麻花艽×狭叶柴胡F1代β-amyrin合成酶基因表达及齐墩果酸含量分析

摘要

[目的]本文旨在分析麻花艽×狭叶柴胡F1代β-amyrin合成酶基因的表达及下游产物齐墩果酸的含量.[方法]首先根据进化树分析亲本及体细胞杂种中β-amyrin合成酶基因与其它物种中同族基因的亲缘关系,然后通过序列分析进行了该基因在杂种中的等位变异研究,最后利用半定量RT-PCR和HPLC方法分析了亲本及杂种中β-amyrin合成酶的基因表达情况及其下游产物齐墩果酸的含量变化.[结果]β-amyrin合成酶基因在杂种ZA中发生了1个碱基突变,在ZB中未发生突变,在ZC中发生了2个碱基突变.虽然本实验证明β-amyrin合成酶基因的表达与齐墩果酸含量呈正相关,但杂种中这些基因的变异与齐墩果酸含量没有对应关系.[结论]本实验表明,β-amyrin合成酶中个别基因的突变对其功能未造成影响.%[Objective] The paper aimed to analyze the expression of β-amyrin synthase gene and the content of oleanolic acid in the biparents and hybrids between Gentiana straminea and Bupleurum scoronerifolium.[Method] First,phylogenetic trees was created to reveal the genetic relationship of β-amyrin synthase gene among different species.Then,the allelic variation of β-amyrin synthase gene in hybrids was studied by sequence analysis.Last,semi quantitative RT-PCR and HPLC were used to analyze the expression of β-amyrin synthase gene and the content of oleanolic acid in the parents and hybrids.[Result] β-amyrin synthase gene in the hybrids was positively correlated with the content of oleanolic acid,which had one base mutation in hybrid ZA,no mutation in ZB and two base mutations in ZC.But there was no corresponding relationship between the variation of these genes and the content of oleanolic acid in the hybrids.[Conclusion] The mutation of individual gene in β-amyrin synthase gene had no effect on the function of β-amyrin synthase.