Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
권호 표지
DOI QR코드

DOI QR Code

Molecular characterization and biological changes caused by Agrobacterium-mediated infiltration of PgTRX1

  • Choi, Seung Hyuk (Department of Bio-Resource Sciences, Kangwon National University) ;
  • Seo, Ji Won (Interdisciplinary Program in Smart Science, Kangwon National University) ;
  • Lee, Jae Geun (Research Institute of Biotechnology, HwajinBioCosmetic) ;
  • Yu, Chang Yeon (Interdisciplinary Program in Smart Science, Kangwon National University) ;
  • Seong, Eun Soo (Department of Medicinal Plant, Suwon Women's University)
  • Received : 2021.05.25
  • Accepted : 2021.07.07
  • Published : 2021.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

In order to test the functionality of Panax ginseng thioredoxin 1 (PgTRX1) isolated from fermented wild ginseng roots, a transient effect on physiological activity were performed over a short time frame using the Agrobacterium infiltration technique. The PgTRX1 gene isolated from fermented wild ginseng was confirmed to have a size of 579 bp, and the expression of PgTRX1 was the highest in the sample after 6 h of fermentation. As a result of constructing this gene and confirming the infiltration reaction mediated by Agrobacterium in tobacco leaves, it was found that the expression of the NbHSR203j gene was also induced as PgTRX1 expression increased. As a result of measuring the biological activity of the infiltration samples, the total phenol content increased by 35.45±1.84 to 49.01±1.84 ㎍ GAE/mL compared to the control, and the total flavonoid amount of 9.52±0.41 to 9.82±0.25 ㎍ QE/mL was slightly high. From these results, Agrobacterium-mediated PgTRX1 appears to be related to the hypersensitive response induction mechanism of plants and the production of secondary metabolites such as phenolic substances.

Keywords

Acknowledgement

This study was supported by the Bioherb Research Institute, Kangwon National University, Republic of Korea.

References

  1. In JG, Yang DC (2004) Current Status and Prospect of Cultured-Root in vitro of the Korean Wild Ginseng. Kor J Plant Resour 17: 1-18
  2. Zhang S, Chen R, Wang C (2006) Ginsenoside Extraction from Panax quinquefolium L. (American ginseng) Root by using Ultrahigh Pressure. J Pharmaceut Biomed Anal 41:57-63. doi: 10.1016/j.jpba.2005.10.043
  3. Attele AS, Wu JA, Yuan CS (1999) Multiple pharmacological effects of ginseng. Biochem Pharmacol 58: 1685-1693 https://doi.org/10.1016/S0006-2952(99)00212-9
  4. Schurmann P, Jacquot JP (2000) Plant thioredoxin systems revisited. Annu Rev Plant Physiol Plant Mol Biol 51: 371-400. doi: 10.1146/annurev.arplant.51.1.371
  5. Kim IS, Kim YS, Yoon HS (2013) Expression of salt-induced 2-Cys Peroxiredoxin from Oryza sativa Increases Stress Tolerance and Fermentation Capacity in Genetically Engineered Yeast Saccharomyces Cerevisiae. Appl Microbiol Biotechnol 97: 3519-3533 https://doi.org/10.1007/s00253-012-4410-8
  6. Clemente T (2006) Nicotiana (Nicotiana tobaccum, Nicotiana benthamiana). In: Wang K (ed) Agrobacterium protocols, 2nd edn. Humana press, New Jersey, pp 143-154. doi: 10.1385/1-59745-130-4:143
  7. Dohm A, Ludwig C, Schiling D, Debener T (2001) Transformation of roses with genes for antifungal proteins. Acta Hortic 547: 27-33 https://doi.org/10.17660/actahortic.2001.547.3
  8. Marchant R, Power JB, Lucas JA, Davey MR (1998) Biolistic transformation of rose (Rosa hybrida L.). Ann Bot 81: 109-114. doi: 10.1006/anbo.1997.0538
  9. Wroblewski T, Tomczak A, Michelmore R (2005) Optimization of Agrobacterium mediated transient expression assays for lettuce, tomato and Arabidopsis. Plant Biotech J 3: 259-273 https://doi.org/10.1111/j.1467-7652.2005.00123.x
  10. Kapila J, Rycke RD, Montagu MV, Angenon G (1997) An Agrobacterium-mediated transient gene expression system for intact leaves. Plant Sci 122: 101-108. doi: 10.1016/S0168-9452(96)04541-4
  11. Yi SY, Kim JH, Joung YH, Lee S, Kim WT, Yu SH, Choi D (2004) The pepper transcription factor CaPF1 confers pathogen and freezing tolerance in Arabidopsis. Plant Physiol 136: 2862-2874. doi: 10.1104/pp.104.042903
  12. Seong ES, Kwon SS, Ghimire BK, Yu CY, Cho DH, Lim JD, Kim KS, Heo K, Lim ES, Chung IM, Kim MJ, Lee YS (2008) LebZIP2 induced by salt and drought stress and transient overexpression by Agrobacterium. BMB Rep 41: 693-698. doi: 10.5483/BMBRep.2008.41.10.693
  13. Seong ES, Yoo JH, Lee JG, Kim HY, Hwang IS, Heo K, Lim JD, Lee DK, Sacks EJ, Yu CY (2013) Transient overexpression of the Miscanthus sinensis glucose-6-phosphate isomerase gene (MsGPI) in Nicotiana benthamiana enhances expression of genes related to antioxidant metabolism. Plant Omics J 6: 408-414
  14. Blois MS (1958) Antioxidant determinations by the use of a stable free radical. Nature 181: 1199-1200 https://doi.org/10.1038/1811199a0
  15. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Evans CR (1999) Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology Med 26: 1231-1237. doi: 10.1016/S0891-5849(98)00315-3
  16. Taga MS, Miller EE, Pratt DE (1984) Chia seeds as a source of natural lipids antioxidants. J Am Oil Chem Soc 61: 928-993 https://doi.org/10.1007/BF02542169
  17. Moreno MIN, Isla MI, Sampietro AR and Vattuone MA (2000) Comparison of the free radical-scavenging activity of propolis from several regions of Argentina. J Ethnopharmacol 71: 109-114. doi: 10.1016/S0378-8741(99)00189-0
  18. Meyer Y, Siala W, Bashandy T, Riondet C, Vignols F, Reichheld JP (2008) Glutaredoxins and thioredoxins in plants. Biochim Biophys Acta 1783: 589-600. doi: 10.1016/j.bbamcr.2007.10.017
  19. Lee KH, Choi HS, Hwang KA, Song JIN (2016) Quality Changes in Doenjang upon Fermentation with Two Different Bacillus subtilis Strains. J East Asia Soc Die Life 26: 163-170. doi: 10.17495/easdl.2016.4.26.2.163
  20. Hwang YS (1993) Characteristics of Cell Walls in Melon Fruits Affected by Abnormal Fermemtation. Hort Environ Biotech 34: 273-278
  21. Zheng L, Liu G, Meng X, Li Y, Wang Y (2012) A versatile Agrobacterium-mediated transient gene expression system for herbaceous plants and trees. Biochem Genet 50: 761-769 https://doi.org/10.1007/s10528-012-9518-0
  22. Andrieu A, Breitler JC, Sire C, Meynard D, Gantet P, Guiderdoni E (2012) An in planta, Agrobacterium-mediated transient gene expression method for inducing gene silencing in rice (Oryza sativa L.) leaves. Rice 5: 23 https://doi.org/10.1186/1939-8433-5-23
  23. Koukol J, Conn EE (1961) The metabolism of aromatic compounds in higher plants. IV. Purification and properties of the phenylalanine deaminase of Hordeum vulgare. J Biol Chem 236: 2692-2698. doi: 10.1016/S0021-9258(19)61721-7
  24. Badawi GH, Kawano N, Yamauchi Y, Shimada E, Sasaki R, Kubo A, Tanaka K (2004) Over-expression of ascorbate peroxidase in tobacco chloroplasts enhances the tolerance to salt stress and water deficit. Physiol Plant 121: 231-238. doi: 10.1111/j.0031-9317.2004.00308.x
  25. Liu R, Xu S, Li J, Hu Y, Lin Z (2006) Expression profile of a PAL gene from Astragalus membranaceus var. mongholicus and its crucial role in flux into flavonoid biosynthesis. Plant Cell Rep 25: 705-710 https://doi.org/10.1007/s00299-005-0072-7
  26. Oh MM, Carey EE, Rajashekar CB (2009) Environmental stresses induce health-promoting phytochemicals in lettuce. Plant Physiol Biochem 47: 578-583. doi: 10.1016/j.plaphy.2009.02.008
  27. Payton P, Allen R, Trolinder N, Holaday A (1997) Over-expression of chloroplast-targeted Mn superoxide dismutase in cotton does not alter the reduction of photosynthesis after short exposures to low temperature and high light intensity. Phytosynth Res 52: 233-244 https://doi.org/10.1023/A:1005873105596
  28. Torsethaugen G, Lynne HP, Zilinskas A, Eva JP (1997) Overproduction of ascorbate peroxidase in the tobacco chloroplast does not provide protection against ozone. Plant Physiol 114: 529-537. doi: 10.1104/pp.114.2.529
  29. Pontier D, Godiard L, Marco Y, Roby D (1994) Hsr203J, a tobacco gene whose activation is rapid, highly localized and specific for incompatible plant/pathogen interactions. The Plant J 5: 507-521. doi: 10.1046/j.1365-313X.1994.05040507.x
  30. Delledonne M, Xia Y, Dixon RA, Lamb C (1998) Nitricoxide functions as a signaling plant disease resistance. Nature, 394: 585-588 https://doi.org/10.1038/29087
  31. Seong ES, Yoo JH, Lee JG, Kim HY, Ghimire BK, Lim JD, Jine DC, Yu CY (2012) Molecular Characterization and Stress Signaling of the Miscanthus sinensis MsCOMT Gene by Transient Assay. Russia J Plant Physiol 59: 626-631 https://doi.org/10.1134/S1021443712050135