Transcriptomic Insight into Terpenoid Biosynthesis and Functional Characterization of Three Diterpene Synthases in Scutellaria barbata

被引:13
作者
Zhang, Huabei [1 ]
Jin, Baolong [1 ,2 ]
Bu, Junling [1 ]
Guo, Juan [1 ]
Chen, Tong [1 ]
Ma, Ying [1 ]
Tang, Jinfu [1 ]
Cui, Guanghong [1 ]
Huang, Luqi [1 ,2 ]
机构
[1] China Acad Chinese Med Sci, Natl Resource Ctr Chinese Mat Med, State Key Lab Breeding Base Dao Di Herbs, Beijing 100700, Peoples R China
[2] Hubei Univ Chinese Med, Coll Pharm, Wuhan 430065, Hubei, Peoples R China
基金
中国国家自然科学基金;
关键词
Scutellaria barbata; transcriptomics analysis; terpenoid backbone biosynthesis; terpene synthase; diterpene synthase; CYTOTOXIC NEOCLERODANE DITERPENOIDS; GENOME SEQUENCE; PLANT; PATHWAY; GENES; PRECURSOR; CLONING; SYSTEM;
D O I
10.3390/molecules23112952
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Scutellaria barbata (Lamiaceae) is an important medicinal herb widely used in China, Korea, India, and other Asian countries. Neo-clerodane diterpenoids are the largest known group of Scutellaria diterpenoids and show promising cytotoxic activity against several cancer cell lines. Here, Illumina-based deep transcriptome analysis of flowers, the aerial parts (leaf and stem), and roots of S. barbata was used to explore terpenoid-related genes. In total, 121,958,564 clean RNA-sequence reads were assembled into 88,980 transcripts, with an average length of 1370 nt and N50 length of 2144 nt, indicating high assembly quality. We identified nearly all known terpenoid-related genes (33 genes) involved in biosynthesis of the terpenoid backbone and 14 terpene synthase genes which generate skeletons for different terpenoids. Three full length diterpene synthase genes were functionally identified using an in vitro assay. SbTPS8 and SbTPS9 were identified as normal-CPP and ent-CPP synthase, respectively. SbTPS12 reacts with SbTPS8 to produce miltiradiene. Furthermore, SbTPS12 was proven to be a less promiscuous class I diterpene synthase. These results give a comprehensive understanding of the terpenoid biosynthesis in S. barbata and provide useful information for enhancing the production of bioactive neo-clerodane diterpenoids through genetic engineering.
引用
收藏
页数:16
相关论文
共 62 条
[1]  
Alvarenga SAV, 2001, PHYTOCHEMISTRY, V56, P583, DOI 10.1016/S0031-9422(00)00424-6
[2]   Expanding the Landscape of Diterpene Structural Diversity through Stereochemically Controlled Combinatorial Biosynthesis [J].
Andersen-Ranberg, Johan ;
Kongstad, Kenneth Thermann ;
Nielsen, Morten Thrane ;
Jensen, Niels Bjerg ;
Pateraki, Irini ;
Bach, Soren Spanner ;
Hamberger, Britta ;
Zerbe, Philipp ;
Staerk, Dan ;
Bohlmann, Jorg ;
Moller, Birger Lindberg ;
Hamberger, Bjorn .
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2016, 55 (06) :2142-2146
[3]   Phylogenomic Mining of the Mints Reveals Multiple Mechanisms Contributing to the Evolution of Chemical Diversity in Lamiaceae [J].
Boachon, Benoit ;
Buell, C. Robin ;
Crisovan, Emily ;
Dudareva, Natalia ;
Garcia, Nicolas ;
Godden, Grant ;
Henry, Laura ;
Kamileen, Mohamed O. ;
Kates, Heather Rose ;
Kilgore, Matthew B. ;
Lichman, Benjamin R. ;
Mavrodiev, Evgeny, V ;
Newton, Linsey ;
Rodriguez-Lopez, Carlos ;
O'Connor, Sarah E. ;
Soltis, Douglas ;
Soltis, Pamela ;
Vaillancourt, Brieanne ;
Wiegert-Rininger, Krystle ;
Zhao, Dongyan .
MOLECULAR PLANT, 2018, 11 (08) :1084-1096
[4]   Trimmomatic: a flexible trimmer for Illumina sequence data [J].
Bolger, Anthony M. ;
Lohse, Marc ;
Usadel, Bjoern .
BIOINFORMATICS, 2014, 30 (15) :2114-2120
[5]   Towards Elucidating Carnosic Acid Biosynthesis in Lamiaceae: Functional Characterization of the Three First Steps of the Pathway in Salvia fruticosa and Rosmarinus officinalis [J].
Bozic, Dragana ;
Papaefthimiou, Dimitra ;
Brueckner, Kathleen ;
de Vos, Ric C. H. ;
Tsoleridis, Constantinos A. ;
Katsarou, Dimitra ;
Papanikolaou, Antigoni ;
Pateraki, Irini ;
Chatzopoulou, Fani M. ;
Dimitriadou, Eleni ;
Kostas, Stefanos ;
Manzano, David ;
Scheler, Ulschan ;
Ferrer, Albert ;
Tissier, Alain ;
Makris, Antonios M. ;
Kampranis, Sotirios C. ;
Kanellis, Angelos K. .
PLOS ONE, 2015, 10 (05)
[6]   Characterization of two genes for the biosynthesis of abietane-type diterpenes in rosemary (Rosmarinus officinalis) glandular trichomes [J].
Brueckner, Kathleen ;
Bozic, Dragana ;
Manzano, David ;
Papaefthimiou, Dimitra ;
Pateraki, Irini ;
Scheler, Ulschan ;
Ferrer, Albert ;
de Vos, Ric C. H. ;
Kanellis, Angelos K. ;
Tissier, Alain .
PHYTOCHEMISTRY, 2014, 101 :52-64
[7]   Discovery and functional characterization of two diterpene synthases for sclareol biosynthesis in Salvia sclarea (L.) and their relevance for perfume manufacture [J].
Caniard, Anne ;
Zerbe, Philipp ;
Legrand, Sylvain ;
Cohade, Allison ;
Valot, Nadine ;
Magnard, Jean-Louis ;
Bohlmann, Joerg ;
Legendre, Laurent .
BMC PLANT BIOLOGY, 2012, 12
[8]   The family of terpene synthases in plants: a mid-size family of genes for specialized metabolism that is highly diversified throughout the kingdom [J].
Chen, Feng ;
Tholl, Dorothea ;
Bohlmann, Joerg ;
Pichersky, Eran .
PLANT JOURNAL, 2011, 66 (01) :212-229
[9]   A (-)-kolavenyl diphosphate synthase catalyzes the first step of salvinorin A biosynthesis in Salvia divinorum [J].
Chen, Xiaoyue ;
Berim, Anna ;
Dayan, Franck E. ;
Gang, David R. .
JOURNAL OF EXPERIMENTAL BOTANY, 2017, 68 (05) :1109-1122
[10]  
Committee C.P., 2015, PHARMACOPOEIA PEOPLE, V1, P134