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Attempts to eradicate graft-transmissible infections through somatic embryogenesis in Citrus ssp. and analysis of genetic stability of regenerated plants

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Abstract

Stigma/style somatic embryogenesis (SE) is proved to be effective in the complete elimination of the main Citrus virus and virus-like diseases in 100 % of analysed cases. In the present research SE was applied on 13 genotypes, belonging to the Algerian germplasm collection of two different Citrus species (Citrus limon and C. sinensis) infected by one or more graft-transmissible agents, to evaluate the genetic stability and the sanitation of regenerants. The infected genotypes were regenerated through stigma/style SE and the obtained plants were tested by serological, molecular and biological assays, 6 and 18 months after plantlet grafting, for assessing the elimination of the virus, viroid and virus-like agents present in the mother plants. No evidence of infections of virus and virus–like agents was present in the regenerated plants; whereas, after 18 months up to 24 % of tested plants were found to be infected by viroids, independently from the species and genotypes of the mother plants. Hop stunt viroid (HSVd) proved to be the most infectious viroid; conversely, Citrus exocortis viroid (CEVd) and Citrus viroid III (CVd-III) were present only in ‘Sécile’ lemon, as mixed infection with HSVd. SE from stigma/style explants was 100 % effective in the elimination of mixed viroid infections in two lemons (‘Lunario’ and ‘Sans pépins’) and one orange (‘Mitidja navel’). To evaluate genetic stability of regenerants, DNA analyses were performed. No somaclonal variability was observed in lemon regenerants. However, the amplification products of ‘Washington navel 251’ (C. sinensis) revealed genetic instability in some of the regenerated plants.

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Author contribution statement

AC, AMD, FM, KD and FC conceived and designed the study; MM, DF, MB and FM developed the methodology and performed the experiments; AC, AMD, FM KD and FC analysed data; MM, AC, AMD, FM, KD and FC prepared the manuscript. All authors discussed the results and commented on the manuscript.

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Authors and Affiliations

  1. Université Hassiba Ben Bouali, Hay Salem, Route Nationale 19 (02000), Chlef, Algérie

    Malika Meziane

  2. Centre International de Hautes Etudes Agronomiques Méditerranéennes (CIHEAM)/Mediterranean Agronomic Institute of Bari (MAIB), Via Ceglie 9, 70010, Valenzano, BA, Italy

    Dajana Frasheri, Anna Maria D’Onghia & Khaled Djelouah

  3. Institute of Biosciences and BioResources, National Research Council (CNR), Research Division of Palermo, Corso Calatafimi 414, 90129, Palermo, Italy

    Angela Carra, Francesco Mercati & Francesco Carimi

  4. Laboratoire des Cultures in vitro, ENS Kouba, Alger, Algeria

    Messaoud Boudjeniba

Authors
  1. Malika Meziane
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  2. Dajana Frasheri
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  3. Angela Carra
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  4. Messaoud Boudjeniba
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  5. Anna Maria D’Onghia
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  6. Francesco Mercati
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  7. Khaled Djelouah
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  8. Francesco Carimi
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Correspondence to Angela Carra.

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Meziane, M., Frasheri, D., Carra, A. et al. Attempts to eradicate graft-transmissible infections through somatic embryogenesis in Citrus ssp. and analysis of genetic stability of regenerated plants. Eur J Plant Pathol 148, 85–95 (2017). https://doi.org/10.1007/s10658-016-1072-x

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  • DOI: https://doi.org/10.1007/s10658-016-1072-x

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