通过体细胞胚胎发生和生物反应器技术快速繁殖两个阿拉比卡杂交咖啡品种 F1 Centroamericano (H1) 和 Mundo Maya (H16) 的研究

N. G. Khong, Thi Nhu Le
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摘要

本文的研究重点是确定一些成分和培养条件,以优化使用生物反应器对两个阿拉比卡 F1 咖啡品种 H1 和 H16 进行体细胞胚胎发生和胚胎再生的 4 个主要步骤,包括:(1) 胚胎发生胼胝体的形成;(2) 胚胎发生细胞的繁殖;(3) 胚胎分化;(4) 使用临时浸泡生物反应器 RITA 1L 进行体细胞胚胎再生。结果表明,在添加了 Kinetin 2 mg/ll 和 Phytagel 4 g/l 的培养基上,H1 和 H16 两个品种的胼胝体形成率分别达到 94.2%和 92.1%。此外,在培养基中添加甘氨酸 20 毫克/升,两个品种的胚性胼胝体形成率均提高了 58.1%。在胚分化培养基中添加 0.03 毫克/升的 Trichostatin A,在培养的前 7 天,鱼雷胚的数量从 1 克初始胼胝体增加到 H1 的 1287 个和 H16 的 1635 个。最后,使用生物反应器 1L-RITA,每 6 小时浸泡 1 分钟,H1 的再生效率达到 75.02%,H16 的再生效率达到 82.03%。
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Study on rapid multiplication of two hybrid Arabica coffee varieties F1 Centroamericano (H1) and Mundo Maya (H16) through somatic embryogenesis and Bioreactor technology
In this paper, the research focuses on determining some components and culture conditions in order to optimise the 4 main steps of somatic embryogenesis and embryo regeneration by Bioreactor of two Arabica F1 coffee varieties H1 and H16, including: (1) formation of embryogenic callus, (2) multiplication of embryogenic cells, (3) embryonic differentiation, and (4) regeneration of somatic embryo using the temporary immersion Bioreactor RITA 1L. The results showed that on the medium supplemented with Kinetin 2 mg/ll and Phytagel 4 g/l, the callus formation rate reached 94.2 and 92.1% for 2 varieties H1 and H16, respectively. Besides, the addition of Glycine 20 mg/l to the culture medium increased the embryogenic callus rate by 58.1% in both varieties. Adding Trichostatin A 0.03 mg/l in the embryo differentiation medium in the first 7 days of culture showed that the number of torpedo embryos was scored from 1g of initial callus up to 1287 in H1 and 1635 in H16. Finally, using Bioreactor 1L-RITA, soaking for 1 min every 6 hours, the regeneration efficiency reached 75.02% in H1 and 82.03% in H16.
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