Enhanced accumulation of high-value deoxymiroestrol and isoflavonoids using hairy root as a sustainable source of Pueraria candollei var. mirifica
- 153 Downloads
Pueraria candollei var. mirifica (PM), a rejuvenating herb of Thailand, has been applied efficaciously to relieve symptoms of estrogen deficiency in various clinical trials. However, there is a shortage of the natural source of the plant because of high demand. A clone of hairy root was produced by genetic transformation of T-DNA from an Agrobacterium rhizogenes ATCC15834 strain. An in vitro elicitation strategy for PM hairy root was successfully established. The hairy root of PM produced a high amount of deoxymiroestrol [71.9 ± 4.67–77.9 ± 6.63 µg g−1 dry weight (DW)] and isoflavonoids (6.57 ± 0.044–7.05 ± 0.534 mg g−1 DW). These amounts are higher than that obtained from the root of its parental intact PM, which contained deoxymiroestrol (15.41 ± 0.71 µg g−1 DW) and isoflavonoids (3.45 ± 0.21 mg g−1 DW). Interestingly, the treatment of the hairy root with methyl jasmonate (50–200 µM) increased the accumulation of deoxymiroestrol (113 ± 11.4–245 ± 12.0 µg g−1 dry weight) and isoflavonoids (9.94 ± 0.365–16.49 ± 0.466 mg g−1 DW). Therefore, this approach markedly improved the production of PM phytoestrogens. The field cultivation of PM takes at least 1 year to produce a substantial amount of deoxymiroestrol and isoflavonoids; moreover, there are geographical and seasonal variations in phytoestrogen content, influencing the limited supply and quality of PM materials. Therefore, this system of PM hairy root culture and elicitation is quite simple and effective for the production of useful phytoestrogens for phytochemical and cosmeceutical industries. Additionally, this process can be scaled up to the industrial level as a sustainable source of PM.
KeywordsPueraria candollei var. mirifica Elicitation Phytoestrogen Deoxymiroestrol Isoflavonoids
The authors thank the Faculty of Pharmaceutical Sciences, Khon Kaen University, Thailand for their support. The authors thank Dr. Chaiyo Chaichantipyuth, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand for providing deoxymiroestrol and kwakhurin.
The experiments were designed and performed by OU. Data analysis, interpretation, and manuscript preparation were performed by OU and GY. WK performed experiments in the part of an untransformed root culture of PM. The molecular marker confirmation of hairy root genetic transformation was performed by LU. TK and TJ provided advice and technical support for plant tissue culture and HPLC analysis. Grant and research facilities were managed by WP. All authors were involved in the editing of all versions for publication.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Boonsnongcheep P, Korsangruang S, Soonthornchareonnon N, Chintapakorn Y, Saralamp P, Prathanturarug S (2010) Growth and isoflavonoid accumulation of Pueraria candollei var. candollei and P. candollei var. mirifica cell suspension cultures. Plant Cell Tissue Org 101(2):119–126. https://doi.org/10.1007/s11240-010-9668-x CrossRefGoogle Scholar
- Chandeying V, Lamlertkittikul S (2007) Challenges in the conduct of Thai herbal scientific study: efficacy and safety of phytoestrogen, Pueraria mirifica (Kwao Keur Kao), phase I, in the alleviation of climacteric symptoms in perimenopausal women. J Med Assoc Thail 90(7):1274–1280Google Scholar
- Chandeying V, Sangthawan M (2007) Efficacy comparison of Pueraria mirifica (PM) against conjugated equine estrogen (CEE) with/without medroxyprogesterone acetate (MPA) in the treatment of climacteric symptoms in perimenopausal women: phase III study. J Med Assoc Thail 90(9):1720–1726Google Scholar
- Danphitsanuparn P, Boonsnongcheep P, Boriboonkaset T, Chintapakorn Y, Prathanturarug S (2012) Effects of Agrobacterium rhizogenes strains and other parameters on production of isoflavonoids in hairy roots of Pueraria candollei Grah. ex Benth. var. candollei. Plant Cell Tissue Org 111(3):315–322. https://doi.org/10.1007/s11240-012-0196-8 CrossRefGoogle Scholar
- Jeong YJ, An CH, Park SC, Pyun JW, Lee J, Kim SW, Kim HS, Kim H, Jeong JC, Kim CY (2018) Methyl jasmonate increases isoflavone production in soybean cell cultures by activating structural genes involved in isoflavonoid biosynthesis. J Agric Food Chem 66(16):4099–4105. https://doi.org/10.1021/acs.jafc.8b00350 CrossRefPubMedGoogle Scholar
- Lamlertkittikul S, Chandeying V (2004) Efficacy and safety of Pueraria mirifica (Kwao Kruea Khao) for the treatment of vasomotor symptoms in perimenopausal women: Phase II Study. J Med Assoc Thail 87(1):33–40Google Scholar
- Suzuki H, Reddy MS, Naoumkina M, Aziz N, May GD, Huhman DV, Sumner LW, Blount JW, Mendes P, Dixon RA (2005) Methyl jasmonate and yeast elicitor induce differential transcriptional and metabolic re-programming in cell suspension cultures of the model legume Medicago truncatula. Planta 220(5):696–707. https://doi.org/10.1007/s00425-004-1387-2 CrossRefPubMedGoogle Scholar
- Udomsuk L, Juengwatanatrakul T, Putalun W, Jarukamjorn K (2012a) Bimodal action of miroestrol and deoxymiroestrol, phytoestrogens from Pueraria candollei var. mirifica, on hepatic CYP2B9 and CYP1A2 expressions and antilipid peroxidation in mice. Nutr Res 32(1):45–51. https://doi.org/10.1016/j.nutres.2011.11.003 CrossRefPubMedGoogle Scholar
- Udomsuk L, Juengwattanatrakul T, Jarukamjorn K, Putalun W (2012b) Increased miroestrol, deoxymiroestrol and isoflavonoid accumulation in callus and cell suspension cultures of Pueraria candollei var. mirifica. Acta Physiol Plant 34(3):1093–1100. https://doi.org/10.1007/s11738-011-0906-6 CrossRefGoogle Scholar
- Yusakul G, Udomsin O, Juengwatanatrakul T, Tanaka H, Chaichantipyuth C, Putalun W (2013a) High performance enzyme-linked immunosorbent assay for determination of miroestrol, a potent phytoestrogen from Pueraria candollei. Anal Chim Acta 785(Supplement C):104–110. https://doi.org/10.1016/j.aca.2013.04.053 CrossRefPubMedGoogle Scholar
- Yusakul G, Udomsin O, Juengwatanatrakul T, Tanaka H, Chaichantipyuth C, Putalun W (2013b) Highly selective and sensitive determination of deoxymiroestrol using a polyclonal antibody-based enzyme-linked immunosorbent assay. Talanta 114(Supplement C):73–78. https://doi.org/10.1016/j.talanta.2013.04.011 CrossRefPubMedGoogle Scholar