a Henan Key Laboratory of Ion-beam Bioengineering, Zhengzhou University, Zhengzhou 450000, China.
b School of Nursing, Zhengzhou University, Zhengzhou 450000, China.
Radiat Res. 2019 Feb;191(2):189-200. doi: 10.1667/RR15125.1. Epub 2018 Nov 30.
MicroRNAs (miRNAs) are a non-coding regulatory RNAs that play significant roles in plant growth and development, especially in the stress response. Low-energy ion radiation, a type of environmental stress, can cause multiple biological effects. To understand the roles of miRNAs in response to low-energy N ion radiation in Oryza sativa, high-throughput sequencing of small RNAs was carried out to detect the expression of miRNAs in the shoots of the rice after 2 × 10 N+/cm irradiation. The differentially expressed 28 known miRNAs were identified, 17 of these identified miRNAs were validated by real-time quantitative fluorescent PCR (q-PCR), including 9 up-regulated miRNAs (miR1320-3p, miR1320-5p, miR156b-3p, miR156c-5p, miR156c-3p/g-3p, miR1561-5p, miR398b and miR6250) and 8 down-regulated miRNAs (miR156a/e/i, miR156k, miR160f-5p, miR166j-5p, miR1846e and miR399d). In addition, 45 novel radiationresponsive miRNAs were predicted, and 8 of them were verified by q-PCR. The target genes of radiation-responsive miRNAs were predicted and gene function enrichment analysis was performed with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). The expression of 9 targets of 4 known miRNA families (miR156, miR399, miR1320 and miR398) and 2 targets of 2 novel miRNAs were quantified by q-PCR, and a strong negative regulation relation between miRNAs and their targets were observed. Those targets including SQUAMOSA promoterbinding-like protein (SPL) genes, copper/zinc superoxide dismutase (Cu/Zn-SOD), copper chaperone for SOD (CCS1) and electron transporter/ heat-shock protein binding protein (HSP), which are involved in growth and defense against various stresses, especially associated with reactive oxygen species (ROS) scavenging. This work provides important information for understanding the ROS generation and elimination mechanisms closely related to miRNAs in rice seedlings after low-energy N radiation exposure.
microRNAs (miRNAs) 是一类非编码调节 RNA，在植物生长发育中发挥重要作用，尤其是在应激反应中。低能离子辐射是一种环境胁迫，可引起多种生物学效应。为了了解低能 N 离子辐射在水稻中的作用，我们对辐射后水稻苗的 shoot 进行了高通量小 RNA 测序，以检测 miRNA 的表达情况。鉴定出 28 个差异表达的已知 miRNA，其中 17 个 miRNA 通过实时定量荧光 PCR (q-PCR) 验证，包括 9 个上调的 miRNA (miR1320-3p、miR1320-5p、miR156b-3p、miR156c-5p、miR156c-3p/g-3p、miR1561-5p、miR398b 和 miR6250) 和 8 个下调的 miRNA (miR156a/e/i、miR156k、miR160f-5p、miR166j-5p、miR1846e 和 miR399d)。此外，还预测了 45 个新的辐射响应 miRNA，其中 8 个通过 q-PCR 验证。对辐射响应 miRNA 的靶基因进行预测，并通过 Gene Ontology (GO) 和 Kyoto Encyclopedia of Genes and Genomes (KEGG) 进行基因功能富集分析。通过 q-PCR 定量检测了 4 个已知 miRNA 家族 (miR156、miR399、miR1320 和 miR398) 的 9 个靶基因和 2 个新 miRNA 的 2 个靶基因的表达，观察到 miRNA 与其靶基因之间存在强烈的负调控关系。这些靶基因包括 SQUAMOSA promoter-binding-like protein (SPL) 基因、铜/锌超氧化物歧化酶 (Cu/Zn-SOD)、铜伴侣蛋白 SOD (CCS1) 和电子转运体/热休克蛋白结合蛋白 (HSP)，它们参与生长和防御各种应激，特别是与活性氧 (ROS) 清除有关。这项工作为了解低能 N 辐射暴露后水稻幼苗中与 miRNA 密切相关的 ROS 生成和消除机制提供了重要信息。
