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Novel allelic variant of Lpa1 gene associated with a significant reduction in seed phytic acid content in rice (Oryza sativa L.)

机译:水稻种子植酸含量显着降低的LPA1基因的新等位基因变体(Oryza sativa L.)

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摘要

In plants, myo-inositol-1,2,3,4,5,6-hexakisphosphate (InsP6), also known as phytic acid (PA), is a major component of organic phosphorus (P), and accounts for up to 85% of the total P in seeds. In rice (Oryza sativa L.), PA mainly accumulates in rice bran, and chelates mineral cations, resulting in mineral deficiencies among brown rice consumers. Therefore, considerable efforts have been focused on the development of low PA (LPA) rice cultivars. In this study, we performed genetic and molecular analyses of OsLpa1, a major PA biosynthesis gene, in Sanggol, a low PA mutant variety developed via chemical mutagenesis of Ilpum rice cultivar. Genetic segregation and sequencing analyses revealed that a recessive allele, lpa1-3, at the OsLpa1 locus (Os02g0819400) was responsible for a significant reduction in seed PA content in Sanggol. The lpa1-3 gene harboured a point mutation (C623T) in the fourth exon of the predicted coding region, resulting in threonine (Thr) to isoleucine (Ile) amino acidsubstitution at position 208 (Thr208Ile). Three-dimensional analysis of Lpa1 protein structure indicated that myo-inositol 3-monophosphate [Ins(3)P1] could bind to the active site of Lpa1, with ATP as a cofactor for catalysis. Furthermore, the presence of Thr208 in the loop adjacent to the entry site of the binding pocket suggests that Thr208Ile substitution is involved in regulating enzyme activity via phosphorylation. Therefore, we propose that Thr208Ile substitution in lpa1-3 reduces Lpa1 enzyme activity in Sanggol, resulting in reduced PA biosynthesis.
机译:在植物中,肌醇1,2,3,4,5,6-六磷酸(InsP6),也被称为肌醇六磷酸(PA),是的有机磷(P)的主要成分,并占高达85 %在种子的总P的。在水稻(Oryza sativa L.),PA主要积聚在米糠,和螯合物矿物阳离子,致使糙米消费者中矿物质缺乏。因此,相当大的努力都集中在低PA(LPA)水稻品种的发展。在这项研究中,我们进行OsLpa1,一个主要的PA合成基因,在Sanggol,通过Ilpum水稻品种的化学诱变开发了低PA突变品种的遗传和分子分析。遗传分离和测序分析显示,隐性等位基因,lpa1-3,在OsLpa1基因座(Os02g0819400)是负责在Sanggol种子PA含量显著降低。所述lpa1-3基因窝藏在预测编码区的外显子第四点突变(C623T),导致苏氨酸(Thr)至异亮氨酸(ILE)氨基acidsubstitution在位置208(Thr208Ile)。 LPA1蛋白质结构的三维分析表明,肌醇-3-磷酸[宏(3)P1]可以结合LPA1的活性位点,与ATP作为用于催化辅因子。此外,Thr208的在环邻近所述结合口袋的入口点的存在表明Thr208Ile取代参与经由磷酸化调节酶活性。因此,我们建议,在lpa1-3 Thr208Ile取代降低LPA1酶活性Sanggol,从而降低PA的生物合成。

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