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首页> 外文期刊>Journal of genetics >Cloning, characterization and expression of $OsFMO_{(t)}$ in rice encoding a flavin monooxygenase
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Cloning, characterization and expression of $OsFMO_{(t)}$ in rice encoding a flavin monooxygenase

机译：水稻中编码黄素单加氧酶的$ OsFMO _ {（t）} $的克隆，表征和表达

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Flavin monooxygenases (FMO) play a key role in tryptophan (Trp)-dependent indole-acetic acid (IAA) biosynthesis in plants and regulate plant growth and development. In this study, the full-length genomic DNA and cDNA of $OsFMO_{(t)}$, a FMO gene that was originally identified from a rolled-leaf mutant in rice, was isolated and cloned from wild type of the rolled-leaf mutant. $OsFMO_{(t)}$ was found to have four exons and three introns, and encode a protein with 422 amino acid residues that contains two basic conserved motifs, with a a€?G?—e??o?—?—Ga€? characteristic structure. OsFMO(t) showed high amino acid sequence identity with FMO proteins from other plants, in particular with YUCCA from Arabidopsis, FLOOZY from Petunia, and OsYUCCA1 from rice. Our phylogenetic analysis showed that OsFMO$_{ ext{(t)}}$ and the homologous FMO proteins belong to the same clade in the evolutionary tree. Overexpression of $OsFMO_{(t)}$ in transformed rice calli produced IAA-excessive phenotypes that showed browning and lethal effects when exogenous auxins such as naphthylacetic acid (NAA) were added to the medium. These results suggested that the OsFMO$_{ ext{(t)}}$ protein is involved in IAA biosynthesis in rice and its overexpression could lead to the malformation of calli. Spatio-temporal expression analysis using RT-PCR and histochemical analysis for GUS activity revealed that expression of $OsFMO_{(t)}$ was totally absent in the rolled-leaf mutant. However, in the wild type variety, this gene was expressed at different levels temporally and spatially, with the highest expression observed in tissues with fast growth and cell division such as shoot apexes, tender leaves and root tips. Our results demonstrated that IAA biosynthesis regulated by $OsFMO_{(t)}$ is likely localized and might play an essential role in shaping local IAA concentrations which, in turn, is critical for regulating normal growth and development in rice.
机译：黄素单加氧酶（FMO）在植物中依赖色氨酸（Trp）的吲哚乙酸（IAA）生物合成中起关键作用，并调节植物的生长发育。在这项研究中，全长基因组DNA和cDNA $ OsFMO _ {（t）} $是一种FMO基因，最初是从水稻的卷叶突变体中鉴定出来的，并从野生型的卷叶中分离并克隆突变体。发现$ OsFMO _ {（t）} $具有四个外显子和三个内含子，并编码具有422个氨基酸残基的蛋白质，该蛋白质包含两个基本的保守基序，并带有aa？G？-e ?? o？-？-Ga €？特征结构。 OsFMO（t）与其他植物的FMO蛋白，尤其是拟南芥的YUCCA，矮牵牛的FLOOZY和水稻的OsYUCCA1，具有很高的氨基酸序列同一性。我们的系统发育分析表明，OsFMO $ _ {ext {（t）}} $和同源FMO蛋白在进化树中属于同一进化枝。在转化的水稻愈伤组织中$ OsFMO _ {（t）} $的过表达产生了IAA过多的表型，当向培养基中添加外源性生长素如萘乙酸（NAA）时，该表型呈现褐变和致死作用。这些结果表明，OsFMO $ _ {ext {（t）}} $蛋白参与了水稻的IAA生物合成，其过表达可能导致愈伤组织畸形。使用RT-PCR的时空表达分析和GUS活性的组织化学分析显示，在卷叶突变体中完全不存在$ OsFMO _ {（t）} $的表达。然而，在野生型品种中，该基因在时间和空间上以不同水平表达，在具有快速生长和细胞分裂的组织（如芽尖，嫩叶和根尖）中观察到最高的表达。我们的结果表明，受$ OsFMO _ {（t）} $调控的IAA生物合成很可能是局部的，并且可能在塑造局部IAA浓​​度方面起着至关重要的作用，而这对调节稻米的正常生长和发育至关重要。

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