Browse > Article
http://dx.doi.org/10.1007/s11816-010-0147-y

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)
Publication Information
Plant Biotechnology Reports / v.4, no.4, 2010 , pp. 269-280 More about this Journal
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.
Keywords
Carotenoid; Ketocarotenoid; Agrobacteriummediated transformation; Lilium ${\times}$ formolongi; Metabolic engineering; Multi-gene construct;
Citations & Related Records

Times Cited By Web Of Science : 0  (Related Records In Web of Science)
연도 인용수 순위
  • Reference
1 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   DOI   ScienceOn
2 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   DOI   ScienceOn
3 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   DOI   ScienceOn
4 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   DOI   ScienceOn
5 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   DOI   ScienceOn
6 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   DOI   ScienceOn
7 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   DOI   ScienceOn
8 Stalberg K, Lindgren O, Ek B, Hoglund AS (2003) Synthesis of ketocarotenoids in the seed of Arabidopsis thaliana. Plant J 36:771-779   DOI   ScienceOn
9 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   DOI   ScienceOn
10 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   DOI   ScienceOn
11 Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight DNA. Nucleic Acids Res 8:4321-4325   DOI   ScienceOn
12 Misawa N (2009) Pathway engineering of plants toward astaxanthin production. Plant Biotechnol 26:93-99   DOI   ScienceOn
13 Misawa N (2010) Carotenoids. In: Mander L, Lui HW (eds) Comprehensive natural products II chemistry and biology, vol 1. Elsevier, Oxford, pp 733-753
14 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   DOI   ScienceOn
15 Morris WL, Ducreux LJ, Fraser PD, Millam S, Taylor MA (2006) Engineering ketocarotenoid biosynthesis in potato tubers. Metab Eng 8:253-263   DOI   ScienceOn
16 Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15:473-497   DOI
17 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   DOI   ScienceOn
18 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   DOI   ScienceOn
19 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   DOI   ScienceOn
20 Gerjets T, Sandmann G (2006) Ketocarotenoid formation in transgenic potato. J Exp Bot 57:3639-3645   DOI   ScienceOn
21 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   DOI   ScienceOn
22 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
23 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   DOI
24 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   DOI   ScienceOn
25 Jayaraj J, Devlin R, Punja Z (2008) Metabolic engineering of novel ketocarotenoid production in carrot plants. Transgenic Res 17:489-501   DOI   ScienceOn
26 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   DOI   ScienceOn
27 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   DOI   ScienceOn
28 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
29 Frank HA, Cogdell RJ (1996) Carotenoids in photosynthesis. Photochem Photobiol 63:257-264   DOI   ScienceOn
30 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   DOI   ScienceOn
31 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   DOI
32 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   DOI   ScienceOn
33 Combs GF Jr (1998) Vitamin A, chapter 5 in the vitamins: fundamental aspects in nutrition and health. Academic, New York, pp 107-153
34 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   DOI   ScienceOn
35 Demmig-Adams B, Adams WW (2002) Antioxidants in photosynthesis and human nutrition. Science 298:2149-2153   DOI   ScienceOn
36 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   DOI   ScienceOn
37 Collins AR (1999) Oxidative DNA damage, antioxidants, and cancer. Bioessays 21:238-246   DOI   ScienceOn
38 Apel W, Bock R (2009) Enhancement of carotenoid biosynthesis in transplastomic tomatoes by induced lycopene-to-provitamin A conversion. Plant Physiol 151:59-66   DOI   ScienceOn
39 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   DOI   ScienceOn
40 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   DOI   ScienceOn
41 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   DOI   ScienceOn
42 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   DOI   ScienceOn
43 Demmig-Adams B, Adams WW (1996) The role of xanthophyll cycle carotenoids in the protection of photosynthesis. Trends Plant Sci 1:21-26   DOI   ScienceOn
44 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   DOI   ScienceOn
45 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   DOI   ScienceOn
46 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   DOI   ScienceOn
47 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   DOI   ScienceOn
48 Sandmann G (2001) Genetic manipulation of carotenoid biosynthesis: strategies, problems and achievements. Trends Plant Sci 6:14-17   DOI   ScienceOn
49 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   DOI   ScienceOn
50 Mann V, Harker M, Pecker I, Hirschberg J (2000) Metabolic engineering of astaxanthin production in tobacco flowers. Nat Biotechnol 18:888-892   DOI   ScienceOn
51 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   DOI   ScienceOn