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茉莉酸甲酯和水杨酸诱导条件的优化提高了睡茄(L.)Dunal 不定根培养物中生物堿的产量。

Optimization of elicitation conditions with methyl jasmonate and salicylic acid to improve the productivity of withanolides in the adventitious root culture of Withania somnifera (L.) Dunal.

机构信息

Department of Biotechnology and Genetic Engineering, School of Biotechnology, Bharathidasan University, Tiruchirappalli 620-024 Tamil Nadu, India.

出版信息

Appl Biochem Biotechnol. 2012 Oct;168(3):681-96. doi: 10.1007/s12010-012-9809-2. Epub 2012 Jul 28.

Abstract

Adventitious root cultures derived from leaf derived callus of Withania somnifera (L.) Dunal were treated with methyl jasmonate and salicylic acid independently. Biomass accumulation, culture age, elicitation period, and culture duration were optimized for higher withanolides production in the two best-responding varieties collected from Kolli hills (Eastern Ghats) and Cumbum (Western Ghats) of Tamil Nadu, India. Between the two elicitors, salicylic acid (SA) improved the production of major withanolides (withanolide A, withanolide B, withaferin A, and withanone) as well as minor constituents (12-deoxy withastramonolide, withanoside V, and withanoside IV) in the Kolli hills variety. Treatment of root biomass (11.70 g FW) on 30-day-old adventitious root cultures with 150 μM SA for 4 h elicitor exposure period resulted in the production of 64.65 mg g(-l) dry weight (DW) withanolide A (48-fold), 33.74 mg g(-l) DW withanolide B (29-fold), 17.47 mg g(-l) DW withaferin A (20-fold), 42.88 mg g(-l) DW withanone (37-fold), 5.34 mg g(-l) DW 12-deoxy withastramonolide (nine fold), 7.23 mg g(-l) DW withanoside V (seven fold), and 9.45 mg g(-l) DW withanoside IV (nine fold) after 10 days of elicitation (40th day of culture) when compared to untreated cultures. This is the first report on the use of elicitation strategy on the significant improvement in withanolides production in the adventitious root cultures of W. somnifera.

摘要

从印度泰米尔纳德邦科利山(东高止山脉)和库姆巴姆(西高止山脉)收集的两个最佳响应品种的叶衍生愈伤组织中衍生的不定根培养物分别用茉莉酸甲酯和水杨酸处理。优化生物量积累、培养年龄、诱导期和培养时间,以提高来自印度泰米尔纳德邦科利山(东高止山脉)和库姆巴姆(西高止山脉)两个品种的不定根培养物中生物总含量的生产。在两种诱导剂中,水杨酸(SA)提高了科利山品种中主要生物总含量(生物总含量 A、生物总含量 B、蛇根碱 A 和蛇根酮)以及次要成分(12-脱氧蛇根碱、羽扇豆苷 V 和羽扇豆苷 IV)的产生。用 150 μM SA 处理 30 天龄的不定根培养物的根生物量(11.70 g FW),诱导剂暴露期 4 小时,可产生 64.65 mg g(-1) 干重(DW)生物总含量 A(48 倍)、33.74 mg g(-1) DW 生物总含量 B(29 倍)、17.47 mg g(-1) DW 蛇根碱 A(20 倍)、42.88 mg g(-1) DW 蛇根酮(37 倍)、5.34 mg g(-1) DW 12-脱氧蛇根碱(9 倍)、7.23 mg g(-1) DW 羽扇豆苷 V(7 倍)和 9.45 mg g(-1) DW 羽扇豆苷 IV(9 倍),与未处理的培养物相比,在诱导后 10 天(第 40 天培养)。这是首次报道在蛇根草不定根培养物中使用诱导策略显著提高生物总含量的生产。

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