State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Agriculture, Guangxi University, Nanning 530004, China.
Int J Mol Sci. 2023 Jul 4;24(13):11087. doi: 10.3390/ijms241311087.
Leaf rolling is a crucial agronomic trait to consider in rice ( L.) breeding as it keeps the leaves upright, reducing interleaf shading and improving photosynthetic efficiency. The () gene plays a key role in regulating leaf rolling, as it encodes a glycosylphosphatidylinositol-anchored protein located on the plasma membrane. In this study, we used CRISPR/Cas9 to target the second and third exons of the gene in the indica rice line GXU103, which resulted in the generation of 14 T transgenic plants with a double-target mutation rate of 21.4%. After screening 120 T generation plants, we identified 26 T-DNA-free homozygous double-target mutation plants. We designated the resulting homozygous double-target knockout as . This line exhibited defects in leaf development, leaf rolling in the mature upright leaves, and a compact nature of the fully grown plants. Compared with the wild type (WT), the T generation of varied in two key aspects: the width of flag leaf (12.6% reduction compared with WT) and the leaf rolling index (48.77% increase compared with WT). In order to gain a deeper understanding of the involvement of in the regulatory network associated with rice leaf development, we performed a transcriptome analysis for the T generation of . A comparison of with WT revealed 459 differentially expressed genes (DEGs), including 388 upregulated genes and 71 downregulated genes. In terms of the function of the DEGs, there seemed to be a significant enrichment of genes associated with cell wall synthesis (, , , , , , , and ) and vacuole-related genes (), which may partially explain the increased leaf rolling in . Furthermore, the significant downregulation of BAHD acyltransferase-like protein gene () could be the main reason for the decreased leaf angle and the compact nature of the mutant plants. In summary, this study successfully elucidated the gene regulatory network in which participates, providing theoretical support for targeting this gene in rice breeding programs to promote variety improvement.
叶片卷曲是水稻（L.）育种中需要考虑的一个重要农艺性状，因为它可以使叶片保持直立，减少叶片间的遮荫，提高光合作用效率。()基因在调节叶片卷曲方面起着关键作用，因为它编码一种位于质膜上的糖基磷脂酰肌醇锚定蛋白。在本研究中，我们使用 CRISPR/Cas9 靶向籼稻 GXU103 中基因的第二和第三外显子，导致产生了 14 株具有 21.4%双靶点突变率的 T 转基因植株。在筛选了 120 株 T 代植株后，我们鉴定出了 26 株 T-DNA 缺失的纯合双靶点突变植株。我们将产生的纯合双靶点敲除命名为。该突变体表现出叶片发育缺陷、成熟直立叶片叶片卷曲以及完全生长植株紧凑的特性。与野生型（WT）相比，T 代的表型在两个关键方面有所不同：旗叶的宽度（与 WT 相比减少了 12.6%）和叶片卷曲指数（与 WT 相比增加了 48.77%）。为了更深入地了解在与水稻叶片发育相关的调控网络中发挥作用，我们对 T 代进行了转录组分析。与 WT 相比，发现 459 个差异表达基因（DEGs），包括 388 个上调基因和 71 个下调基因。就 DEGs 的功能而言，似乎与细胞壁合成（、、、、、、和）和液泡相关基因（）相关的基因显著富集，这可能部分解释了在中叶片卷曲增加的原因。此外，BAHD 酰基转移酶样蛋白基因（）的显著下调可能是突变体叶片角度减小和植株紧凑的主要原因。综上所述，本研究成功阐明了中参与的基因调控网络，为在水稻育种计划中靶向该基因以促进品种改良提供了理论支持。
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