Cloning Of ZDS And LYCE Related To Floral Colour From Narcissus Tazetta L. Var. Chinensis Roem

Narcissus tazetta L. var. chinensis Roem, a kind of perennial plant with beautiful color and sweet fragrance, belongs to the family Amaryllidaceae. Zhangzhou which is in FuJian province is the most important produce area of Chinese narcissus and the Zhangzhou narcissus have gotten a National Geographical Indication of well-known trademarks in 2008. But the simplex color have become a main obstacle for Chinese narcissus's development. So it's imperative to develop new varieties with modern breeding methods for the narcissus industry.In this research, we isolated the full length cDNA of Zeta-carotene desaturase gene (ZDS) and a partial sequence of Lycopene epsilon cyclase gene(LYCE) from Chinese narcissus petals by RT-PCR and RACE and tried to express ZDS in E.coli. This laid a foundation for its application on subsequent genetic transformation study, which aims to provide a new way to the genetic improvement of the color of Chinese narcissus.1,The RNA extracted from Narcissus flower petal was used for the isolation of ZDS by RT-PCR (GenBank accession number: EU573238). The results of analysis of the coding sequences of the genes demonstrated that the gene of ZDS is 2189bp long. It contains an ORF encoding 499 amino acid residues(Mw=63.6kDa), a 5'untranslated region of 69bp and a 3'untranslated region of 395bp. Compared with ZDSs of other plants, Chinese narcissus's amino acid sequence had 97%, 85%, 83% identity with that of Narcissus pseudonarcissus, Vitis vinifera and Citrus unshiu.2,The ZDS gene cDNA was insert into the vevtor pET-28a degisted by EcoR I and Xoh I. The expression construct was transformed into BL21 and was indued. The result displayed that the ZDS was expressed in E.coli. And the SDS-PAGE experiments show that the actual value of ZDS molecular weight agrees well with the theoretical one.3,The partial sequence of LYCE isolated from Chinese narcissus by RACE is 1235bp long, encoding 371 amino acid residues. It's amino acid sequence had 80%, 76%, 76%, 75% identity with that of Daucus carota, Citrus maxima, Citrus sinensis and Citrus unshiu.