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Analysis of propagation of Bacopa monnieri (L.) from hairy roots, elicitation and Bacoside A contents of Ri transformed plants

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Abstract

Agrobacterium rhizogenes mediated transformation has been experimented in leaf explants of the memory herb Bacopa monnieri in order to assess the regeneration potential of hairy roots (HR) followed by the elicitation of transformed plants for increased Bacoside A production. Out of the four strains tested, A4 and MTCC 532 derived HR exhibited regrowth in MS basal medium while MTCC 2364 derived HR showed regeneration in MS medium supplemented with suitable phyto hormones. R1000 derived HR possessed no regeneration potential. Comparable to A4, MTCC 532 derived HR displayed maximum regrowth frequency of about 85.71 ± 1.84 % with an increase in biomass to threefold. Therefore, five HR plant lines (MTCC 532 derived) were generated and maintained in MS basal liquid medium in which HR3 topped the others in producing a huge biomass of about 67.09 ± 0.66 g FW. PCR amplification and southern hybridization analysis of rol A gene (280 bp) has been performed in order to confirm the transformation process. Moreover, HR3 plant line has accumulated highest total phenolic content of about 165.68 ± 0.82 mg GAE/g DW and highest total flavonoid content of about 497.78 ± 0.57 mg QRE/g DW when compared to other lines and untransformed controls. In addition, HR3 plant extract showed 85.58 ± 0.14 % of DPPH (2, 2-diphenyl-1-picryl hydrazyl) inhibition displaying its reliable anti oxidant potential. Further on elicitation with 10 mg/L chitosan for 2 weeks, HR3 has produced 5.83 % of Bacoside A which is fivefold and threefold increased production when compared to untransformed and transformed unelicited controls respectively. This is the first report on eliciting HR plants for increased metabolite accumulation in B. monnieri.

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Acknowledgments

The first and the second author greatly acknowledge the University Grants Commission (UGC) for awarding financial support in the form of Basic Science Research Fellowship (UGC order no. F.4-1/2006 (BSR)/7-326/2011(BSR), dated 25.02.2013). We sincerely thank Prof. A. Ganapathy, Department of biotechnology, Bharathidasan University, Tiruchirapalli for providing us the A. rhizogenes strains used in this study. All authors thank the Department of Biotechnology (DBT), Government of India (Grant No. BT/BI/25/001/2006) for the Computational and Bioinformatics Facility provided by the Alagappa University Bioinformatics Infrastructure Facility.

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Correspondence to Manikandan Ramesh.

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Largia, M.J.V., Satish, L., Johnsi, R. et al. Analysis of propagation of Bacopa monnieri (L.) from hairy roots, elicitation and Bacoside A contents of Ri transformed plants. World J Microbiol Biotechnol 32, 131 (2016). https://doi.org/10.1007/s11274-016-2083-7

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