Regeneration of Algerian Citrus germplasm by stigma/style somatic embryogenesis

Stigma/style somatic embryogenesis is one of the efficient methods in plant regeneration of most Citrus ssp., without inducing somaclonal variations. Furthermore, somatic embryogenesis from style/stigma proved to be effective in the elimination of the main citrus virus and virus-like diseases. This technique was applied on Algerian citrus collection. Different Citrus species [Citrus sinensis (L.) Osbeck, C. limon (L.) Burm, C. reticulata Blanco, C. paradisi Macfad, C. reshni Hort. ex Tan., C. jambhiri Lush and C. maxima (Burm.) Merrill] were chosen and tested for the presence of the main virus and virus-like agents. Most of the genotypes showed to be infected, mainly by viroid agents. Closed flowers were collected and in vitro cultured on a Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine. All explants produced callus about 4 to 9 days after culture initiation, whereas embryogenesis occurred after 38 to 150 days in most of the cultured genotypes. Formed embryos were cultured in a single tube before in vivo acclimatization. After sanitary assays, regenerated plants were shown to be free from the agents detected in the mother trees.