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Metabolic engineering of Lilium × formolongi using multiple genes of the carotenoid biosynthesis pathway

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Abstract

Lilium × 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 μg/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 μg/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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Acknowledgments

The authors thank to Dr. Masaharu Kuroda (Hokuriku Research Center, National Agricultural Research Center) for kindly providing binary vector pZH2B, which was used in plasmid pCrtZW-N8idi-crtEBIY. This work was supported by Ministry of Education, Culture, Sport, Science and Technology of the Japanese Government (MONBUKAGAKUSHO) and as a part of the project P02001 by the New Energy and Industrial Technology Development Organization (NEDO). P. Azadi is the recipient of a PhD fellowship from Ministry of Education, Science and Culture of Japan.

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Authors and Affiliations

  1. Laboratory of Plant Cell Technology, Graduate School of Horticulture, Chiba University, 648 Matsudo, Matsudo, Chiba, 271-8510, Japan

    Pejman Azadi, Ntui Valentaine Otang, Dong Poh Chin, Ikuo Nakamura & Masahiro Mii

  2. Central Laboratories for Frontier Technology, i-BIRD, 3-570 Suematsu, Nonoichi-machi, Ishikawa, 921-8836, Japan

    Masaki Fujisawa, Hisashi Harada & Norihiko Misawa

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  1. Pejman Azadi
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  2. Ntui Valentaine Otang
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  4. Ikuo Nakamura
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  5. Masaki Fujisawa
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  8. Masahiro Mii
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Correspondence to Masahiro Mii.

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Azadi, P., Otang, N.V., Chin, D.P. et al. Metabolic engineering of Lilium × formolongi using multiple genes of the carotenoid biosynthesis pathway. Plant Biotechnol Rep 4, 269–280 (2010). https://doi.org/10.1007/s11816-010-0147-y

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  • DOI: https://doi.org/10.1007/s11816-010-0147-y

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