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1 November 2003 SOMATIC EMBRYOGENESIS FROM IMMATURE ZYGOTIC EMBRYOS OF ANNATTO (BIXA ORELLANA L.)
VESPASIANO B. PAIVA NETO, MARCEL N. BOTELHO, ROSANE AGUİAR, ELDO A. M. SILVA, WAGNER C. OTONI
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Abstract

In order to establish a protocol for somatic embryogenesis of annatto, Bixa orellanaL., seeds (70 d after anthesis) from field-grown orchards had their coats dissected off, and immature zygotic embryos were excised aseptically from immature seeds collected from field-grown trees and used as explants. Embryos were cultured onto MS medium supplemented with or without different combinations of plant growth regulators and activated charcoal. Direct somatic embryogenesis was induced on explants incubated either in Murashige and Skoog (MS), 2,4-dichlorophenoxyacetic acid (2,4-D), and/or kinetin-supplemented media after 25 d of culture. The highest frequencies of embryogenesis and embryos per explant were obtained on medium containing 2.26 μM 2,4-D, 4.52 μM kinetin, and 1.0 g l−1 activated charcoal. The presence of charcoal was critical in increasing embryos per explant, to reduce the time to obtain somatic embryos, and mainly to prevent callus proliferation and subsequent indirect somatic embryogenesis. No embryogenic response was achieved when mature embryos were used. It was also observed that embryogenic response was significantly affected by genotype. Histological investigations revealed that primary direct somatic embryos differentiated exclusively from the protodermis or together with the outer ground meristem cell layers of the zygotic embryo axis, and from the protodermis of zygotic cotyledons. Diverse morphological differences, including malformed embryos, were observed among somatic embryos. In spite of the high frequencies of histodifferentiation of all embryo stages, a very low conversion frequency to normal plants from somatic embryos was observed.

VESPASIANO B. PAIVA NETO, MARCEL N. BOTELHO, ROSANE AGUİAR, ELDO A. M. SILVA, and WAGNER C. OTONI "SOMATIC EMBRYOGENESIS FROM IMMATURE ZYGOTIC EMBRYOS OF ANNATTO (BIXA ORELLANA L.)," In Vitro Cellular and Developmental Biology - Plant 39(6), 629-634, (1 November 2003). https://doi.org/10.1079/IVP2003465
Received: 20 August 2002; Accepted: 1 May 2003; Published: 1 November 2003
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KEYWORDS
Activated charcoal
annatto
embryogenic response
Histology
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