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Identification and characterization of terpene synthase genes accounting for volatile terpene emissions in flowers of Freesia x hybrida

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Identification and characterization of terpene synthase gene...

中国植物学会第十六次全国会员代表大会暨85周年学术年会

作者： Fengzhan Gao Baofeng Liu Min Li Xiaoyan Gao Qiang Fang Chang Liu Hui Ding Li Wang Xiang Gao Key Laboratory of Molecular Epigenetics of MOE and Institute of Genetics ＆ Cytology Northeast Normal University

The development of flower scents was a crucial event in biological evolution,providing olfactory signals by which plants can attract *** this study,bioinformatics,metabolomics,and biochemical and molecular methodologies were integrated to investigate the candidate genes involved in the biosynthesis of volatile components in two cultivars of Freesia x hybrida,Red River? and Ambiance,which release different categories of *** found that terpene synthase（TPS） genes were the pivotal genes determining spatiotemporal release of volatile compounds in both *** FhTPS genes were isolated,and they were phylogenetically clustered into different subgroups with different amino acid motifs and had divergent subcellular *** vitro biochemical analysis of the FhTPS proteins was performed to characterize their functions and six of them were found to be functional: FhTPS1 was a single-product enzyme catalyzing the formation of linalool,whereas the other four FhTPS proteins were multi-product enzymes,among which FhTPS4,FhTPS6,and FhTPS7 could recognize geranyl diphosphate and farnesyl diphosphate ***,the main catalytic products of the FhTPS proteins were also found in transgenic tobacco *** FhTPS enzymatic products closely matched the volatile terpenes emitted from flowers,and significant correlations were found between release of volatile terpenes and FhTPS gene *** addition,Enzyme kinetic parameter analysis demonstrated that FPP was the preferred substrate for FhTPS7 in the cultivar *** on these results,graphical models are proposed that summarize the biosynthesis of Freesia floral volatile *** characterization of FhTPS genes paves the way to decipher their roles in the speciation and fitness of Freesia,and this knowledge could also be used to introduce or enhance scent in other plants.

关键词： Emission pattern fitness flower scent Freesia ornamental plant speciation substrate selectivity terpene synthase

Molecular Basis for Sesterterpene Diversity Produced by Plant terpene synthases

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Plant Communications 2020年第5期1卷 41-52页

作者： Qingwen Chen Jianxu Li Zhixi Liu Takaaki Mitsuhashi Yuting Zhang Haili Liu Yihua Ma Juan He Tetsuro Shinada Tsutomu Sato Yong Wang Hongwei Liu Ikuro Abe Peng Zhang Guodong Wang State Key Laboratory of Plant Genomics and National Center for Plant Gene Research Institute of Genetics and Developmental BiologyThe Innovative Academy of Seed DesignChinese Academy of SciencesBeijing 100101China National Key Laboratory of Plant Molecular Genetics Center for Excellence in Molecular Plant SciencesShanghai Institute of Plant Physiology and EcologyChinese Academy of SciencesShanghai 200032China Graduate School of Pharmaceutical Sciences University of Tokyo7-3-1 HongoBunkyo-kuTokyo 113-0033Japan State Key Laboratory of Mycology Institute of MicrobiologyChinese Academy of SciencesBeijing 100101China Key Laboratory of Synthetic Biology Institute of Plant Physiology and EcologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200032China University of Chinese Academy of Sciences Beijing 100039China Graduate School of Science Osaka City University3-3-138 SugimotoSumiyoshiOsaka 558-8585Japan Department of Applied Biological Chemistry Faculty of AgricultureGraduate School of Science and TechnologyNiigata University8050 Ikarashi-2Niigata 950-2181Japan

Class I terpene synthase(TPS)generates bioactive terpenoids with diverse *** synthase(sester-TPS,C25),a branch of class I TPSs,was recently identified in ***,the catalytic mechanisms of sester-TPSs are not fully ***,we first identified three nonclustered functional sester-TPSs(AtTPS06,AtTPS22,and AtTPS29)in Arabidopsis ***06 utilizes a type-B cyclization mechanism,whereas most other sester-TPSs produce various sesterterpene backbones via a type-A cyclization *** then determined the crystal structure of the AtTPS18–FSPP complex to explore the cyclization mechanism of plant *** used structural comparisons and site-directed mutagenesis to further elucidate the mechanism:(1)mainly due to the outward shift of helix G,plant sester-TPSs have a larger catalytic pocket than do mono-,sesqui-,and di-TPSs to accommodate GFPP;(2)type-A sester-TPSs have more aromatic residues(five or six)in their catalytic pocket than classic TPSs(two or three),which also determines whether the type-A or type-B cyclization mechanism is active;and(3)the other residues responsible for product fidelity are determined by interconversion of AtTPS18 and its close ***,this study improves our understanding of the catalytic mechanism of plant sester-TPS,which ultimately enables the rational engineering of sesterterpenoids for future applications.

关键词： terpenoid terpene synthase sesterterpene cyclization mechanism crystal structure

A chromosome-scale genome assembly of Artemisia argyi reveals unbiased subgenome evolution and key contributions of gene duplication to volatile terpenoid diversity

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Plant Communications 2023年第3期4卷 228-243页

作者： Hongyu Chen Miaoxian Guo Shuting Dong Xinling Wu Guobin Zhang Liu He Yuannian Jiao Shilin Chen Li Li Hongmei Luo Key Laboratory of Chinese Medicine Resources Conservation State Administration of Traditional Chinese Medicine of the People’s Republic of ChinaInstitute of Medicinal Plant DevelopmentChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100193China Shandong University of Traditional Chinese Medicine Jinan 250355China State Key Laboratory of Crop Biology Shandong Agricultural UniversityTaian 271018China College of Agronomy Shandong Agricultural UniversityTaian 271018China State Key Laboratory of Systematic and Evolutionary Botany Institute of BotanyChinese Academy of SciencesBeijing 100093China University of the Chinese Academy of Sciences Beijing 100049China Institute of Chinese Materia Medica China Academy of Chinese Medical SciencesBeijing 100700China Chengdu University of Traditional Chinese Medicine Chengdu 611137China Robert W.Holley Center for Agriculture and Health USDA-ARSCornell UniversityIthacaNY 14853USA Plant Breeding and Genetics Section School of Integrative Plant ScienceCornell UniversityIthacaNY 14853USA

Artemisia argyi Le´*** Vant.,a perennial Artemisia herb with an intense fragrance,is widely used in traditional medicine in China and many other Asian ***,we present a chromosome-scale genome assembly of *** comprising 3.89 Gb assembled into 17 *** and comparative genomic analyses revealed that *** underwent a recent lineage-specificwhole-genomeduplication(WGD)event after divergence fromArtemisia annua,resulting in two *** the diploid ancestral genome of ***,and unbiased subgenome evolution was *** recent WGD led to a large number of duplicated genes in the *** *** of the terpene synthase(TPS)gene family through various types of gene duplication may have greatly contributed to the diversity of volatile terpenoids in *** particular,we identified a typical germacrene D synthase gene cluster within the expanded TPS gene *** entire biosynthetic pathways of germacrenes,(+)-borneol,and(+)-camphor were elucidated in *** addition,partial deletion of the amorpha-4,11-diene synthase(ADS)gene and loss of function of ADS homologs may have resulted in the lack of artemisinin production in *** study provides newinsights into the genome evolution of Artemisia and lays a foundation for further improvement of the quality of this important medicinal plant.

关键词： Artemisia argyi subgenome evolution gene duplication terpene synthase germacrene synthase nonartemisinin production

The chromosome-level genome of double-petal phenotype jasmine provides insights into the biosynthesis of floral scent

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Horticultural Plant Journal 2024年第1期10卷 259-272页

作者： Xiangyu Qi Huadi Wang Shuyun Liu Shuangshuang Chen Jing Feng Huijie Chen Ziyi Qin Quanming Chen Ikram Blilou Yanming Deng Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement Institute of Leisure AgricultureJiangsu Academy of Agricultural SciencesNanjingJiangsu 210014China School of Life Sciences Jiangsu UniversityZhenjiangJiangsu 212013China College of Horticulture Nanjing Agricultural UniversityNanjingJiangsu 210014China Shanghai Personal Biotechnology Company Limited Shanghai 200231China Biological and Environmental Sciences and Engineering King Abdullah University of Science and TechnologyThuwal 23955-6900Saudi Arabia

Jasmine(Jasminum sambac Aiton)is a well-known cultivated plant species for its fragrant flowers used in the perfume industry and ***,the genetic basis of its floral scent is largely *** this study,using PacBio,Illumina,10×Genomics and highthroughput chromosome conformation capture(Hi-C)sequencing technologies,a high-quality chromosome-level reference genome for *** was obtained,exploiting a double-petal phenotype cultivar‘Shuangbanmoli’(JSSB).The results showed that the final assembled genome of JSSB is 580.33 Mb in size(contig N50=1.05 Mb;scaffold N50=45.07 Mb)with a total of 39618 predicted protein-coding *** analyses revealed that the JSSB genome has undergone an ancient whole-genome duplication(WGD)event at 91.68 million years ago(Mya).It was estimated that *** diverged from the lineage leading to Olea europaea and Osmanthus fragrans about 28.8 *** the basis of a combination of genomic,transcriptomic and metabolomic analyses,a range of floral scent volatiles and genes were identified involved in the benzenoid/phenylpropanoid and terpenoid biosynthesis *** results provide new insights into the molecular mechanism of its fragrance biosynthesis in jasmine.

关键词： Jasminum sambac Aiton Oleaceae Genome evolution Floral scent terpene synthase

Engineering Nannochloropsis oceanica for the production of diterpenoid compounds

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mLife 2023年第4期2卷 428-437页

作者： Zhi-Yan Du Wajid W.Bhat Eric Poliner Sean Johnson Conor Bertucci Eva Farre Bjoern Hamberger Department of Molecular Biosciences and Bioengineering University of Hawaii at ManoaHonoluluHawaiiUSA Department of Biochemistry and Molecular Biology Michigan State UniversityEast LansingMichiganUSA Department of Plant Biology Michigan State UniversityEast LansingMichiganUSA New England Biolabs Inc. 240 County Roadlpswich01938 MAUSA

Photosynthetic microalgae like Nannochloropsis hold enormous potential as sustainable,light-driven biofactories for the production of high-value natural products such as *** oceanica is distinguished as a particularly robust host with extensive genomic and transgenic resources *** capacity to grow in wastewater,brackish,and sea waters,coupled with advances in microalgal metabolic engineering,genome editing,and synthetic biology,provides an excellent *** the present work,we demonstrate how *** can be engineered to produce the diterpene casbene—an important intermediate in the biosynthesis of pharmacologically relevant macrocyclic *** accumulated after stably expressing and targeting the casbene synthase from Daphne genkwa(DgTPS1)to the algal *** engineered strains yielded production titers of up to 0.12 mg g^(−1) total dry cell weight(DCW)*** overexpression and chloroplast targeting of two upstream rate-limiting enzymes in the 2-C-methyl-d-erythritol 4-phosphate pathway,Coleus forskohlii 1-deoxy-d-xylulose-5-phosphate synthase and geranylgeranyl diphosphate synthase genes,further enhanced the yield of casbene to a titer up to 1.80 mg g^(−1) *** results presented here form a basis for further development and production of complex plant diterpenoids in microalgae.

关键词： casbene microalgae plastid targeting synthetic biology terpene synthase

Functional characterization of seven terpene synthases from Ophiopogon japonicus via engineered Escherichia coli

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Science of Traditional Chinese Medicine 2024年第4期2卷 312-318页

作者： Wen Xu Qing Ma Tong Chen Haiyan Zhang Guanglei Ma Sibongile Mafu Juan Guo Xiaohui Fan Guanghong Cui Baolong Jin State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs National Resource Center for Chinese Materia MedicaChina Academy of Chinese Medical SciencesBeijingChina College of Pharmaceutical Sciences Zhejiang UniversityHangzhouChina National Key Laboratory of Chinese Medicine Modernization Innovation Center of Yangtze River DeltaZhejiang UniversityJiaxingChina Department of Biochemistry and Molecular Biology University of Massachusetts-AmherstAmherstMAUnited States

Background:Ophiopogon japonicus(L.f)*** in Zhejiang is recognized as the Dao-di medicinal herb for the production of Ophiopogonis ***-7-O-[β-D-apiofuranosyl-(1→6)]-β-D-glucopyranoside,a prominent pharmacologically active compound,serves as a marker distinguishing *** in Zhejiang from those in other geographical *** is synthesized from borneol through glycosylation,with terpene synthase(TPS)being the critical enzyme catalyzing the conversion of terpene precursors into ***:The aim of the study was to define key genes involved in biosynthesis of borneol in ***:The candidate terpene synthase genes were identified from the root and leaf transcriptome data of *** in Zhejiang and the functions of these enzymes were identified using engineered Escherichia ***:This study developed a rapid expression system for monoterpene and sesquiterpene synthases based on engineered *** terpene synthase genes(OjTPS1 to OjTPS7)were identified in different terpene synthase subfamilies,including 2 from TPS-a,4 from TPS-b,and 1 from *** analysis using an engineered system *** demonstrated that all the 7 terpene synthases produced monoterpenes,and OjTPS3,OjTPS5,and OjTPS6 also yielded ***:These 7 terpene synthases produced 13 monoterpenes and 8 ***,OjTPS1 produced borneol establishing the groundwork for elucidating the biosynthetic pathways of borneol-7-O-[β-D-apiofuranosyl-(1→6)]-β-D-glucopyranoside and other volatile oil components.

关键词： Ophiopogon japonicus Ophiopogon japonicus in Zhejiang terpene synthase Borneol

A Genome-Wide Scenario of terpene Pathways in Camellia sinensis

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A Genome-Wide Scenario of Terpene Pathways in Camellia sinen...

第七届长三角植物科学研讨会暨青年学术报告会

作者： Jieyang Jin Mingyue Zhao Miaojing Xu Shangrui Zhang Binbin Cai Yongxian Chen Na Zhang Tingting Jing Chuan-kui Song 安徽农业大学茶学国家重点实验室

The volatile aromas developed by chemical and biochemical transformations during its cultivation,production,and processing are important criterion in the evaluation of tea *** this study,we propose an integrated approach of pyrosequencing,metabolomics and functional certification to characterize genome-wide terpene biosynthesis in tea bud,leaves and *** terpene synthase gene CsCPS/HUS-2 and CsMYS/OCS-1 were recovered from tea plants,transgenic tobacco studies and a gene-specific antisense oligodeoxynucleotide suppression applied in tea leaves indicated that both transgenic tobacco of CsCPS/HUS-2 and CsMYS/OCS-1 can promote the produce of geraniol,citral,linalool,farnesol and *** oligodeoxynucleotide suppression further prove the characters of CsCPS/HUS-2 and CsMYS/OCS-1 in tea plants,indicate that integrated approach of pyrosequencing,metabolomics is feasible to dig those uncharted genes.

关键词： Camellia sinensis integrated approach terpene synthase transgenic

A tandem array of ent-kaurene synthases in maize with roles in gibberellin and more specialized metabolism

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A tandem array of ent-kaurene synthases in maize with roles ...

2016年全国植物生物学大会

作者： Jingye Fu Fei Ren Hongjie Mao Qiang Wang Institute of Ecological Agriculture Sichuan Agricultural University

While most commonly associated with its role in gibberellin(GA) phytohormone biosynthesis,entkaurene also serves as an intermediate in more specialized diterpenoid metabolism,as exemplified by the more than 800 known derived natural products. Among these are the maize kauralexins. However,no ent-kaurene synthases(KSs) have been identified from maize. The maize GA-deficient dwarf-5(d5) mutant has been associated with a loss of KS activity. The relevant genetic lesion has been previously mapped,and was found here to correlate with the location of the KS-like gene Zm KSL3. Intriguingly,this forms part of a tandem array with two other terpene synthases(TPSs). Although one of these,Zm TPS1,has been previously reported to encode a sesquiterpene synthase,and both Zm TPS1 and that encoded by the third gene,Zm KSL5,have lost the N-terminal ?-domain prototypically associated with KS(L)s,all three genes fall within the KS(L) or TPS-e sub-family. Here it is reported that all three genes encode enzymes that are targeted to the plastid in planta,where diterpenoid biosynthesis is initiated,and which all readily catalyze the production of ent-kaurene. Consistent with the closer phylogenetic relationship of Zm KSL3 with previously identified KSs from cereals,only transcription of this gene is affected in d5 plants. On the other hand,the expression of all three of these genes is inducible,suggesting a role in more specialized metabolism,such as that of the kauralexins. Thus,these results clarify not only gibberellin phytohormone,but also diterpenoid phytoalexin biosynthesis in this important cereal crop plant.

关键词： gibberellin phytoalexin kaurene terpene synthase gene duplication