Engineering of tomato type VI glandular trichomes for trans-chrysanthemic acid biosynthesis,the acid moiety of natural pyrethrin insecticides |
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| Institution: | 1. School of Life Sciences, Chongqing University, Chongqing, 401331, China;2. Center of Plant Functional Genomics, Institute of Advanced Interdisciplinary Studies, Chongqing University, Chongqing, 401331, China;3. State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 100101, Beijing, China;4. Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, USA;1. Department of Chemical Engineering (BK21 FOUR Integrated Engineering Program), Kyung Hee University, Yongin-si, Gyeonggi-do, 17104, South Korea;2. Department of Chemical and Biomolecular Engineering, Sogang University, Seoul, 04107, South Korea;1. National Bioenergy Center, National Renewable Energy Laboratory, Golden, CO, 80401, USA;2. Bioscience Division, MS M888, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA;3. Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN, 37831, USA;1. Chongqing Key Laboratory of Plant Resource Conservation and Germplasm Innovation, SWU-TAAHC Medicinal Plant Joint R&D Centre, School of Life Sciences, Southwest University, Chongqing, 400715, China;2. Chongqing Academy of Science and Technology, Chongqing, 401123, China;3. College of Life Sciences, Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), Guizhou University, Guiyang, 550025, China;4. TAAHC-SWU Medicinal Plant Joint R&D Centre, Xizang Agricultural and Husbandry College, Nyingchi of Tibet, 860000, China;5. College of Pharmaceutical Sciences, Key Laboratory of Luminescent and Real-Time Analytical Chemistry (Ministry of Education), Southwest University, Chongqing, 400715, China;6. Citrus Research Institute, Southwest University/Chinese Academy of Agricultural Sciences, Chongqing, 400712, China;1. Charité – Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Department of Rheumatology and Clinical Immunology, Berlin, Germany;2. German Rheumatism Research Centre (DRFZ) Berlin, a Leibniz Institute, Berlin, Germany;3. Berlin Institute of Health and Max-Delbrück-Centrum for Molecular Medicine, Germany;4. Charité – Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Center for Musculoskeletal Surgery, Berlin, Germany;5. Berlin Institute of Health at Charité – Universitätsmedizin Berlin, BIH Biomedical Innovation Academy, BIH Charité Clinician Scientist Program, Berlin, Germany;6. Julius Wolff Institute, Berlin Institute of Health at Charité – Universitätsmedizin, Berlin, Germany;7. Berlin Institute of Health Center for Regenerative Therapies, Berlin Institute of Health at Charité – Universitätsmedizin Berlin, Berlin, Germany;1. Department of Chemical and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan;2. Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan |
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| Abstract: | Glandular trichomes, known as metabolic cell factories, have been proposed as highly suitable for metabolically engineering the production of plant high-value specialized metabolites. Natural pyrethrins, found only in Dalmatian pyrethrum (Tanacetum cinerariifolium), are insecticides with low mammalian toxicity and short environmental persistence. Type I pyrethrins are esters of the monoterpenoid trans-chrysanthemic acid with one of the three rethrolone-type alcohols. To test if glandular trichomes can be made to synthesize trans-chrysanthemic acid, we reconstructed its biosynthetic pathway in tomato type VI glandular trichomes, which produce large amounts of terpenoids that share the precursor dimethylallyl diphosphate (DMAPP) with this acid. This was achieved by coexpressing the trans-chrysanthemic acid pathway related genes including TcCDS encoding chrysanthemyl diphosphate synthase and the fusion gene of TcADH2 encoding the alcohol dehydrogenase 2 linked with TcALDH1 encoding the aldehyde dehydrogenase 1 under the control of a newly identified type VI glandular trichome-specific metallocarboxypeptidase inhibitor promoter. Whole tomato leaves harboring type VI glandular trichomes expressing all three aformentioned genes had a concentration of total trans-chrysanthemic acid that was about 1.5-fold higher (by mole number) than the levels of β-phellandrene, the dominant monoterpene present in non-transgenic leaves, while the levels of β-phellandrene and the representative sesquiterpene β-caryophyllene in transgenic leaves were reduced by 96% and 81%, respectively. These results suggest that the tomato type VI glandular trichome is an alternative platform for the biosynthesis of trans-chrysanthemic acid by metabolic engineering. |
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| Keywords: | Natural pyrethrin pesticides Trans-Chrysanthemic acid Specialized metabolism Trichome-specific metabolic engineering Type VI glandular Trichome Metallocarboxypeptidase inhibitor promoter |
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