Niu Xiangli, Tang Wei, Huang Weizao, Ren Guangjun, Wang Qilin, Luo Di, Xiao Yingyong, Yang Shimei, Wang Feng, Lu Bao-Rong, Gao Fangyuan, Lu Tiegang, Liu Yongsheng
Ministry of Education Key Laboratory for Bio-resource and Eco-environment, College of Life Science, Sichuan University, Chengdu 610064, PR China.
BMC Plant Biol. 2008 Oct 8;8:100. doi: 10.1186/1471-2229-8-100.
Aromatic rice is popular worldwide because of its characteristic fragrance. Genetic studies and physical fine mapping reveal that a candidate gene (fgr/OsBADH2) homologous to betaine aldehyde dehydrogenase is responsible for aroma metabolism in fragrant rice varieties, but the direct evidence demonstrating the functions of OsBADH2 is lacking. To elucidate the physiological roles of OsBADH2, sequencing approach and RNA interference (RNAi) technique were employed to analyze allelic variation and functions of OsBADH2 gene in aroma production. Semi-quantitative, real-time reverse transcription-polymerase chain reaction (RT-PCR), as well as gas chromatography-mass spectrometry (GC-MS) were conducted to determine the expression levels of OsBADH2 and the fragrant compound in wild type and transgenic OsBADH2-RNAi repression lines, respectively.
The results showed that multiple mutations identical to fgr allele occur in the 13 fragrant rice accessions across China; OsBADH2 is expressed constitutively, with less expression abundance in mature roots; the disrupted OsBADH2 by RNA interference leads to significantly increased 2-acetyl-1-pyrroline production.
We have found that the altered expression levels of OsBADH2 gene influence aroma accumulation, and the prevalent aromatic allele probably has a single evolutionary origin.
香稻因其独特的香气而在全球广受欢迎。遗传研究和物理精细定位表明,与甜菜碱醛脱氢酶同源的一个候选基因(fgr/OsBADH2)负责香稻品种的香气代谢,但缺乏直接证明OsBADH2功能的证据。为阐明OsBADH2的生理作用,采用测序方法和RNA干扰(RNAi)技术分析OsBADH2基因在香气产生中的等位变异和功能。分别通过半定量、实时逆转录-聚合酶链反应(RT-PCR)以及气相色谱-质谱联用(GC-MS)来测定野生型和转基因OsBADH2-RNAi抑制系中OsBADH2的表达水平和香气化合物。
结果表明,中国13个香稻品种中出现了与fgr等位基因相同的多个突变;OsBADH2组成型表达,在成熟根中的表达丰度较低;RNA干扰导致OsBADH2功能破坏,使2-乙酰-1-吡咯啉的产量显著增加。
我们发现OsBADH2基因表达水平的改变影响香气积累,且普遍存在的香气等位基因可能有单一的进化起源。