Yiqiong Chen , Yumin Lin , Yingying Qiu , Wanying Li , Yanting Shen , Lili Huang
{"title":"Pogostemon cablin (Blanco) Benth.中二萜合成酶家族的鉴定和功能表征","authors":"Yiqiong Chen , Yumin Lin , Yingying Qiu , Wanying Li , Yanting Shen , Lili Huang","doi":"10.1016/j.plaphy.2024.109190","DOIUrl":null,"url":null,"abstract":"<div><div><em>Pogostemon cablin</em> (Blanco) Benth (Patchouli) is an aromatic herb extensively used in pharmaceutical and cosmetic industries. Sesquiterpenes are the characteristic constitutes in patchouli which are synthesized in the glandular trichomes on leaves and stems. Gibberellic acid (GA), a tetracyclic diterpenoid, plays a crucial role in the formation of glandular trichome. However, the diterpene biosynthesis remains largely unknown in patchouli. Here we identified a small diterpene synthases (diTPSs) family comprising three class II diTPSs (PatCPS1-3) and three class I diTPSs (PatKSL1 and PatGLS1-2). These diTPSs are functionally characterized using a yeast heterologous expression system. PatCPS1 was identified as an <em>ent</em>-copalyl diphosphate synthase (<em>ent</em>-CPS), in combination with PatKSL1, yield <em>ent</em>-kaurene, the precursor of GA, indicating their involvement in primary metabolism. PatCPS2 converted GGPP into (+)-8, 13-copalyl diphosphate (CPP). No activity was detected for PatCPS3, PatGLS1 and PatGLS2. Three ohnologs of <em>PatCPS1</em> were further characterized to explore the possible functional differentiation of <em>ent</em>-CPS during the evolution of tetraploid hybrid patchouli genome. GC-MS analysis showed all ohnologs are functional <em>ent</em>-CPSs, demonstrating the functional conservation of PatCPS1 during evolution. Expression profiling by qRT-PCR showed <em>PatCPS1</em> and <em>PatKSL1</em> are ubiquitously expressed in all tissues, consistent with their involvement in primary metabolism. Conversely, <em>PatCPS2</em> and <em>PatCPS3</em> were predominantly expressed in the above ground parts, indicating a role in specialized metabolism. In summary, these findings clarify the early stages of GA biosynthesis in patchouli and provide gene elements for further metabolic engineering of sesquiterpenes via diterpenoids.</div></div>","PeriodicalId":20234,"journal":{"name":"Plant Physiology and Biochemistry","volume":"216 ","pages":"Article 109190"},"PeriodicalIF":6.1000,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Identification and functional characterization of the diterpene synthase family in Pogostemon cablin (Blanco) Benth\",\"authors\":\"Yiqiong Chen , Yumin Lin , Yingying Qiu , Wanying Li , Yanting Shen , Lili Huang\",\"doi\":\"10.1016/j.plaphy.2024.109190\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div><em>Pogostemon cablin</em> (Blanco) Benth (Patchouli) is an aromatic herb extensively used in pharmaceutical and cosmetic industries. Sesquiterpenes are the characteristic constitutes in patchouli which are synthesized in the glandular trichomes on leaves and stems. Gibberellic acid (GA), a tetracyclic diterpenoid, plays a crucial role in the formation of glandular trichome. However, the diterpene biosynthesis remains largely unknown in patchouli. Here we identified a small diterpene synthases (diTPSs) family comprising three class II diTPSs (PatCPS1-3) and three class I diTPSs (PatKSL1 and PatGLS1-2). These diTPSs are functionally characterized using a yeast heterologous expression system. PatCPS1 was identified as an <em>ent</em>-copalyl diphosphate synthase (<em>ent</em>-CPS), in combination with PatKSL1, yield <em>ent</em>-kaurene, the precursor of GA, indicating their involvement in primary metabolism. PatCPS2 converted GGPP into (+)-8, 13-copalyl diphosphate (CPP). No activity was detected for PatCPS3, PatGLS1 and PatGLS2. Three ohnologs of <em>PatCPS1</em> were further characterized to explore the possible functional differentiation of <em>ent</em>-CPS during the evolution of tetraploid hybrid patchouli genome. GC-MS analysis showed all ohnologs are functional <em>ent</em>-CPSs, demonstrating the functional conservation of PatCPS1 during evolution. Expression profiling by qRT-PCR showed <em>PatCPS1</em> and <em>PatKSL1</em> are ubiquitously expressed in all tissues, consistent with their involvement in primary metabolism. Conversely, <em>PatCPS2</em> and <em>PatCPS3</em> were predominantly expressed in the above ground parts, indicating a role in specialized metabolism. In summary, these findings clarify the early stages of GA biosynthesis in patchouli and provide gene elements for further metabolic engineering of sesquiterpenes via diterpenoids.</div></div>\",\"PeriodicalId\":20234,\"journal\":{\"name\":\"Plant Physiology and Biochemistry\",\"volume\":\"216 \",\"pages\":\"Article 109190\"},\"PeriodicalIF\":6.1000,\"publicationDate\":\"2024-10-10\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Plant Physiology and Biochemistry\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0981942824008581\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"PLANT SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Plant Physiology and Biochemistry","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0981942824008581","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
Identification and functional characterization of the diterpene synthase family in Pogostemon cablin (Blanco) Benth
Pogostemon cablin (Blanco) Benth (Patchouli) is an aromatic herb extensively used in pharmaceutical and cosmetic industries. Sesquiterpenes are the characteristic constitutes in patchouli which are synthesized in the glandular trichomes on leaves and stems. Gibberellic acid (GA), a tetracyclic diterpenoid, plays a crucial role in the formation of glandular trichome. However, the diterpene biosynthesis remains largely unknown in patchouli. Here we identified a small diterpene synthases (diTPSs) family comprising three class II diTPSs (PatCPS1-3) and three class I diTPSs (PatKSL1 and PatGLS1-2). These diTPSs are functionally characterized using a yeast heterologous expression system. PatCPS1 was identified as an ent-copalyl diphosphate synthase (ent-CPS), in combination with PatKSL1, yield ent-kaurene, the precursor of GA, indicating their involvement in primary metabolism. PatCPS2 converted GGPP into (+)-8, 13-copalyl diphosphate (CPP). No activity was detected for PatCPS3, PatGLS1 and PatGLS2. Three ohnologs of PatCPS1 were further characterized to explore the possible functional differentiation of ent-CPS during the evolution of tetraploid hybrid patchouli genome. GC-MS analysis showed all ohnologs are functional ent-CPSs, demonstrating the functional conservation of PatCPS1 during evolution. Expression profiling by qRT-PCR showed PatCPS1 and PatKSL1 are ubiquitously expressed in all tissues, consistent with their involvement in primary metabolism. Conversely, PatCPS2 and PatCPS3 were predominantly expressed in the above ground parts, indicating a role in specialized metabolism. In summary, these findings clarify the early stages of GA biosynthesis in patchouli and provide gene elements for further metabolic engineering of sesquiterpenes via diterpenoids.
期刊介绍:
Plant Physiology and Biochemistry publishes original theoretical, experimental and technical contributions in the various fields of plant physiology (biochemistry, physiology, structure, genetics, plant-microbe interactions, etc.) at diverse levels of integration (molecular, subcellular, cellular, organ, whole plant, environmental). Opinions expressed in the journal are the sole responsibility of the authors and publication does not imply the editors'' agreement.
Manuscripts describing molecular-genetic and/or gene expression data that are not integrated with biochemical analysis and/or actual measurements of plant physiological processes are not suitable for PPB. Also "Omics" studies (transcriptomics, proteomics, metabolomics, etc.) reporting descriptive analysis without an element of functional validation assays, will not be considered. Similarly, applied agronomic or phytochemical studies that generate no new, fundamental insights in plant physiological and/or biochemical processes are not suitable for publication in PPB.
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