大豆查尔酮还原酶基因(Gmchr4)的克隆与功能鉴定

Isolation and functional characterization of soybean chalcone reductase gene Gmchr4

大豆查尔酮还原酶(chalcone reductase,CHR)是大豆苷元合成途径中必需的关键酶之一。在大豆基因组中分离到新的查尔酮还原酶基因Gmchr4,克隆基因cDNA片段长度为1 410bp(Gen Bank登录号:KF938604),含开发阅读框966bp,分析了Gmchr4在大豆基因组的进化关系。通过实时荧光定量PCR技术(Q-PCR)和高效液相色谱技术(HPLC)分析了基因的表达水平和催化产物异甘草素的含量。实验结果证明得到一种新的大豆查尔酮还原酶蛋白基因,为深入探索大豆苷元等异黄酮类化合物的生物合成途径,尤其是相关基因间的表达调控提供了参考。

Chalcone reductase （CHR） is a key enzyme in the biological synthesis of daidzein, which is the main composition of isoflavones. In this study, chalcone reductase gene Gmchr4 （ GenBank code ： KF938604） was isolated from the soybean Jinong 17. This gene consisted of a 1 410bp fragment including an open reading frame of 969bp. Phylogenetic tree of CHR protein was performed by using MEGAd. 0 program. The Gmchr4 gene was transformed into tobacco, and the presence of isoliquiritigenin in tobacco was confirmed by high - performance liquid chromatography （HPLC）. Expression of Gmchr4 gene under the control of 35S CaMV promoter was also examined by real -time fluorescence quantitative PCR （Q -PCR）. Cloning and characterization of Gmchr4 gene were beneficial to elucidate the biological synthesis and regulation of soybean isoflavones, and to manipulate the phenylpro- panoid pathway generating isoflavonoid phytoalexin.