Cloning Of DXPS,DXR And PAL Related To Floral Fragrance From Narcissus Tazetta L. Var. Chinensis Roem

Narcissus tazetta L. var. chinensis Roem, a variant of polyanthous daffodils, is a kind of perennial plants and belongs to the family Amaryllidaceae. Zhangzhou which is a city of FuJian province is the most important produce area of Chinese narcissus. Zhangzhou narcissus has a rich aromas and gets people's love.Now, Some high-quality ornamental plants lack fragrance and can not completely meet the market demand. It's a value to the floral gene clone.In this study, we clone the full length cDNA of 1-deoxyxylu-lose-5-phosphate synthase(DXPS) and 1-deoxyxylu-lose-5-phosphate reduetoisomerase(DXR) and a partial sequence of Phenylalanine ammonia-lyase (PAL) from Chinese narcissus petals by RT-PCR and RACE. It becomes a possibility to improve lack of fragrance plant genetic on subsequent genetic transformation study. The results were as follows:1. The full length cDNA of 1-deoxyxylu-lose-5-phosphate synthase (DXPS) is obtained ( GenBank accession number:GU574804 ) .The RNA extracted from Narcissus flower was used for the isolation of DXPS by RT-PCR. It's a key gene in the methylerythritol Synthetic approach. The results of analysis of the coding sequences of the gene demonstrated that the cDNA of DXPS is 2338 bp long,including a 5' untranslated region of 191bp and a 3'untranslated region of 242bp. It contains an ORF encoding 634 amino acid residues. Chinese narcissus's amino acid sequence had 92%, 75% and 75% identity with that of Narcissus pseudonarcissus, Hevea brasiliensis, Adonis palaestina.2. The full length cDNA of 1-deoxyxylu-lose-5-phosphate reduetoisomerase(DXR) (GenBank accession number:GU574805)is obtained.It's also a key gene in the methylerythritol Synthetic approach. The full-length cDNA of DXR is 1606 bp long, including a 5'untranslated region of 119bp and a 3'untranslated region of 77bp. It contains an ORF encoding470 amino acid residues. Chinese narcissus's amino acid sequence had 81%,75%,77% and76% identity with that of Vanda hybrid, Hordeum vulgare, Amomum villosum,Mentha. 3.Using the same method, we clone a partial sequence of Phenylalanineammonia-lyase (PAL) from Chinese narcissus(GenBank accession number:GU574806).The Length of the cDNA is 1971bp.It contains 586 amino acid residues. Chinese narcissus's amino acid sequence had 91%, 84% and 83% identity with that of Lycoris radiata, Musa acuminata, Vitis vinifera.