Functional Characterization of Nine Norway Spruce TPS Genes and Evolution of Gymnosperm Terpene Synthases of the TPS-d Subfamily  

Abstract

Constitutive and induced terpenoids are important defense compounds for many plants against potential herbivores and pathogens. In Norway spruce (Picea abies L. Karst), treatment with methyl jasmonate induces complex chemical and biochemical terpenoid defense responses associated with traumatic resin duct development in stems and volatile terpenoid emissions in needles. The cloning of (1)-3-carene synthase was the first step in characterizing this system at the molecular genetic level. Here we report the isolation and functional characterization of nine additional terpene synthase (TPS) cDNAs from Norway spruce. These cDNAs encode four monoterpene synthases, myrcene synthase, (2)-limonene synthase, (2)-a/bpinene synthase, and (2)-linalool synthase; three sesquiterpene synthases, longifolene synthase, E,E-a-farnesene synthase, and E-a-bisabolene synthase; and two diterpene synthases, isopimara-7,15-diene synthase and levopimaradiene/abietadiene synthase, each with a unique product profile. To our knowledge, genes encoding isopimara-7,15-diene synthase and longifolene synthase have not been previously described, and this linalool synthase is the first described from a gymnosperm. These functionally diverse TPS account for much of the structural diversity of constitutive and methyl jasmonate-induced terpenoids in foliage, xylem, bark, and volatile emissions from needles of Norway spruce. Phylogenetic analyses based on the inclusion of these TPS into the TPS-d subfamily revealed that functional specialization of conifer TPS occurred before speciation of Pinaceae. Furthermore, based on TPS enclaves created by distinct branching patterns, the TPS-d subfamily is divided into three groups according to sequence similarities and functional assessment. Similarities of TPS evolution in angiosperms and modeling of TPS protein structures are discussed.

Identification of Syn-Pimara-7,15-Diene Synthase Reveals Functional Clustering of Terpene Synthases Involved in Rice Phytoalexin/Allelochemical Biosynthesis

“…2B) demonstrated only weak similarity (,27% identity) to those not involved in labdanerelated diterpenoid biosynthesis (Dudareva et al, 1996;Wildung and Croteau, 1996;Bohlmann et al, 1998b). Slightly higher similarity (approximately 30% identity) was found with the identified gymnosperm bifunctional class II/I diterpene synthases, which are involved in labdane-related biosynthesis (Stofer Vogel et al, 1996;Schepmann et al, 2001;Martin et al, 2004). Further, the sequence is moderately similar (39%-42% identity) to the known ent-kaurene synthases involved in gibberellin biosynthesis (Yamaguchi et al, 1996(Yamaguchi et al, , 1998Richman et al, 1999).…”

Studies on Biosynthetic Genes and Enzymes of Isoprenoids Produced by Actinomycetes

“…Besides Abies grandis, dozens of isoprenoid cyclase genes have been cloned from plants such as castor bean [31] Norway spruce [32,33], maize [34ϳ36], pine [37], ginkgo [38], snapdragon [39], grapevine [40], lotus [41], and tobacco [42]; some of these genes were isolated to investigate the relationship between isoprenoid production and their defensive effects against potential herbivores and pathogens at the molecular genetic level. Moreover, it was proved that among putative isoprenoid synthase genes discovered by whole genome sequence analysis and in silico analysis in Arabidopsis thaliana, at least six genes, namely, At3g25810, At1g61680, At4g16740, At2g24210, At3g25820, and At3g25830, encoded monoterpene synthases [43ϳ46].…”

Variation of Herbivore-Induced Volatile Terpenes among Arabidopsis Ecotypes Depends on Allelic Differences and Subcellular Targeting of Two Terpene Synthases, TPS02 and TPS03

“…S5). Several other TPSs producing (E)-b-ocimene or (E,E)-a-farnesene or both of these terpenes have been identified previously from gymnosperms (TPS-d; Martin et al, 2004) and in the TPS-a, -b, -f, and -g subfamilies of angiosperms (Dudareva et al, 2003;Arimura et al, 2004;Mercke et al, 2004;Nieuwenhuizen et al, 2009; Supplemental Fig. S5).…”