Abstract

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Title:: Cloning and Expression Analysis of CHS Gene in Freshcut Chinese Waterchestnut

Author(s):: SONG Mubo1; 2; SHUAI Liang1; 2; LI Lin1; FANG Fang1; 2; CHEN Zhenlin1; 2; DUAN Zhenhua1; 2; (1.Institute of Food Science and Engineering Technology,Hezhou University,Hezhou,Guangxi 542899;2.Guangxi Talent Highland of Preservation and Deep Processing Research in Fruit and Vegetables,Hezhou,Guangxi 542899)

Keywords:: freshcut Chinese waterchestnut; chalcone synthase; gene cloning; expression analysis

Abstract:: Freshcut Chinese waterchestnut was used as material,the full cDNA sequence of CHS was obtained by using the RTPCR and RACE methods，the bioinformatics and expression of CHS were analyzed in order to supply reference for gene function study.The results showed that the cDNA sequence of CHS was obtained,and it was named CwCHS.CwCHS consisted of 1 519 bp,and encoding a polypeptide of 394 amino acids.The formula of CwCHS was C.1 927H.3 072N.526O.568S.21,molecular weight was 43.37 kDa.CwCHS contained three conserved domains.Besides,CwCHS included casein kinase II phosphorylation sites for five,protein kinase C phosphorylation sites for one,Nmyristoylation site for six and Nglycosylation site for one.Alpha helix was the maximum structural part in the protein secondary structure of CwCHS.The multiple sequence alignment showed that CwCHS had the highly conservative enzyme activity sites,and was highly homologous to other CHS proteins from different plants.Phylogenetic tree analysis showed that CwCHS was very closely related to CHS protein of Ananas comosus.The analysis of realtime PCR showed that the expression level of CwCHS in freshcut Chinese waterchestnut increased dramatically,however,salicylic acid treatment significantly inhibited the expression of CwCHS.These results suggested that expression of CwCHS closely related to surface etiolation phenomenon of freshcut Chinese waterchestnut.