Molecular cloning, expression pattern, and putative cis-acting elements of a 4-coumarate:CoA ligase gene in bamboo ( Neosinocalamus affinis )|
Cao, Ying; Hu, Shang-lian; Huang, Sheng-xiong; Ren, Peng & Lu, Xue-qin
Background: 4-coumarate:CoA ligase (4CL) plays an important role at the divergence point from
general phenylpropanoid metabolism to several branch pathways. Although 4CL sin higher plants have
been extensively studied, little has known about the 4CL gene of bamboo. Results: In current study, a
Na4CL gene putative encoding 4-coumarate:CoA ligase (4CL) and its 5’-flanking region were isolated
from bamboo (Neosinocalamus affinis) by RACE-PCR and genomic DNA walker, respectively. Na4CL
encodes a predicted protein of 557 amino acids, with conserved motifs of adenylate-forming enzymes.
Phylogenetic analysis showed that Na4CL shared 62~85% identity with other known plant 4CLs, and
cluster closely with some known 4CLs in monocots. Sequence analysis revealed conserved cis-acting
elements (Box A and AC-II element) present in the Na4CL promoter. Additionally, a Na4CL RNAi
construct was transformed into tobacco. Transgenic tobaccos displayed significant down-expression of
endogenesis 4CL and reduced lignin contents. Conclusion:These results contribute to the knowledge
of the presence of Na4CL gen and its possible role in phenylpropanoid metabolism.
expression pattern; Neosinocalamus affinis; promoter; RACE; RNAi