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MOLECULAR CLONING OF CAFFEOYL-COA 3-O-METHYLTRANSFERASE GENE FROM

CHAMAECYPARIS FORMOSENSIS AND ITS EFFECTS ON LIGNIN CONTENT IN TRANSGENIC TOBACCO PLANTS

Yan-Liang Lin1 Sheng-Yang Wang2 Fang-Hua Chu3

!Received: February 25, 2008; Accepted: July 27, 2008.

1 !"#$%&'()*+,-&./01

Master, School of Forestry and Resource Conservation, National Taiwan University. 2 !"23%&'(&.4561

Associate Professor, School of Forestry, National Chung Hsing University. 3 !"#$%&'()*+,-&.78569:;<=1

Assistant Professor, School of Forestry and Resource Conservation, National Taiwan University, Corresponding Author, No.1, Sec. 4, Roosevelt Road, 106 Taipei, Taiwan.

'324' YZ Caffeoyl-CoA 3-O-methyltransferase Be;]^lGCBeÀ¨*+,��

ÌAbstractÍLignin is the second abundant biopolymer and less then cellulose present in the plant cell wall. O-Methyltransferase (OMT) plays as a key enzyme for stepwise biosynthesis of lignin. In this study, 526 base pair DNA fragment of OMT were obtained from Chamae-cyparis formosensis. Then the 5’ and 3’ RACE (Rapid Amplify of cDNA End) coupled with the Genome walking were used to clone the full length of CCoAOMT, named CfCCoAOMT. After combining the sequence data, there is a total of 1721 bps in full length including pro-moter sequence. The coding region of this DNA fragment was 750 bps, which encodes a pro-tein of 249 amino acids. Moreover, the developing xylem from C. formosensis has the most abundance expression pattern by Northern blotting analysis. The CfCCoAOMT was con-structed into the E. coli to produce a CfCCoAOMT recombinant protein and then the poly-clonal antibody was prepared. Finally, the CfCCoAOMT’s coding region of C. formosensis was transformed into tobacco, a type of herbaceous plant, by using Agrobacterium-mediated transgenic method. Due to the biosynthesis pathways of lignin in woody plant and herbaceous plant are quite different, there is no significant difference in the total lignin content in sense transgenic tobacco, but in antisense transgenic line had slightly increased. On the other hand, the G/S ratio increased dramatically in both transgenic lines.

ÌKey wordsÍ Chamaecyparis formosensis, G/S ratio, Lignin, O-methyltransferase, Trans-genic tobacco

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