Shanghai Key Laboratory of Plant Molecular Sciences, College of Life Sciences, Shanghai Normal University, Shanghai 200234, China.
Section of Cell and Developmental Biology, University of California San Diego, La Jolla, California 92093-0116.
Plant Physiol. 2019 May;180(1):453-464. doi: 10.1104/pp.19.00127. Epub 2019 Mar 13.
Maternal cells play a critical role in ensuring the normal development of embryos, endosperms, and seeds. Mutations that disrupt the maternal control of embryogenesis and seed development are difficult to identify. Here, we completely deleted four () genes in Arabidopsis () using a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein9 (Cas9) genome-editing technology. We found that plants with a deletion of phenocopied plants overexpressing miRNA167-resistant versions of () or , two miRNA167 targets. Both the mutant and the overexpression lines were defective in anther dehiscence and ovule development. Serendipitously, we found that the (♀) × wild type (♂) crosses failed to produce normal embryos and endosperms, despite the findings that embryos with either or genotypes developed normally when plants were self-pollinated, revealing a central role of in maternal control of seed development. The phenotype is 100% penetrant, providing a great genetic tool for studying the roles of miRNAs and auxin in maternal control. Moreover, we found that mutants flowered significantly later than wild-type plants, a phenotype that was not observed in the overexpression lines. We show that the reproductive defects of mutants were suppressed by a decrease of activities of , , or both. Our results clearly demonstrate that is the predominant member in regulating Arabidopsis reproduction and that acts as a maternal gene that functions largely through and .
母细胞在确保胚胎、胚乳和种子的正常发育方面起着关键作用。那些破坏母细胞对胚胎发生和种子发育的控制的突变很难被识别。在这里,我们使用了一种成簇的、规律间隔的短回文重复序列(CRISPR)/CRISPR 相关蛋白 9(Cas9)基因组编辑技术,完全删除了拟南芥中的四个()基因。我们发现,缺失的植物表现出类似于过表达 miRNA167 抗性版本的()或()的表型,这两个都是 miRNA167 的靶基因。()突变体和过表达系在花药开裂和胚珠发育方面都有缺陷。偶然的是,我们发现(♀)×野生型(♂)杂交未能产生正常的胚胎和胚乳,尽管发现当()植物自花授粉时,具有或基因型的胚胎正常发育,这表明()在母细胞对种子发育的控制中起着核心作用。()表型是 100%的外显率,为研究 miRNA 和生长素在母细胞控制中的作用提供了一个很好的遗传工具。此外,我们发现()突变体比野生型植物开花晚得多,而在过表达系中没有观察到这种表型。我们表明,()突变体的生殖缺陷被()、()或两者活性的降低所抑制。我们的结果清楚地表明,()是调控拟南芥生殖的主要()成员,()作为一个母性基因,主要通过()和()发挥作用。