Terpene Biosynthesis in Red Algae Is Catalyzed by Microbial Type But Not Typical Plant Terpene Synthases

Guo Wei; Qidong Jia; Xinlu Chen; Tobias G. Köllner; Debashish Bhattacharya; Gane Ka-Shu Wong; Jonathan Gershenzon; Feng Chen

doi:10.1104/pp.18.01413

Differential gene expression associated with a floral scent polymorphism in the evening primrose Oenothera harringtonii (Onagraceae)

10.1101/2021.01.12.426409 ◽

2021 ◽

Author(s):

Lindsey L. Bechen ◽

Matthew G. Johnson ◽

Geoffrey T. Broadhead ◽

Rachel A. Levin ◽

Rick P. Overson ◽

...

Keyword(s):

Gene Expression ◽

Differential Gene Expression ◽

Floral Scent ◽

Terpene Synthase ◽

Rna Seq ◽

Loss Of Function ◽

Terpene Biosynthesis ◽

Terpene Synthases ◽

Evening Primrose ◽

Differential Gene

AbstractBackgroundPlant volatiles play an important role in both plant-pollinator and plant-herbivore interactions. Intraspecific polymorphisms in volatile production are ubiquitous, but studies that explore underlying differential gene expression are rare. Oenothera harringtonii populations are polymorphic in floral emission of the monoterpene (R)-(-)-linalool; some plants emit (R)-(-)-linalool (linalool+ plants) while others do not (linalool-plants). However, the genes associated with differential production of this floral volatile in Oenothera are unknown. We used RNA-Seq to broadly characterize differential gene expression involved in (R)-(-)-linalool biosynthesis. To identify genes that may be associated with the polymorphism for this trait, we used RNA-Seq to compare gene expression in six different Oenothera harringtonii tissues from each of three linalool+ and linalool-plants.ResultsThree clusters of differentially expressed genes were enriched for terpene synthase activity: two were characterized by tissue-specific upregulation and one by upregulation only in plants with flowers that produce (R)-(-)-linalool. A molecular phylogeny of all terpene synthases identified two putative (R)-(-)-linalool synthase transcripts in Oenothera harringtonii, a single allele of which is found exclusively in linalool+ plants.ConclusionsBy using a naturally occurring polymorphism and comparing different tissues, we were able to identify genes putatively involved in the biosynthesis of (R)-(-)-linalool. Expression of these genes in linalool-plants suggests a regulatory polymorphism, rather than a population-specific loss-of-function allele. Additional terpene biosynthesis-related genes that are up-regulated in plants that emit (R)-(-)-linalool may be associated with herbivore defense, suggesting a potential economy of scale between plant reproduction and defense.

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Emission and biosynthesis of volatile terpenoids from the plasmodial slime mold Physarum polycephalum

Beilstein Journal of Organic Chemistry ◽

10.3762/bjoc.15.281 ◽

2019 ◽

Vol 15 ◽

pp. 2872-2880

Author(s):

Xinlu Chen ◽

Tobias G Köllner ◽

Wangdan Xiong ◽

Guo Wei ◽

Feng Chen

Keyword(s):

Physarum Polycephalum ◽

Slime Mold ◽

Terpene Biosynthesis ◽

Terpene Synthases ◽

Slime Molds ◽

Geranyl Diphosphate ◽

Monoterpene Synthase ◽

Cellular Slime ◽

Volatile Terpenoids

Terpene synthases (TPSs) are pivotal enzymes for the production of diverse terpenes, including monoterpenes, sesquiterpenes, and diterpenes. In our recent studies, dictyostelid social amoebae, also known as cellular slime molds, were found to contain TPS genes for making volatile terpenes. For comparison, here we investigated Physarum polycephalum, a plasmodial slime mold also known as acellular amoeba. Plasmodia of P. polycephalum grown on agar plates were found to release a mixture of volatile terpenoids consisting of four major sesquiterpenes (α-muurolene, (E)-β-caryophyllene, two unidentified sesquiterpenoids) and the monoterpene linalool. There were no qualitative differences in terpenoid composition at two stages of young plasmodia. To understand terpene biosynthesis, we analyzed the transcriptome and genome sequences of P. polycephalum and identified four TPS genes designated PpolyTPS1–PpolyTPS4. They share 28–73% of sequence identities. Full-length cDNAs for the four TPS genes were cloned and expressed in Escherichia coli to produce recombinant proteins, which were tested for sesquiterpene synthase and monoterpene synthase activities. While neither PpolyTPS2 nor PpolyTPS3 was active, PpolyTPS1 and PpolyTPS4 were able to produce sesquiterpenes and monoterpenes from the respective substrates farnesyl diphosphate and geranyl diphosphate. By comparing the volatile profile of P. polycephalum plasmodia and the in vitro products of PpolyTPS1 and PpolyTPS4, it was concluded that most sesquiterpenoids emitted from P. polycephalum were attributed to PpolyTPS4. Phylogenetic analysis placed the four PpolyTPSs genes into two groups: PpolyTPS1 and PpolyTPS4 being one group that was clustered with the TPSs from the dictyostelid social amoeba and PpolyTPS2 and PpolyTPS3 being the other group that showed closer relatedness to bacterial TPSs. The biological role of the volatile terpenoids produced by the plasmodia of P. polycephalum is discussed.

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Terpene synthase genes in eukaryotes beyond plants and fungi: Occurrence in social amoebae

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1610379113 ◽

2016 ◽

Vol 113 (43) ◽

pp. 12132-12137 ◽

Cited By ~ 47

Author(s):

Xinlu Chen ◽

Tobias G. Köllner ◽

Qidong Jia ◽

Ayla Norris ◽

Balaji Santhanam ◽

...

Keyword(s):

Early Stage ◽

Terpene Synthase ◽

Functional Study ◽

Terpene Biosynthesis ◽

Terpene Synthases ◽

Multicellular Development ◽

Stage Of Development ◽

The Social ◽

Volatile Terpenes ◽

Social Amoebae

Terpenes are structurally diverse natural products involved in many ecological interactions. The pivotal enzymes for terpene biosynthesis, terpene synthases (TPSs), had been described only in plants and fungi in the eukaryotic domain. In this report, we systematically analyzed the genome sequences of a broad range of nonplant/nonfungus eukaryotes and identified putative TPS genes in six species of amoebae, five of which are multicellular social amoebae from the order of Dictyosteliida. A phylogenetic analysis revealed that amoebal TPSs are evolutionarily more closely related to fungal TPSs than to bacterial TPSs. The social amoeba Dictyostelium discoideum was selected for functional study of the identified TPSs. D. discoideum grows as a unicellular organism when food is abundant and switches from vegetative growth to multicellular development upon starvation. We found that expression of most D. discoideum TPS genes was induced during development. Upon heterologous expression, all nine TPSs from D. discoideum showed sesquiterpene synthase activities. Some also exhibited monoterpene and/or diterpene synthase activities. Direct measurement of volatile terpenes in cultures of D. discoideum revealed essentially no emission at an early stage of development. In contrast, a bouquet of terpenes, dominated by sesquiterpenes including β-barbatene and (E,E)-α-farnesene, was detected at the middle and late stages of development, suggesting a development-specific function of volatile terpenes in D. discoideum. The patchy distribution of TPS genes in the eukaryotic domain and the evidence for TPS function in D. discoideum indicate that the TPS genes mediate lineage-specific adaptations.

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First Draft Genome of a Brazilian Atlantic Rainforest Burseraceae reveals commercially-promising genes involved in terpenic oleoresins synthesis

10.1101/467720 ◽

2018 ◽

Author(s):

Luana Ferreira Afonso ◽

Danielle Amaral ◽

Marcela Uliano-Silva ◽

André Luiz Quintanilha Torres ◽

Daniel Reis Simas ◽

...

Keyword(s):

De Novo ◽

Pfam Domain ◽

Draft Genome ◽

Atlantic Rainforest ◽

Volatile Fraction ◽

Terpene Biosynthesis ◽

Terpene Synthases ◽

Limonene Synthase ◽

Molecular Bases

BackgroundProtium species produce abundant aromatic oleoresins composed mainly of different types of terpenes, which are highly sought after by the flavor and fragrance industry.ResultsHere we present (i) the first draft genome of an endemic tree of the Brazil’s Atlantic Rainforest (Mata Atlântica), Protium kleinii Cuatrec., (ii) a first characterization of its genes involved in the terpene pathways, and (iii) the composition of the resin’s volatile fraction. The de novo draft genome was assembled using Illumina paired-end-only data, totalizing 407 Mb in size present in 229,912 scaffolds. The N50 is 2.60 Kb and the longest scaffold is 52.26 Kb. Despite its fragmentation, we were able to infer 53,538 gene models of which 5,434 were complete. The draft genome of P. kleinii presents 76.67 % (62.01 % complete and 14.66 % partial) of plant-core BUSCO genes. InterProScan was able to assign at least one Gene Ontology annotation and one Pfam domain for 13,629 and 26,469 sequences, respectively. We were able to identify 116 enzymes involved in terpene biosynthesis, such as monoterpenes α-terpineol, 1,8-cineole, geraniol, (+)-neomenthol and (+)-(R)-limonene. Through the phylogenetic analysis of the Terpene Synthases gene family, three candidates of limonene synthase were identified. Chemical analysis of the resin’s volatile fraction identified four monoterpenes: terpinolene, limonene, α-pinene and α-phellandrene.ConclusionThese results provide resources for further studies to identify the molecular bases of the main aroma compounds and new biotechnological approaches to their production.

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Diversifying Isoprenoid Platforms via Atypical Carbon Substrates and Non-model Microorganisms

Frontiers in Microbiology ◽

10.3389/fmicb.2021.791089 ◽

2021 ◽

Vol 12 ◽

Author(s):

David N. Carruthers ◽

Taek Soon Lee

Keyword(s):

Gene Expression ◽

Feedback Inhibition ◽

Heterologous Gene Expression ◽

High Titer ◽

Model Organisms ◽

Terpene Biosynthesis ◽

Terpene Synthases ◽

Methylerythritol Phosphate ◽

Commercial Viability ◽

Carbon Substrates

Isoprenoid compounds are biologically ubiquitous, and their characteristic modularity has afforded products ranging from pharmaceuticals to biofuels. Isoprenoid production has been largely successful in Escherichia coli and Saccharomyces cerevisiae with metabolic engineering of the mevalonate (MVA) and methylerythritol phosphate (MEP) pathways coupled with the expression of heterologous terpene synthases. Yet conventional microbial chassis pose several major obstacles to successful commercialization including the affordability of sugar substrates at scale, precursor flux limitations, and intermediate feedback-inhibition. Now, recent studies have challenged typical isoprenoid paradigms by expanding the boundaries of terpene biosynthesis and using non-model organisms including those capable of metabolizing atypical C1 substrates. Conversely, investigations of non-model organisms have historically informed optimization in conventional microbes by tuning heterologous gene expression. Here, we review advances in isoprenoid biosynthesis with specific focus on the synergy between model and non-model organisms that may elevate the commercial viability of isoprenoid platforms by addressing the dichotomy between high titer production and inexpensive substrates.

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RNA sequencing on Amomum villosum Lour. induced by MeJA identifies the genes of WRKY and terpene synthases involved in terpene biosynthesis

Genome ◽

10.1139/gen-2017-0142 ◽

2018 ◽

Vol 61 (2) ◽

pp. 91-102 ◽

Cited By ~ 3

Author(s):

Xueying He ◽

Huan Wang ◽

Jinfen Yang ◽

Ke Deng ◽

Teng Wang

Keyword(s):

Rna Sequencing ◽

Terpene Synthase ◽

Wrky Transcription Factors ◽

Terpene Biosynthesis ◽

Terpene Synthases ◽

New Information ◽

Volatile Terpenes ◽

Highly Correlated ◽

Amomum Villosum ◽

Chinese Medicinal Plant

Amomum villosum Lour. is an important Chinese medicinal plant that has diverse medicinal functions, and mainly contains volatile terpenes. This study aims to explore the WRKY transcription factors (TFs) and terpene synthase (TPS) unigenes that might be involved in terpene biosynthesis in A. villosum, and thus providing some new information on the regulation of terpenes in plants. RNA sequencing of A. villosum induced by methyl jasmonate (MeJA) revealed that the WRKY family was the second largest TF family in the transcriptome. Thirty-six complete WRKY domain sequences were expressed in response to MeJA. Further, six WRKY unigenes were highly correlated with eight deduced TPS unigenes. Ultimately, we combined the terpene abundance with the expression of candidate WRKY TFs and TPS unigenes to presume a possible model wherein AvWRKY61, AvWRKY28, and AvWRKY40 might coordinately trans-activate the AvNeoD promoter. We propose an approach to further investigate TF unigenes that might be involved in terpenoid biosynthesis, and identified four unigenes for further analyses.

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Terpene synthases of oregano (Origanum vulgare L.) and their roles in the pathway and regulation of terpene biosynthesis

Plant Molecular Biology ◽

10.1007/s11103-010-9636-1 ◽

2010 ◽

Vol 73 (6) ◽

pp. 587-603 ◽

Cited By ~ 86

Author(s):

Christoph Crocoll ◽

Julia Asbach ◽

Johannes Novak ◽

Jonathan Gershenzon ◽

Jörg Degenhardt

Keyword(s):

Origanum Vulgare ◽

Terpene Biosynthesis ◽

Terpene Synthases

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A molecular perspective on terpene variation in Australian Myrtaceae

Australian Journal of Botany ◽

10.1071/bt07146 ◽

2008 ◽

Vol 56 (3) ◽

pp. 197 ◽

Cited By ~ 61

Author(s):

Andras Keszei ◽

Curt L. Brubaker ◽

William. J. Foley

Keyword(s):

Chemical Diversity ◽

Oil Yield ◽

Biosynthesis Pathway ◽

Ecological Interactions ◽

Molecular Variation ◽

Ecological Context ◽

Terpene Biosynthesis ◽

Terpene Synthases ◽

Genetic Studies ◽

Molecular Information

The terpenoid-dominated essential oils in Australian Myrtaceae mediate many ecological interactions and are important industrially. Of all the significant essential oil-producing families, Myrtaceae is the only one for which there is no molecular information on terpene biosynthesis. Here we summarise available knowledge on terpene biosynthesis and its relevance to the Myrtaceae to provide a foundation for ecological and genetic studies of chemical diversity. There are several steps in the terpene biosynthesis pathway that have potential for influencing the oil yield, profile and composition of leaf oils in Myrtaceae. The biochemical steps that influence oil yield in Myrtaceae probably occur in the steps of the pathway leading up to the synthesis of the terpene backbone. Qualitative differences in oil profiles are more likely to be due to variation in terpene synthases and terpene-modifying enzymes. Most of the information on molecular variation in terpene biosynthesis is based on the analysis of artificially derived mutants but Australian Myrtaceae can provide examples of the same mechanisms in an ecological context.

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