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In vitro induction of hairy root from isoflavones-producing Korean wild arrowroot Pueraria lobata

  • Kim, Soo-Jung (Department of Horticulture, Daegu University) ;
  • Cha, Min-Seok (Department of Horticulture, Daegu University) ;
  • Lee, Eun-Ji (Department of Horticulture, Daegu University) ;
  • Kim, In-Hye (Functional Food & Nutrition Division, National Academy of Agricultural Science, RDA) ;
  • Kwon, Jung-Eun (Department of Life Science, Gachon University) ;
  • Kang, Se-Chan (Department of Life Science, Gachon University) ;
  • Park, Tae-Ho (Department of Horticulture, Daegu University)
  • Received : 2012.09.19
  • Accepted : 2012.09.21
  • Published : 2012.09.30

Abstract

Pueraria lobata is a perennial legume plant, widely distributed in the countries of East Asia. It is a medicinally important leguminous plant and produces various isoflavones such as puerarin, daidzein etc which have potential for preventing several chronic diseases including osteoporosis, cardiovascular disease and cancer. In this study, we tried to induce hairy roots in vitro from Korean wild arrowroot P. lobata and investigated the effects of hormones and light conditions. Initially leaf and stem segments were infected with Agrobacterium rhizogenes and incubated in different conditions. Hairy roots were induced from only stem segments and the induction was best at dark condition and the presence of IBA during incubation. Secondary roots were also significantly much more induced at the dark condition than at the 16 hours light condition. Among plant growth regulators of auxin, IBA was best for secondary root formation while 2,4-D, IAA and NAA produced callus or less hairy roots. The presence of the foreign gene rolC transferred by A. rhizogenes that plays a major role in hairy root induction was confirmed by PCR. The accumulation of isoflavones such as puerarin and daidzin was also confirmed. These results will facilitate mass production of hairy root and can be used for the production of functional substances from wild arrowroots.

Keywords

Acknowledgement

Supported by : Rural Development Administration

References

  1. Barbara T (2003) In vitro propagation of isoflavone-producting Pueraria lobata (Willd.) Ohwi. Plant Sci 165:1123-1128 https://doi.org/10.1016/S0168-9452(03)00320-0
  2. Bonhomme V, Laurain-Mattar D, Lacoux J, Fliniaux M-A, Jacquin-Dubreuil A (2000) Tropane alkaloid production by hairy roots of Atropa belladonna obtained after transformation with Agrobacterium rhizogenes 15834 and Agrobacterium tumefaciens containing rol A, B, C genes only. J Biotechnol 81:151-158 https://doi.org/10.1016/S0168-1656(00)00287-X
  3. Boonsnongcheep P, Korsangruang S, Soonthornchareonnon N, Chintapakorn Y, Saralamp P, Prathanthurarug S (2010) Growth and isoflavonoid accumulation of Pueraria candollei var. candollei and P. candollei var. mirifica cell suspension cultures. Plant Cell Tiss Cult 101:119-126 https://doi.org/10.1007/s11240-010-9668-x
  4. Bousquet J, Simon L, Lalonde M (1990) DNA amplification from vegetative and sexual tissues of trees using polymerase chain reaction. Can J For Res 20:254-257 https://doi.org/10.1139/x90-037
  5. Bulgakov VP, Veselova MV, Tchernoded GK, Kiselev KV, Fedoreyev SA, Zhuravlev YN (2005) Inhibitory effect of the Agrobacterium rhizogenes rolC gene on rabdosiin and rosmarinic acid production in Eritrichium sericeum and Lithospermum erythrorhizon transformed cell cultures. Plant 221:471-478 https://doi.org/10.1007/s00425-004-1457-5
  6. Cardarelli M, Mariotii D, Pomponi M, Spanó L, Capone I, Costantino P (1987) Agrobacterium rhizogenes T-DNA gene capable of inducting hairy root phenotype. Mol Gen Genet 209:475-480 https://doi.org/10.1007/BF00331152
  7. Chansakaow S, Ishikawa T, Sekine K, Okada M, Higushi Y, Kudo M, Chaichantipyuth C (2000) Isoflavonoids from Pueraria mirifica and their estrogenic activity. Planta Med 66: 572-574 https://doi.org/10.1055/s-2000-8603
  8. Chen G, Li L (2007) Nutrient consumption and production of isoflavones in bioreactor cultures of Pueraria lobata (Willd). J Environ Biol 28:321-326
  9. Cherdshewasart W, Sriwatcharakul S (2008) Metabolic activation promotes estrogenic activity of the phytoestrogen-rich plant. Maturitas 59:128-136 https://doi.org/10.1016/j.maturitas.2008.01.002
  10. Cherdshewasart W, Subtang S, Dahlan W (2007) Major isoflavonoid contents of the phytoestrogen rich-herb Pueraria mirifica in comparison with Pueraria lobata. J Pharm Biomed Anal 43:428-434 https://doi.org/10.1016/j.jpba.2006.07.013
  11. Crane C, Wright E, Dixon RA, Wang Z-Y (2006) Transgenic Medicago truncatula plants obtained from Agrobacterium tumefaciens-transformed roots and Agrobacterium rhizogenes-transformed hairy roots. Planta 223:1344-1354 https://doi.org/10.1007/s00425-006-0268-2
  12. Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13-15
  13. Giri A, Narasu ML (2000) Transgenic hairy roots: recent trends and applications. Biotechnol Adv 18:1-22 https://doi.org/10.1016/S0734-9750(99)00016-6
  14. Goyal S, Ramawat KG (2007) Effect of chemical factors on production of isoflavones in Pueraria tuberose (Roxb.ex. Willd.) DC suspension culture. Indian J Exp Biol 45:1063-1067
  15. Goyal S, Ramawat KG (2008) Increased isoflavonids accumulation in cell suspension cultures of Pueraria tuberose by elicitors. Indian J Biotechnol 7:378-382
  16. Hwang SJ (2006) Baicalin production in transformed hairy root clones of Scutellaria baicalensis. Biotechnol Bioprocess Eng 11:849-853
  17. Kim C, Shin S, Ha H, Kim JM (2003) Study of substance changes in flowers of Pueraria thunbergiana Benth. Duing storage. Arch Pharm Res 26:210-213 https://doi.org/10.1007/BF02976832
  18. Kim D-K, Jang DS, Kim JS, Kim J-H (2009) Genetic variations and phylogenetic relationship of and Pueraria lobata Ohwi (Fabaceae) and related taxa by RAPD makers. Korean J Plant Res 22:446-453
  19. Kim JS, Oh EJ, Lee SK (2010) Influence of different strains of Agrobacterium rhizogenes on hairy root induction andn rosmarinic acid production in Agastache rugosa Kuntze. Korean J Plant Res 23:14-18
  20. Kinjo J-E, Furusawa J-I, Baba J, Takeshita T, Yamasaki M, Nohara T (1987) Studies on the constituents of Pueraria lobata. III: Isoflavonoids and related compounds in the roots and the voluble stems. Chem Pharm Bull 35:4846-4850 https://doi.org/10.1248/cpb.35.4846
  21. Kinjo J, Ikeda T, Okawa M, Udayama M, Hirakawa T, Shii Y, Nohara T (2000) Hepatoprotective and hepatotoxic activities of Soporadiol analogs on rat primary liver cell cultures. Biol Chem Bull 23:1118-1121
  22. Ko SM, In DS, Chung H-J, Choi D-W, Liu JR (2007) Mass production of gain-of-function mutants of hair roots in Ginseng. J Plant Biotechnol 34:285-591 https://doi.org/10.5010/JPB.2007.34.4.285
  23. Korsangruang S, Soonthornchareonnon N, Chintapakorn Y, Saralamp P, Prathanturarug S (2010) Effects of abiotic and biotic elicitors on growth and isoflvonoid accumulation in Pueraria candollei var. candollei and P. candollei var. mirifica cell suspension cultures. Plant Cell Tiss Organ Cult 103: 333-342 https://doi.org/10.1007/s11240-010-9785-6
  24. Limpens E, Ramos J, Franken C, Raz V, Compaan B, Franssen H, Bisseling T, Geurts R (2004) RNA interference in Agrobacterium rhizogenes-transformed roots of Arabidopsis and Medicago truncatula. J Exp Bot 55:983-992 https://doi.org/10.1093/jxb/erh122
  25. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  26. Ono NN, Bandaranayake PCG, Tian L (2012) Establishment of pomegranate (Punica granatum) hairy root cultures for genetic interrogation of the hydrolysable tannin biosynthetic pathway. Planta 236:931-941 https://doi.org/10.1007/s00425-012-1706-y
  27. Pandey N, Chaurasia JK, Tiwari OP, Triphati YB (2007) Antioxidant properties of different fractions of tubers from Pueraria tuberose Linn. Food Chem 105:219-222 https://doi.org/10.1016/j.foodchem.2007.03.072
  28. Pappert RA, Hamrich JL, Donovan LA (2000) Genetic variation in Pueraria lobata (Fabaceae), an introduced, clonal, invasive plant of the southeastern United States. Am J Bot 87:1240-1245 https://doi.org/10.2307/2656716
  29. Park T-H, Jones JDG (2009) A new resistance gene to powdery mildew identified in Solanum neorossii has been localized on the short arm of potato chromosome 6. Euphytica 166: 331-339 https://doi.org/10.1007/s10681-008-9811-5
  30. Prasain JK, Reppert A, Jones K, Moore DR, Barnes S, Lila MA (2007) Identification of isoflavones glycosides in Pueraria lobata cultures by tandem mass spectrometry. Phytochem Anal 18:50-59 https://doi.org/10.1002/pca.951
  31. Rathore MS, Shekhawat NS (2009) Micropropagation of Pueraria tuberose (Roxb. Ex Willd.) and determination of puerarin content in different tissues. Plant Cell Tiss Organ Cult 99:327-334 https://doi.org/10.1007/s11240-009-9608-9
  32. Reppert A, Yousef GG, Rogers RB, Lila MA (2008) Isolation of radiolabeled isoflavones from Kudzu (Pueraria lobata) root cultures. J Agric. Food Chem 56:7860-7865 https://doi.org/10.1021/jf801413z
  33. Santos-Gomes PC, Seabra RM, Andrade PB, Fernandes-Ferreira M (2002) Phenolic antioxidant compounds produced by in vitro shoots of sage (Salvia officinalis L.). Plant Sci 162:981-987 https://doi.org/10.1016/S0168-9452(02)00052-3
  34. Sato T, Kawamoto A, Tamura A, Tatsumi Y, Fujii T (1992) Mechanism of antioxidant action of pueraria glycoside (PG)-1 (an isoflavonoid) and mangiferin (a xanthonoid). Chem Pharm Bull 40:721-724 https://doi.org/10.1248/cpb.40.721
  35. Sharma V, Goyal S, Ramawat KG (2009) Scale up production of isoflavonoids in cell suspension cultures of Pueraria tuberose grown in shake flasks and bioreactor. Eng Life Sci 9:267-271 https://doi.org/10.1002/elsc.200800114
  36. Sharma V, Goyal S, Ramawat KG (2011) Increased puerarin biosynthesis during in vitro shoot formation in Pueraria tuberose grown in growtek bioreactor with aeration. Physiol Mol Biol Plants 17:87-92 https://doi.org/10.1007/s12298-011-0049-7
  37. Shi H.-P and Kintzios S (2003) Genetic transformation of Pueraria phaseoloides with Agrobacterium rhizogenes and puerarin production in hairy roots. Plant Cell Rep 21:1103-1107 https://doi.org/10.1007/s00299-003-0633-6
  38. Swain SS, Sabu L, Pal A, Barik D.P, Pradhan C, Chand P.K (2012) Hairy root cultures of butterfly pea (Clitoria ternatea L.): Agrobacterium ${\times}$ plant factors influencing transformation. World J Microbiol Biotechnol 28:729-739 https://doi.org/10.1007/s11274-011-0869-1
  39. Thanonkeo S, Panichajakul S (2006) Production of isoflavones, daidzein and genistein in callus cultures of Pueraria candollei Wall. Ex Benth. Var. mirifica. Songklanakarin J Sci Technol 28:45-53
  40. Thiem B (2003) In vitro propagation of isolavone-producing Pueraria lobata (Willd.) Ohwi. Plant Sci 165:1123-1128 https://doi.org/10.1016/S0168-9452(03)00320-0
  41. Uddin MR, Li X, Won OJ, Park SU, Pyon JY (2012) Herbicidal activity of phenolic compounds from hairy root cultures of Fagopyrum tataricum. Weed Res 52:25-33 https://doi.org/10.1111/j.1365-3180.2011.00894.x
  42. Udomsuk L, Jarukamjorn K, Tanaka H, Putalun W (2011) Improved isoflavonoid production in Pueraria candollei hairy root cultures using elicitation. Biotechnol Lett 33: 369-374 https://doi.org/10.1007/s10529-010-0417-3
  43. Vaishnav K, Goyal S, Ramawat KG (2006) Isoflavonoids production in callus culture of Pueraria tuberose, the Indian kudzu. Indian J Exp Biol 44:1012-1017
  44. Verma P, Singh D, Rahman L, Gupta MM, Banerjee S (2002) In vitro-studies in Plumbago zeylanica: rapid micropropagation and establishment of higher plumbagin yielding hairy root cultures. J Plant Physiol 159:547-552 https://doi.org/10.1078/0176-1617-00518
  45. Villalobos-Amador E, Rodriguez-Hernandez G, Perez-Molphe-Balch E (2002) Organogenesis and Agrobacterium rhizogenesinduced rooting in Pinus maximarinezii Rzedowsky and P. pinceana Gordon. Plant Cell Rep 20:779-785 https://doi.org/10.1007/s00299-001-0396-x

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