Overexpression of the lemon basil alpha-zingiberene synthase gene increases both mono- and sesquiterpene contents in tomato fruit.
alpha-Zingiberene synthase (ZIS), a sesquiterpene synthase gene that was isolated from lemon basil (Ocimum basilicum L.), encodes an enzyme that catalyzes the formation of alpha-zingiberene, and other sesquiterpenes, from farnesyl diphosphate. Transgenic tomato fruits overexpressing ZIS under the control of the fruit ripening-specific tomato polygalacturonase promoter (PG) accumulated high levels of alpha-zingiberene (224-1000 ng g(-1) fresh weight) and other sesquiterpenes, such as alpha-bergamotene, 7-epi-sesquithujene, beta-bisabolene and beta-curcumene, whereas no sesquiterpenes were detected in non-transformed control fruits. The ZIS-transgenic fruits also produced monoterpenes, such as alpha-thujene, alpha-pinene, beta-phellandrene and gamma-terpinene (1-22 ng g(-1) fresh weight), which were either not detected or were found only in minute concentrations in control fruits. Recombinant ZIS overexpressed in Escherichia coli catalyzed the formation of these monoterpenes from geranyl diphosphate. As the ZIS protein apparently lacks a transit peptide, and is localized in the cytosol, the production of monoterpenes in the transgenic tomatoes suggests that a pool of geranyl diphosphate is available in the cytosol. The phenotype of the ZIS-transgenic tomatoes was the same as that for wild-type tomatoes, with regard to plant vigor and shape, but transgenic plants exhibited a small decrease in lycopene content. This study thus showed that the synthesis of both mono- and sesquiterpenes can be enhanced by the ectopic expression of a single transgene in tomato fruit, and it further demonstrated the interconnection between the pools of terpenoid precursors in the plastids and the cytosol.
Davidovich-Rikanati
Rachel
R
Department of Vegetable Crops, Newe Ya'ar Research Center, Agricultural Research Organization, P.O. Box 1021, Ramat Yishay 30095, IsraelDepartment of Life Sciences, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, IsraelDepartment of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USAPresent address: Department of Biotechnology Research, Kazusa DNA Research Institute, 2-5-23 Kazusa-Kamatari, Kisarazu, Chiba 292-0818, JapanThe Jacob Blaustein Institutes for Desert Research, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, Israel.
Lewinsohn
Efraim
E
Department of Vegetable Crops, Newe Ya'ar Research Center, Agricultural Research Organization, P.O. Box 1021, Ramat Yishay 30095, IsraelDepartment of Life Sciences, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, IsraelDepartment of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USAPresent address: Department of Biotechnology Research, Kazusa DNA Research Institute, 2-5-23 Kazusa-Kamatari, Kisarazu, Chiba 292-0818, JapanThe Jacob Blaustein Institutes for Desert Research, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, Israel.
Bar
Einat
E
Department of Vegetable Crops, Newe Ya'ar Research Center, Agricultural Research Organization, P.O. Box 1021, Ramat Yishay 30095, IsraelDepartment of Life Sciences, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, IsraelDepartment of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USAPresent address: Department of Biotechnology Research, Kazusa DNA Research Institute, 2-5-23 Kazusa-Kamatari, Kisarazu, Chiba 292-0818, JapanThe Jacob Blaustein Institutes for Desert Research, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, Israel.
Iijima
Yoko
Y
Department of Vegetable Crops, Newe Ya'ar Research Center, Agricultural Research Organization, P.O. Box 1021, Ramat Yishay 30095, IsraelDepartment of Life Sciences, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, IsraelDepartment of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USAPresent address: Department of Biotechnology Research, Kazusa DNA Research Institute, 2-5-23 Kazusa-Kamatari, Kisarazu, Chiba 292-0818, JapanThe Jacob Blaustein Institutes for Desert Research, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, Israel.
Pichersky
Eran
E
Department of Vegetable Crops, Newe Ya'ar Research Center, Agricultural Research Organization, P.O. Box 1021, Ramat Yishay 30095, IsraelDepartment of Life Sciences, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, IsraelDepartment of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USAPresent address: Department of Biotechnology Research, Kazusa DNA Research Institute, 2-5-23 Kazusa-Kamatari, Kisarazu, Chiba 292-0818, JapanThe Jacob Blaustein Institutes for Desert Research, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, Israel.
Sitrit
Yaron
Y
Department of Vegetable Crops, Newe Ya'ar Research Center, Agricultural Research Organization, P.O. Box 1021, Ramat Yishay 30095, IsraelDepartment of Life Sciences, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, IsraelDepartment of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USAPresent address: Department of Biotechnology Research, Kazusa DNA Research Institute, 2-5-23 Kazusa-Kamatari, Kisarazu, Chiba 292-0818, JapanThe Jacob Blaustein Institutes for Desert Research, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, Israel.
D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism*; D002338:Carotenoids / Q000378:metabolism; D003001:Cloning, Molecular; D011756:Diphosphates / Q000378:metabolism; D004224:Diterpenes / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005638:Fruit / Q000201:enzymology / Q000235:genetics; D008401:Gas Chromatography-Mass Spectrometry; D005822:Genetic Vectors; D018551:Solanum lycopersicum / Q000201:enzymology* / Q000235:genetics; D000081222:Monocyclic Sesquiterpenes; D039821:Monoterpenes / Q000378:metabolism; D018649:Ocimum basilicum / Q000201:enzymology* / Q000235:genetics; D009822:Oils, Volatile / Q000378:metabolism; D010840:Phytosterols / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics; D011106:Polyisoprenyl Phosphates / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics; D012717:Sesquiterpenes / Q000378:metabolism*; D014170:Transformation, Genetic
D011756:Diphosphates; D004224:Diterpenes; D000081222:Monocyclic Sesquiterpenes; D039821:Monoterpenes; D009822:Oils, Volatile; D010840:Phytosterols; D011106:Polyisoprenyl Phosphates; D018749:RNA, Plant; D012717:Sesquiterpenes; C511282:geranyl diphosphate; D002338:Carotenoids; C004808:farnesyl pyrophosphate; C477983:zingiberene; D019883:Alkyl and Aryl Transferases; C108877:terpene synthase