The establishment of cocoa embryogenic cell lines in liquid medium starting from high frequency somatic embryogenesis (HFSE) callus is described. The growth kinetics of the cultures during the multiplication and the expression steps conducted in 250 mL Erlenmeyer flasks were described for three genotypes selected for their agronomical traits (EET95, EET96, and EET103). The glucose and dissolved oxygen concentrations and the absorption of Murashige and Skoog medium macronutrients (nitrate, ammonium, potassium, sulfate, calcium, phosphorus, and magnesium) were monitored. The multiplication of the embryogenic calluses in a medium containing 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) at 1 mg L-1, initiated with an inoculation density of 20 g L-1 of callus, was achieved. The growth rate was characterized by two phases, with the second being concomitant with a depletion of phosphorus and magnesium, and a decrease in the embryogenic potential of the callus. The expression of the callus embryogenic capacity was conducted in an auxin-free medium. The embryo production starting from 1 and 5 g L-1 inoculation densities was compared. When placed in the optimal expression conditions in flasks, 1 g of callus produced 1000 to 1500 embryos within 5 to 7 wk. Finally, two paths for improving the plantlet regenerative capacities of cocoa SE produced in liquid medium were identified. Supplementing the expression medium with myo-inositol used as an osmotic agent at a concentration of 50 g L-1 increased the embryo-to-plantlet conversion rate from 13-16% to 40-48%. A 6-wk culture of the embryos on a maturation medium in Petri dishes optimized their subsequent development into plantlets.

中文翻译：

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可可液在液体培养基中的间接体细胞胚胎发生和提高胚胎到小植株的转化率。

描述了从高频体细胞胚发生（HFSE）愈伤组织开始在液体培养基中建立可可胚发生细胞系。描述了在250 mL锥形瓶中进行繁殖和表达的过程中培养物的生长动力学，描述了三种根据农艺性状选择的基因型（EET95，EET96和EET103）。监测葡萄糖和溶解氧的浓度以及Murashige和Skoog培养基中大量营养素（硝酸盐，铵，钾，硫酸盐，钙，磷和镁）的吸收。在含有20 g L-1愈伤组织的接种密度下，实现了在1 mg L-1的含2,4,5-三氯苯氧基乙酸（2,4,5-T）的培养基中繁殖胚性愈伤组织。 。增长率分为两个阶段：第二种伴随着磷和镁的消耗，以及愈伤组织的胚发生潜能的降低。在无生长素的培养基中进行愈伤组织胚发生能力的表达。比较了从1和5 g L-1接种密度开始的胚胎产量。当置于最佳表达条件的烧瓶中时，1 g愈伤组织在5至7周内产生1000至1500个胚胎。最后，确定了改善液体培养基中产生的可可SE植株再生能力的两条途径。向表达培养基中添加浓度为50 g L-1的肌醇作为渗透剂，可使胚间转化率从13-16％提高至40-8％。