Rice Science Center and Rice Gene Discovery, Kasetsart University Kamphaeng Saen Campus, Nakhon Pathom, Thailand.
Plant Biotechnol J. 2011 Jan;9(1):75-87. doi: 10.1111/j.1467-7652.2010.00533.x.
2-Acetyl-1-pyrroline (2AP), the volatile compound that provides the 'popcorn-like' aroma in a large variety of cereal and food products, is widely found in nature. Deficiency in amino aldehyde dehydrogenase (AMADH) was previously shown to be the likely cause of 2AP biosynthesis in rice (Oryza sativa L.). In this study, the validity of this mechanism was investigated in soybeans (Glycine max L.). An assay of AMADH activity in soybeans revealed that the aromatic soybean, which contains 2AP, also lacked AMADH enzyme activity. Two genes, GmAMADH1 and GmAMADH2, which are homologous to the rice Os2AP gene that encodes AMADH, were characterized. The transcription level of GmAMADH2 was lower in aromatic varieties than in nonaromatic varieties, whereas the expression of GmAMADH1 did not differ. A double nucleotide (TT) deletion was found in exon 10 of GmAMADH2 in all aromatic varieties. This variation caused a frame-shift mutation and a premature stop codon. Suppression of GmAMADH2 by introduction of a GmAMADH2-RNAi construct into the calli of the two nonaromatic wild-type varieties inhibited the synthesis of AMADH and induced the biosynthesis of 2AP. These results suggest that deficiency in the GmAMADH2 product, AMADH, plays a similar role in soybean as in rice, which is to promote 2AP biosynthesis. This phenomenon might be a conserved mechanism among plant species.
2-乙酰基-1-吡咯啉(2AP)是一种挥发性化合物,赋予了许多谷物和食品产品“爆米花般”的香气,广泛存在于自然界中。先前的研究表明,氨基酸醛脱氢酶(AMADH)的缺乏可能是导致水稻(Oryza sativa L.)中 2AP 生物合成的原因。在本研究中,我们研究了该机制在大豆(Glycine max L.)中的有效性。对大豆中 AMADH 活性的测定表明,含有 2AP 的芳香型大豆也缺乏 AMADH 酶活性。两个基因,GmAMADH1 和 GmAMADH2,与编码 AMADH 的水稻 Os2AP 基因同源,被鉴定出来。GmAMADH2 的转录水平在芳香型品种中低于非芳香型品种,而 GmAMADH1 的表达没有差异。在所有芳香型品种中,GmAMADH2 的第 10 外显子中发现了一个双核苷酸(TT)缺失。这种变异导致移码突变和提前终止密码子。通过将 GmAMADH2-RNAi 构建体导入两个非芳香型野生型品种的愈伤组织中,抑制了 GmAMADH2 的表达,从而抑制了 AMADH 的合成并诱导了 2AP 的生物合成。这些结果表明,GmAMADH2 产物 AMADH 的缺乏在大豆中与在水稻中发挥相似的作用,即促进 2AP 的生物合成。这种现象可能是植物物种之间的一种保守机制。