Institute of Plant Biology and Department of Life Science, National Taiwan University, Taipei, Taiwan.
Department of Life Sciences, National Chung Hsing University, Taichung, Taiwan.
J Exp Bot. 2019 Jan 7;70(2):469-483. doi: 10.1093/jxb/ery381.
MiRNAs play diverse roles in plant development and defense responses by binding to their mRNA targets based on sequence complementarity. Here, we investigated a wound-related miR408 and its target genes in sweet potato (Ipomoea batatas) by small RNA deep sequencing and transcriptome analysis. The expression patterns of miR408 and the miR408 precursor were significantly repressed by wounding and jasmonate (JA). In contrast, expression of the putative target genes IbKCS (3-ketoacyl-CoA synthase 4), IbPCL (plantacyanin), and IbGAUT (galacturonosyltransferase 7-like) of miR408 was increased following wounding, whereas only IbKCS was increased after JA treatment. Target cleavage site mapping and Agrobacterium-mediated transient assay demonstrated that IbKCS, IbPCL, and IbGAUT were the targets of miR408. The expression of miR408 target genes was repressed in transgenic sweet potatoes overexpressing miR408. These data indicated a relationship between miR408 and its target genes. Notably, miR408-overexpressing plants showed a semi-dwarf phenotype and attenuated resistance to insect feeding, while transgenic plants overexpressing IbKCS exhibited more insect resistance than plants overexpressing only the empty vector. Collectively, sweet potato reduces the abundance of miR408 upon wounding to elevate the expression of IbKCS, IbPCL, and IbGAUT. The expression of IbKCS enhances the defense system against herbivore wounding.
miRNAs 通过与基于序列互补性的 mRNA 靶标结合,在植物发育和防御反应中发挥多种作用。在这里,我们通过小 RNA 深度测序和转录组分析研究了甘薯(Ipomoea batatas)中与伤口相关的 miR408 及其靶基因。miR408 和 miR408 前体的表达模式在受到伤害和茉莉酸(JA)处理后显著受到抑制。相比之下,miR408 的假定靶基因 IbKCS(3-酮酰基辅酶 A 合酶 4)、IbPCL(植物血蓝蛋白)和 IbGAUT(半乳糖基转移酶 7 样)的表达在受到伤害后增加,而只有 IbKCS 在 JA 处理后增加。靶标切割位点映射和农杆菌介导的瞬时测定表明,IbKCS、IbPCL 和 IbGAUT 是 miR408 的靶标。miR408 靶基因的表达在过表达 miR408 的转基因甘薯中受到抑制。这些数据表明 miR408 与其靶基因之间存在关系。值得注意的是,过表达 miR408 的植物表现出半矮化表型和对昆虫取食的抗性减弱,而过表达 IbKCS 的转基因植物比仅过表达空载体的植物表现出更强的抗虫性。总之,甘薯在受伤时减少 miR408 的丰度,以提高 IbKCS、IbPCL 和 IbGAUT 的表达。IbKCS 的表达增强了植物对草食性昆虫伤害的防御系统。