Improved Agrobacterium rhizogenes-mediated hairy root culture system of Withania somnifera (L.) Dunal using sonication and heat treatment

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• 作者：Chandrasekaran Thilip ; Chellappan Soundar Raju ; Kandhan Varutharaju…
• 关键词：Agrobacterium rhizogenes ; Withania somnifera ; Hairy roots ; Withanolides ; Sonication ; Heat treatment
• 刊名：3 Biotech
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：5
• 期：6
• 页码：949-956
• 全文大小：1,274 KB
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• 作者单位：Chandrasekaran Thilip (1)
  Chellappan Soundar Raju (1)
  Kandhan Varutharaju (1)
  Abubakker Aslam (1)
  Appakan Shajahan (1)
  
  1. Plant Molecular Biology Laboratory, Department of Botany, Jamal Mohamed College, Tiruchirappalli, 620 020, Tamil Nadu, India
  
• 刊物主题：Biotechnology; Agriculture; Cancer Research; Bioinformatics; Stem Cells; Biomaterials;
• 出版者：Springer Berlin Heidelberg
• ISSN：2190-5738

文摘

An improved Agrobacterium rhizogenes-mediated genetic transformation of Withania somnifera (L.) Dunal was developed using the bacterial strain R1000 with leaf segment explants of in vitro raised plantlets. Out of the three strains used (R1000, MTCC 2364 and MTCC 532), the strain R1000 proved to be more efficient than others. Among the different conditions tested, the highest (93.3 %) transformation rate was observed after 3 weeks when the explants were subjected to sonication (15 s) and heat treatment (41 °C for 5 min). Transgenic status of the hairy roots was confirmed by PCR using rol B-specific primers. HPLC analysis showed the ability of hairy roots to synthesize withaferin A and withanolide A, both steroidal lactones of medicinal value. This protocol offers new avenue in A. rhizogenes-mediated hairy root induction and is useful for large-scale production of these bioactive compounds from W. somnifera. Keywords Agrobacterium rhizogenes Withania somnifera Hairy roots Withanolides Sonication Heat treatment