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The effects of five carbon sources (glucose, fructose, maltose, sorbitol, and sucrose) and two explant types (petals and staminodes) on cacao somatic embryogenesis was studied. No growth was observed on both types of explants cultured on sorbitol containing media and slow growth was obtained on media supplemented with maltose. Depending on the genotype, the percentage of explants producing one or more embryos ranged from 6% to 99%, 18% to 98%, and 3% to 82% on media containing glucose, fructose and sucrose respectively. Explants cultured continuously on maltose or sorbitol-containing media failed to produce embryos. Staminode explants produced 3 to 10 times more somatic embryos than petals. A strong genotypic effect on somatic embryogenesis was observed. Staminode explants of the Forastero clones Laranja and PSUSca 6 produced 2 to 30 times more somatic embryos than the Trinitarios UF 613 and ICS 16. During embryo maturation and conversion, no significant differences were observed among glucose, fructose, maltose, or sucrose for embryo weight, total shoot and root production. However, we found that all plantlets produced on glucose had shoots with normal cacao leaves while the other carbon sources sometimes produced plantlets with cotyledon-like leaves.
Twelve cacao (Theobroma cacao) clones propagated by grafting and orthotropic rooted cuttings of somatic embryo-derived plants were grown on an Ultisol soil at Corozal, Puerto Rico, and evaluated for 6 years of production under intensive management. Year, variety, year × variety, and propagation treatment × variety interactions indicated significant effects for dry bean yield, number of pods produced, pod index, plant height, and stem diameter. Propagation treatments had a significant effect on dry bean yield and pod index but not on number of pods produced. Average yield across varieties for both propagation treatments was 2087.9 kg·ha−1 per year of dry beans. There was a highly significant variety effect. ‘UF-668’ was the top yielder averaging 2536.7 kg·ha−1 per year of dry beans; however, this yield was not significantly different from the average yield of varieties ‘TARS-30’, ‘TARS-1’, ‘TARS-13’, ‘TARS-14’, and ‘TARS-2’, which averaged 2427.0 kg·ha−1 per year. Except for ‘UF-668’, the TARS varieties were released in 2009 as high-yielding selections. Propagation treatments had a significant effect on dry bean yield. Dry bean yield of varieties propagated by grafting was 7% higher (2166.7 kg·ha−1 per year) than those propagated by orthotropic rooted cuttings of somatic embryo-derived plants (2009.2 kg·ha−1 per year). This yield difference could not be attributed to grafted plants being more vigorous nor by differences in root architecture. The lowest pod index value in both propagation treatments was obtained by ‘UF-668’; however, pod index for this variety did not differ significantly from values for ‘TARS-2’ and ‘TARS-23’ in grafted plants and from ‘TARS-2’, ‘TARS-23’, and ‘TARS-1’ in plants propagated by orthotropic rooted cuttings of somatic embryo-derived plants. With few exceptions, flavor characteristics were not significantly affected by propagation treatments. Although there were significant differences between plant propagation treatments for some of the variables measured in this study, these were not of a magnitude that would preclude the use of somatic embryogenesis as a viable propagation system for cacao.
During plant starch biosynthesis, starch branching enzymes (SBE) catalyze a-1,6 branch point formation in starch, and thus are responsible for many properties of the starch polymer. Recently we have cloned cDNAs encoding the two major branching enzymes in developing maize endosperm, SBEI and SBEII. These genes are being used to alter starch biosynthesis via genetic engineering strategies. Transgenic tobacco plants with sense and antisense constructs of SBEI and SBEII have been produced. No major difference in the phenotypes of control and transgenic plants have been observed. Initial experiments demonstrated the transcription of the introduced genes. Enzyme levels and the molecular properties of the starch in the transgenic plants will be determined. These experiments will provide us with information as to the role of starch branching enzymes in starch biosynthesis, the feasibility of creating novel starch, and the effect altered starch has on plastid development and photosynthesis.