Project description:Agarwood is an expensive resinous heartwood derived from the wounded Aquilaria plants. To identify the primary genes that maybe related to agarwood formation, we sequenced 2 cDNA libraries generated from healthy and wounded A. sinensis (Lour.) Gilg. A total of 89,137 unigenes with an average length of 678.65 bp were obtained, and they were annotated in detail at bioinformatics levels. Of those associated with agarwood formation, 30 putatively encoded enzymes in the sesquiterpene biosynthesis pathway, a handful of transcription factors, and protein kinases related to wound signal transduction. Three full-length cDNAs of sesquiterpene synthases (ASS1-3) were cloned and expressed in Escherichia coli, and enzyme assays revealed that they are active enzymes, with the major products being M-NM-4-guaiene. A methyl jasmonate (MJ) induction experiment revealed that the expression of ASS was significantly induced by MJ, and the production of sesquiterpenes was elevated accordingly. The expression of some transcription factors and protein kinases, especially MYB4, WRKY4, MPKK2 and MAPK2, was also induced by MJ and coordinated with ASS expression, suggesting they maybe positive regulators of ASS. This study provides extensive transcriptome information for Aquilaria spp. and valuable clues for elucidating the mechanism of wound-induced agarwood sesquiterpenes biosynthesis and their regulation. healthy and wounded stems of three-year-old A. sinensis trees
Project description:Agarwood is a kind of precious traditional Chinese medicine, the scarity of natural agarwood limits its wide application and resulted in the high price of agarwood. Agarwood formed when Aquilaria sinensis was suffered from chemical stimulus, mechanical damage and fungal infection, however, the molecular mechanism of which has not been fully elucidated. In this study, the quantitative proteomic analysis using iTRAQ technology combined with transcriptome analysis on chemically induced A. sinensis were firstly performed to elucidate the agarwood formation mechanism by formic acid stimulus. 3,029 proteins were detected, 504 differential proteins that showed at least 2-fold differences in their expression levels were selected ,