Baldi Ashish, Srivastava Ashok K, Bisaria Virendra S
Plant Cell Culture Laboratory, Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, New Delhi, India.
Biotechnol J. 2008 Oct;3(9-10):1256-63. doi: 10.1002/biot.200800173.
Various cell and hairy root cultures of L. album were developed and analyzed for podophyllotoxin content. Transformed callus and hairy root cultures developed from infection of stem portions of in vitro-germinated L. album plant with Agrobacterium rhizogenes NCIM 5140 strain were selected on the basis of high podophyllotoxin content and growth. Based on the integration of Ri T(L)-DNA and T(R)-DNA, integration of only the ags and not the rol gene in transformed cell culture indicated fragmented integration pattern. The effect of different cultivation media and carbon source on growth and podophyllotoxin production were studied in shake-flask suspension cultures. Detailed batch growth and production kinetics with sugar consumption profile were also established. Maximum volumetric productivity of 4.40 and 2.75 mg/L per day was obtained in cell suspension and hairy root cultures, respectively.
开发了白刺花的各种细胞和毛状根培养物，并分析了鬼臼毒素含量。基于高鬼臼毒素含量和生长情况，选择了由发根农杆菌NCIM 5140菌株感染体外萌发的白刺花植株茎段而产生的转化愈伤组织和毛状根培养物。基于Ri T(L)-DNA和T(R)-DNA的整合，转化细胞培养物中仅ags基因而非rol基因的整合表明存在片段化整合模式。在摇瓶悬浮培养中研究了不同培养基和碳源对生长和鬼臼毒素产生的影响。还建立了详细的分批生长、生产动力学以及糖消耗曲线。细胞悬浮培养和毛状根培养中每天的最大体积产率分别为4.40和2.75 mg/L。
