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Genetic instability in calamondin (Citrus madurensis Lour.) plants derived from somatic embryogenesis induced by diphenylurea derivatives

  • Genetics and Genomics
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Abstract

Somatic embryos were regenerated in vitro from calamondin style–stigma explants cultured in the presence of N 6-benzylaminopurine (BAP) cytokinin and three synthetic phenylurea derivatives, N-(2-chloro-4-pyridyl)-N-phenylurea (4-CPPU), N-phenyl-N′-benzothiazol-6-ylurea (PBU) and N,N′-bis-(2,3-methilendioxyphenyl)urea (2,3-MDPU). The phenylurea derivative compounds tested at micromolar level (12 μM) were able to induce a percentage of responsive explants significantly higher from that obtained with BAP and hormone-free (HF) conditions. In order to verify the genetic stability of the regenerants, 27 plants coming from different embryogenic events were randomly selected from each different culture condition and evaluated for somaclonal variations using inter-simple sequence repeat and random amplified polymorphic DNA analyses. We observed that 2,3-MDPU and PBU gave 3.7% of somaclonal mutants, whereas 4-CPPU gave 7.4% of mutants. No somaclonal variability was observed when plantlets were regenerated in BAP or HF medium. Although diphenylurea derivatives show a higher embryogenic potential as compared to BAP, they induce higher levels of somaclonal variability. This finding should be taken in consideration when new protocols for clonal propagation are being developed.

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Abbreviations

BAP:

N 6-benzylaminopurine

4-CPPU:

N-(2-chloro-4-pyridyl)-N-phenylurea

HF:

Hormone-free

ISSR:

Inter-simple sequence repeats

2,3-MDPU:

N,N′-bis-(2,3-methilendioxyphenyl)urea

NO:

Nitric oxide

PBU:

N-phenyl-N′-benzothiazol-6-ylurea

RAPD:

Random amplified polymorphic DNA

ROS:

Reactive oxygen species

TDZ:

Thidiazuron

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Acknowledgments

This research was financed in part by the Regione Siciliana, Assessorato Agricoltura e Foreste, project “Coltura in vitro per la conservazione del germoplasma vegetale siciliano minacciato da erosione genetica”.

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Authors and Affiliations

  1. Institute of Plant Genetics (Section Palermo), National Research Council, Corso Calatafimi 414, 90129, Palermo, Italy

    Mirko Siragusa, Angela Carra, Fabio De Pasquale & Francesco Carimi

  2. Developmental and Cellular Biology Department, University of Palermo, Viale delle Scienze, 90128, Palermo, Italy

    Lidia Salvia & Anna Maria Puglia

Authors
  1. Mirko Siragusa
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  2. Angela Carra
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  3. Lidia Salvia
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  4. Anna Maria Puglia
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  5. Fabio De Pasquale
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  6. Francesco Carimi
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Correspondence to Francesco Carimi.

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Communicated by D.A. Somers.

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Siragusa, M., Carra, A., Salvia, L. et al. Genetic instability in calamondin (Citrus madurensis Lour.) plants derived from somatic embryogenesis induced by diphenylurea derivatives. Plant Cell Rep 26, 1289–1296 (2007). https://doi.org/10.1007/s00299-007-0326-7

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