Srivastava Mrinalini, Sharma Swati, Misra Pratibha
Tissue Culture and Transformation Lab, Council of Scientific and Industrial Research - National Botanical Research Institute, Lucknow, Uttar Pradesh, India; Department of Biosciences, Integral University, Lucknow, Uttar Pradesh, India.
Department of Biosciences, Integral University, Lucknow, Uttar Pradesh, India.
Pharmacogn Mag. 2016 May;12(Suppl 3):S315-20. doi: 10.4103/0973-1296.185726.
Rauwolfia serpentina and Solanum khasianum are well-known medicinally important plants contained important alkaloids in their different parts. Elicitation of these alkaloids is important because of associated pharmaceutical properties. Targeted metabolites were ajmaline and ajmalicine in R. serpentina; solasodine and α-solanine in S. khasianum.
Enhancement of secondary metabolites through biotic and abiotic elicitors in hairy root cultures of R. serpentina and S. khasianum.
In this report, hairy root cultures of these two plants were established through Agrobacterium rhizogenes mediated transformation by optimizing various parameters as age of explants, duration of preculture, and co-cultivation period. NaCl was used as abiotic elicitors in these two plants. Cellulase from Aspergillus niger was used as biotic elicitor in S. khasianum and mannan from Saccharomyces cerevisiae was used in R. serpentina.
First time we have reported the effect of biotic and abiotic elicitors on the production of important metabolites in hairy root cultures of these two plants. Ajmalicine production was stimulated up to 14.8-fold at 100 mM concentration of NaCl after 1 week of treatment. Ajmaline concentration was also increased 2.9-fold at 100 mg/l dose of mannan after 1 week. Solasodine content was enhanced up to 4.0-fold and 3.6-fold at 100 mM and 200 mM NaCl, respectively, after 6 days of treatments.
This study explored the potential of the elicitation strategy in A. rhizogenes transformed cell cultures and this potential further used for commercial production of these pharmaceutically important secondary metabolites.
Hairy roots of Rauwolfia serpentina were subjected to salt (abiotic stress) and mannan (biotic stress) treatment for 1 week. Ajmaline and ajmalicine secondary metabolites were quantified before and after stress treatmentAjmalicine yield was enhanced up to 14.8-fold at 100 mM concentration of NaCl. Ajmaline content was also stimulated 2.9-fold at 100 mg/l dose of mannan after 1 weekHairy roots of Solanum khasianum were treated with cellulase (biotic elicitor) and salt (abiotic stress)Solasodine content was improved up to 4.0-fold and 3.6-fold at 100 mM and 200 mM NaCl, respectively, after 6.days of treatmentsThe α-solanine content increased to 1.6-fold after 24 h of treatment at 100 μg/mL cellulase concentration. Abbreviations used: MS medium: Murashige and Skoog medium, B5 medium: Gamborg B5 medium, OD: Optical Density, NaCl: Sodium Chloride.
蛇根木和卡西茄是著名的具有重要药用价值的植物,其不同部位含有重要生物碱。由于这些生物碱具有相关的药用特性,诱导其产生具有重要意义。蛇根木中的目标代谢产物是阿吗灵和阿吗碱;卡西茄中的是茄解碱和α-茄碱。
通过生物和非生物诱导剂提高蛇根木和卡西茄毛状根培养物中次生代谢产物的含量。
在本报告中,通过发根农杆菌介导的转化,优化外植体年龄、预培养时间和共培养时间等各种参数,建立了这两种植物的毛状根培养物。氯化钠用作这两种植物的非生物诱导剂。黑曲霉的纤维素酶用作卡西茄的生物诱导剂,酿酒酵母的甘露聚糖用作蛇根木的生物诱导剂。
我们首次报道了生物和非生物诱导剂对这两种植物毛状根培养物中重要代谢产物产量的影响。处理1周后,在100 mM氯化钠浓度下,阿吗碱产量提高了14.8倍。处理1周后,在100 mg/l甘露聚糖剂量下,阿吗灵浓度也增加了2.9倍。处理6天后,在100 mM和200 mM氯化钠浓度下,茄解碱含量分别提高了4.0倍和3.6倍。
本研究探索了发根农杆菌转化细胞培养物中诱导策略的潜力,该潜力可进一步用于这些具有重要药用价值的次生代谢产物的商业生产。
蛇根木的毛状根接受盐(非生物胁迫)和甘露聚糖(生物胁迫)处理1周。在胁迫处理前后对阿吗灵和阿吗碱次生代谢产物进行定量。在100 mM氯化钠浓度下,阿吗碱产量提高了14.8倍。处理1周后,在100 mg/l甘露聚糖剂量下,阿吗灵含量也提高了2.9倍。卡西茄的毛状根用纤维素酶(生物诱导剂)和盐(非生物胁迫)处理。处理6天后,在100 mM和200 mM氯化钠浓度下,茄解碱含量分别提高了4.0倍和3.6倍。在100 μg/mL纤维素酶浓度下处理24小时后,α-茄碱含量增加到1.6倍。使用的缩写:MS培养基:Murashige和Skoog培养基,B5培养基:Gamborg B5培养基,OD:光密度,NaCl:氯化钠。