de Carvalho.

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Clarindo, W.R., de Carvalho, C.R., Araújo, F.S. et al. Recovering polyploid papaya in vitro regenerants as screened by flow cytometry. Plant Cell Tiss Organ Cult 92, 207–214 (2008). https://doi.org/10.1007/s11240-007-9325-1

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  • DOI: https://doi.org/10.1007/s11240-007-9325-1

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