Cultivation Optimization and Biological Active Substances Activation Under In Vitro Callus Culture Conditions of Basil (Ocimum Basilicum)

Sergey L. Tikhonov, Maxim N. Harapaev
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Abstract

Callus cell cultures have a wide range of applications in the food industry, pharmacology and pharmacy, agriculture and biotechnology. The study aim is to develop an optimal technology for the cultivation and biological substances activation under in vitro callus culture conditions of basil (Ocimum basilicum). At the first research stage, a man developed a nutrient medium for culturing callus culture including, by weight %: ammonium nitrate – 4.75; potassium nitrate – 5.47; calcium chloride 2-aqueous – 1.27; magnesium sulfate 7-aqueous – 1.06; potassium dihydroorthophosphate – 0.49; EDTA disodium salt – 0.1; iron sulfate 7-aqueous – 0.08; boric acid – 0.02; magnesium sulfate 4-aqueous – 0.06; zinc sulfate 7-aqueous – 0.02; potassium iodide – 0.002; sodium molybdate 2-aqueous – 0.001; copper sulfate 5-aqueous – 0.001; cobalt chloride 2-aqueous – 0.001; glycine – 0.01; mesoinosite – 0.3; nicotinic acid – 0.001; pyridoxine – 0.002; sucrose – 86.35; benzylaminopurine – 0.01; naphthylacetic acid – 0.003. The second stage consists of a technology development for activating biological active substances of callus culture when exposed to biomass with blue light intensity of 1500 lux (Samsung 281b Quantum Line LEDs) in light/dark mode 24/24 h. As a result, the basil biomass yield was (24.44 ± 1.00) g/l. The number of secondary metabolites was, mg/g of dry weight: rosemary acid (54.5 ± 2.0); chicory acid (64.4 ± 1.9); eugenol (0.50 ± 0.100); caffeic acid (0.42 ± 0.1). The developed technology of cultivation and biological substances activation enables to obtain callus culture samples of high-quality ordinary basil regardless of various environmental factors, geographical restrictions and seasonal climate changes. The practical research results are of interest to the food industry.
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罗勒(Ocimum Basilicum)愈伤组织培养条件下的培养优化及生物活性物质活化
愈伤组织细胞培养在食品工业、药理学、药学、农业和生物技术等领域有着广泛的应用。本研究旨在探索罗勒愈伤组织体外培养条件下的最佳培养工艺及生物物质的活化。在第一个研究阶段,一个人开发了一种培养愈伤组织的营养培养基,按重量%计:硝酸铵- 4.75;硝酸钾- 5.47;氯化钙2-水溶液- 1.27;硫酸镁7水- 1.06;正磷酸二氢钾- 0.49;EDTA二钠- 0.1;硫酸铁7水- 0.08;硼酸- 0.02;硫酸镁- 0.06;硫酸锌水溶液- 0.02;碘化钾- 0.002;钼酸钠2-水溶液- 0.001;硫酸铜5-水溶液- 0.001;氯化钴2-水溶液- 0.001;甘氨酸- 0.01;中肌酶- 0.3;烟酸- 0.001;吡哆醇- 0.002;蔗糖- 86.35;苄氨基嘌呤- 0.01;萘乙酸- 0.003。第二阶段是在光/暗模式下,在蓝光强度为1500 lux (Samsung 281b Quantum Line LEDs)的生物质照射下,24/24小时激活愈伤组织培养的生物活性物质的技术开发。结果表明,罗勒的生物量产量为(24.44±1.00)g/l。次生代谢物的数量为,mg/g干重:迷迭香酸(54.5±2.0);菊苣酸(64.4±1.9);丁香酚(0.50±0.100);咖啡酸(0.42±0.1)。发达的培养和生物物质活化技术,使普通罗勒的愈伤组织培养样品不受各种环境因素、地理限制和季节气候变化的影响。实际研究成果对食品工业具有重要意义。
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