Al-Shara Baker, Taha Rosna Mat, Mohamad Jamaludin, Elias Hashimah, Khan Asif
Institute of Biological Sciences, Faculty of Science, University Malaya, Kuala Lumpur 50603, Malaysia.
Faculty of Resource Science and Technology, University Malaysia Sarawak (UNIMAS), Kota Samarahan 94300, Sarawak, Malaysia.
Plants (Basel). 2020 Mar 12;9(3):360. doi: 10.3390/plants9030360.
A highly efficient protocol for regeneration of L. cv. Eksotika somatic embryos from immature zygotic embryos was developed. This study was designed to overcome the obstacles in regeneration of somatic embryos from immature zygotic embryos of "Eksotika", especially problems associated with formation of better root quality and callus formation at the base of somatic embryos. Somatic embryos were generated by incubation of immature zygotic embryos in half-strength salt Murashige and Skoog (MS) medium with full-strength vitamins supplemented with 7.5 mg L 2,4-D, 100 mg L L-glutamine, 50 mg L -inositol, 45 mg L adenine sulphate, 0.33% gelrite, and 6% sucrose, followed by transfer to maturation medium consisting of ½ MS medium supplemented with 5 mg L phloroglucinol, 100 mg L L-glutamine, 100 mg L -inositol, 68 mg L adenine sulphate, 0.38% gelrite, and 3% sucrose. After that, well-formed somatic embryos were transferred to MS medium containing 3% sucrose and 0.8% agar for shoot production. The embryos were elongated in MS medium supplemented with 1 mg L gibberellic acid, 0.5 mg L indole-3-butyric acid, 100 mg L -inositol, and 3.76 mg L riboflavin. Root regeneration was achieved on MS medium containing 7.9 mg L phloroglucinol and supported with vermiculite after 4 days of cultivation on ½ MS medium with 2 mg L indole-3-butyric acid. After the rooting phase, in vitro plantlets were acclimatized in peat moss soil.
开发了一种从未成熟合子胚高效再生“Eksotika”番荔枝体细胞胚的方案。本研究旨在克服从“Eksotika”未成熟合子胚再生体细胞胚过程中的障碍，特别是与形成更好的根质量以及体细胞胚基部愈伤组织形成相关的问题。通过将未成熟合子胚在含有全量维生素的半强度盐Murashige和Skoog（MS）培养基中培养来产生体细胞胚，该培养基添加了7.5 mg/L 2,4 -二氯苯氧乙酸、100 mg/L L -谷氨酰胺、50 mg/L肌醇、45 mg/L硫酸腺嘌呤、0.33%吉丽特和6%蔗糖，随后转移至成熟培养基，该成熟培养基由添加了5 mg/L间苯三酚、100 mg/L L -谷氨酰胺、100 mg/L肌醇、68 mg/L硫酸腺嘌呤、0.38%吉丽特和3%蔗糖的½ MS培养基组成。之后，将发育良好的体细胞胚转移至含有3%蔗糖和0.8%琼脂的MS培养基上进行芽的诱导。胚在添加了1 mg/L赤霉素、0.5 mg/L吲哚 - 3 -丁酸、100 mg/L肌醇和3.76 mg/L核黄素的MS培养基中伸长。在含有7.9 mg/L间苯三酚的MS培养基上，经过在含有2 mg/L吲哚 - 3 -丁酸的½ MS培养基上培养4天后，在蛭石的支持下实现了根的再生。生根阶段结束后，将试管苗在泥炭藓土壤中进行驯化。
