摘要
根据GenBank发表的丁香酚合成酶(EGS)基因的蛋白保守序列设计引物,利用RT-PCR结合RACE-PCR技术,从古老月季(Rosa chinensis)品种‘月月粉’盛开期的花瓣中获得了1个新的丁香酚合成酶基因RcEGS1,GenBank登录号为JQ522949。该基因全长为1171bp,开放阅读框(ORF)951bp,编码317个氨基酸;蛋白质理论分子量为35.64kD,等电点为7.36。氨基酸同源性分析表明,RcEGS1与矮牵牛PhIGS1的氨基酸同源性达到70%,与罗勒ObEGS1的同源性为59%,与月季RhEGS1的同源性仅为48.2%。运用实时荧光定量PCR法分析RcEGS1在不同组织部位和不同开放时期花瓣中的表达,发现其在叶片和茎段中没有表达,在花瓣和萼片中有表达,在盛开期的花瓣中表达量最高。
Using PCR degenerate primers designed based on GenBank published the conserved protein sequences of eugenol synthase genes to amplify cDNA fragments by RT-PCR and RACE-PCR, a new eugenol synthase gene named RcEGS1 was cloned from the flower petal of Rosa chinensis ＇Pallida＇ , and its GenBank accession number is JQ522949. The full cDNA was 1 171 bp in length with an open reading frame （ORF） of 951 bp encoding a protein of 317 amino acids. Molecular weight and isoelectric point of the protein was 35.64 kD and 7.36, respectively. Amino acids homology analysis indicated that the RcEGS1 and RhEGS1 proteins are 47.7% identical to each other, and had 70% and 59% homologies with CbEGS2 in Clarkia breweri and PhEGS1 in Petunia hybirida, respectively. The expression profile of RcEGS1 in different tissues and different developmental stages of rose flower was analyzed by quantitative Real-time PCR. The results showed that RcEGS1 expressed specifically in the flower petals and the sepals, and it had the highest transcript level in the petal. In additional, RcEGS1 was higher transcript level in the developmental blooming stage, and lower levels in that of bud and senescence stage.
出处
《园艺学报》
CAS
CSCD
北大核心
2012年第7期1387-1394,共8页
Acta Horticulturae Sinica
基金
国家自然科学基金项目(31060268)
云南省青年基金项目(2010CD097)
云南省科技创新强省计划项目(2007C0004Z)
