万寿菊类胡萝卜素裂解双加氧酶基因CCD1克隆与表达分析

doi:10.3864/j.issn.0578-1752.2017.10.018

Abstract

Abstract: 【Objective】Carotenoid Cleavage Dioxygenase 1 gene of Tagetes erecta L. ‘Scarletade’(TeCCD1) was cloned for bioinformatics and gene expression analysis, which can help clarifying its biological functions in carotenoid degradation pathway and providing a theoretical foundation to further clarify the mechanism of African marigold flower color formation.【Method】According to the transcriptome of African marigold flower bud, the full-length cDNA of TeCCD1 had been obtained, and gene expression profile of ray florets at developmental stages of closed bud, semi-open bud, open flower and fully open flower was studied by Real-time PCR.【Result】The full-length sequence of CCD1 cDNA obtained from African marigold is 1 746 bp (GenBank accession number: KX557488), with a coding region length of 1 626 bp, putatively encoding 541 amino acids. Protein analysis indicated that TeCCD1 is an unstable protein and has no signal peptide, which belongs to the RPE65 superfamily (GenBank accession number is PF03055) having the same conserved domain of CCD family, and it is mainly located in the cytoplasm. CCD1 nucleic acid sequence of African marigold is 89% homologous to that of Pyrethrum. Amino acid sequence analysis suggested that CCD1 of African marigold is 93% homologous to that of Pyrethrum, and 75%-83% homologous to that of 19 different species, indicating that TeCCD1 is highly conserved gene. Phylogenetic analysis showed that the evolution of TeCCD1 is basically in accordance with the evolution law of plant taxonomy and has obvious characteristics of species, which has a closest relationship with that of the species in Compositae. The results of Real-time PCR demonstrated that expression of TeCCD1 increased along with the development of ray floretsand reached the maximum value at S4 stage.【Conclusion】CCD1 homolog was cloned in Tagetes erecta L. ‘Scarletade’ identified to be a typical member of the CCD family, which is a highly conserved gene located in the cytoplasm. The color fading of ray florets during the late development phase is possibly caused by the increase of expression of TeCCD1, which contributes to a decrease in carotenoid content.

Key words: Tagetes erecta L., carotenoid cleavage dioxygenase 1 (CCD1), flower color, gene expression

LIU XiaoCong, ZENG Li, LIU GuoFeng, PENG YongZheng, TAO YiWei, ZHANG YaoYue, WANG MengRu. Cloning and Expression Analysis of Carotenoid Cleavage Dioxygenase 1 (CCD1) Gene in Tagetes erecta L.

[J].Scientia Agricultura Sinica, 2017, 50(10): 1930-1940.

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Cloning and Expression Analysis of Carotenoid Cleavage Dioxygenase 1 (CCD1) Gene in Tagetes erecta L.

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