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Molecular and cellular study of flavonoid metabolism and detection of relating genes in soybean

大豆黄酮代谢的分子细胞研究及相关基因检测

基本信息

批准号：
19780089
负责人：
TODA Kyoko
金额：
$ 2.53万
依托单位：
National Agricultural Research Organization
依托单位国家：
日本
项目类别：
Grant-in-Aid for Young Scientists (B)
财政年份：
2007
资助国家：
日本
起止时间：
2007 至 2009
项目状态：
已结题

项目摘要

Chilling tolerance is an important trait of soybeans (Glycine max (L.) Merr.) produced in cool climates. We previously isolated a soybean flavonoid 3' hydroxylase (F3' H) gene corresponding to the T locus, which controls pubescence and seed coat color. A genetic link between the T gene and chilling tolerance has been reported, although the exact underlying mechanisms remain unclear. Using the soybean near-isogenic lines (NILs) To7B (TT) and To7G (tt), we examined the relationship between chilling injury, antioxidant activity and flavonoid profiles associated with chilling treatment (15℃). Chilling injury was more severe in the second trifoliate leaves of To7G than in those of To7B. Lipid peroxidation was enhanced by chilling in To7G. Chilling-induced enhancement of antioxidant activity was more prominent in To7B than in To7G. High performance liquid chromatography (HPLC) analysis indicated that the contents of quercetin glycosides and isorhamnetin glycosides (3',4'-dihydroxylated flavonol derivatives) increase in the second trifoliate leaves of To7B after chilling treatment, whereas the same treatment increased kaempferol glycoside (4'-monohydroxylated flavonol derivatives) content in the corresponding leaves of To7G. Histochemical staining also demonstrated chilling-induced flavonoid accumulation. Microarray analysis and real-time reverse transcription-PCR (RT-PCR) demonstrated that the transcript levels of soybean F3' H are upregulated by chilling. The differences in chilling injury, antioxidant activity and flavonoid species between the two NILs support the notion that soybean F3' H affects chilling tolerance by increasing antioxidant activity via production of 3',4'-dihydroxylated flavonol derivatives.

耐冷性是大豆（Glycine max（L.）梅尔）在凉爽的气候中生产。我们先前分离了一个大豆类黄酮3'羟化酶（F3' H）基因，该基因对应于T基因座，该基因控制短柔毛和种皮颜色。T基因和耐冷性之间的遗传联系已被报道，尽管确切的潜在机制仍不清楚。以大豆近等基因系To 7 B（TT）和To 7 G（tt）为材料，研究了低温处理（15℃）对大豆抗氧化活性和类黄酮含量的影响。To 7 G的第二片三叶冷害比To 7 B严重。低温处理增强了To 7 G的膜脂过氧化作用。低温诱导的抗氧化活性增强在To 7 B中比在To 7 G中更突出。高效液相色谱（HPLC）分析表明，低温处理后，To 7 B的第二片三叶中槲皮素苷和异鼠李素苷（3 '，4'-二羟基黄酮醇衍生物）含量增加，而To 7 G相应叶片中山奈酚苷（4 '-单羟基黄酮醇衍生物）含量增加。组织化学染色也证明了低温诱导的类黄酮积累。基因芯片分析和实时荧光定量RT-PCR（real-time reverse transcription-PCR，RT-PCR）结果表明，大豆F3' H基因的转录水平受到低温处理的上调。两个近等基因系在冷害、抗氧化活性和黄酮类化合物种类上的差异支持了大豆F3' H通过产生3'，4 '-二羟基黄酮醇衍生物提高抗氧化活性从而影响耐冷性的观点。