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以能够产生抗虫皂苷的高抗小菜蛾资源G型欧洲山芥(Barbarea vulgaris R.Br.)B44为材料,利用RACE技术克隆出皂苷合成关键酶beta–香树脂合成酶的基因(Barbarea vulgaris beta-amyrin synthase,Bv-beta-AS)。该基因编码区序列长为2289bp(GenBank登录号JQ172795),推导其编码762个氨基酸;在基因组水平上长度为4107bp(GenBank登录号JQ172796),含有15个内含子。Bv-beta-AS编码的氨基酸具有beta–香树脂合成酶基因家族的保守序列,即DCTAE序列和QW特征序列,氨基酸多序列比对和进化树分析表明,该基因与拟南芥beta–香树脂合成酶基因的相似性最高,为74%。利用荧光定量PCR对欧洲山芥在小菜蛾诱导下该基因的表达进行研究,结果表明,该基因受虫害诱导时上调表达,但是上升到12h达顶峰后随时间推移呈回归的趋势。从序列特征和表达模式上推测,Bv-beta-AS可能是抗虫皂苷合成途径的一个关键酶的基因。
The gene Barbasea vulgaris beta, a key enzyme in the synthesis of saponin, was cloned by RACE technique from Barbarea vulgaris R. Br. B44, -amyrin synthase, Bv-beta-AS). The coding sequence of this gene was 2289bp in length (GenBank accession number JQ172795). The deduced amino acid sequence of the deduced amino acid sequence was 762 amino acids. At the genome level, it was 4107bp in length (GenBank accession number JQ172796) and contained 15 introns. The amino acids encoded by Bv-beta-AS have conserved sequences of the beta-fragrant resin synthase gene family, ie, the DCTAE sequence and the QW signature sequence. Amino acid sequence alignment and phylogenetic tree analysis show that this gene binds to Arabidopsis beta- The highest similarity of the synthetase genes was 74%. Fluorescent quantitative PCR was used to study the expression of the gene in European horse mustard (Brassica ciliate). The results showed that the gene was up-regulated when induced by pest but rose to the peak at 12h and then regressed with time. From the sequence characteristics and expression patterns speculated that Bv-beta-AS may be a key enzyme in the synthesis of insect-resistant saponin genes.