Through the hybridization of blue flower line X red flower line, and the screening of offspring, we have obtained a few stable blue anther lines from model plant Petunia hybrida. For investigating the mechanism of blue anther formation, the relativity between flavanone- 3'5'-hydroxylase (F3'5'H)and blue anther was studied in this article, which is the key enzyme in the synthesis pathway of 3'5'-hydroxylated anthocyanin (blue anthocyanidin). The PCR amplification results and the sequences of genomic DNA, and the expression characters in the third stage of flower development were analyzed. The results showed that, the genomic amplified model of F3'5'H gene in blue anthers plant were similar to its CK(yellow anthers plant), there are two encoded loci, Hf1 and Hf2 . In blue anther Petunia hybrida , Hf1 contain two genes, one is same as the republished cDNA of Hf1 , the other is a pseudogene, which can not encode the complete protein; Hf2 also include two genes, the 5'-terminal sequences of one gene show 96% identity with a republished Hf2 sequence, and there have a non specific amplification band. Moreover, the expression analysis indicated that Hf1 can be transcribed in anther tissue but only in blue one, and Hf2 couldn't express in anther. The results sugges- ted that there must be some relation between Hf1 and blue color quality of anther in Petunia, which laid the foundation for the further study of blue anther formation mechanism in Petunia hybrida.%通过品种间杂交和后代筛选,从模式植物矮牵牛中筛选出具有蓝花冠蓝花药的稳定株系.为了明确蓝色花药形成的分子机制,对蓝色花色素苷合成途径中的关键酶黄烷酮3′,5′-羟化酶(F3 ′5′H)进行研究,比较不同花药着色的矮牵牛植株中,F3′5′H基因组DNA的PCR扩增模式、基因组序列,以及花发育第3阶段的花药组织中基因表达差异.结果表明,蓝色花药植株与对照材料间的F3＇5＇H基因组DNA扩增模式基本相同,都存在两个基因编码位点Hf1和Hf2.在蓝色花药植株中,Hf1位点扩增出两个片段,一个与已公布的HF1力的序列完全一致,另一个是不能编码完整蛋白的假基因；Hf2基因位点也扩增出两个基因片段,其中有一个片段的5′-末端序列与已公布的Hf2序列有96％同源性,另一个是非特异性扩增产物.此外,Hf1基因可以在花药组织中表达,但仅在蓝色花药中有转录,而Hf2基因在花药组织中没有表达.根据上述结果认为,矮牵牛植株中,编码F3′5′H的Hf1基因与蓝色花药的形成相关,这一结论为进一步揭示矮牵牛蓝色花药形成的分子机制奠定了基础.
展开▼
