Isolation and characterization of terpene synthases in cotton (Gossypium hirsutum) |
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| Affiliation: | 1. National Key Laboratory of Plant Molecular Genetics and National Center for Plant Gene Research, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China;2. Plant Science Research Center, Shanghai Chenshan Botanical Garden, Shanghai 201602, China;1. Program of Plant Metabolism and Metabolic Engineering, Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Campus UAB, Bellaterra, E-08193, Barcelona, Spain;2. Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, 08028 Barcelona, Spain;1. Department of Agriculture, Food and Environment, University of Pisa, Via del Borghetto 80, I-56124 Pisa, Italy;2. Nutraceuticals and Food for Health – Nutrafood, University of Pisa, Via del Borghetto 80, I-56124 Pisa, Italy;1. Centro de Ciências do Mar, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal;2. Centre for Mediterranean Bioresources and Food (MeditBio), Faculdade de Ciências e Tecnologia, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal;3. Centro de Estudos do Ambiente e do Mar Lisboa, Faculdade de Ciências, Universidade de Lisboa, CBV, DBV, 1749‐016 Lisboa, Portugal;4. Center of Electronics, Optoelectronics and Telecommunications, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal;1. Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune 411008, Maharashtra, India;2. Division of Organic Chemistry, CSIR-National Chemical Laboratory, Pune 411008, Maharashtra, India;3. Department of Biotechnology, Savitribai Phule Pune University, Pune 411007, Maharashtra, India;4. Biospheres, Eshwari, 52/403, Lakshminagar, Parvati, Pune 411 009, Maharashtra, India;5. CSIR- Institute of Genomics and Integrative Biology, Mall Road, New Delhi 110007, India |
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| Abstract: | Cotton plants accumulate gossypol and related sesquiterpene aldehydes, which function as phytoalexins against pathogens and feeding deterrents to herbivorous insects. However, to date little is known about the biosynthesis of volatile terpenes in this crop. Herein is reported that 5 monoterpenes and 11 sesquiterpenes from extracts of a glanded cotton cultivar, Gossypium hirsutum cv. CCRI12, were detected by gas chromatography–mass spectrometry (GC–MS). By EST data mining combined with Rapid Amplification of cDNA Ends (RACE), full-length cDNAs of three terpene synthases (TPSs), GhTPS1, GhTPS2 and GhTPS3 were isolated. By in vitro assays of the recombinant proteins, it was found that GhTPS1 and GhTPS2 are sesquiterpene synthases: the former converted farnesyl pyrophosphate (FPP) into β-caryophyllene and α-humulene in a ratio of 2:1, whereas the latter produced several sesquiterpenes with guaia-1(10),11-diene as the major product. By contrast, GhTPS3 is a monoterpene synthase, which produced α-pinene, β-pinene, β-phellandrene and trace amounts of other monoterpenes from geranyl pyrophosphate (GPP). The TPS activities were also supported by Virus Induced Gene Silencing (VIGS) in the cotton plant. GhTPS1 and GhTPS3 were highly expressed in the cotton plant overall, whereas GhTPS2 was expressed only in leaves. When stimulated by mechanical wounding, Verticillium dahliae (Vde) elicitor or methyl jasmonate (MeJA), production of terpenes and expression of the corresponding synthase genes were induced. These data demonstrate that the three genes account for the biosynthesis of volatile terpenes of cotton, at least of this Upland cotton. |
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| Keywords: | Malvaceae Terpene synthase VIGS |
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