Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Molecular Cloning and Characterization of Wound-inducible Beta-amyrin Synthase from Soybean

콩으로부터 상처 유도 beta-amyrin synthase 유전자의 동정 및 발현분석

  • Park, Seong-Whan (Faculty of Natural Resources and Life science, Dong-A university) ;
  • Lee, Jai-Heon (Faculty of Natural Resources and Life science, Dong-A university)
  • 박성환 (동아대학교 생명자원과학부) ;
  • 이재헌 (동아대학교 생명자원과학부)
  • Published : 2002.06.01
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Abstract

Suppression subtractive hybridization (SSH) was used to isolate wound-induced cDNAs from wounded soybean. One of wound-induced cDNA, gmwi33 showed high homology with genes encoding $\beta$-amyrin synthase. The full length cDNA of gmwi33, designated GmAMS1, is 2416 bp long and contains an open reading frame consisted of 739 amino acids. GmAMS1 protein showed 89% identity with licorice GgbAS1 and 86% identity with pea OSCPSY. In 5 day-old, dark-grown seedlings, the expression of GmAMS1 was most strongly induced by light and weakly induced by methyl jasmonate and by low temperature. However, GmAMS1 was not induced by elicitor or UV-B treatment. Such expression pattern might be closely related with the oxygen-radical scavenging activity of soyasaponin.

Suppression subtractive hybridization (SSH)를 통해 상처에 의해 발현이 유도되는 cDNA들을 분리하였고, 그 중 하나인 gmwi33은 $\beta$-amyrin synthase 유전자들과 높은 유사성을 보였다. gmwi33의 전장 cDNA인 GmAMS1은 2416 bp 길이에 739개 아미노산으로 구성된 긴 open reading frame(ORF)를 포함하고 있었다. GmAMS1 단백질은 감초의 $\beta$-amyrin synthase인 GgbAS와 89%, 완두의 OSCPSY와 86%의 유사성을 보였다. 암조건 하에 5일간 기른 콩나물에서, GmAMS1는 빛을 쪼여주었을 때 가장 강하게 발현되었고 methyl jasmonate 처리와 저온처리 시에도 발현이 유도된 반면, UV-B나 elicitor를 처리하였을 때는 발현이 유도되지 않았다. 이러한 GmAMS1의 발현양상은 사포닌의 활성산소 제거기능과 밀접한 연관이 있을 것으로 추측된다.

Keywords

References

  1. Abe I, Rohmer M, Prestwich GD (1993) Enzymatic cydization of squalene and oxidosqualene to sterols and triterpenes. Chem Rev 93:2189-2206 https://doi.org/10.1021/cr00022a009
  2. Abe I, Prestwich GC (1994) Active site mapping of affinity rabeledrat oxidosqualene cyclase. J Biol Chem 269:802-804
  3. Abe I, Prestwich GC (1995) Molecular cloning, characterization, and functional expression of rat oxidosqualene cylase cDNA. Proc Nat'l Acad Sci USA 92:9274-9278 https://doi.org/10.1073/pnas.92.20.9274
  4. Anderson RL, Wolf WJ (1995) Compositional changes in trypsin inhibitors, Phytic acid, saponins and isoflavones related to soybean proccessing. J Nutr 125:581-588
  5. Bestwick CS, Brown IR, Mansfield JW (1998) Localized changes in peroxidase activity accompany hydrogen peroxide generation during the development of a non-host hypersensitive reaction in lettuce. Plant Physiol 118:1067-1078 https://doi.org/10.1104/pp.118.3.1067
  6. Buntel. CJ, Griffin JH (1992) Nucleotide sequence and deduced amino acid sequences of the oxidosqualene cyclase from Candida atbicans. J Am Chem Soc 114:9711-9713 https://doi.org/10.1021/ja00050a090
  7. Corey EJ, Matsuda SPT, Bartel B (1993) Isolation of an Arabidopsis thaliana gene encoding cycloartenol synthase by functional expression in a yeast mutant lacking lanosterol synthase by the use of a chromatographic screen. Proc Nat'l Acad Sci USA 90:11628-11632 https://doi.org/10.1073/pnas.90.24.11628
  8. Church GM, Gilbert (1984) Genomic sequencing. Proc Natl Acad Sci USA 81:1991-1995 https://doi.org/10.1073/pnas.81.7.1991
  9. Hayashi H, Huang P, Kirakosyan A, Inoue K, Hiraoka N, Ikeshiro Y, Kushiro T, Shibuya M, Ebizuka Y (2001) Cloning and characterization of a cDNA encoding beta-amyrin synthase involved in glycyrrhizin and soyasaponin biosyntheses in licorice. Biol Pharm Bull 24:912-916 https://doi.org/10.1248/bpb.24.912
  10. Hendrich S, Lee KW, Xu X, Wang HJ, Murphy PA (1994) Defining food components as new nutrients. J Nutr 124:1789-1792 https://doi.org/10.1093/jn/124.suppl_9.1789S
  11. Herrera JBR, Bartel B, Wilson WK, Matsuda SPT (1998) Cloning and characterization of the Arabidopsis thaliana lupeol synthase gene. Phytochemistiy 49:1905-1911 https://doi.org/10.1016/S0031-9422(98)00366-5
  12. Hodges DM, Andrews CJ, Johnson DA, Hamilton Rl (1997) Antioxidant enzyme and compound responses to chilling stress and their combining abilities in differentially sensitive maize hybrids. Crop science 37:857-863 https://doi.org/10.2135/cropsci1997.0011183X003700030027x
  13. Husselstein T, Schaller H, Benveniste P (2001) Molecular cloning and expression in yeast of 2,3-oxidosqualene-triterpenoid cyclases from Arabidopsis thaIiana. Plant Mol Biol 45:75-92 https://doi.org/10.1023/A:1006476123930
  14. Jabs T (1999) Reactive oxygen intermediates as mediators of programmed cell death in plants and animals. Biochem Pharmacol 57:231-245 https://doi.org/10.1016/S0006-2952(98)00227-5
  15. Kushiro T, Shibuya M, Ebizuka Y (1998a) $\beta$-amyrin synthase. Cloning of oxidosqualene cyclase that catalyzes the formation of the most popular triterpene among higher plant. Eur J Biochem 256:238-244 https://doi.org/10.1046/j.1432-1327.1998.2560238.x
  16. Kushiro T, Shibuya M, Ebizuka Y (1998b) Molecular cloning of cDNA from Panax ginseng. The isogene that encodes $\beta$-amyrin synthase. In: Ageta H, Aimi N, Ebizuka Y, Fujita T, Honda G, (eds.) Towards Natural Medicine Research in the 21st Centry. Excepta Medica International Congress Series 1157. Elsevier Science, Amsterdam, pp 421-428
  17. Messina MJ, Persky V, Setchell KD, Barnes S (1994) Soy intake and cancer risk: a review of the in vitro and in vivo data. Nutr Cancer 21:113-31 https://doi.org/10.1080/01635589409514310
  18. Morita M, Shibuya M, Kushiro T, Masuda K, Ebizuka Y (2000) Molecular cloning and functional expression of triterpene synthases from pea (Pisum sativum) new alpha-amyrin-producing enzyme is a multifunctional triterpene synthase. Eur J Biochem 267:3453-3460 https://doi.org/10.1046/j.1432-1327.2000.01357.x
  19. Nakashima H, Okubo K, Honda Y, Tamura T, Matsuda S, Yamamoto N (1989) Inhibitory effect of glycosides like saponin from soybean on the infectivity of HIV in vitro. AIDS 3:655-658 https://doi.org/10.1097/00002030-198910000-00007
  20. Oh YJ, Sung MK (2001) Soybean saponins inhibit cell proliferation by suppressing PKC activation and induce differentiation of HT-29 human colon adenocarcinoma cells. Nutr Cancer 39:132-138 https://doi.org/10.1207/S15327914nc391_18
  21. Okubo K, Yoshiki Y (1996) Oxygen-radical-scavenging activity of DDMP-conjugated saponins and physiological role in leguminous plant. Adv Exp Med Biol 405:141-54 https://doi.org/10.1007/978-1-4613-0413-5_12
  22. Okubo K, Yoshiki Y (2000) The role of triterpenoid on reactive oxygen scavenging system: approach from the new chemilum inescence system (XYZ system). Biofactors 13:219-223 https://doi.org/10.1002/biof.5520130134
  23. Papadopoulou K, Melton RE, Leggett M, Daniels MJ, Osbourn AE (1999) Compromised disease resistance in saponin-dedicient plants. Proc Hat'l Acad Sci USA 96:12923-12928 https://doi.org/10.1073/pnas.96.22.12923
  24. Pinhero RG, Rao MV, Paliyath G, Murr DP, Fletcher RA (1997) Changes in activities of antioxidant enzymes and their relationship to genetic and paclobutrazol-induced chilling tolerance of maize seedlings. Plant Physiol 114:695-704 https://doi.org/10.1104/pp.114.2.695
  25. Poralla K, Hewelt A, Prestwich GD, Abe I, Reipen I, Sprenger G (1994) A specific amino acid repeat in squalene and oxidos qualene cyclases. Trends Biochem Sci 19:157-158 https://doi.org/10.1016/0968-0004(94)90276-3
  26. Sambrook J, Russell DW (2000) Molecular cloning: A laboratory mannual (3rd ed). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
  27. Shi Z, Buntel CJ, Griffin JH (1994) Isolation and characterization of the gene encoding 2,3-oxidosqualene-lanosterol cyclase from Saccharomyces cerevisiae. Proc Nat' I Acad Sci USA 91:7370-7374 https://doi.org/10.1073/pnas.91.15.7370
  28. Shiraiwa M, Kudo S, Shimoyamada M, Harada K, Okubo K (1991) Composition and structure of 'group A saponin' in soybean [Glycin max] seed. Agri Biol Chem 55:315-322 https://doi.org/10.1271/bbb1961.55.315
  29. Shure M, Wessler S, Fedoroff N (1983) Molecular identification and isolation of Waxy locus in maize. Cell 35:225-233 https://doi.org/10.1016/0092-8674(83)90225-8
  30. Van der Heijden R, Threlfall DR, Verpoorte R, Whitehead IM (1989) Regulation and enzymology of pentacyclic triterpenoid phytoalexin biosynthesis in cell suspension cultures of Tabernaemontana divaricata. Phytochemistry 28:2981-2988 https://doi.org/10.1016/0031-9422(89)80264-X
  31. Verwoerd TC, Dekker BM, Hoekema A (1989) A small-scale procedure for rapid isolation of plant RNAs. Nucl Acid Res 17:2362 https://doi.org/10.1093/nar/17.6.2362