Lakhssassi Naoufal, Colantonio Vincent, Flowers Nicholas D, Zhou Zhou, Henry Jason, Liu Shiming, Meksem Khalid
Department of Plant, Soil, and Agricultural Systems, Southern Illinois University, Carbondale, Illinois 62901.
Department of Microbiology, Southern Illinois University, Carbondale, Illinois 62901.
Plant Physiol. 2017 Jul;174(3):1531-1543. doi: 10.1104/pp.16.01929. Epub 2017 May 1.
Stearoyl-acyl carrier protein desaturase (SACPD-C) has been reported to control the accumulation of seed stearic acid; however, no study has previously reported its involvement in leaf stearic acid content and impact on leaf structure and morphology. A subset of an ethyl methanesulfonate mutagenized population of soybean () 'Forrest' was screened to identify mutants within the gene. Using a forward genetics approach, one nonsense and four missense mutants were identified to have high levels of seed, nodule, and leaf stearic acid content. Homology modeling and in silico analysis of the GmSACPD-C enzyme revealed that most of these mutations were localized near or at conserved residues essential for diiron ion coordination. Soybeans carrying mutations at conserved residues showed the highest stearic acid content, and these mutations were found to have deleterious effects on nodule development and function. Interestingly, mutations at nonconserved residues show an increase in stearic acid content yet retain healthy nodules. Thus, random mutagenesis and mutational analysis allows for the achievement of high seed stearic acid content with no associated negative agronomic characteristics. Additionally, expression analysis demonstrates that nodule leghemoglobin transcripts were significantly more abundant in soybeans with deleterious mutations at conserved residues of GmSACPD-C. Finally, we report that mutations cause an increase in leaf stearic acid content and an alteration of leaf structure and morphology in addition to differences in nitrogen-fixing nodule structure.
硬脂酰 - 酰基载体蛋白去饱和酶(SACPD - C)已被报道可控制种子硬脂酸的积累;然而,此前尚无研究报道其与叶片硬脂酸含量有关,以及对叶片结构和形态的影响。对用甲磺酸乙酯诱变的大豆()‘Forrest’群体的一个子集进行筛选,以鉴定基因内的突变体。采用正向遗传学方法,鉴定出一个无义突变体和四个错义突变体,它们的种子、根瘤和叶片中的硬脂酸含量较高。对GmSACPD - C酶进行同源建模和计算机分析表明,这些突变大多位于二价铁离子配位所必需的保守残基附近或其上。在保守残基处携带突变的大豆硬脂酸含量最高,且发现这些突变对根瘤发育和功能有有害影响。有趣的是,非保守残基处的突变显示硬脂酸含量增加,但根瘤仍保持健康。因此,随机诱变和突变分析能够实现高种子硬脂酸含量,且无相关负面农艺性状。此外,表达分析表明,在GmSACPD - C保守残基处有有害突变的大豆中,根瘤豆血红蛋白转录本明显更丰富。最后,我们报道,突变除了导致固氮根瘤结构差异外,还会使叶片硬脂酸含量增加,并改变叶片结构和形态。