Dataset Information

An Integrative Volatile Terpenoid Profiling and Transcriptomics Analysis for Gene Mining and Functional Characterization of AvBPPS and AvPS Involved in the Monoterpenoid Biosynthesis in Amomum villosum.

ABSTRACT: Amomum villosum, also known as Fructus Amomi, has been used to treat digestive diseases such as abdominal pain, vomiting, and dysentery. Volatile terpenoids are the active metabolites in the essential oil of Fructus Amomi. Nevertheless, downstream genes responsible for activating metabolites biosynthesis in A. villosum still remain unclear. Here, we report the use of an integrative volatile terpenoid profiling and transcriptomics analysis for mining the corresponding genes involved in volatile terpenoid biosynthesis. Ten terpene synthase (TPS) genes were discovered, and two of them were cloned and functionally characterized. AvTPS1 (AvPS: pinene synthase) catalyzed GPP to form ?-pinene and ?-pinene; AvTPS3 (AvBPPS: bornyl diphosphate synthase) produced bornyl diphosphate as major product and the other three monoterpenoids as minor products. Metabolite accumulation and gene expression pattern combined with AvPS biochemical characterization suggested that AvPS might play a role in biotic defense. On the other hand, the most active ingredient, bornyl acetate, was highly accumulated in seeds and was consistent with the high expression of AvBPPS, which further indicated that AvBPPS is responsible for the biosynthesis of bornyl acetate, the final metabolite of bornyl diphosphate in A. villosum. This study can be used to improve the quality of A. villosum through metabolic engineering, and for the sustainable production of bornyl acetate in heterologous hosts.

SUBMITTER: Wang H

PROVIDER: S-EPMC6020762 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

An Integrative Volatile Terpenoid Profiling and Transcriptomics Analysis for Gene Mining and Functional Characterization of AvBPPS and AvPS Involved in the Monoterpenoid Biosynthesis in Amomum villosum.

Wang Hong H Ma Dongming D Yang Jinfen J Deng Ke K Li Meng M Ji Xiaoyu X Zhong Liting L Zhao Haiying H

Frontiers in plant science 20180620

Amomum villosum, also known as Fructus Amomi, has been used to treat digestive diseases such as abdominal pain, vomiting, and dysentery. Volatile terpenoids are the active metabolites in the essential oil of Fructus Amomi. Nevertheless, downstream genes responsible for activating metabolites biosynthesis in A. villosum still remain unclear. Here, we report the use of an integrative volatile terpenoid profiling and transcriptomics analysis for mining the corresponding ...[more]

PMID: 29973947

Similar Datasets

Project description:The fruits of Amomum villosum and Amomum longiligulare are both used medicinally as Fructus Amomi the famous traditional Chinese medicine, however, the medicinal quality of A. villosum is better than that of A. longiligulare. Volatile terpenoids in the seeds, especially bornyl acetate and borneol, are the medicinal components of Fructus Amomi. The volatile terpenoids and transcriptome of developing seeds of A. villosum and A. longiligulare were compared in this study. The result revealed that the bornyl acetate and borneol contents were higher in A. villosum than in A. longiligulare. Additionally, six terpenoid synthase genes (AlTPS1-AlTPS6) were screened from the transcriptome of A. longiligulare, and AlTPS2 and AlTPS3 were found to share 98 and 83% identity with AvTPS2 and AvBPPS (bornyl diphosphate synthase) from A. villosum, respectively. BPPS is the key enzyme for the biosynthesis of borneol and bornyl acetate. Biochemical assays improved that AlTPS2 had an identical function to AvTPS2 as linalool synthase; however, AlTPS3 produced camphene as the major product and bornyl diphosphate (BPP) as the secondary product, whereas AvBPPS produced BPP as its major product. There was only one different amino acid between AlTPS3 (A496) and AvBPPS (G495) in their conserved motifs, and the site-directed mutation of A496G in DTE motif of AlTPS3 changed the major product from camphene to BPP. Molecular docking suggests that A496G mutation narrows the camphene-binding pocket and decreases the BPP-binding energy, thus increases the product BPP selectivity of enzyme. In addition, the expression level of AvBPPS was significantly higher than that of AlTPS3 in seeds, which was consistent with the related-metabolites contents. This study provides insight into the TPS-related molecular bases for the biosynthesis and accumulation differences of the bioactive terpenoids between A. villosum and A. longiligulare. BPPS, the key gene involved in the biosynthesis of the active compound, was identified as a target gene that could be applied for the quality-related identification and breeding of Fructus Amomi.

Project description:BackgroundThe essential oil is one of the main active ingredients of Amomum villosum Lour. However, volatile compounds are easily lost during the drying, storage and even sample preparation procedure. Therefore, using fresh samples can obtain more accurately data for qualitative and comparative analysis.MethodsIn this study, the volatile compounds in different parts of fresh A. villosum from different origins were systemic analyzed and compared by using cryogenic grinding combined HS-SPME-GC-MS for the first time. GC-MS analyses were performed on a 6890 Series GC instrument coupled to a 5973 N mass spectrometer. The volatile compounds were extracted by the SPME fiber (100 μm PDMS). Analytes separation was achieved on a HP-5MS capillary column. The oven temperature was initially programmed at 70 °C, then raised 4 °C/min to reach 125 °C and then programmed at 0.5 °C/min to 133 °C, then at 6 °C/min to 170 °C and finally, at 20 °C/min to 280 °C held for 2 min. The temperatures of the injection port, ion source and transfer line were set at 250 °C, 230 °C and 280 °C, respectively.ResultsForty-eight main compounds were identified in different parts of fresh A. villosum. The most abundant components in fresh fruit samples were camphor (3.91%), bornyl acetate (10.53%), caryophyllene (8.70%), β-bisabolene (11.50%), (E)-nerolidol (14.82%) and cubenol (10.04%). This is quite different with that of dried samples analyzed in our previous work. As different parts of the same plant, many common components with biological activities were detected in fruit and other parts. In principle components analysis (PCA) and hierarchical clustering analysis (HCA), four parts of A. villosum were divided into different groups clearly. Additionally, fruit and root samples also could be divided into two subgroups (HCA) in accordance with their regions.ConclusionThe developed method was successfully used for qualitative and comparative analysis of volatile compounds in fresh A. villosum samples. Additionally, using fresh samples can obtain much more information which is helpful for their performance in the fields of functional foods, agriculture and biomedical industry. Furthermore, our research is helpful for comprehensive utilization and quality control of A. villosum.

Dataset Information

An Integrative Volatile Terpenoid Profiling and Transcriptomics Analysis for Gene Mining and Functional Characterization of AvBPPS and AvPS Involved in the Monoterpenoid Biosynthesis in Amomum villosum.

Publications

An Integrative Volatile Terpenoid Profiling and Transcriptomics Analysis for Gene Mining and Functional Characterization of AvBPPS and AvPS Involved in the Monoterpenoid Biosynthesis in <i>Amomum villosum</i>.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets