水芹咖啡酰辅酶A-O-甲基转移酶基因的克隆与表达特性分析.

[Objectives]The study aimed to investigate the sequence and expression characteristics of caffeoyl-CoA O-methyltransferase gene in Oenanthe javanica(OjCCoAOMT) and to understand its role in lignin metabolism. [Methods]The OjCCoAOMT gene was cloned from O.javanica using RT-PCR technique. The bioinformatics methods were used to analyze putative amino acid sequence and RT-qPCR method was used to study the expression profiles of OjCCoAOMT gene in different tissues of O.javanica and under different storage conditions. [Results]Sequence analysis result showed that OjCCoAOMT gene contained an open reading frame(ORF) with length of 726 bp,encoded 241 amino acids. The relative molecular mass of OjCCoAOMT protein was 27.111×103. Theoretical isoelectric point was 5.38. The protein belonged to hydrophobic protein and the ratio of its unordered sequence was 12.45%. Tertiary structure of OjCCoAOMT protein mainly consisted of eight α-helix and seven β-sheet. The analysis results of multiple alignment and phylogenetic tree showed that OjCCoAOMT had the nearest phylogeny with CCoAOMT protein from the same family species Petroselinum crispum. RT-qPCR analysis showed that the expression profiles of OjCCoAOMT gene were significantly different in different tissues(leaf blade and petiole) of ‘Baguazhoushuiqin’ and ‘Nanxuanbaguazhouzishuiqin’. The expression levels of OjCCoAOMT gene were different under various storage conditions,such as room temperature(25℃),low temperature(4℃) and darkness at room temperature(25℃). The expression level was peak at 4℃. Moreover,significant differences also existed between two varieties. [Conclusions]OjCCoAOMT protein has high conservation. The relative expression level of OjCCoAOMT gene has tissue and material specialties under different treatments. Moreover,OjCCoAOMT gene has high expression level under the treatment of 4℃. [ABSTRACT FROM AUTHOR]