Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Molecular and functional characterization of a Brmecp gene encoding 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase from Brassica rapa

배추 유래 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase 관련 Brmecp 유전자의 발현 및 분자적 특성

  • Jung, Yu-Jin (Institute of Genetic Engineering, Hankyong National University) ;
  • Choi, Jang-Sun (National Institute of Horticultural & Herbal Science) ;
  • Sun, Ju-Nam (Department of Horticulture, Hankyong National University) ;
  • Nou, Ill-Sup (Department of Horticulture, Sunchon National University) ;
  • Cho, Yong-Gu (Department of Crop Science, Chungbuk National University) ;
  • Kang, Kwon-Kyoo (Institute of Genetic Engineering, Hankyong National University)
  • Received : 2012.08.30
  • Accepted : 2012.09.10
  • Published : 2012.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

In plants, the fifth step of the plastidial 2-Cmethyl-D-erythritol 4-phosphate (MEP) pathway is catalyzed by 2-C-Methyl-D-erythritol 2,4-cyclodiphosphate synthase (MECP; EC: 4. 6. 1. 12), an enzyme proposed to play a key role in the regulation of isoprenoid biosynthesis. Here we report the isolation and functional characterization of a 823 bp Brassica rapa MECP (Brmecp) cDNA encoding a deduced polypeptide of 230 amino acid residues. Transcription levels of Brmecp were two-fold higher in petal compared to leaves. In addition, Brmecp expression in cabbage seedlings treated with ABA, $H_2O_2$ and drought was higher than control seedlings. These results were consistent with changes in chlorophyll contents in transgenic Arabidopsis. Thus, the Brmecp may contribute to the production of primary (chlorophylls and carotenoids) isoprenoid end-products in chloroplasts.

Keywords

References

  1. Adam KP, Zapp J (1998) Biosynthesis of the isoprene units of chamomile sesquiterpenes; Phytochemistry 48:953-959 https://doi.org/10.1016/S0031-9422(97)00992-8
  2. Adam P, Hecht S, Eisenreich W, Kaiser J, GRAWERT T, Arigoni BA, Rohdish F (2002) Biosynthesis of terpenes: Studies on 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase. Proc Natl Acard Sci 99:12108-12113 https://doi.org/10.1073/pnas.182412599
  3. Araki N, Kusumi K, Masamoto K, lba K (2000) Temperature sensitive Arabidopsis mutant defective in 1-deoxy-D-xyluose-5-phosphate synthase with in the plastitinon-mevinoplte pathway of is terepenoitibiosynthesis. Physiol Plant 108:19-24
  4. Bechtold N, Pelletier G (1998) In planta Agrobacterium-mediated transformation of adult Arabidopsis thaliana plants by vacuum infiltration. Methods Mol Biol 82:259-66
  5. Bernal C, Palacin C, Boronat A, Imperial S (2005) A colorimetric assay for the determination of 4-diphosphocytidyl-2-C-methyl-D-erythritol 4-phosphate synthase activity. Anal Biochem 337:55-61 https://doi.org/10.1016/j.ab.2004.10.011
  6. Bouvier F, d''Harlingue A, Suire C, Backhaus RA, Camara B (1998) Dedicated roles of plastid transketolase during the early onset of isoprenoid biogenesis in pepper fruits; Plant Physiol 117:1423-1431 https://doi.org/10.1104/pp.117.4.1423
  7. Campos N, Rodriguez-Concepcion M, Seemann M, Rohmer M, Boronat A (2001) Identification of gcpE as a novel gene of the 2-C-methyl-D-erythritol 4-phosphate pathway for isoprenoid biosynthesis in Escherichia coli. FEBS Lett 488:170-173. https://doi.org/10.1016/S0014-5793(00)02420-0
  8. Dubey VS, Bhalla R, Luthra R (2003) An overview of the non-mevalonate pathway for terpenoid biosynthesis in plants. J Biosci 40:637-646
  9. Gabrielsen M, Rohdich F, Eisenreich W, Grawert T, Hech, S, Bacher A, Hunter, WN (2004) Biosynthesis of isoprenoids: a bifunctional IspDF enzyme from Campylobacter jejuni. Eur J Biochem 271:3028-3035 https://doi.org/10.1111/j.1432-1033.2004.04234.x
  10. Gabrielsen M, Kaiser J, Rohdich F, Eisenreich W, Laupitz R, Bacher A, Bond CS. Hunter WN (2006) The crystal structure of a plant 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase exhibits a distinct quaternary structure compared to bacterial homologues and a possible role in feedback regulation for cytidine monophosphate. FEBS J 273:1065-1073 https://doi.org/10.1111/j.1742-4658.2006.05133.x
  11. Gao S, Lin J, Liu X, Deng Z, Li Y, Sun X, Tang K (2006) Molecular cloning, characterization and functional analysis of a 2-C-methyl- D-erythritol 2, 4-cyclodiphosphate synthase gene from ginkgo biloba. J. Biochem Mol Biol 39: 502-510 https://doi.org/10.5483/BMBRep.2006.39.5.502
  12. Han YS, Roytrakul S, Verberne MC, van der Heijden R, Linthorst HJM. Verpoorte R (2003) Cloning of a cDNA encoding 1-deoxy-D-xylulose 5-phosphate synthase from Morinda citrifolia and analysis of its expression in relation to anthraquinone accumulation. Plant Sci 164:911-917 https://doi.org/10.1016/S0168-9452(02)00362-X
  13. Hirai N, Yoshida R, Todoroki Y, Ohigashi H (2000) Biosynthesis of abscisic acid by the non-mevalonate pathway in plants, and by the mevalonate pathway in fungi. Biosci Biotechnol Biochem 64:1448-1458 https://doi.org/10.1271/bbb.64.1448
  14. Hunter WN (2007) The non-mevalonate pathway of isoprenoid precursor biosynthesis J Biochem 282:21573-21577
  15. Julliard J H, Douce R (1991) Biosynthesis of the thiazole moiety of thiamin (vitamin B1) in higher plant chloroplasts. Proc Natl Acad Sci USA 88:2042-2045 https://doi.org/10.1073/pnas.88.6.2042
  16. Kumar S, Tamura K, Jakobsen IB, Nei M (2001) MEGA2:molecular evolutionary genetics analysis software. Bioinformatics 17:1244-1245 https://doi.org/10.1093/bioinformatics/17.12.1244
  17. Lange BM, Croteau R (1999a) Isoprenoid biosynthesis via a mevalonate-independent pathway in plants: cloning and heterologous expression of 1-deoxy-D-xylulose-5-phosphate reductoisomerase from peppermint. Arch Biochem Biophys 365:170-174 https://doi.org/10.1006/abbi.1999.1168
  18. Lange BM, Croteau R (1999b) Isopentenyl diphosphate biosynthesis via a mevalonate-independent pathway: isopentenyl monophosphate kinase catalyses the terminal enzymatic step Proc Natl Acad Sci 96:13714-13719 https://doi.org/10.1073/pnas.96.24.13714
  19. Lichtenthaler HK, Schwender J, Disch A, Rohmer M (1997) Biosynthesis of isoprenoids in higher plant chloroplasts proceeds via a mevalonate-independent pathway. FEBS Lett 400:271-274 https://doi.org/10.1016/S0014-5793(96)01404-4
  20. Livak K.J, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the $2^{-{\Delta}{\Delta}Ct}$ method. Methods 25:402-408 https://doi.org/10.1006/meth.2001.1262
  21. Milborrow BV, Lee HS (1998) Endogenous biosynthetic precursors of (+) abscisic acid. VI. Carotenoids and abscisic acid formed by the non-mevalonate triose-pyruvate pathway in chloroplasts. Aust J Plant Physiol 25:507-512 https://doi.org/10.1071/PP98006
  22. Shi W, Feng J, Zhang M, Lai X, Xu S, Zhang X, Wang H (2007) Biosynthesis of Isoprenoids: Characterization of a Functionally Active Recombinant 2-C-methyl-D-erythritol 4-phosphate Cytidyltransferase (IspD) from Mycobacterium tuberculosis H37Rv. J Biochem Mol Biol 40:911-920 https://doi.org/10.5483/BMBRep.2007.40.6.911
  23. Steinbacher S, Kaiser J, Wungsintaweekul J, Hecht S, Eisenreich W, Gerhardt S, Bacher A, Rohdich F (2002) Structure of 2-Cmethyl-d-erythritol-2,4-cyclodiphosphate synthase involved in mevalonate-independent biosynthesis of isoprenoids. J Mol Biol 316:79-88 https://doi.org/10.1006/jmbi.2001.5341