Cloning and Characterization of Cinnamaldehyde and Linalool Synthase Gene from Cinnamomoum osmophloeum

• Main Authors: Wen-Ke Huang, 黃文科
• Other Authors: 曲芳華
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/28907939300601212831

碩士 === 臺灣大學 === 森林環境暨資源學研究所 === 98 === Phenylalanine ammonia-lyase (PAL), trans-cinnamate- 4-hydroxylase (C4H), 4-coumarate: coenzyme A ligase 1 and 3 (4CL1 and 4CL3), cinnamoyl-CoA reductase (CCR) and linalool synthase (LIS) in Cinnamomum osmophloeum were cloned. The PAL has a 11 bp 5’ UTR, a 2127 bp ORF predicting 908 amino acid, and a 192 bp 3’UTR. The C4H has a 76 bp 5’UTR, a 1518 bp ORF predicting 505 amino acid, and a 129 bp 3’UTR. The 4CL1 has a 69 bp 5’UTR, a 1620 bp ORF predicting 539 amino acid, and a 378 bp 3’UTR. The 4CL3 has a 232 bp 5’UTR, a 1698 bp ORF predicting 565 amino acid, and a 293 bp 3’UTR. The CCR has a 95 bp 5’UTR, a 993 bp ORF predicting 330 amino acid, and a 325 bp 3’UTR. The LIS has a 34 bp 5’UTR, a 1755 bp ORF predicting 585 amino acid, and a 209 bp 3’UTR. Then these genes (PAL, C4H, 4CL1, 4CL3 and CCR) were transformed into Arabidopsis using pBI121 vector. After the genomic DNA and RNA of these transgenic plants were certificated, the volatile compounds were analyzed by solid phase microextration (SPME) and gas chromatography-mass spectrometry (GC-MS). These results demonstrate that the expressions of C4H, 4CL1, 4CL3 and CCR were correlated with cinnamaldehyde biosynthesis. The expression level of genes involved in cinnamaldehyde and linalool synthsis from 14 different sources were analyzed by RT-PCR. The result indicates the major compound of chemotype have no relationship with RNA expression. Comparison the amino acid sequence of 4CL3, CCR, and LIS genes on various type of Cinnamomum osmophloeum, indicated some specific residues of 4CL3 were strongly related with cinnamaldehyde type and some nucleotides of LIS gene were correlated with specific chemotype.