Isolation and characterization of terpene synthases in cotton (Gossypium hirsutum).
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.
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.
Chang-Qing
CQ
Yang
Xiu-Ming
XM
Wu
Ju-Xin
JX
Ruan
Wen-Li
WL
Hu
Yin-Bo
YB
Mao
Xiao-Ya
XY
Chen
Ling-Jian
LJ
Wang
D000085:Acetates / Q000378:metabolism; D019883:Alkyl and Aryl Transferases / Q000302:isolation & purification* / Q000378:metabolism*; D000081008:Bicyclic Monoterpenes; D001643:Bridged Bicyclo Compounds / Q000378:metabolism; D000081005:Cyclohexane Monoterpenes; D053138:Cyclohexenes / Q000378:metabolism; D003517:Cyclopentanes / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D003368:Gossypium / Q000201:enzymology* / Q000235:genetics; D019753:Intramolecular Lyases / Q000378:metabolism; D000081222:Monocyclic Sesquiterpenes; D039821:Monoterpenes / Q000378:metabolism; D054883:Oxylipins / Q000378:metabolism; D000081223:Polycyclic Sesquiterpenes; D011106:Polyisoprenyl Phosphates / Q000378:metabolism; D012717:Sesquiterpenes / Q000378:metabolism; D045789:Sesquiterpenes, Guaiane / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism; D055549:Volatile Organic Compounds / Q000378:metabolism; D000094162:Phytoalexins
D000085:Acetates; D000081008:Bicyclic Monoterpenes; D001643:Bridged Bicyclo Compounds; D000081005:Cyclohexane Monoterpenes; D053138:Cyclohexenes; D003517:Cyclopentanes; D000081222:Monocyclic Sesquiterpenes; D039821:Monoterpenes; D054883:Oxylipins; D000081223:Polycyclic Sesquiterpenes; D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; D045789:Sesquiterpenes, Guaiane; D013729:Terpenes; D055549:Volatile Organic Compounds; C501248:guaia-1(10),11-diene; C058582:beta-phellandrene; C010789:beta-pinene; C042686:humulene; C015234:geranyl pyrophosphate; C004808:farnesyl pyrophosphate; C072239:methyl jasmonate; C024714:caryophyllene; D019883:Alkyl and Aryl Transferases; C108877:terpene synthase; D019753:Intramolecular Lyases; C040265:pinene cyclase I; C005451:alpha-pinene; D000094162:Phytoalexins
Cotton (Gossypium hirsutum); Malvaceae; Terpene synthase; VIGS