Bornyl-diphosphate synthase from Lavandula angustifolia: A major monoterpene synthase involved in essential oil quality

Yang

et al. 2018

Front. Plant Sci.

Amomum villosum, also known as Fructus Amomi, has been used to treat digestive diseases such as abdominal pain, vomiting, and dysentery. Volatile terpenoids are the active metabolites in the essential oil of Fructus Amomi. Nevertheless, downstream genes responsible for activating metabolites biosynthesis in A. villosum still remain unclear. Here, we report the use of an integrative volatile terpenoid profiling and transcriptomics analysis for mining the corresponding genes involved in volatile terpenoid biosynthesis. Ten terpene synthase (TPS) genes were discovered, and two of them were cloned and functionally characterized. AvTPS1 (AvPS: pinene synthase) catalyzed GPP to form α-pinene and β-pinene; AvTPS3 (AvBPPS: bornyl diphosphate synthase) produced bornyl diphosphate as major product and the other three monoterpenoids as minor products. Metabolite accumulation and gene expression pattern combined with AvPS biochemical characterization suggested that AvPS might play a role in biotic defense. On the other hand, the most active ingredient, bornyl acetate, was highly accumulated in seeds and was consistent with the high expression of AvBPPS, which further indicated that AvBPPS is responsible for the biosynthesis of bornyl acetate, the final metabolite of bornyl diphosphate in A. villosum. This study can be used to improve the quality of A. villosum through metabolic engineering, and for the sustainable production of bornyl acetate in heterologous hosts.

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

An Integrative Volatile Terpenoid Profiling and Transcriptomics Analysis for Gene Mining and Functional Characterization of AvBPPS and AvPS Involved in the Monoterpenoid Biosynthesis in Amomum villosum

Yang

et al. 2018

Front. Plant Sci.

“…Monoterpenes were important components of leaf essential oil in C. porrectum, and the study of monoterpenes biosynthesis was of great significance. Monoterpene synthase was the key enzyme in monoterpene biosynthesis, and now many monoterpene synthases have been isolated from plants [22][23][24][25][26][27][28][29]. Linalool synthase (LIS) gene was isolated from different source of Cinnamomum osmophloeum [26].…”

Section: Introductionmentioning

confidence: 99%

“…The recombinant LIS protein can produce s-(+)-linalool from GPP and (E) -Neroli from FPP. Bornyl diphosphate synthase (LaBPPS) was cloned from Lavandula angustifolia [27], which can catalyze the production of Bornyl or camphor. Through bioinformatics analysis, a monoterpene synthetase was identified from Laurus nobilis, and mainly catalyzes the formation of 1,8-cineole [28].…”

Section: Introductionmentioning

confidence: 99%

Full-Length Transcriptome Sequencing and Different Chemotype Expression Profile Analysis of Genes Related to Monoterpenoid Biosynthesis in Cinnamomum porrectum

Qiu

Zheng

et al. 2019

IJMS

Leaves of C. porrectum are rich in essential oils containing monoterpenes, sesquiterpenes and aromatic compounds, but the molecular mechanism of terpenoid biosynthesis in C. porrectum is still unclear. In this paper, the differences in the contents and compositions of terpenoids among three chemotypes were analyzed using gas chromatography mass spectrometry (GC/MS). Furthermore, the differential expression of gene transcripts in the leaf tissues of the three C. porrectum chemotypes were analyzed through a comparison of full-length transcriptomes and expression profiles. The essential oil of the three C. porrectum chemotypes leaves was mainly composed of monoterpenes. In the full-length transcriptome of C. porrectum, 104,062 transcripts with 306,337,921 total bp, an average length of 2944 bp, and an N50 length of 5449 bp, were obtained and 94025 transcripts were annotated. In the eucalyptol and linalool chemotype, the camphor and eucalyptol chemotype, and the camphor and linalool chemotype comparison groups, 21, 22 and 18 terpene synthase (TPS) unigenes were identified respectively. Three monoterpene synthase genes, CpTPS3, CpTPS5 and CpTPS9, were upregulated in the eucalyptol chemotype compared to the linalool chemotype and camphor chemotype. CpTPS1 was upregulated in the camphor chemotype compared to the linalool chemotype and the eucalyptol chemotype. CpTPS4 was upregulated in the linalool chemotype compared to the camphor chemotype and the eucalyptol chemotype. Different unigenes had different expression levels among the three chemotypes, but the unigene expression levels of the 2-C-methyl-D-erythritol 4phosphate (MEP) pathway were generally higher than those of the mevalonate acid (MVA) pathway. Quantitative reverse transcription PCR(qRT-PCR) further validated these expression levels. The present study provides new clues for the functional exploration of the terpenoid synthesis mechanism and key genes in different chemotypes of C. porrectum.

“…Condensation of one DMAPP and one IPP catalyzed by geranyl diphosphate synthase (GPPS) leads to the formation of geranyl diphosphate (GPP) in the plastids, whereas the condensation of one DMAPP and two IPP molecules catalyzed by farnesyl diphosphate synthase (FPPS) leads to the formation of farnesyl diphosphate (FPP) in the cytosol (Despinasse et al, 2017). GPP and FPP serve as substrates for terpene synthases (TPSs) for synthesizing mono- and sesqui-terpenes, respectively (Yahyaa et al, 2015; Despinasse et al, 2017), during which the synthesis of monoterpenes is initiated by GPP dephosphorylation and ionization to geranyl carbocation, while the synthesis of sesquiterpene starts with FPP ionization to a farnesyl cation (Degenhardt et al, 2009; Huang et al, 2010). This is then followed by a series of complex chemical mechanisms involving isomerizations, cyclizations, and rearrangements catalyzed by TPSs, which finally generate structurally diverse terpenoids (Keszei et al, 2010; Koo et al, 2016; Piechulla et al, 2016) ( Figure 1 ).…”

Section: Introductionmentioning

confidence: 99%

De novo Transcriptome Characterization of Rhodomyrtus tomentosa Leaves and Identification of Genes Involved in α/β-Pinene and β-Caryophyllene Biosynthesis

Yang

et al. 2018

Front. Plant Sci.

Plant-derived terpenes are effective in treating chronic dysentery, rheumatism, hepatitis, and hyperlipemia. Thus, understanding the molecular basis of terpene biosynthesis in some terpene-abundant Chinese medicinal plants is of great importance. Abundant in mono- and sesqui-terpenes, Rhodomyrtus tomentosa (Ait.) Hassk, an evergreen shrub belonging to the family Myrtaceae, is widely used as a traditional Chinese medicine. In this study, (+)-α-pinene and β-caryophyllene were detected to be the two major components in the leaves of R. tomentosa, in which (+)-α-pinene is higher in the young leaves than in the mature leaves, whereas the distribution of β-caryophyllene is opposite. Genome-wide transcriptome analysis of leaves identified 138 unigenes potentially involved in terpenoid biosynthesis. By integrating known biosynthetic pathways for terpenoids, 7 candidate genes encoding terpene synthase (RtTPS1-7) that potentially catalyze the last step in pinene and caryophyllene biosynthesis were further characterized. Sequence alignment analysis showed that RtTPS1, RtTPS3 and RtTPS4 do not contain typical N-terminal transit peptides (62–64aa), thus probably producing multiple isomers and enantiomers by terpenoid isomerization. Further enzyme activity in vitro confirmed that RtTPS1-4 mainly produce (+)-α-pinene and (+)-β-pinene, as well as small amounts of (−)-α-pinene and (−)-β-pinene with GPP, while RtTPS1 and RtTPS3 are also active with FPP, producing β-caryophyllene, along with a smaller amount of α-humulene. Our results deepen the understanding of molecular mechanisms of terpenes biosynthesis in Myrtaceae.