Secondary Metabolite Contents and Antioxidant Enzyme Activities of Cichorium intybus Hairy Roots in Response to Zinc

Abstract

Hairy root systems are formed by transforming plant tissues with the “natural genetic engineer” Agrobacterium rhizogenes. In most plants such as Cichorium intybus L., hairy root cultures have proven to be an efficient system for secondary metabolites production. The effect of Zinc (ZnSO4), a heavy metal, was investigated at different concentrations (0, 1, 5 and 10 mM) on some secondary metabolite contents at three time course levels (24, 48 and 72 h).The treated hairy roots of chicory were compared with control and with each other in growth rate, phenol flavonoid and chicoric acid production rate. In addition, antioxidant enzyme activities were determined. Results showed decreased hairy roots weights and increased phenol, flavonoid, chicoric acid and antioxidant enzyme activities in response to higher concentrations of Zinc at higher time courses. Also, an increase in chicoric acid release into the culture media was observed that is important for industrial uses. 

Keywords


1. Velayuthaml P, Ranjitha kumari BD, Baskaran P. An efficient in vitro plant regeneration system for Cichorium intybus L.–an important medicinal plant. J Agric Technol. 2006; 2:287-298.

2. Malarz J, Stojakowska A, Kisiel W .Sesquiterpene lactones in a hairy root culture of Cichorium intybus. Zeitschrift fur Naturforsch C-J Biosci. 2002;57:994-997.

3. Nandagopal S, Ranjitha Kumari BD. Effectiveness of auxin induced in vitro root culture in chicory. J Cent Eur Agric. 2007;8:73-80.

4. Nandagopad S, Ranjitha Kumari BD. Phytochemical and antibacterial studies of chicory (Cichorium intybus L.)–A multipurpose medicinal plant. Adv Biol Res. 2007;1:17-21.

5. Grzegorczyk  I, Krolicka A, Wysokinska H. Establishment of Salvia officinalis L. hairy root cultures for the production of rosmarinic acid. Zeitschrift fur Naturforschung. 2006;61:351-356.

6.  Bais HP, Sudha G, George J , Ravishankar GA. Influence of exogenous hormones on growth and secondary metabolite production in hairy root cultures of Cichorium intybus L. cv. Lucknow local. In vitro Cell Develop Biol Plant. 200;37:293-299.

7. Ercan AG, Taskin KM, Turgut K, Yuce S. Agrobacterium rhizogenes-mediated hairy root formation in some Rubia tinctorum L. populations grown in Turkey.Turk. J Bot. 1999;23:373-377.

8. Pawar  PK, Maheshwari VL. Agrobacterium rhizogenes mediated hairy root induction in two medicinally important members of family Solanaceae. Indian J Biotechnol. 2004;3:414-417.

9. Georgiev MI, Pavlov AI, Bley T. Hairy root type plant in vitro systems as sources of bioactive substances. Appl Microbiol Biotechnol. 2007;74:1175-1185.

10. Kumar V, Sharma A, Narasimha Prasad BC, Gururaj HB, Ravishankar GA. Agrobacterium rhizogenes mediated genetic transformation resulting in hairy root formation is enhanced by ultrasonication and acetosyringone treatment. Elect J Biotechnol. 2006;9:349-357.

11. Kamada H, Ono A, Saitou A and Harada H. No requirement of vernalization for flower formation in ri-transformed Cichorium plants. Plant Tissue Cult Lett. 1992;9:206-208.

12. Chabaud M, Boisson-Dernier A, Zhang J, Taylor CG, Yu O, Barker DG. Agrobacterium rhizogenes-mediated root transformation.The Medicago truncatula Handbook. 2006; pp. 1-8.

13. Michalak  A. Phenolic compounds and their antioxidant activity in plants growing under heavy metal stress. Polish J Environ Stud. 2006;15:523-530.

14. Michael  PI, Krishnaswamy M .The effect of Zinc stress combined with high irradiance stress on membrane damage and antioxidative response in bean seedlings. Environ Exp Bot. 2011;74:171-177.

15. Luo  ZB, He XJ, Chen L, Tang L,Gao SH, Chen F. Effects of Zinc on growth and antioxidant responses in Jatropha curcas seedlings. Int J Agric Biol. 2010;12:119-124.

16. Conforti F, Sosa S, Marrelli M, Menichini F, Statti GA, Uzunov D, Tubaro A, Menichini F. The protective ability of Mediterranean dietary plants against the oxidative damage: The role of radical oxygen species in inflammation and the polyphenol, flavonoid and sterol contents. Food Chem. 2009;112:587-594.

17. Deglinnoocenti E, Pardossi A, Tattini M, Guidi L. Phenolic compounds and antioxidant power in minimally processed salad. J Food Biochem. 2008;32:642-653.

18. Hafsi C. Romero-Puertas MC, Rio LA. Abdelly CH. Sandalio LM. Antioxidative response of Hordeum maritimum L. to potassium deficiency. Acta Physiol Plantar. 2011;33:193-202.

19. Dahajipour Heidarabadi M, Ghanati F, Fujiwara T. Interaction between boron and aluminum and their effects on phenolic metabolism of Linum usitatissimum L. roots. Plant Physiol Biochem. 2011;49:1377-1383.

20. Ali  MB, Singh N, Shohael AM, Hahn EJ, Paek KY. Phenolics metabolism and lignin synthesis in root suspension cultures of Panax ginseng in response to copper stress. Plant Sci. 2006;171:147-154.

21. Chkhikvishvili  ID, Kharebava GI. Chicoric and chlorogenic acids in plant species from Georgia. Appl Biochem Microbiol. 2001;37:188-191.

22. Lee J, Scagel CF. Chicoric acid levels in commercial basil (Ocimum basilicum) and Echinacea purpurea products. J Funct Foods 2010;2:77.

23. Nuissier G, Rezzonico B, Grignon-Dubois M. Chicoric acid from Syringodium filiforme. Food Chem. 2010;120:783-788.

24. Lee J. Caffeic acid derivatives in dried Lamiaceae and Echinacea purpurea products. J Funct Foods 2010;2:158-162.

25. Heimler  D, Isolani L, Vignolini P, Romani A. Polyphenol content and antiradical activity of Cichorium intybus L. from biodynamic and conventional farming. Food Chem. 2009;114:765-770.

26. Llorach  R, Martinez-Sanchez A, Tomas-Barberan FA, Gil MI, Ferreres F. Characterisation of polyphenols and antioxidant properties of five lettuce varieties and escarole. Food Chem. 2008;108:1028-1038.

27. Bradford  M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248-254.

28. Giannopolitis  CN, Ries SK. Superoxide dismutases I. Occurrence in higher plants. Plant Physiol. 1977;59:309-314.

29. Kar M, Mishra D. Catalase, peroxidase, and polyphenoloxidase activities during rice leaf senescence. Plant Physiol. 1976;57:315-319.

30. Cherif J, Derbel N, Nakkach M, Bergmann HV, Jemal F, Lakhdar ZB. Analysis of in vivo chlorophyll fluorescence spectra to monitor physiological state of tomato plants growing under Zinc stress. J Photochem Photobiol B: Biology 2010;101:332-339.

31. Khudsar T, Mahmooduzzafar, Iqbal M, Sairam RK. Zinc-induced changes in morpho-physiological and biochemical parameters in Artemisia annua. Biol Plantar. 2004;48:255-260.

32. Prasad K, Paradha Saradhi P, Sharmila P. Concerted action of antioxidant enzymes and curtailed growth under Zinc toxicity in Brassica juncea. Environ Exp Bot. 1999;42:1-10.

33. Mithofer A, Schulze B, Boland W. Biotic and heavy metal stress response in plants: evidence for common signals. FEBS Lett. 2004;566:1-5.

 34. Diaz J, Bernal A, Pomar F, Merino F. Induction of shikimate dehydrogenase and peroxidase in pepper (Capsicum annuum L.) seedlings in response to copper stress and its relation to lignification. Plant Sci. 2001;161:179-188.

35. Winkel-Shirley B. Biosynthesis of flavonoids and effects of stress. Current Opin Plant Biol. 2002;5:218-223.

36. Wakabayashi K, Hoson T, Kamisaka S. Osmotic stress suppresses cell wall stiffening and the increase in cell wall-bound ferulic and diferulic acids in wheat coleoptiles. Plant Physiol. 1997;113:967-973.

 37. Camacho-Cristobal  JJ, Anzellotti D, Gonzalez-Fontes A. Changes in phenolic metabolism of tobacco plants during short-term boron deficiency. Plant Physiol Biochem. 2002;40:997-1002.

 38. Moran JF, Klucas RV, Grayer RJ, Abian J, Becana M. Complexes of Iron with phenolic compounds from soybean nodules and other legume tissues: prooxidant and antioxidant properties. Free Rad Biol Med. 1997;22:861-870.

39. Hakkinen SH, Torronen AR. Content of flavonols and selected phenolic acids in strawberries and Vaccinium species: influence of cultivar, cultivation site and technique. Food Res Int. 2000;33:517-524.