Expression of genes encoding terpenoid biosynthesis enzymes during leaf development of Eucalyptus camaldulensis

Abstract

- gas chromatography-mass spectrometry; GO - gene ontology; HMGCS - hydroxymethylglutaryl-CoA synthase; IPP - isoprene phosphate; KEGG - Kyoto encyclopedia of genes and genomes; ME-CPP - 2-C-methyl-D-erythritol- 2,4-cyclodiphosphate synthase; MEP - methylerythritol 4-phosphate; MVA - mevalonate; RIN - RNA integrity number; RPKM - reads per kb per million reads; TPS - terpene synthase; TFs - transcription factors. Abstract To reveal the regulation mechanism of terpenoid biosynthesis in the leaves of Eucalyptus camaldulensis , the content of volatiles in eucalyptus leaves and the transcriptome databases of young and mature leaves were analyzed. The results showed that E. camaldulensis contains 92 and 89 kinds of volatile substances in the young and mature leaves, respectively. Among them, the content of 1,8-cineole, β-pinene, and other substances was significantly different in young and mature leaves. A total of 99 802 unigenes were obtained from the transcriptome database of young and mature leaves of E. camaldulensis and 18 441 genes displayed obviously differential expressions during both developmental stages. There were 6 982 up-regulated unigenes and 11 461 down-regulated unigenes in the young leaf stage compared to the mature leaf stage. The key genes for terpenoid biosynthesis, including limonene synthase-10, limonene synthase-11, myrcene synthase-1, α- pinene synthase-2, and 1,8-cineole synthase-2 , were selected for further analysis to explore the mechanism of gene regulation and genetic transformation. The expressions of key genes were validated by RT-qPCR, and their expressions were consistent with RNA-seq data. WRKY, MYB, NAC , and bHLH transcription factors (TFs) displayed important regulatory effects on the above key genes. Thus, a regulatory network model of terpenoid biosynthesis was constructed using target genes and TFs during leaf development in E. camaldulensis . These results provide theoretical evidence for understanding the terpenoid biosynthesis in plants and reference for terpenoids utilization by genetic engineering methods in E. camaldulensis .