Plant Biotechnology Reports

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Metabolic engineering of Lilium ${\times}$ formolongi using multiple genes of the carotenoid biosynthesis pathway

  • Azadi, Pejman (Laboratory of Plant Cell Technology, Graduate School of Horticulture, Chiba University) ;
  • Otang, Ntui Valentaine (Laboratory of Plant Cell Technology, Graduate School of Horticulture, Chiba University) ;
  • Chin, Dong Poh (Laboratory of Plant Cell Technology, Graduate School of Horticulture, Chiba University) ;
  • Nakamura, Ikuo (Laboratory of Plant Cell Technology, Graduate School of Horticulture, Chiba University) ;
  • Fujisawa, Masaki (Central Laboratories for Frontier Technology, i-BIRD) ;
  • Harada, Hisashi (Central Laboratories for Frontier Technology, i-BIRD) ;
  • Misawa, Norihiko (Central Laboratories for Frontier Technology, i-BIRD) ;
  • Mii, Masahiro (Laboratory of Plant Cell Technology, Graduate School of Horticulture, Chiba University)
  • Received : 2010.07.08
  • Accepted : 2010.07.31
  • Published : 2010.12.30
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Abstract

Lilium ${\times}$ formolongi was genetically engineered by Agrobacterium-mediated transformation with the plasmid pCrtZW-N8idi-crtEBIY, which contains seven enzyme genes under the regulation of the CaMV 35S promoter. In the transformants, ketocarotenoids were detected in both calli and leaves, which showed a strong orange color. In transgenic calli, the total amount of carotenoids [133.3 ${\mu}g/g$ fresh weight (FW)] was 26.1-fold higher than in wild-type calli. The chlorophyll content and photosynthetic efficiency in transgenic orange plantlets were significantly lowered; however, after several months of subculture, they had turned into plantlets with green leaves that showed significant increases in chlorophyll and photosynthetic efficiency. The total carotenoid contents in leaves of transgenic orange and green plantlets were quantified at 102.9 and 135.2 ${\mu}g/g$ FW, respectively, corresponding to 5.6- and 7.4-fold increases over the levels in the wild-type. Ketocarotenoids such as echinenone, canthaxanthin, 3'-hydroxyechinenone, 3-hydroxyechinenone, and astaxanthin were detected in both transgenic calli and orange leaves. A significant change in the type and composition of ketocarotenoids was observed during the transition from orange transgenic plantlets to green plantlets. Although 3'-hydroxyechinenone, 3-hydroxyechinenone, astaxanthin, and adonirubin were absent, and echinenone and canthaxanthin were present at lower levels, interestingly, the upregulation of carotenoid biosynthesis led to an increase in the total carotenoid concentration (+31.4%) in leaves of the transgenic green plantlets.

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References

  1. Apel W, Bock R (2009) Enhancement of carotenoid biosynthesis in transplastomic tomatoes by induced lycopene-to-provitamin A conversion. Plant Physiol 151:59-66 https://doi.org/10.1104/pp.109.140533
  2. Azadi P, Chin DP, Kuroda K, Khan RS, Mii M (2010) Macro elements in inoculation and co-cultivation medium strongly affect the efficiency of Agrobacterium-mediated transformation in Lilium. Plant Cell Tissue Organ Cult 101:201-209 https://doi.org/10.1007/s11240-010-9677-9
  3. Choi SK, Nishida Y, Matsuda S, Adachi K, Kasai H, Peng X, Komemushi S, Miki W, Misawa N (2005) Characterization of $\beta-carotene$ ketolases, CrtW, from marine bacteria by complementation analysis in Escherichia coli. Mar Biotechnol (NY) 7:515-522 https://doi.org/10.1007/s10126-004-5100-z
  4. Clotault J, Peltier D, Berruyer R, Thomas M, Briard M, Geoffriau E (2008) Expression of carotenoid biosynthesis genes during carrot root development. J Exp Bot 59:3563-3573 https://doi.org/10.1093/jxb/ern210
  5. Collins AR (1999) Oxidative DNA damage, antioxidants, and cancer. Bioessays 21:238-246 https://doi.org/10.1002/(SICI)1521-1878(199903)21:3<238::AID-BIES8>3.0.CO;2-3
  6. Combs GF Jr (1998) Vitamin A, chapter 5 in the vitamins: fundamental aspects in nutrition and health. Academic, New York, pp 107-153
  7. Dall'Osto L, Cazzaniga S, North H, Marion-Poll A, Bassi R (2007) The Arabidopsis aba4-1 mutant reveals a specific function for neoxanthin in protection against photooxidative stress. Plant Cell 19:1048-1064 https://doi.org/10.1105/tpc.106.049114
  8. Demmig-Adams B, Adams WW (1996) The role of xanthophyll cycle carotenoids in the protection of photosynthesis. Trends Plant Sci 1:21-26 https://doi.org/10.1016/S1360-1385(96)80019-7
  9. Demmig-Adams B, Adams WW (2002) Antioxidants in photosynthesis and human nutrition. Science 298:2149-2153 https://doi.org/10.1126/science.1078002
  10. Diretto G, Tavazza R, Welsch R, Pizzichini D, Mourgues F, Papacchioli V, Beyer P, Giuliano G (2006) Metabolic engineering of potato tuber carotenoids through tuber-specific silencing of lycopene epsilon cyclase. BMC Plant Biol 6:13 https://doi.org/10.1186/1471-2229-6-13
  11. Diretto G, Al-Babili S, Tavazza R, Papacchioli V, Beyer P, Giuliano G (2007a) Metabolic engineering of potato carotenoid content through tuber-specific overexpression of a bacterial minipathway. PLoS ONE 2:e350 https://doi.org/10.1371/journal.pone.0000350
  12. Diretto G, Welsch R, Tavazza R, Mourgues F, Pizzichini D, Beyer P, Giuliano G (2007b) Silencing of $\beta-carotene$ hydroxylase increases total carotenoid and $\beta-carotene$ levels in potato tubers. BMC Plant Biol 7:11 https://doi.org/10.1186/1471-2229-7-11
  13. Ducreux LJ, Morris WL, Hedley PE, Shepherd T, Davies HV, Millam S, Taylor MA (2005) Metabolic engineering of high carotenoid potato tubers containing enhanced levels of $\beta-carotene$and lutein. J Exp Bot 56:81-89
  14. Frank HA, Cogdell RJ (1996) Carotenoids in photosynthesis. Photochem Photobiol 63:257-264 https://doi.org/10.1111/j.1751-1097.1996.tb03022.x
  15. Fraser PD, Pinto ME, Holloway DE, Bramley PM (2000) Technical advance: application of high-performance liquid chromatography with photodiode array detection to the metabolic profiling of plant isoprenoids. Plant J 24:551-558 https://doi.org/10.1046/j.1365-313x.2000.00896.x
  16. Fraser PD, Romer S, Shipton CA, Mills PB, Kiano JW, Misawa N, Drake RG, Schuch W, Bramley PM (2002) Evaluation of transgenic tomato plants expressing an additional phytoene synthase in a fruit specific manner. Proc Natl Acad Sci USA 99:1092-1097 https://doi.org/10.1073/pnas.241374598
  17. Fujisawa M, Watanabe M, Choi SK, Teramoto M, Ohyama K, Misawa N (2008) Enrichment of carotenoids in flaxseed (Linum usitatissimum) by metabolic engineering with introduction of bacterial phytoene synthase gene crtB. J Biosci Bioeng 105:636-641 https://doi.org/10.1263/jbb.105.636
  18. Fujisawa M, Takita E, Harada H, Sakurai N, Suzuki H, Ohyama K, Shibata D, Misawa N (2009) Pathway engineering of Brassica napus seeds using multiple key-enzyme genes involved in ketocarotenoid formation. J Exp Bot 60:1319-1332 https://doi.org/10.1093/jxb/erp006
  19. Gerjets T, Sandmann G (2006) Ketocarotenoid formation in transgenic potato. J Exp Bot 57:3639-3645 https://doi.org/10.1093/jxb/erl103
  20. Gerjets T, Sandmann M, Zhu C, Sandmann G (2007) Metabolic engineering of ketocarotenoid biosynthesis in leaves and flowers of tobacco species. Biotechnol J 2:1263-1269 https://doi.org/10.1002/biot.200700040
  21. Hamill JD, Rounsley S, Spencer A, Todd G, Rhodes MJC (1991) The use of the polymerase chain reaction in plant transformation studies. Plant Cell Rep 10:221-224
  22. Harada H, Fujisawa M, Teramoto M, Sakurai N, Suzuki H, Shibata D, Misawa N (2009) Simple functional analysis of key genes involved in astaxanthin biosynthesis using Arabidopsis cultured cells. Plant Biotechnol 26:81-92 https://doi.org/10.5511/plantbiotechnology.26.81
  23. Hasunuma T, Miyazawa SI, Yoshimura S, Shinzaki Y, Tomizawa KI, Shindo K, Choi SK, Misawa N, Miyake C (2008) Biosynthesis of astaxanthin in tobacco leaves by transplastomic engineering. Plant J 55:857-868 https://doi.org/10.1111/j.1365-313X.2008.03559.x
  24. Jayaraj J, Devlin R, Punja Z (2008) Metabolic engineering of novel ketocarotenoid production in carrot plants. Transgenic Res 17:489-501 https://doi.org/10.1007/s11248-007-9120-0
  25. Krapp A, Hofmann B, Schafer C, Stitt M (1993) Regulation of the expression of rbcS and other photosynthetic genes by carbohydrates: a mechanism for the ''sink-regulation'' of photosynthesis? Plant J 3:817-828 https://doi.org/10.1111/j.1365-313X.1993.00817.x
  26. Krinsky NI, Landrum JT, Bone RA (2003) Biologic mechanisms of the protective role of lutein and zeaxanthin in the eye. Annu Rev Nutr 23:171-201 https://doi.org/10.1146/annurev.nutr.23.011702.073307
  27. Mann V, Harker M, Pecker I, Hirschberg J (2000) Metabolic engineering of astaxanthin production in tobacco flowers. Nat Biotechnol 18:888-892 https://doi.org/10.1038/78515
  28. Mii M, Yuzawa Y, Suetomi H, Motegi T, Godo T (1994) Fertile plant regeneration from protoplasts of a seed-propagated cultivar of Lilium ${\times}$ formolongi by utilizing meristematic nodular cell clumps. Plant Sci 100:221-226 https://doi.org/10.1016/0168-9452(94)90079-5
  29. Misawa N (2009) Pathway engineering of plants toward astaxanthin production. Plant Biotechnol 26:93-99 https://doi.org/10.5511/plantbiotechnology.26.93
  30. Misawa N (2010) Carotenoids. In: Mander L, Lui HW (eds) Comprehensive natural products II chemistry and biology, vol 1. Elsevier, Oxford, pp 733-753
  31. Misawa N, Yamano S, Linden H, de Felipe MR, Lucas M, Ikenaga H, Sandmann G (1993) Functional expression of the Erwinia uredovora carotenoid biosynthesis gene crtI in transgenic plants showing an increase of $\beta-carotene$ biosynthesis activity and resistance to the bleaching herbicide norflurazon. Plant J 4:833-840 https://doi.org/10.1046/j.1365-313X.1993.04050833.x
  32. Misawa N, Masamoto K, Hori T, Ohtani T, Boger P, Sandmann G (1994) Expression of an Erwinia phytoene desaturase gene not only confers multiple resistance to herbicides interfering with carotenoid biosynthesis but also alters xanthophyll metabolism in transgenic plants. Plant J 6:481-489 https://doi.org/10.1046/j.1365-313X.1994.6040481.x
  33. Morris WL, Ducreux LJ, Fraser PD, Millam S, Taylor MA (2006) Engineering ketocarotenoid biosynthesis in potato tubers. Metab Eng 8:253-263 https://doi.org/10.1016/j.ymben.2006.01.001
  34. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15:473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  35. Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight DNA. Nucleic Acids Res 8:4321-4325 https://doi.org/10.1093/nar/8.19.4321
  36. Naqvi S, Zhu C, Farre G, Ramessar K, Bassie L, Breitenbach J, Perez Conesa D, Ros G, Sandmann G, Capell T, Christou P (2009) Transgenic multivitamin corn through biofortification of endosperm with three vitamins representing three distinct metabolic pathways. Proc Natl Acad Sci USA 106:7762-7767 https://doi.org/10.1073/pnas.0901412106
  37. Ogaki M, Furuichi Y, Kuroda K, Chin DP, Ogawa Y, Mii M (2008) Importance of co-cultivation medium pH for successful Agrobacterium- mediated transformation of Lilium ${\times}$ formolongi. Plant Cell Rep 27:699-705 https://doi.org/10.1007/s00299-007-0481-x
  38. Paine JA, Shipton CA, Chaggar S, Howells RM, Kennedy MJ, Vernon G, Wright SY, Hinchliffe E, Adams JL, Silverstone AL, Drake R (2005) Improving the nutritional value of Golden Rice through increased pro-vitamin A content. Nat Biotechnol 23:482-487 https://doi.org/10.1038/nbt1082
  39. Park SH, Lee BM, Salas MG, Srivatanakul M, SmithRH (2000) Shorter T-DNA or additional virulence genes improve Agrobactrium-mediated transformation. Theor Appl Genet 101:1015-1020 https://doi.org/10.1007/s001220051575
  40. Ralley L, Enfissi EM, Misawa N, Schuch W, Bramley PM, Fraser PD (2004) Metabolic engineering of ketocarotenoid formation in higher plants. Plant J 39:477-486 https://doi.org/10.1111/j.1365-313X.2004.02151.x
  41. Rosati C, Aquilani R, Dharmapuri S, Pallara P, Marusic C, Tavazza R, Bouvier F, Camara B, Giuliano G (2000) Metabolic engineering of betacarotene and lycopene content in tomato fruit. Plant J 24:413-419 https://doi.org/10.1046/j.1365-313x.2000.00880.x
  42. Sandmann G (2001) Genetic manipulation of carotenoid biosynthesis: strategies, problems and achievements. Trends Plant Sci 6:14-17 https://doi.org/10.1016/S1360-1385(00)01817-3
  43. Shabala SN, Shabala SI, Martynenko AI, Babourina O, Newman IA (1998) Salinity effect on bioelectric activity, growth, Na+ accumulation and chlorophyll fluorescence of maize leaves: a comparative survey and prospects for screening. Australian J Plant Physiol 25:609-616 https://doi.org/10.1071/PP97146
  44. Shewmaker CK, Sheehy JA, Daley M, Colburn S, Ke DY (1999) Seed-specific overexpression of phytoene synthase: increase in carotenoids and other metabolic effects. Plant J 20:401-412 https://doi.org/10.1046/j.1365-313x.1999.00611.x
  45. Stalberg K, Lindgren O, Ek B, Hoglund AS (2003) Synthesis of ketocarotenoids in the seed of Arabidopsis thaliana. Plant J 36:771-779 https://doi.org/10.1046/j.1365-313X.2003.01919.x
  46. Suzuki S, Nishihara M, Nakatsuka T, Misawa N, Ogiwara I, Yamamura S (2007) Flower color alteration in Lotus japonicus by modification of the carotenoid biosynthetic pathway. Plant Cell Rep 26:951-959 https://doi.org/10.1007/s00299-006-0302-7
  47. Yamagishi M, Kishimoto S, Nakayama M (2010) Carotenoid composition and changes in expression of carotenoid biosynthetic genes in tepals of Asiatic hybrid lily. Plant Breed 129:100-107 https://doi.org/10.1111/j.1439-0523.2009.01656.x
  48. Ye X, Al-Babili S, Kloti A, Zhang J, Lucca P, Beyer P, Potrykus I (2000) Engineering the provitamin A ($\beta-carotene$) biosynthetic pathway into (carotenoid-free) rice endosperm. Science 287:303-305 https://doi.org/10.1126/science.287.5451.303
  49. Yuan JP, Chen F (1999) Hydrolysis kinetics of astaxanthin esters and stability of astaxanthin of Haematococcus pluvialis during saponification. J Agric Food Chem 47:31-35 https://doi.org/10.1021/jf980465x
  50. Zhu C, Gerjets T, Sandmann G (2007) Nicotiana glauca engineered for the production of ketocarotenoids in flowers and leaves by expressing the cyanobacterial crtO ketolase gene. Transgenic Res 16:813-821 https://doi.org/10.1007/s11248-007-9151-6
  51. Zhu C, Naqvia S, Breitenbach J, Sandmann G, Christoua P, Capella T (2008) Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize. Proc Natl Acad Sci USA 105:18232-18237 https://doi.org/10.1073/pnas.0809737105

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