漳州水仙ZDS 基因cDNA克隆及其表达分析

摘要

采用RACE技术从漳州水仙‘金盏银台’Narcissus tazetta var. chinensis花器官总RNA中克隆ZDS的cDNA序列，用相关软件进行序列分析，并利用实时荧光定量技术检测ZDS基因在各水仙品种不同花器官的表达。序列分析表明，ZDS基因cDNA全长2189 bp，其中包含69 bp 5'非编码区，395 bp 3'非编码区，1725 bp编码区（编码574个氨基酸，分子量约63.6 kDa），命名为Ntzds （GenBank登录号：EU573238），其编码的氨基酸序列（ACB87206.1）与喇叭水仙（CAA12062.1）、葡萄（XP_002277348.21）和温州蜜柑（ABC33728.1） ZDS基因编码产物的相似性分别为97%、85%和83%。实时荧光定量PCR分析表明，Ntzds基因在漳州水仙‘金盏银台’、‘Minnow’、‘Fortissimo’中均有表达，且同一品种中副冠的表达量高于花瓣；随着花色的加深，其转录水平也逐渐趋高。%　　The ZDS gene was cloned from total RNA of flower in Narcissus tazetta var. chinensis by RACE technology. The expression analysis of ZDS gene was predicted with some related software tools, and the expression of ZDS gene in different floral organ of different varieties was analysis with real-time PCR. The sequence analysis indicated that the length of ZDS is 2189 bp, it contains an ORF encoding 499 amino acid residues(Mw=63.6 kDa), as well as 5' non-coding region with 69 bp and 3' non-coding region with 395 bp, named as Ntzds (GenBank accession number EU573238). Compared with ZDSs of other plants, Ntzds amino acid sequence has 97%, 85%, 83% identity with that of Narcissus pseudonarcissus, Vitis vinifera and Citrus unshiu respectively. Expressions of Ntzds in petals and corona from N. tazetta var. chinensis, ‘Minnow’, ‘Fortissimo’ were investigated by real-time qPCR. The results showed that Ntzds gene expression was different in three varieties. The expression level in corona was higher than in petals in same variety; we concluded that the transcription level of Ntzds was developing with the deepening of flower color.