High-frequency transgenic plant regeneration and plumbagin production through methyl jasmonate elicitation from hairy roots of <Emphasis Type="Italic">Plumbago indica</Emphasis> L. |
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Authors: | Kottackal Poulose Martin Aneta Sabovljevic Joseph Madassery |
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Institution: | (1) Medicinal Plant Laboratory, Department of Botany, Bose Institute, 93/1 APC Road, Kolkata, West Bengal, 700009, India;(2) Plant Molecular and Cellular Genetics, Bose Institute, Centenary Campus, P 1/12 CIT Road, Scheme VIIM, Kolkata, West Bengal, 700054, India;(3) Department of Genetics, Bidhan Chandra Krishi Visyavidyalaya, Mohonpur, Nadia, West Bengal, 741252, India;(4) Present address: Department of Bioscience and Biotechnology, Bansthali Vidyapith, Banasthali, Rajasthan, 304022, India |
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Abstract: | High-frequency transgenic plant regeneration and production of plumbagin were accomplished from hairy roots induced by Agrobacterium rhizogenes strain A4M70GUS on Plumbago indica L. Of the two types of hairy roots developed on Murashige and Skoog (MS) basal medium, Type I was long and thick with lesser
branches and root hairs, and Type II was highly-branched short and slender with tufts of root hairs. Of the different lines
grown in half-strength MS liquid basal media, root line 5 (R5) of the Type I yielded the highest plumbagin (0.92% DW) and
was significantly different to that of the in vitro control. R5 showed a stable production of plumbagin (1.09% DW) in subsequent cultures. Elicitation of R5 with 50 μM methyl
jasmonate for 48 h increased the yield of plumbagin to 5.0% DW, and was superior to 100 μM acetylsalicylic acid (3.8% DW).
Plumbagin yield was five times that of twoyear-old ex vitro roots. Histochemical assay and PCR analysis using the primers of uidA coding region confirmed the transformation. Hairy root segments cultured on MS medium containing 8.8 μM benzyladenine and
2.5 μM indole-3-butyric acid induced a mean of 9.1 shoots. Subsequent culture of the isolated shoots developed more than 50
normal shoots per culture. The root-free shoots were rooted on half-strength MS basal medium. The plantlets transferred in
field conditions grew normally and exhibited 90% survival. Transgenic plant regeneration and hairy root induction in P. indica serves as reliable source of plumbagin which in turn cut off the mass destruction of the plant species. |
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