Maheshwari Priti, Songara Bharti, Kumar Shailesh, Jain Prachi, Srivastava Kamini, Kumar Anil
School of Biotechnology, Devi Ahilya University, Indore, India.
Biotechnol J. 2007 Aug;2(8):1026-32. doi: 10.1002/biot.200700033.
Fast-growing callus, cell suspension and root cultures of Vernonia cinerea, a medicinal plant, were analyzed for the presence of alkaloids. Callus and root cultures were established from young leaf explants in Murashige and Skoog (MS) basal media supplemented with combinations of auxins and cytokinins, whereas cell suspension cultures were established from callus cultures. Maximum biomass of callus, cell suspension and root cultures were obtained in the medium supplemented with 1 mg/L alpha-naphthaleneacetic acid (NAA) and 5 mg/L benzylaminopurine (BA), 1.0 mg/L NAA and 0.1 mg/L BA and 1.5 mg/L NAA, respectively. The 5-week-old callus cultures resulted in maximum biomass and alkaloid contents (750 microg/g). Cell suspension growth and alkaloid contents were maximal in 20-day-old cultures and alkaloid contents were 1.15 mg/g. A 0.2-g sample of root tissue regenerated in semi-solid medium upon transfer to liquid MS medium containing 1.5 mg/L NAA regenerated a maximum increase in biomass of 6.3-fold over a period of 5 weeks. The highest root growth and alkaloid contents of 2 mg/g dry weight were obtained in 5-week-old cultures. Maximum alkaloid contents were obtained in root cultures in vitro compared to all others including the alkaloid content of in vivo obtained with aerial parts and roots (800 microg/g and 1.2 mg/g dry weight, respectively) of V. cinerea.
对药用植物伤寒草的快速生长愈伤组织、细胞悬浮培养物和根培养物进行了生物碱检测。愈伤组织和根培养物是由幼叶外植体在添加了生长素和细胞分裂素组合的Murashige和Skoog(MS)基础培养基中建立的,而细胞悬浮培养物则是从愈伤组织培养物中建立的。愈伤组织、细胞悬浮培养物和根培养物的最大生物量分别在添加1 mg/Lα-萘乙酸(NAA)和5 mg/L苄氨基嘌呤(BA)、1.0 mg/L NAA和0.1 mg/L BA以及1.5 mg/L NAA的培养基中获得。5周龄的愈伤组织培养物产生了最大生物量和生物碱含量(750μg/g)。细胞悬浮培养物的生长和生物碱含量在20日龄培养物中最高,生物碱含量为1.15 mg/g。将0.2 g在半固体培养基中再生的根组织转移到含有1.5 mg/L NAA的液体MS培养基中,在5周内生物量最大增加了6.3倍。在5周龄培养物中获得了最高的根生长和2 mg/g干重的生物碱含量。与所有其他部分相比,包括伤寒草地上部分和根的体内生物碱含量(分别为800μg/g和1.2 mg/g干重),体外根培养物获得了最大生物碱含量。