Identification and functional characterization of the diterpene synthase family in Pogostemon cablin (Blanco) Benth

IF 6.1 2区 生物学 Q1 PLANT SCIENCES
Yiqiong Chen , Yumin Lin , Yingying Qiu , Wanying Li , Yanting Shen , Lili Huang
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引用次数: 0

Abstract

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.
Pogostemon cablin (Blanco) Benth.中二萜合成酶家族的鉴定和功能表征
Pogostemon cablin (Blanco) Benth(广藿香)是一种芳香草本植物,广泛用于制药和化妆品行业。倍半萜是广藿香的特征成分,由叶和茎上的腺毛合成。赤霉素(GA)是一种四环二萜类化合物,在腺毛的形成过程中起着至关重要的作用。然而,广藿香中的二萜生物合成在很大程度上仍然未知。在这里,我们发现了一个小型二萜合成酶(diTPSs)家族,包括三个 II 类 diTPSs(PatCPS1-3)和三个 I 类 diTPSs(PatKSL1 和 PatGLS1-2)。利用酵母异源表达系统对这些 diTPSs 进行了功能鉴定。PatCPS1 被鉴定为ent-copalyl diphosphate synthase(ent-CPS),与 PatKSL1 结合产生 GA 的前体 ent-kaurene,表明它们参与了初级代谢。PatCPS2 将 GGPP 转化为 (+)-8,13-醛基二磷酸(CPP)。没有检测到 PatCPS3、PatGLS1 和 PatGLS2 的活性。为了探索四倍体杂交广藿香基因组进化过程中 ent-CPS 的可能功能分化,对 PatCPS1 的三个同源物进行了进一步鉴定。GC-MS 分析表明,所有的外源基因都是功能性的 ent-CPS,证明了 PatCPS1 在进化过程中的功能保护。通过 qRT-PCR 进行的表达谱分析显示,PatCPS1 和 PatKSL1 在所有组织中普遍表达,这与它们参与初级代谢的情况一致。相反,PatCPS2 和 PatCPS3 主要在地上部分表达,表明它们在特化代谢中发挥作用。总之,这些发现澄清了广藿香中 GA 生物合成的早期阶段,并为进一步通过二萜进行倍半萜类化合物的代谢工程提供了基因元素。
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来源期刊
Plant Physiology and Biochemistry
Plant Physiology and Biochemistry 生物-植物科学
CiteScore
11.10
自引率
3.10%
发文量
410
审稿时长
33 days
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. Plant Physiology and Biochemistry publishes several types of articles: Reviews, Papers and Short Papers. Articles for Reviews are either invited by the editor or proposed by the authors for the editor''s prior agreement. Reviews should not exceed 40 typewritten pages and Short Papers no more than approximately 8 typewritten pages. The fundamental character of Plant Physiology and Biochemistry remains that of a journal for original results.
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