Characterisation of an (S)-linalool synthase from kiwifruit (Actinidia arguta) that catalyses the first committed step in the production of floral lilac compounds

2010 ◽  
Vol 37 (3) ◽  
pp. 232 ◽  
Author(s):  
Xiuyin Chen ◽  
Yar-Khing Yauk ◽  
Niels J. Nieuwenhuizen ◽  
Adam J. Matich ◽  
Mindy Y. Wang ◽  
...  
Keyword(s):  
Terpene Synthase ◽  
Pcr Analysis ◽  
In Planta ◽  
Terpene Synthases ◽  
Actinidia Arguta ◽  
Cdna Sequences ◽  
Key Enzyme ◽  
Enantioselective Gas Chromatography ◽  
Actinidia Spp ◽  
Linalool Synthase

Kiwifruit (Actinidia spp. Lindl.) flowers and fruit contain many compounds of interest to the flavour and fragrance industries. In particular, Actinidia arguta (Sieb. et Zucc.) Planch. ex Miq. flowers produce β-linalool and important derivatives thereof, including linalool oxides, lilac aldehydes, alcohols and alcohol epoxides. Dynamic headspace sampling of whole A. arguta flowers showed that the peak emission rate of linalool, lilac alcohols and lilac aldehydes occurred around 0800 hours. After solvent extraction, linalool levels remained constant throughout the day and night, but lilac alcohol levels peaked at noon. In whole flowers, linalool was found predominantly in pistils and petals, and the lilac compounds were found mainly in petals. Two highly homologous (96.6% nucleotide identity) terpene synthase cDNA sequences, AaLS1 and ApLS1, were isolated from A. arguta and Actinidia polygama (Sieb. et Zucc.) Maxim flower EST libraries respectively. Real-time PCR analysis revealed that AaLS1 was expressed constitutively throughout the day and night, and primarily in petal tissue. Functional analysis in Escherichia coli showed that AaLS1 and ApLS1 each encoded a linalool synthase which was confirmed by transient expression in planta. Enantioselective gas chromatography revealed that both terpene synthases produced only (S)-(+)-linalool. AaLS1, therefore, is likely to be the key enzyme producing the (S)-linalool precursor of the lilac alcohols and aldehydes in A. arguta flowers.

scholarly journals A Wheat β-Patchoulene Synthase Confers Resistance against Herbivory in Transgenic Arabidopsis

Genes ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 441 ◽  
Author(s):  
Qingyu Pu ◽  
Jin Liang ◽  
Qinqin Shen ◽  
Jingye Fu ◽  
Zhien Pu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Catalytic Mechanism ◽  
Transgenic Arabidopsis ◽  
Triticum Aestivum L ◽  
Site Directed Mutagenesis ◽  
Beet Armyworm ◽  
Terpene Synthase ◽  
In Planta ◽  
Terpene Synthases ◽  
Sesquiterpene Synthase

Terpenoids play important roles in plant defense. Although some terpene synthases have been characterized, terpenoids and their biosynthesis in wheat (Triticum aestivum L.) still remain largely unknown. Here, we describe the identification of a terpene synthase gene in wheat. It encodes a sesquiterpene synthase that catalyzes β-patchoulene formation with E,E-farnesyl diphosphate (FPP) as the substrate, thus named as TaPS. TaPS exhibits inducible expression in wheat in response to various elicitations. Particularly, alamethicin treatment strongly induces TaPS gene expression and β-patchoulene accumulation in wheat. Overexpression of TaPS in Arabidopsis successfully produces β-patchoulene, verifying the biochemical function of TaPS in planta. Furthermore, these transgenic Arabidopsis plants exhibit resistance against herbivory by repelling beet armyworm larvae feeding, thereby indicating anti-herbivory activity of β-patchoulene. The catalytic mechanism of TaPS is also explored by homology modeling and site-directed mutagenesis. Two key amino acids are identified to act in protonation and stability of intermediates and product formation. Taken together, one wheat sesquiterpene synthase is identified as β-patchoulene synthase. TaPS exhibits inducible gene expression and the sesquiterpene β-patchoulene is involved in repelling insect infestation.

scholarly journals Transcriptome and proteome of the corm, leaf and flower of Hypoxis hemerocallidea (African potato)

PLoS ONE ◽  
2021 ◽  
Vol 16 (7) ◽  
pp. e0253741
Author(s):  
Mihai-Silviu Tomescu ◽  
Selisha Ann Sooklal ◽  
Thuto Ntsowe ◽  
Previn Naicker ◽  
Barbara Darnhofer ◽  
...  
Keyword(s):  
De Novo ◽  
Differential Expression Analysis ◽  
Terpene Synthase ◽  
Future Research ◽  
Terpene Synthases ◽  
Illumina Hiseq ◽  
Prostate Hyperplasia ◽  
Inflammatory Conditions ◽  
Urinary Infections ◽  
Hypoxis Hemerocallidea

The corm of Hypoxis hemerocallidea, commonly known as the African potato, is used in traditional medicine to treat several medical conditions such as urinary infections, benign prostate hyperplasia, inflammatory conditions and testicular tumours. The metabolites contributing to the medicinal properties of H. hemerocallidea have been identified in several studies and, more recently, the active terpenoids of the plant were profiled. However, the biosynthetic pathways and the enzymes involved in the production of the terpene metabolites in H. hemerocallidea have not been characterised at a transcriptomic or proteomic level. In this study, total RNA extracted from the corm, leaf and flower tissues of H. hemerocallidea was sequenced on the Illumina HiSeq 2500 platform. A total of 143,549 transcripts were assembled de novo using Trinity and 107,131 transcripts were functionally annotated using the nr, GO, COG, KEGG and SWISS-PROT databases. Additionally, the proteome of the three tissues were sequenced using LC-MS/MS, revealing aspects of secondary metabolism and serving as data validation for the transcriptome. Functional annotation led to the identification of numerous terpene synthases such as nerolidol synthase, germacrene D synthase, and cycloartenol synthase amongst others. Annotations also revealed a transcript encoding the terpene synthase phytoalexin momilactone A synthase. Differential expression analysis using edgeR identified 946 transcripts differentially expressed between the three tissues and revealed that the leaf upregulates linalool synthase compared to the corm and the flower tissues. The transcriptome as well as the proteome of Hypoxis hemerocallidea presented here provide a foundation for future research.

scholarly journals Identification of Sesquiterpene Synthases from Nostoc punctiforme PCC 73102 and Nostoc sp. Strain PCC 7120

2008 ◽  
Vol 190 (18) ◽  
pp. 6084-6096 ◽  
Author(s):  
Sean A. Agger ◽  
Fernando Lopez-Gallego ◽  
Thomas R. Hoye ◽  
Claudia Schmidt-Dannert
Keyword(s):  
Functional Expression ◽  
Terpene Synthase ◽  
Putative Hybrid ◽  
Nostoc Punctiforme ◽  
Terpene Synthases ◽  
E Coli ◽  
Reductase Gene ◽  
Pcc 7120 ◽  
Defense Compound

ABSTRACT Cyanobacteria are a rich source of natural products and are known to produce terpenoids. These bacteria are the major source of the musty-smelling terpenes geosmin and 2-methylisoborneol, which are found in many natural water supplies; however, no terpene synthases have been characterized from these organisms to date. Here, we describe the characterization of three sesquiterpene synthases identified in Nostoc sp. strain PCC 7120 (terpene synthase NS1) and Nostoc punctiforme PCC 73102 (terpene synthases NP1 and NP2). The second terpene synthase in N. punctiforme (NP2) is homologous to fusion-type sesquiterpene synthases from Streptomyces spp. shown to produce geosmin via an intermediate germacradienol. The enzymes were functionally expressed in Escherichia coli, and their terpene products were structurally identified as germacrene A (from NS1), the eudesmadiene 8a-epi-α-selinene (from NP1), and germacradienol (from NP2). The product of NP1, 8a-epi-α-selinene, so far has been isolated only from termites, in which it functions as a defense compound. Terpene synthases NP1 and NS1 are part of an apparent minicluster that includes a P450 and a putative hybrid two-component protein located downstream of the terpene synthases. Coexpression of P450 genes with their adjacent located terpene synthase genes in E. coli demonstrates that the P450 from Nostoc sp. can be functionally expressed in E. coli when coexpressed with a ferredoxin gene and a ferredoxin reductase gene from Nostoc and that the enzyme oxygenates the NS1 terpene product germacrene A. This represents to the best of our knowledge the first example of functional expression of a cyanobacterial P450 in E. coli.

Combined transcriptome sequencing and prokaryotic expression to investigate the key enzyme in the 2-C-methylerythritol-4-phosphate pathway of Osmanthus fragrans

2020 ◽  
Vol 47 (10) ◽  
pp. 945
Author(s):  
Rui Xiong ◽  
Zhu Chen ◽  
Weiyu Wang ◽  
Li Jiang ◽  
Yan Xiang ◽  
...  
Keyword(s):  
Prokaryotic Expression ◽  
Expression Vectors ◽  
Pcr Analysis ◽  
Subcellular Localisation ◽  
Reaction Products ◽  
Osmanthus Fragrans ◽  
Qrt Pcr ◽  
Key Enzyme ◽  
Series Of Experiments ◽  
Main Components

Terpenoids are one of the main components of plant aromas. In the present study we investigated these compounds in Osmanthus fragrans Lour., which is a fragrant plant widely used for the production of essential oils. Quantitative real-time PCR (qRT-PCR) results of enzymes associated with the 2-C-methylerythritol-4-phosphate pathway confirmed that the TPS is a key enzyme for terpenoid synthesis in O. fragrans. In a series of experiments, we identified the TPS candidate genes in O. fragrans and revealed the underlying catalytic activity and subcellular localisation of the encoded proteins. Because there is no available O. fragrans reference genome, we sequenced and analysed its transcriptome and identified two putative TPS genes, OfTPS1 and OfTPS2. According to qRT-PCR analysis, both genes were most highly expressed at the full-bloom stage, suggesting that OfTPS1 and OfTPS2 contribute to O. fragrans terpenoid synthesis. To verify this hypothesis, we constructed prokaryotic expression vectors to obtain protein. In order to study the function of OfTPS1 and OfTPS2 in the synthesis of monoterpenes, the obtained proteins were reacted with geranyl pyrophosphate. As a result, two kinds of monoterpenes, (E)-β-ocimene and linalool, were detected from reaction products, respectively. In conclusion, OfTPS1 and OfTPS2 are both monoterpene synthases.

scholarly journals A Comprehensive Survey on the Terpene Synthase Gene Family Provides New Insight into Its Evolutionary Patterns

2019 ◽  
Vol 11 (8) ◽  
pp. 2078-2098 ◽  
Author(s):  
Shu-Ye Jiang ◽  
Jingjing Jin ◽  
Rajani Sarojam ◽  
Srinivasan Ramachandran
Keyword(s):  
Gene Family ◽  
Large Scale ◽  
Family Members ◽  
Terpene Synthase ◽  
Limited Information ◽  
Terpene Synthases ◽  
Genome Wide ◽  
A Genome ◽  
Family Expansion ◽  
Insight Into

Abstract Terpenes are organic compounds and play important roles in plant growth and development as well as in mediating interactions of plants with the environment. Terpene synthases (TPSs) are the key enzymes responsible for the biosynthesis of terpenes. Although some species were employed for the genome-wide identification and characterization of the TPS family, limited information is available regarding the evolution, expansion, and retention mechanisms occurring in this gene family. We performed a genome-wide identification of the TPS family members in 50 sequenced genomes. Additionally, we also characterized the TPS family from aromatic spearmint and basil plants using RNA-Seq data. No TPSs were identified in algae genomes but the remaining plant species encoded various numbers of the family members ranging from 2 to 79 full-length TPSs. Some species showed lineage-specific expansion of certain subfamilies, which might have contributed toward species or ecotype divergence or environmental adaptation. A large-scale family expansion was observed mainly in dicot and monocot plants, which was accompanied by frequent domain loss. Both tandem and segmental duplication significantly contributed toward family expansion and expression divergence and played important roles in the survival of these expanded genes. Our data provide new insight into the TPS family expansion and evolution and suggest that TPSs might have originated from isoprenyl diphosphate synthase genes.

scholarly journals Scenarios of Genes-to-Terpenoids Network Led to the Identification of a Novel α/β-Farnesene/β-Ocimene Synthase in Camellia sinensis

2020 ◽  
Vol 21 (2) ◽  
pp. 655
Author(s):  
Jieyang Jin ◽  
Shangrui Zhang ◽  
Mingyue Zhao ◽  
Tingting Jing ◽  
Na Zhang ◽  
...  
Keyword(s):  
Function Analysis ◽  
Pearson Correlation ◽  
Integrated Approach ◽  
In Planta ◽  
Terpene Synthases ◽  
Gene Transcripts ◽  
Pearson Correlation Analysis ◽  
Tea Plants ◽  
And Function

Terpenoids play vital roles in tea aroma quality and plants defense performance determination, whereas the scenarios of genes to metabolites of terpenes pathway remain uninvestigated in tea plants. Here, we report the use of an integrated approach combining metabolites, target gene transcripts and function analyses to reveal a gene-to-terpene network in tea plants. Forty-one terpenes including 26 monoterpenes, 14 sesquiterpenes and one triterpene were detected and 82 terpenes related genes were identified from five tissues of tea plants. Pearson correlation analysis resulted in genes to metabolites network. One terpene synthases whose expression positively correlated with farnesene were selected and its function was confirmed involved in the biosynthesis of α-farnesene, β-ocimene and β-farnesene, a very important and conserved alarm pheromone in response to aphids by both in vitro enzymatic assay in planta function analysis. In summary, we provided the first reliable gene-to-terpene network for novel genes discovery.

scholarly journals Agasicles hygrophila attack increases nerolidol synthase gene expression in Alternanthera philoxeroides, facilitating host finding

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Yuanxin Wang ◽  
Yanhong Liu ◽  
Xingchun Wang ◽  
Dong Jia ◽  
Jun Hu ◽  
...  
Keyword(s):  
Host Plants ◽  
Molecular Mechanisms ◽  
Host Finding ◽  
Alternanthera Philoxeroides ◽  
Terpene Synthase ◽  
Mechanical Wounding ◽  
Plant Volatile ◽  
Egg Deposition ◽  
Key Enzyme ◽  
Plant Volatile Compounds

Abstract Herbivorous insects use plant volatile compounds to find their host plants for feeding and egg deposition. The monophagous beetle Agasicles hygrophila uses a volatile (E)-4,8-dimethyl-1,3,7-nonanetriene (DMNT) to recognize its host plant Alternanthera philoxeroides. Alternanthera philoxeroides releases DMNT in response to A. hygrophila attack and nerolidol synthase (NES) is a key enzyme in DMNT biosynthesis; however, the effect of A. hygrophila on NES expression remains unclear. In this study, the A. philoxeroides transcriptome was sequenced and six putative NES genes belonging to the terpene synthase-g family were characterized. The expression of these NES genes was assayed at different times following A. hygrophila contact, feeding or mechanical wounding. Results showed that A. hygrophila contact and feeding induced NES expression more rapidly and more intensely than mechanical wounding alone. This may account for a large release of DMNT following A. hygrophila feeding in a previous study and subsequently facilitate A. hygrophila to find host plants. Our research provides a powerful genetic platform for studying invasive plants and lays the foundation for further elucidating the molecular mechanisms of the interaction between A. philoxeroides and its specialist A. hygrophila.

scholarly journals Promiscuous terpene synthases from Prunella vulgaris highlight the importance of substrate and compartment switching in terpene synthase evolution

2019 ◽  
Vol 223 (1) ◽  
pp. 323-335 ◽  
Author(s):  
Sean R. Johnson ◽  
Wajid Waheed Bhat ◽  
Radin Sadre ◽  
Garret P. Miller ◽  
Alekzander Sky Garcia ◽  
...  
Keyword(s):  
Terpene Synthase ◽  
Prunella Vulgaris ◽  
Terpene Synthases

PI-PLCbeta1 Expression in Patients with High-Risk Myelodysplastic Syndromes Is Affected by Azacitidine Treatment.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2448-2448
Author(s):  
Carlo Finelli ◽  
Matilde Y. Follo ◽  
Costanza Bosi ◽  
Sara Mongiorgi ◽  
Cristina Clissa ◽  
...  
Keyword(s):  
High Risk ◽  
Myelodysplastic Syndromes ◽  
Erythroid Differentiation ◽  
Response To Treatment ◽  
Hl60 Cell ◽  
Pcr Analysis ◽  
Cellular Processes ◽  
Healthy Donors ◽  
Key Enzyme ◽  
Hl60 Cell Line

Abstract Phosphoinositide-specific phospholipase C (PI-PLC) beta1 is a key enzyme in nuclear signal transduction, and it is involved in many cellular processes, such as proliferation and differentiation (Manzoli L et al, Prog Lipid Res, 2005). In particular, the involvement of the PI-PLCbeta1 gene in erythroid differentiation lead us to investigate this gene in patients affected by high-risk Myelodysplastic Syndromes (MDS). It is still unclear what is the pathogenesis of the evolution of MDS into Acute Myeloid Leukemia (AML), even if the presence of a mono-allelic and cryptic deletion of the PI-PLCbeta1 gene, as well as an impaired regulation of the PI3K/Akt/mTOR axis, have been recently hypothesized to be implicated in mechanisms related to the disease progression (Lo Vasco VR et al, Leukemia, 2004;Follo MY et al, Cancer Res, 2007). In the present study, we performed a relative quantification, by Real-Time Polymerase Chain Reaction (PCR) analysis, on high-risk MDS patients, at baseline and during treatment with azacitidine. Furthermore, we evaluated the expression of the PI-PLCbeta1 gene on healthy donors and the HL60 cell line, which is useful for testing the accuracy of the technology because of its low expression of PI-PLCbeta1. To analyze and quantify the levels of the two different splicing variants of the PI-PLCbeta1 gene (a and b), we used a TaqMan isoform specific approach. We studied 8 patients with high-risk MDS (IPSS risk high or intermediate-II) treated with azaciditine, 5 of them showing a favourable response to treatment (1 patient: complete remission; 2 patients: partial remission; 2 patients: haematologic improvement). During the treatment with azacitidine, the non responsive patients (3/8) did not show any significant change in the levels of PI-PLCbeta1 mRNAs, whilst all the responders showed a marked increase of the PI-PLCbeta1 mRNA as compared with their baseline amount. Interestingly, the responsive patients showed fluctuations of PI-PLCbeta1 levels that could be related to their clinical status.Our data show a correlation between azacitidine treatment and PI-PLCbeta1 signalling in high-risk MDS, hinting at the likelihood that azacitidine could affect the transcriptional activity of PI-PLCbeta1, which is indeed a key player in the control of cell cycle.

The terpene synthase gene family in Gossypium hirsutum harbors a linalool synthase GhTPS12 implicated in direct defence responses against herbivores

2017 ◽  
Vol 41 (1) ◽  
pp. 261-274 ◽  
Author(s):  
Xin-Zheng Huang ◽  
Yu-Tao Xiao ◽  
Tobias G. Köllner ◽  
Wei-Xia Jing ◽  
Jun-Feng Kou ◽  
...  
Keyword(s):  
Gossypium Hirsutum ◽  
Gene Family ◽  
Terpene Synthase ◽  
Defence Responses ◽  
Direct Defence ◽  
Linalool Synthase
Sign in / Sign up

Export Citation Format

Share Document