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옥수수 유식물 신초에서 Brassinosteroids의 항상성 조절을 위반 C-26 탈메틸 반응의 중요성

Importance of C-26 Demethylation for Homeostatic Regulation of Brassinosteroids in Seedling Shoots of Zea mays L

  • 발행 : 2006.03.01

초록

옥수수 유식물 줄기에서 중요 BRs의 함량조절 기작을 옥수수 유식물 줄기로부터 얻어진 효소원을 이용하여 조사하였다. 먼저 활성형 BR인 CS의 대사를 [$^2H_0$]-와[$^2H_6$]-CS를 기질로 사용하여 실험한 결과 [$^2H_0$]- 와 [$^2H_6$]-CS는 각각 [$^2H_0$]-26-norCS와 [$^2H_3$]-28-norCS로 전환됨을 GC-MS 분석을 통해 확인하였으며, 이러한 두 가지의 대사과정 중 C-26 탈메틸 반응에 의한 CS에서 26-norCS로의 전환만이 생체 내에서 일어나는 반응임을 확인하였다. 이와 함께 주요 생합성 전구물질인 6-deoxoTE와 6-deoxoTY에 대해서도 같은 효소원을 이용하여 C-26 탈메틸 반응에 의한 대사를 조사한 결과 6-deoxoTE는 6-deoxo-3-dehydroTE와 6-deoxoTY로, 6-deoxoTY는 6-deoxo-3-dehydroTE와 6-deoxoTE로 전환됨을 확인함과 동시에, 6-deoxoTE는 6-deoxo-26-norTE 로, 6-deoxo-3-DHT는 3-dehydro-6-deoxo-26-norTE, 6-deoxoTY는 6-deoxo-26-norTY로 전환됨을 확인하였다. 이러한 결과들은 옥수수 유식물 줄기에서 중요 BRs가 모두 C-26 탈메틸 반응이 일어날 수 있음을 나타내는 결과로서 BRs의 C-26 탈메틸 반응이 활성형 BR뿐만 아니라 그 생합성 전구물질에도 중요한 함량조절 기작임을 확인 할 수 있었다.

Regulatory mechanism for endogenous levels of castasterone (CS) and its biosynthetic precursors in shoots of maize was investigated by the use of enzyme solution prepared from the plant tissue. When [$^2H_0$]- and [$^2H_6$]-CS was used as substrates, [$^2H_0$]-26-norCS and [$^2H_3$]-28-norCS were identified as products, indicating that [$^2H_0$]- and [$^2H_6$]-CS are differently metabolized into [$^2H_0$]-26-norCS and [$^2H_3$]-28-norCS by C-26 and C-28 demethylation, respectively. This suggests that both C-26 and C-28 demethylation can be involved in CS catabolism. In fact that C-28 demethylation only occurred when isotope labeled substrate was used, however, C-26 demethylation is thought be a natural reaction occurred in the maize shoots. When 6-deoxoteasterone (6-deoxoTE) was used, 6-deoxo-26-norTE and 3-dehydro-6-deoxo-26-norTE as well as 6-deoxo-3-dehydroTE and 6-deoxotyphasterol (6-deoxoTY) were identified as enzyme products. When 6-deoxoTY was added, 6-deoxo-26-norTY as well as 6-deoxo-3-dehydroTE and 6-deoxoTE was identified as products. These indicate that C-26 demethylation of 6-deoxoTE, 6-deoxo-3-dehydroTE and 6-deoxoTY as well as a reversible C-3 epimerization from 6-deoxoTE to 6-deoxoTY intermediated by 6-deoxo-3-dehydroTE are operative in the maize shoots, demonstrating that endogenous levels of biosynthetic precursors of CS are also controlled by C-26 demethylation. Therefore, it is thought that C-26 demethylation is an important and a common deactivation process which functions to maintain steady state levels of endogenous brassinosteroids in the maize shoots.

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참고문헌

  1. Adam G, Petzold U (1994) Brassinosteroids: a new phytohormone group. Naturwissenchaften 81:210-217
  2. Arima M, Yokota T, Takahashi N (1984) Identification and quantification of brassinolide-related steroids in the insect gall and healthy tissue of the chesnut plant. Phytochemistry 23: 1587 -1592 https://doi.org/10.1016/S0031-9422(00)83445-7
  3. Azpiroz R, Wu Y, LoCascio JC, Feldmann KA (1998) An Arabidopsis brassinosteroid-dependent mutant is blocked in cell elongation. Plant Cell 10:219-230 https://doi.org/10.1105/tpc.10.2.219
  4. Bishop GJ, Nomura T, Yokota T, Harrison K, Noguchi T, Fujioka S, Takatsuto S, Jones JDG, Kamiya Y (1999) The tomato DWARF enzyme catalyses C-6 oxidation in brassinosteroid biosynthesis. Proc Natl Acad Sci USA 96:1761-1776
  5. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of utilizing the principle of protein-dye binding. Anal Biochem 72:248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  6. Choe S, Dilkes BP, Fujioka S, Takatsuto S, Sakurai A, Feldmann KA (1998) The DWF4 gene of Arabidopsis encodes a cytochrome P450 that mediates multiple 22 a -hydroxylation steps in brassinosteroid biosynthesis. Plant Cell 10:231-243 https://doi.org/10.1105/tpc.10.2.231
  7. Choi YH, Fujioka S, Nomura T, Harada A, Yokota T, Takatsuto S, Sakurai A (1997) An alternative brassinolide biosynthetic pathway via late C-6 oxidation. Phytochemistry 44:609-613 https://doi.org/10.1016/S0031-9422(96)00572-9
  8. Chory J, Catterjee M, Cook R (1996) From seed germination to flowering, light controls plant development via the pigment phytochrome. Proc Natl Acsd Sci USA 93; 12066-12071
  9. Clouse SD, Feldmann KA (1999) Molecular genetics of brassinosteroids action. In: Sakurai A, Yokota T, Clouse SD, eds, Brassinosteroids. Springer-Verlag, Tokyo, pp 163-190
  10. Fujioka S (1999) Natural occurrence of brassinosteroids in the plant kingdom. In: Sakurai A, Yokota T, Clouse SD, eds, Brassinosteroids. Springer-Verlag, Tokyo, pp 21-45
  11. Fujioka S, Li J, Choi YH, Seto H, Takatsuto S, Noguchi T, Watanabe T, Kuriyama H, Yokota T, Chory J, Sakurai A (1997) The Arabidopsis deetiolated2 mutant is blocked early in brassinosteroid biosynthesis. Plant Cell 9: 1951-1962 https://doi.org/10.1105/tpc.9.11.1951
  12. Fujioka S, Sakurai, A (1997) Biosynthesis and metabolism of brassinosteroids. Physiol Plant 100: 710-715 https://doi.org/10.1111/j.1399-3054.1997.tb03078.x
  13. Grove MD, Spencer GF, Rohwedder WK, Mandava N, Worley JF, Warthen JD Jr, Steffen GL, Flippen-Anderson JL, Cook JC Jr (1979) Brassinolide, a plant growth promoting steroid isolated from Brassica napus pollen. Nature 281 :216-217 https://doi.org/10.1038/281216a0
  14. Kang MW, Kim YS, Kim SK (2003) Identification and biosynthetic pathway of brassinosteroids in seedling shoots of Zea mays L.. Korean journal of Plant Biotechnology 30:411-419 https://doi.org/10.5010/JPB.2003.30.4.411
  15. Kim SK, Chang SC, Lee EJ, Chung WS, Kim YS, Hwang S, Lee JS (2000) Involvement of brassinosteroids in the gravitropic response of primary root of maize. Plant Physiol 123:997-1004 https://doi.org/10.1104/pp.123.3.997
  16. Kim SK (1991) Natural occurrences of brassinosteroids. In: Cutler HG, Yokota T, Adam G, eds, Brassinosteroids: Chemistry, Bioactivity, and Application, ACS Symposium Series 474. Amer Chem Soc, Washington DC, pp 26-35
  17. Kim TW, Chang SC, Choo J, Watanabe T, Takatsuto S, Yokota T, Lee JS, Kim SY, Kim SK (2000a) Brassinolide and [26, 28-2H6]brassinolide are differently demethylated by loss of C-26 and C-28, respectively, in Marchantia polymorpha. Plant Cell Physiol 42: 1171-1174 https://doi.org/10.1093/pcp/pce004
  18. Kim TW, Chang SC, Lee JS, Takatsuto S. Yokota T, Kim SK (2004b) Novel biosynthetic pathway of castasterone from cholesterol in tomato. Plant Physiology 135: 1231-1242 https://doi.org/10.1104/pp.104.043588
  19. Kim TW, Chung WS, Kim YS, Kim SK (2004b) 6-Deoxocastasterone and its biosynthetic precursors from primary roots of maize. Bull Korean Chem Soc 25: 1099-1102 https://doi.org/10.5012/bkcs.2004.25.7.1099
  20. Kim YS, Kim TW, Kim SK (2003) Conversion of 6-deoxocastasterone to brassinolide in a liverwort, Marchantia polymorpha. Bull Korean Chem Soc 24: 1385-1388 https://doi.org/10.5012/bkcs.2003.24.9.1385
  21. Marquardt V, Adam G (1991) Recent advances in brassinosteroid research. In: Boerner H, Martin D, Sjut V, eds, chemistry of Plant Protection, Vol 7: Herbicide Resistance-Brassinosteroids, Gibberellins, Plant Growth Regulators. Springer-Verlag, Berlin, pp 103-139
  22. Mathur J, Molnar G, Fujioka S, Takatsuto S, Sakurai A, Yokota T, Adam G, Voigt B, Nagy F, Maas C, et al. (1998) Transcription of the Arabidopsis CPD gene, encoding a steroidogenic cytochrome P450, is negatively controlled by brassinosteroids. Plant J 14:593-602 https://doi.org/10.1046/j.1365-313X.1998.00158.x
  23. Meudt WJ (1987) Chemical and biological aspects of brassinolide. In: Fuller G, Nes WD, eds, Ecology and Metabolism of Plant Lipids. ACS Symp Ser 325, Amer Chem Soc, Washington DC, pp 53-75
  24. Nakajima N, Fujioka S, Tanaka T, Takatsuto S, Yoshida S (2002) Biosynthesis of cholesterol in higher plants. Phytochemistry 60:275-279 https://doi.org/10.1016/S0031-9422(02)00113-9
  25. Neff MM, Nguyen SM, Malancharuvil EJ, Fujioka S, Noguchi T, Sete H, Tsubuki M, Honda T, Takatsuto S, Yoshida S, Chory J (1999) BAS1: a gene regulation brassinosteroid levels and light responsiveness in Arabidopsis. Proc Natl Acad Sci USA 96:15316-15323
  26. Noguchi T, Fujioka S, Choe S, Takatsuto S, Tax FE, Yoshida S, Feldmann KA (2000) Biosynthetic pathways of brassinolide in Arabidopsis. Plant Physiol 124:201-209 https://doi.org/10.1104/pp.124.1.201
  27. Nomura T, Nakayama M, Reid JB, Takeuchi Y, Yokota T (1997) Blockage of brassinosteroid biosynthesis and sensitivity causes dwarfism in garden pea. Plant Physiol 113:31-37 https://doi.org/10.1104/pp.113.1.31
  28. Sakurai A, Fujioka S (1993) The current status of physiology and biochemistry of brassinosteroids. Plant Growth Regul 13:147-159 https://doi.org/10.1007/BF00024257
  29. Sakurai A, Fujioka S (1997) Studies on biosynthesis of brassinosteroids. Biosci Biotec Biochem 61:757-762 https://doi.org/10.1271/bbb.61.757
  30. Sasse JM (1991) Brassinosteroids-induced elongation. In: Culter HG, Yokota T, Adam G, eds, Brassinosteroids; Chemistry, Bioactivity and Application, ACS Symp Ser 474, Amer Chem Soc, Washington DC, pp 255-264
  31. Sekimoto H, Hoshi M, Nomura T, Yokota T (1997) Zinc deficiency affects the levels of endogenous gibberellins in Zea mays L. Plant Cell Physiol 38: 1087 -1090 https://doi.org/10.1093/oxfordjournals.pcp.a029276
  32. Suzuki Y, Yamaguchi I, Yokota T, Takahashi N (1986) Identification of castasterone, typhasterol and teasterone from the pollen of Zea mays. Agric Biol Chem 50: 3133-3188 https://doi.org/10.1271/bbb1961.50.3133
  33. Suzuki H, Fujioka S, Takatsuto S, Yokota T, Murofushi N, Sakurai A (1993) Biosynthesis of brassinolide from castasterone in cultured cells of Catharanthus roseus. Plant Growth Regul 12: 10 1-106 https://doi.org/10.1007/BF00193241
  34. Suzuki H, Inoue T, fujioka S, Saito T, Takatsuto S, Yokota T, Murofushi N, Yanagisawa T, Sakurai A (1995) Conversion of 24-methylcholesterol to 6-oxo-24-methylcholestanol, a putative intermediate of the biosynthesis of brassinosteroids, in cultured cells of Catharanthus rose us. Phytochemistry 40: 1391-1397 https://doi.org/10.1016/0031-9422(95)00579-V
  35. Szekeres M, Nemeth K, koncz-Kalman Z, Mathur J, Kauschmann A, Altmann T, Redei GP, Nagy F, Schell J, Koncz C (1996) Brassinosteroids rescue the deficiency of CYP 90, a cytochrome P450, controlling cell elongation and de-etiolation in Arabidopsis. Cell 85: 1 71-182 https://doi.org/10.1016/S0092-8674(00)81094-6
  36. Takahashi T, Gasch A, Nishizawa N, Chua NH (1995) The DIMINUTO gene of Arabidopsis is involved in regulating cell elongation. Genes Dev 9:97-107 https://doi.org/10.1101/gad.9.1.97
  37. Wang ZY, Seto H, Fujioka S, Yoshida S, Chory J (2001) BRII is a critical component of a plasma-membrane receptor for plant steroids. Nature 410:380-383 https://doi.org/10.1038/35066597
  38. Winter J, Schneider B, Strack D, Adam G (1997) Role of a cytochrome P450-dependent monooxygenase in the hydroxylation of 24-epi-brassinolide. Phytochemistry 45:233-237 https://doi.org/10.1016/S0031-9422(96)00827-8
  39. Yamamoto R, Fujioka S, Demura T, Takatsuto S, Yoshida S, Fukada H (2001) Brassinosteroid levels increase drastically prior to mophogenesis of tracheary elements. Plant Physiol 125:556-563 https://doi.org/10.1104/pp.125.2.556
  40. Yokota T (1997) The structure, biosynthesis and function of brassinosteroids. Elsevier Trends Jurnals 2:137-143
  41. Yokota T (1999) Brassinosteroids. In: Hooykaas MA, Hall MA, Libbenga KR (Eds), Biochemistry and Molecular Biology of Plant Hormones. Elsevier Science, Amsterdam pp. 277-293
  42. Yokota T, Sato T, Takeuchi Y, Nomura T, Uno K, Watanabe T, Takatsuto S (2001) Roots and shoots of tomato produce 6-deoxo-28-norcathasterone, 6-deoxo-28-nortyphasterol and 6-deoxo-28-norcastasterone, possible precursors of 28-norcastasterone. Phytochemistry 58:233-238 https://doi.org/10.1016/S0031-9422(01)00237-0
  43. Yokota T, Watanabe S, Ogino Y, Yamaguchi I, Takahashi N (1990) Radioimmunoassay for brassinosteroids and its use for comparative analysis of brassinosteroids in stems and seeds of Phaseolus vulgaris. J Plant Growth Regul 9:151-159 https://doi.org/10.1007/BF02041955