Abstract
The marshmallow plant (Althaea officinalis L.) has been used for centuries in medicine and other applications. Valuable secondary metabolites have previously been identified in Agrobacterium rhizogenes-generated transgenic ‘hairy’ roots in this species. In the present study, transgenic roots were produced in A. officinalis using A. rhizogenes. In addition to wild-type lines, roots expressing the anti-human immunodeficiency virus microbicide candidate, cyanovirin-N (CV-N), were generated. Wild-type and CV-N root lines were transferred to liquid culture and increased in mass by 49 and 19 % respectively over a 7 day culture period. In the latter, the concentration of CV-N present in the root tissue was 2.4 μg/g fresh weight, with an average secretion rate into the growth medium of 0.02 μg/ml/24 h. A. officinalis transgenic roots may therefore in the future be used not only as a source of therapeutic secondary metabolites, but also as an expression system for the production of recombinant pharmaceuticals.
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References
Basch E, Ulbricht C, Hammerness P, Vora M (2003) Marshmallow (Althaea officinalis L.) monograph. J Herbal Pharmacother 3:71–81
Boyd MR, Gustafson KR, McMahon JB, Shoemaker RH, Okeefe BR, Mori T, Gulakowski RJ, Wu L, Rivera MI, Laurencot CM, Currens MJ, Cardellina JH, Buckheit RW, Nara PL, Pannell LK, Sowder RC, Henderson LE (1997) Discovery of cyanovirin-N, a novel human immunodeficiency virus-inactivating protein that binds viral surface envelope glycoprotein gp120: potential applications to microbicide development. Antimicrob Agents Ch 41:1521–1530
Buffa V, Stieh D, Mamhood N, Hu Q, Fletcher P, Shattock RJ (2009) Cyanovirin-N potently inhibits human immunodeficiency virus type 1 infection in cellular and cervical explant models. J Gen Virol 90:234–243
Drake PMW, Barbi T, Sexton A, McGowan E, Stadlmann J, Navarre C, Ma J-K (2009) Development of rhizosecretion as a production system for recombinant proteins from hydroponic cultivated tobacco. FASEB J 23:3581–3589
Elghabi Z, Karcher D, Zhou F, Ruf S, Bock R (2011) Optimization of the expression of the HIV fusion inhibitor cyanovirin-N from the tobacco plastid genome. Plant Biotechnol J 9:599–608
Ghanem ME, Han RM, Classen B, Quetin-Leclerq J, Mahy G, Ruan CJ, Qin P, Perez-Alfocea F, Lutts S (2010) Mucilage and polysaccharides in the halophyte plant species Kosteletzkya virginica: localization and composition in relation to salt stress. J Plant Physiol 167:382–392
Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general-method for transferring genes into plants. Science 227:1229–1231
Hu Q, Mahmood N, Shattock RJ (2007) High-mannose-specific deglycosylation of HIV-1 gp120 induced by resistance to cyanovirin-N and the impact on antibody neutralization. Virology 368:145–154
Ionkova I (1992) Alternative ways for the production of biologically active substances from Althaea officinalis var. russalka. CR Acad Bulg Sci 45:137–140
Ionkova I, Hu Z, Alfermann A (1993) Polysaccharide production by hairy root cultures of some higher medicinal plants. Planta Med 59:A658–A659
Khidyrova N, Van EV, Shakhidoyatov R, Bobakulov K, Abdullaev N, Shakhidoyatov K (2012) Polyprenols from leaves and stems of the plant Althaea officinalis. Chem Nat Compd 48:358–360
Mori T, Barrientos LG, Han ZZ, Gronenborn AM, Turpin JA, Boyd MR (2002) Functional homologs of cyanovirin-N amenable to mass production in prokaryotic and eukaryotic hosts. Protein Express Purif 26:42–49
Murashige T, Skoog F (1962) A revised medium for rapid growth bioassays with tobacco tissue cultures. Plant Physiol 15:473–497
Naz R, Anis M (2012) Acceleration of adventitious shoots by interaction between exogenous hormone and adenine sulphate in Althaea Officinalis L. Appl Biochem Biotech 168:1239–1255
Sexton A, Drake PM, Mahmood N, Harman SJ, Shattock RJ, Ma JK-C (2005) Transgenic plant production of cyanovirin-N, an HIV microbicide. FASEB J 20:356–358
Shah S, Akhtar N, Akram M, Shah PA, Saeed T, Ahmed K, Asif H (2011) Pharmacological activity of Althaea officinalis L. J Med Plants Res 5:5662–5666
Shenoy SR, O’Keefe BR, Bolmstedt AJ, Cartner LK, Boyd MR (2001) Selective interactions of the human immunodeficiency virus-inactivating protein cyanovirin-N with high-mannose oligosaccharides on gp120 and other glycoproteins. J Pharmacol Exp Ther 297:704–710
Tsai CC, Emau P, Jiang YH, Tian BP, Morton WR, Gustafson KR, Boyd MR (2003) Cyanovirin-N gel as a topical microbicide prevents rectal transmission of SHIV89.6P in macaques. Aids Res Human Retrovir 19:535–541
Tsai CC, Emau P, Jiang YH, Agy MB, Shattock RJ, Schmidt A, Morton WR, Gustafson KR, Boyd MR (2004) Cyanovirin-N inhibits AIDS virus infections in vaginal transmission models. Aids Res Human Retrovir 20:11–18
Witvrouw M, Fikkert V, Hantson A, Pannecouque C, O’Keefe BR, McMahon J, Stamatatos L, de Clercq E, Bolmstedt A (2005) Resistance of human immunodeficiency virus type 1 to the high-mannose binding agents cyanovirin N and concanavalin A. J Virol 79:7777–7784
Acknowledgments
We thank Nigel Dungey for supplying Althaea officinalis seed and Giorgio De Guzman for supplying A. rhizogenes LBA 9402. This project was jointly funded by the Comopharm European Union FP7 project grant, EU COST ACTION FA0804 and the Sir Joseph Hotung Endowment.
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Pascal M. W. Drake and Luisa de Moraes Madeira contributed equally to this work.
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Drake, P.M.W., de Moraes Madeira, L., Szeto, T.H. et al. Transformation of Althaea officinalis L. by Agrobacterium rhizogenes for the production of transgenic roots expressing the anti-HIV microbicide cyanovirin-N. Transgenic Res 22, 1225–1229 (2013). https://doi.org/10.1007/s11248-013-9730-7
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DOI: https://doi.org/10.1007/s11248-013-9730-7