Evaluation of Endogenous Sugars, Chlorogenic Acid and Caffeine Associated with Direct Somatic Embryogenesis of Coffee (Coffea arabica L.)

D. K. Isutsa, Rose N Mayoli, A. Nyende, C. Mweu
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Abstract

Coffee is one of the most important cash crops cultivated in the world with great economic importance. During the induction of somatic embryogenesis, there are different endogenous compounds involved in the success or failure of the somatic embryogenesis response and these compounds determine the specificity of cellular responses. This present experiment identified and quantified endogenous sugars, chlorogenic acid and caffeine present during somatic embryogenesis of ‘Ruiru 11’. Laboratory experiments were set up at Coffee Research Institute, Ruiru-Kenya between 2014 and 2016. Third leaf pair explants were excised from 8-monthold greenhouse-grown mother plants and cultured in half strength Murashige and Skoog basal salts augmented with Thidiazuron. Once embryos had developed, the cultures were analysed for endogenous sugars, caffeine and chlorogenic acid using HPLC. Generally, green embryogenic cultures contained more and higher quantities of the compounds. Glucose and fructose were highest (38.95 mg/g and 45.43 mg/g respectively) in leaf discs of brown non-embryogenic cultures. Sucrose was highest (62.15 mg/g) in embryos of green embryogenic cultures. Embryos of green embryogenic cultures had the highest chlorogenic acid (5.3 mg/g), whereas caffeine was highest (0.55 mg/g) in embryos of brown embryogenic cultures. Endogenous fructose and glucose inhibited embryogenesis, while sucrose, chlorogenic acids and caffeine promoted embryogenesis and are potential biomarkers for embryogenesis. Other biochemical compounds such as organic acids should be identified and their role in coffee somatic embryogenesis determined.
内源糖、绿原酸和咖啡因对咖啡直接体细胞胚胎发生的影响
咖啡是世界上最重要的经济作物之一,具有重要的经济意义。在体细胞胚胎发生的诱导过程中,有不同的内源性化合物参与体细胞胚胎发生反应的成功或失败,这些化合物决定了细胞反应的特异性。本试验对‘瑞如11号’体细胞胚胎发生过程中存在的内源糖、绿原酸和咖啡因进行了鉴定和定量。2014年至2016年,在肯尼亚瑞鲁咖啡研究所进行了实验室实验。从8个月大的温室栽培母本上摘取第三对叶外植体,在添加了噻唑脲的半强度Murashige和Skoog基础盐中培养。一旦胚胎发育,就用高效液相色谱分析培养物的内源性糖、咖啡因和绿原酸。一般来说,绿色胚性培养物含有更多和更高数量的化合物。棕色非胚性培养的叶片中葡萄糖和果糖含量最高,分别为38.95 mg/g和45.43 mg/g。在绿色胚性培养物中,蔗糖含量最高,为62.15 mg/g。绿色胚的绿原酸含量最高(5.3 mg/g),棕色胚的咖啡因含量最高(0.55 mg/g)。内源性果糖和葡萄糖抑制胚胎发生,而蔗糖、绿原酸和咖啡因促进胚胎发生,是潜在的胚胎发生生物标志物。其他生化化合物,如有机酸,应该被鉴定出来,并确定它们在咖啡体细胞胚胎发生中的作用。
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