Effect of elicitors on chromene and total isoflavonoid accumulation in p. candollei var. mirifica hairy root culture

Main Article Content

Orapin Udomsin
Thaweesak Juengwatanatrakul
Waraporn Putalun

Abstract

Introduction: Pueraria candollei Wall, ex Benth. (White Kwao Krua) is belong to the family Leguminosae and commonly known as a Thai herbal medicine having been used for rejuvenation. In Thailand there are two plants varieties, p. candollei var. candollei (PC) and p. candollei var. mirifica (PM) which have similar botanical characteristic but both vary in chemical component containing. Their tuberous roots contain chemical compounds, known as phytoestrogen, such as isoflavonoids, chromenes, etc. In the present study, we established a hairy root culture of p. candollei var. mirifica and optimal conditions for growth, and investigated the effects of plants elicitors (methyl jasmonate and yeast extract) on enhancing isoflavonoid and chromene accumulation. We also comparative studied on isoflavonoid and chromene production between hairy root and native root of both varieties of p. candollei. Materials and methods: Chromene analysis was performed using HPLC method (mobile phase consisted of 20% acetonitrile containing 1.5% acetic acid at 1.0 mL/min flow rate and 254 nm detection wavelengths). Total isoflavonid analysis was investigated using competitive ELISA (anti-puerarin and anti-daidzin polyclonal antibodies). The data were analyzed using one-way analysis of variance (ANOVA). Results: The results found 200 µM methyl jasmonate and 0.5 mg/L yeast extract significantly increased chromene production after 3 days and 6 days of elicitation, respectively (17.03 and 17.47 µg/g dry wt, 2-fold higher than control, respectively). They also increased total isoflavonoid production after 3 days of elicitation. (16.06 and 14.65 mg/g dry wt, 2-fold higher than control, respectively) For the comparative analysis of each phytoestrogen between hairy root and native root, chromene content in PM hairy root, PC hairy root, PM root and PC root were 5.51, 4.62, 18.86 and 17.10 µg/g dry wt., respectively. Total isoflavonoid content in PM hairy root, PC hairy root, PM root and PC root were 7.72, 9.10, 6.64 and 7.05 mg/g dry wt., respectively. Conclusion: Methyl jasmonate and yeast extract can enhance both total chromene and isoflavonoid accumulation in p. candollei var. mirifica hairy root culture. The comparative analysis of hairy root and native root of both varieties of p. candollei found that both native roots produced chromenes higher than hairy roots about 4-fold. เท contrast total isoflavonoid content in both hairy roots were higher than native roots.

Article Details

Section
Appendix

References

Kerr A. A reputed rejuVenator. J Siam Soc. (Natural History Suppl.). 1932; 8: 336-338.

Matsumura A, Ghosh A, Pope G.s, Darbre P.D. Comparative study of oestrogenic properties of eight phytoestrogens in MCF7 human breast cancer cells. J. steroid Biochem. & Mol. Biol. 2005; 94: 431-443.

Yusakul G, Putalun W, Udomsin o, Juengwatanatrakul T, Chaichantipyuth c. Comparative analysis of the chemical constituents of two varieties of Pueraria candollei. Fitoterapia. 2011; 82(2): 203-207.

Chansakaow ร, Ishikawa T, Sekine K, Okeda M, Higuchii Y, Kudo M and Chaichantipyuth. Isoflavonoids from Pueraria mirifica and their estrogenic activity. Planta Med. 2000; 66: 572-575.

Taylor M. Alternative to HRT: An evidence-based review. IntJFertil Menopausal stud. 2003; 48(2):64-68.

Urasopan N, Hamada Y, Cherdshewasart W, Malaivijitnond S. Preventive effects of Pueraria mirifica on bone loss in ovariectomized rats. Maturitas. 2008; 59(2): 137-148.

Cherdshewasart W and Sutjit W. Correlation of antioxidant activity and major isoflavonoid contents of the phytoestrogen-rich Pueraria mirifica and Pueraria lobata tubers. Phytomedicine. 2008; 15(1-2): 38¬43.

Khitkal B, Kupittayanant S, Rangsriwatananon K, Manakasema Y. Antioxidant properties of puerarin and genistein from White Kwao Krua induced by elicitors and their antihyperglycemic effect on rats. Suranaree J. Sci. Technol. 2009; 17(1):27-37.

Cherdshewasart W, Subtang S and Dahlan พ. Major isoflavonoid contents of the phytoestrogen rich¬herb Pueraria mirifica in comparison with Pueraria lobata. J Pharm Biomed Anal. 2007; 43(2):428- 434.

Udomsuk L, Jarukamjorn K, Tanaka H and Putalun W. Isoflavonoid production in hairy roots culture of Pueraria candollei. Zeitschrift fur Naturforschung Section C-A Journal of Biosciences. 2009; 64(9¬10): 687-691.

Boonsnongcheep P, Korsangruang S, Soonthornchareon- non N, Chintapakorn Y, Saralamp p, Prathanturarug S. Growth and isoflavonoid accumulation of Pueraria candollei var. candollei and p. candollei var. mirifica cell suspension cultures. Plant Cell Tiss Organ Cult. 2010; 101:119-126.

Korsangruang S, Soonthornchareonnon N, Chintapakorn Y, Saralamp p, Prathanturarung S. Effects of abiotic and biotic elicitors on growth and isoflavonoid accumulation in Pueraria candollei var. candollei and p. candollei var. mirifica cell suspension culture. Plant Cell Tiss Organ Cult. 2010; 103: 333-342.

Udomsuk L, Jarukamjorn K, Tanaka H, Putalun W. Improved isoflavonoid production in Pueraria candollei hairy root cultures using elicitation. Biotechnology Letters. 2011; 33(2): 369-374.

Chansakaow S, Ishikawa T, Sekine K, Okeda M, Higuchi Y and Chaichantipyuth. Identification of deoxymiroestrol as the actual rejuvenating principle of “kwao keur”, Pueraria mirifica. The known miroestrol may be an artifact. J. Nat Prod. 2000; 63(2): 173-175.

Pongkitwitoon B, Sakamoto ร, Tanaka H, Tsuchihashi R, Kinjo J, Morimoto S and Putalun W. Enzyme- linked immunosorbent assay for total isoflavonoids in Pueraria candollei using anti-puerarin and anti- daidzin polyclonal antibodies. Planta Medica. 2010; 76: 831-836.

Sasaki K, Mito K, Ohara K, Yamamoto H, Yazaki K. Cloning and characterization of naringenin 8- prenyltransferase, a flavonoid-specific prenyltransferase of Sophora flavescens. Plant Physiol. 2008; 146: 1075-1084.

Akashi T, Sasaki K, Aoki T, Ayabe S, Yazaki K. Molecular cloning and characterization of a cDNA for pterocarpan 4-dimethylallyltransferase catalyzing the key prenylation step in the biosynthesis of glyceollin, a soybean phytoalexin. Plant Physiol. 2009; 149: 683-693.

Yu o and McGonigle B. Metabolic engineering of isoflavone biosynthesis. Adv Agr. 2005; 86:147¬190

Dewick PM. Medicinal natural products: a biosynthetic approach. UK: Wiley; 2009.