Analysis Of Genes Related To Terpenoid Synthesis In Cinnamomum Burmanii Based On Transcriptome Data

D-borneol is a bicyclic monoterpenoid compound,which has biological activities such as antibacterial,anti-inflammatory,opening blood-brain barrier,promoting drug absorption and so on.It has been widely used in the fields of medicine,food,spices and so on.The natural Dborneol is mainly extracted from Dipterocarpaceae plants directly,and China has been mainly relying on imports to meet its demand.In order to solve this problem,it is urgent to find and develop new natural resources of natural D-borneol.In this study,the genes related to terpenoid synthesis in Cinnamomum burmanni(C.burmanni),one of the main plant sources of natural D-borneol found in China,were studied by transcriptome sequencing.It is of great significance to improve the utilization rate of the C.burmanni plant resources and to cultivate new varieties.1.Two different chemical types of volatile oil of C.burmanii were extracted by Soxhlet extraction method,and the chemical components in the volatile oil were identified by gas chromatography/mass spectrometry(GC-MS).A total of 41 different compounds were identified,including 5 monoterpenoids and 8 sesquiterpene and a diterpenoid.Among them,there are 34 kinds of high-content D-borneol-type C.burmanii(M),and 19 different compounds in low-content D-borneol-type C.burmanii(Y).In the two chemical types of C.burmanii volatile oil,D-borneol is the main chemical component.Through quantitative analysis of the content of D-borneol in the two chemical types,it was found that the average content of Dborneol in the M group was about 3 times that of the Y group.2.The full-length transcriptome of M1-1 was sequenced and analyzed.After quality control of the original data,a total of 17,114 gene sequences were obtained,with an average length of 2506.84 bp,N50 of 2749 bp,and GC content of 44.04%.Aligned with the Nr,Swissprot,COG/KOG and KEGG databases,16481,14544,11690 and 8127 gene sequences were compared.In the metabolic pathway comparison,76 genes related to the synthesis of terpene compounds were compared,including 57 related genes involved in the carbon skeleton synthesis and metabolism pathway,4 related genes involved in the monoterpenoid synthesis and metabolism pathway,and 8 involved Diterpenoids synthesis and metabolism pathway related genes and 7 related genes involved in triterpenoid and sesquiterpene synthesis metabolism pathway.Searching for sequences with high similarity to the 4 monoterpene synthase sequences,and performing molecular phylogenetic tree analysis,found that these 4sequences are related to short-chain dehydrogenase sequences,neomenthol dehydrogenase and salutaridine reductase in high similarity,and also found the four sequences can be clustered with the short-chain dehydrogenase in Cinnamomum sibiricum(branching reliability is 100%),indicating that they may be homologous genes.3.Transcriptome sequencing analysis was performed on 6 samples of two different chemical types of C.burmanii.Using the three-generation full-length transcriptome sequencing results as the reference transcript Isoform,the second-generation transcriptome sequencing results were compared to Isoform.The comparison rates were respectively 79.11%,77.20%,75.21%,73.72%,72.73% and 74.66%,respectively.Differential expression analysis was performed on the gene expression levels in the two chemical types,and a total of 250 differentially expressed genes were analyzed under the condition of FDR<0.05 and |log2FC|>1.Compared with the M group,there were 26 in the Y group.Genes showed an upward-regulated trend,and 224 genes showed a downward-regulated trend.Differentially expressed genes have been annotated into 6 related genes in the terpenoid synthesis pathway,including 3 carbon skeleton synthesis related genes(Isoform 10235,Isoform 10365,Isoform 10560)and 3diterpenoid synthesis related genes(Isoform 2236,Isoform 2236,Isoform 10560).14050,Isoform 16988).Twelve differentially expressed genes were randomly selected for q RT-PCR,and the experimental results were basically consistent with the sequencing results,which proved the reliability of the transcriptome sequencing data.