Isolation and characterization of soybean chalcone reductase cDNA, which encodes the key enzyme for the biosynthesis of 4,2′,4′-trihydroxychalcone in legumes
Isolation and characterization of soybean chalcone reductase cDNA, which encodes the key enzyme for the biosynthesis of 4,2′,4′-trihydroxychalcone in legumes
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DOI：
10.1007/s11032-014-0169-1
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发表时间：
2014-09
影响因子：
3.1
通讯作者：
Zhuo Zhang;Yongping Fu;Jian Ma;Chao Zhang-;Pi-wu Wang
中科院分区：
文献类型：
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作者：
Zhuo Zhang;Yongping Fu;Jian Ma;Chao Zhang-;Pi-wu Wang
在植物中，病原菌攻击诱导的植物抗毒素在抗病性中起着重要作用。大豆[Glycine maxL.](稳定)]，病原菌的攻击诱导异黄酮合成，尤其是大豆苷元。查尔酮还原酶（CHR）是大豆苷元生物合成的关键酶。它与查尔酮合成酶一起催化异质大豆苷元的形成，异质大豆苷元是大豆苷元生物合成的必要底物。本研究从大豆品种济农17中分离到一个CHR基因Gmchr2（GenBank代码：KF758395）。该cDNA由1417 bp的片段组成，其中包括969 bp的开放阅读框。该基因位于大豆基因组的第9号染色体上。将大豆疫霉菌（Phytophthora sojaee）接种到大豆根部，采用实时荧光定量PCR和高效液相色谱（HPLC）技术分别研究了CHR基因在不同大豆组织中的转录水平和催化活性的变化。结果表明，接种病原菌后，根中异异黄酮含量显著增加。将gmchr2基因转化到烟草中，用高效液相色谱法证实了转化体中存在异尿素。在35S CaMV启动子的控制下，gmchr2基因的表达也得到证实。编码大豆CHR的achrgene的这一特性有助于揭示大豆异黄酮的生物合成和调控，并将有助于操纵导致异黄酮植物抗毒素的苯丙素途径。
In plants, phytoalexins induced by pathogen attack play an important role in disease resistance. In soybean [Glycine maxL. (Merr.)], attack by pathogenic bacteria induces the synthesis of isoflavonoids, especially daidzein. Chalcone reductase (CHR) is the key enzyme in the biosynthesis of daidzein. Along with chalcone synthase, it catalyzes the formation of isoliquiritigenin, which is a necessary substrate for daidzein biosynthesis. In this study, a CHR gene,Gmchr2(GenBank code: KF758395), was isolated from the soybean cultivar Jinong 17. The cDNA consisted of a 1,417-bp fragment that included an open reading frame of 969 bp. The gene is located on chromosome 9 of the soybean genome.Phytophthora sojaewas inoculated onto soybean roots, and changes in the transcript levels of thechrgenes and the catalytic activity of CHR were investigated in different soybean tissues by real-time fluorescence quantitative PCR and high-performance liquid chromatography (HPLC), respectively. The results showed that the isoliquiritigenin content in roots significantly increased after pathogen inoculation. TheGmchr2gene was transformed into tobacco, and the presence of isoliquiritigenin in the transformants was confirmed by HPLC. Expression of theGmchr2gene under the control of the 35S CaMV promoter was also confirmed. This characterization of achrgene encoding a soybean CHR helps to shed light on the biological synthesis and regulation of soybean isoflavones and will be useful for manipulating the phenylpropanoid pathway leading to isoflavonoid phytoalexins.