Functional characterization of SsNES responsible for nerolidol biosynthesis in Senecio scandens.
10.19540/j.cnki.cjcmm.20181204.009
- Author:
Qin-Qin SHEN
1
;
Li-Ping WANG
1
;
Jin LIANG
1
;
Li-Jun LIU
1
;
Qiang WANG
1
Author Information
1. Institute of Ecological Agriculture, Sichuan Agricultural University Chengdu 611130, China.
- Publication Type:Journal Article
- Keywords:
functional identification;
gene expression;
homology modeling;
metabolic engineering;
nerolidol
- MeSH:
Escherichia coli;
Genes, Plant;
Phylogeny;
Senecio;
enzymology;
Sesquiterpenes;
metabolism
- From:
China Journal of Chinese Materia Medica
2019;44(7):1334-1340
- CountryChina
- Language:Chinese
-
Abstract:
A short terpene synthase gene was obtained by screening the transcriptome data of Senecio scandens. The phylogenetic tree and sequence alignment putatively identified this gene as a nerolidol synthase gene, named SsNES(GenBank MH518312). Protein homology modeling indicated that SsNES contained a complete conserved domain and folded correctly. SsNES was cloned and successfully expressed in Escherichia coli as soluble protein. The biochemical function of SsNES was characterized by E. coli metabolic engineering, which showed that SsNES catalyzed formation of trans-nerolidol with(E, E)-farnesyl diphosphate as the substrate. Nerolidol was also detected in stems and leaves of S. scandens, indicating that SsNES might act as the nerolidol synthase in plant. RT-PCR analysis indicated that SsNES was mainly expressed in stem, flowers and leaves, and no expression was observed in roots. After the treatment of SA, MeJA or Ala, SsNES was induced significantly at 6 h, indicating involvement in the defense response of S. scandens. The identification of SsNES not only clarified biosynthesis of nerolidol in S. scandens, but also provided diversity of sesquiterpene synthase, as well as theoretical basis for disease and pest defense mediated by the terpene metabolites.
