High temperature and nib acidification during cacao-controlled fermentation improve cadmium transfer from nibs to testa and the liquor's flavor.

Migration of nib Cd to the testa during fermentation can be achieved with high temperatures (> 45 °C) and low nib pH values (< 5.0) using spontaneous fermentation. However, this low pH can lead to low flavor quality. This study used three controlled temperature fermentation treatments on three cacao genotypes (CCN 51, ICS 95, and TCS 01) to test its effects on the nib pH, the migration of nib Cd to the testa, and the liquor flavor quality. All treatments were effective in reducing the total nib Cd concentration. Nevertheless, the treatment with the higher mean temperature (44.25 °C) and acidification (pH 4.66) reached the highest mean nib Cd reductions throughout fermentation, a 1.37 factor in TCS 01, promoting the development of fine-flavor cocoa sensorial notes. In unfermented beans, the Cd concentration of nibs was higher than that of the testa, and the Cd migration proceeded down the total concentration gradient. However, Cd migration was observed against the concentration gradient (testa Cd > nib Cd) from the fourth day. Cd migration could increase by extensive fermentation until the sixth day in high temperatures and probably by the adsorbent capacity of the testa. Genotype-by-treatment interactions were present for the nib Cd reduction, and a universal percentage of decrease of Cd for each genotype with fermentation cannot be expected. Selecting genotypes with highly adsorbent testa combined with controlled temperatures would help reduce the Cd concentration in the cacao raw material, improving its safety and quality.

Microsporogénesis y ultraestructura de los granos de polen en la planta del cacao, Theobroma cacao (Malvaceae)

Introducción: No conocemos estudios sobre la microsporogénesis de la planta de cacao, y poco se sabe sobre la ultraestructura de sus granos de polen. Objetivo: Describir la microsporogénesis y ultraestructura de los granos de polen en T. cacao. Métodos: Procesamos más de 30 flores para cada etapa floral, teñidas con Safranina-Azul Alcian, PAS-Amidoblack y Lacmoid. Para la microscopía de transmisión procesamos las muestras en resina y las teñimos con azul de toluidina. Para microscopía electrónica de barrido, fijamos y deshidratamos en 2.2-dimetoxipropano, secamos hasta un punto crítico y recubrimos con oro. Resultados: Anteras diferenciadas por una masa celular en los extremos distales a los filamentos estaminales. Durante el desarrollo la pared de las anteras presenta varios estratos celulares y al madurar se reducen a la epidermis y al endotecio. Las células madre de microsporas se dividen por meiosis para formar tétradas. El tapete es secretor e intacto hasta que se liberan los granos, para luego degenerar. Los granos de polen son isopolares, esferoidales, pequeños, tricolpados. La ultraestructura presenta una esporodermis semitectada, con ornamentación reticulada, y un retículo heterobrochado con el muri sin ornamentación. La exina se deposita antes que la intina. Los orbículos son individuales, lisos y de tamaño variado. Hay abundante polenkit en el tectum y entre las columelas. La intina es delgada, pero se desarrolla ampliamente en las áreas del colpo, formando una intina interna compacta y una intina externa inusual con una apariencia columelada. Conclusión: La estructura y el desarrollo de las anteras siguen el patrón de las angiospermas. La microsporogénesis simultánea y la deposición centrípeta de la esporodermis se conocen de Malvaceae, pero los caracteres de la intina son nuevos para la familia.

Introduction: We know of no studies on the microsporogenesis of the cocoa plant, and little is known about the ultrastructure of its pollen grains. Objective: To describe microsporogenesis and ultrastructure of pollen grains in T. cacao. Methods: We processed over 30 flowers for each floral stage and stained with Safranin-Alcian Blue, PAS-Amidoblack and Lacmoid. For transmission microscopy we processed samples on resin and stained with toluidine blue. For scanning electron microscopy, we fixed and dehydrated in 2.2-dimethoxypropane, critically dried and coated with gold. Results: Anthers differentiated by a cellular mass at the ends distal to the staminal filaments. During development, the anther wall has several cellular layers reduced, at maturity, to the epidermis and endothecium. Microspore mother cells divide by meiosis to form tetrads. The tapetum is secretory and intact until the grains are released, to later degenerate. Pollen grains are isopolar, spheroidal, small, tricolpate. Ultrastructure has a semi-tectate sporodermis, with reticulate ornamentation, and heterobrochated reticulum with the muri without ornamentation. Exine is deposited before intine. The orbicles are individual, smooth, and varied in size. There is abundant pollenkitt on the tectum and between the columellae. The intine is thin, but develops widely in the colpus areas, forming a compact internal intine and an unusual external intine with a columellated appearance. Conclusion: Anther structure and development follows the angiosperm pattern. Simultaneous microsporogenesis and centripetal deposition of the sporodermis are known from Malvaceae, but intine characters are novel for the family.