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采用RT-PCR和RACE技术从观赏向日葵‘闽葵3号’黄色花瓣中克隆到类胡萝卜素合成途径关键基因HaPDS的cDNA,该cDNA全长2 017 bp,具有一个1 710 bp的完整开放阅读框(ORF),编码一个570个氨基酸的蛋白质。序列分析表明,HaPDS编码的氨基酸序列与其他植物的PDS蛋白具有很高的同源性,在N-端有一个辅助因子结合结构域,C-端有一个类胡萝卜素结合域。系统进化树分析显示,观赏向日葵HaPDS与万寿菊、菊花蛋白亲缘关系较近。实时荧光定量RT-PCR技术分析表明,HaPDS基因在花发育的盛花期表达量最高;不同组织中的表达量舌状花瓣>苞片>叶片>绿色管状花>黑色管状花;随着基因表达量的增加,花色由白色到黄色、金黄色转变。
The cDNA of the key gene of carotenoid synthesis, HaPDS, was cloned by RT-PCR and RACE from the ornamental sunflower ’Min-3’ yellow petal. The full-length cDNA was 2017 bp with a complete open reading frame of 1 710 bp (ORF), encoding a 570 amino acid protein. Sequence analysis showed that the amino acid sequence encoded by HaPDS has high homology with other plant PDS proteins, a cofactor binding domain at the N-terminus, and a carotenoid binding domain at the C-terminus. Phylogenetic tree analysis showed that ornamental sunflower HaPDS is closely related to marigold and chrysanthemum proteins. Real-time fluorescent quantitative RT-PCR analysis showed that the expression level of HaPDS gene was the highest in the flowering stage of flower development. The expression of HaPDS gene in different tissues was higher than that of tongue-shaped petals> bracts> leaves> green tubular flowers> black tubular flowers. With the increase of gene expression The increase from the color from white to yellow, golden yellow change.