Cloning and Expression Analysis AcNCEDs in Kiwifruit
[Objective]9-cis-epoxy carotenoid dioxygenase(NCED)is an enzyme involving in carot-enoid degrading and a key enzyme in the synthesis of abscisic acid(ABA).To explore the expression characteristics of all members of the AcNCED gene family during kiwifruit development.[Method]Four NCED genes were cloned from kiwifruit by RT-PCR,named AcNCED1-4,and their expression in vari-ousduring tissues and fruit developmental stages and in different tissues of both varieties was analyzed by qRT-PCR.[Result]The open reading frames(ORF)of AcNCED1,AcNCED2,AcNCED3 and AcNCED4 are 1 755,1 830,1 797 and 1 788 bp,respectively,which encode 585,610,599 and 596 amino acids,respectively.It was predicted that AcNCED1,AcNCED3 and AcNCED4 proteins are localized in chloro-plast,while AcNCED2 protein in cytoplasm,all of which lacking signal peptide and belong to non-secreted proteins.The qRT-PCR analysis showed that AcNCEDs has the highest expression level in the stem,the lowest in the roots,and high expression during early fruit development and maturity.[Conclusion]The proteins encoded by all four genes are highly conserved and exhibit tissue-specific ex-pression,which lays the foundation for further exploring the biological function of AcNCED.
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