Response of Flavonoids Production to Some Elicitors in Callus and Cell Suspension Culture of Ginkgo (Ginkgo biloba L.)

S. Sherif, M. Essam, Reem H. I. Hassan
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Abstract

Aims: This study was conducted to investigate how to maximize the production of flavonoids from ginkgo leaves callus or cell suspension cultures. Study Design: The experiment was implemented in a completely randomized design with 3 replicates. Place and Duration of Study: The study was carried out at the Research Laboratory of Tissue Culture and Preservation of Germplasm, Horticulture Research Institute, Agric. Res. Center, Egypt from 2020 to 2022.  Methodology: Adenine sulfate and pyruvic acid were added at concentrations of 0.1, 0.2, and 0.3 mg/l, for each. Cadmium and lead acetate were added (2 and 4 mg/l for each) to the cell suspension. Jasmonic acid (15, 50, and 100 µM), AgNO3 (25, 50, and 75 µM) and tryptophan (50, 100, and 200 mg/l) were used in the hard callus culture. Results: Results indicated that the maximum cell suspension fresh weight (6.50 g) was obtained from (0.5 g) cell suspension grown on MS medium supplemented with adenine 0.2 mg/l while lead acetate at 2.0 mg/l increased dry weight (0.89g) after the second subculture. The maximum packed cell volume (PCV) (24.33%) was obtained from lead acetate at 2.0 mg/l in the second cycle of growth. The best flavonoid production (91.29 mg/100 g DW) was produced in the second cycle of growth from callus treated with AgNO3 at 25 µM. Treating Ginkgo biloba callus cultured with jasmonic acid at 100 µM increased the inhibition of AChE in the serum of AD patients from 235.2 U/l (untreated serum) to 76.08 U/l at 38.71 ug/ml compared with the Ginkgo biloba ® at 260 mg which inhibit the activity from 235.2 U/l to 55.73 U/l at 26.00 ug/ml at amount 25.00 ul for all. Conclusion: Ginkgo biloba is a valuable source of anticholinesterase agents and could be used in pharmaceutical preparations by using callus culture technique treated with jasmonic acid and AgNO3.
银杏(Ginkgo biloba L.)愈伤组织和细胞悬浮培养中黄酮类化合物产生对某些激发子的响应
目的:研究银杏叶片愈伤组织和细胞悬浮培养中黄酮类化合物的最大产量。研究设计:试验采用完全随机设计,设3个重复。研究地点和时间:本研究在农业部园艺研究所种质资源组织培养与保存研究实验室进行。从2020年到2022年,埃及的Res. Center。方法:分别加入浓度为0.1、0.2、0.3 mg/l的硫酸腺嘌呤和丙酮酸。细胞悬浮液中分别加入2 mg/l和4 mg/l的镉和醋酸铅。茉莉酸(15、50和100µM), AgNO3(25、50和75µM)和色氨酸(50、100和200 mg/l)分别用于硬愈伤组织培养。结果:在腺嘌呤浓度为0.2 mg/l的MS培养基上培养的细胞悬浮液(0.5 g)鲜重最大(6.50 g),而在2.0 mg/l的乙酸铅浓度下,二次传代后细胞干重增加(0.89g)。2.0 mg/l的乙酸铅在第2次循环中获得了最大的堆积细胞体积(PCV)(24.33%)。25µM AgNO3处理后,愈伤组织生长第二周期的黄酮类化合物产量最高,为91.29 mg/100 g DW。用100µM茉莉酸处理银杏叶愈伤组织培养的AD患者血清中AChE的抑制作用从235.2 U/l(未处理血清)增加到76.08 U/l (38.71 ug/ml),而用260 mg银杏叶酸处理的AD患者血清中AChE的抑制作用从235.2 U/l增加到55.73 U/l (26.00 ug/ml, 25.00 ul)。结论:银杏叶是抗胆碱酯酶活性物质的重要来源,茉莉酸和AgNO3处理银杏可用于银杏愈伤组织培养技术。
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