Cloning and Expression Analysis of Carotenoid Cleavage Dioxygenase Gene AcCCD in Kiwifruit
[Objective]The carotenoid cleavage dioxygenase(CCD)gene was cloned and analyzed for physicochemical properties and expression.[Method]The CCD candidate genes were identified from the kiwifruit gene database,the full length of three genes were cloned,and the expression levels of these genes in different tissues and fruits of kiwifruit were analyzed by fluorescence quantitative PCR.[Result]Three CCD genes were cloned and named AcCCD1.1,AcCCD1.2 and AcCCD7.Their open reading frames(ORF)were 1 890,1 632 and 1 818 bp,encoding 629,543 and 605 amino acids,respectively.Predic-tion of subcellular location showed that the three AcCCD proteins were located in the cytoplasm and do not have signaling peptides,suggesting they did not belong to secreted proteins.According to the analy-sis,AcCCD1.1 and AcCCD1.2 proteins belong to the CCD1 protein family,while AcCCD7 proteins be-long to the CCD7 family.The phylogenetic tree showed that these were highly similar to plant proteins such as the wild tea tree.The results of qRT-PCR showed that AcCCD1.1 was highly expressed in roots and minimally in leaves,while AcCCD1.2 was highly expressed in late flower and fruit development.AcCCD7 has generally low expression but is higher in roots compared to stems,leaves,flowers and fruits.[Conclusion]It is speculated that AcCCD1 gene is important for the formation of kiwi fruit color and flavor.AcCCD7 gene plays an important role in regulating the root growth of kiwifruit.
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