Somatic embryogenesis and plant regeneration in carob (Ceratonia siliqua L.) |
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Authors: | Jorge M Canhoto Sandra C Rama Gil S Cruz |
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Institution: | (1) Instituto do Ambiente e Vida, Departamento de Botanica, Faculdade de Ciências e Tecnologia, Universidade de Coimbra, 3001-455 Coimbra, Portugal |
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Abstract: | Summary Somatic embryos of carob (Ceratonia siliqua L.) were induced from cotyledonary segments excised from immature seeds when cultured on Murashige and Skoog media supplemented
with several combinations of 6-benzylaminopurine (BA) and indole-3-butyric acid (IBA). The best frequencies of induction (33.8%)
were obtained when 4.4 μM BA and 0.5 μM IBA were used. Shoots were also sporadically formed in the same media. When IBA was replaced by other auxins in the induction
media, only α-naphthaleneacetic acid (NAA) and indole-3-acetic acid (IAA) could induce somatic embryogenesis, although at
lower rates than IBA. 2,4-Dichlorophenoxyacetic acid and 4-amino-3,5,6-trichloropicolinic acid were completely ineffective.
Besides culture media composition, the developmental stage of the explants at the time of culture showed a strong influence
on somatic embryogenesis induction, with cotyledons from stage II pods providing the highest levels of induction. By contrast,
the genotype of the explant did not determine a significant role in the induction process. Attempts to achieve somatic embryo
germination were mostly unsuccessful, since only shoot development was observed; the highest frequencies of development occurred
on media containing only gibberellic acid (3.0 μM). For plant regeneration, the developed shoots were further rooted on IBA-supplemented media, and the plantlets obtained
were transferred to soil, where c. 88% of them survived. Histological observations showed the presence of morphologically normal and abnormal somatic embryos,
the latter displaying an abnormal pattern of vascular bundles. Ultrastructural analysis showed that the cells of the globular
embryos had a dense cytoplasm, whereas those not involved in somatic embryo formation showed signs of senescence. Histological
studies were also used to distinguish between somatic embryos and shoots originated in the same media. |
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Keywords: | Fabaceae genotype histology rooting ultrastructure |
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