外源生长素对月季mirna及其靶基因调控关系的高通量测序分析

Yiqing Wang, Rong Ding, Kezhong Zhang, Jinteng Cui
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摘要

玫瑰(Rosa hybrida L.)是世界上主要的切花之一。研究生长素调控生长的分子机制,对于提高对月季生长发育过程的认识,为月季生产中外源生长素的准确应用提供依据具有重要意义。然而,玫瑰对mirna介导的生长素信号转导的应答机制尚不清楚。本研究对玫瑰植株进行IAA处理,通过小RNA测序鉴定出75个已知mirna和168个新mirna。其中,19个已知mirna和42个差异表达mirna。许多不同的mirna表现出对生长素治疗的分阶段反应。分析了玫瑰中miRNA与生长素调控的关键转录因子的靶向关系,筛选了ARF家族和AUX/IAA家族的靶基因。通过实时荧光定量PCR(qRT-PCR)验证生长素信号转导通路调控miRNA及其靶基因的表达模式,我们发现miR156a、miR160a、miR164a、miR167d、miR396b-3p、novel_miR_189、novel_miR_74、novel_miR_8和novel_miR_207与ARF家族呈抗相关,miR390a-3p和novel_miR_101与AUX/IAA家族呈抗相关。这些结果为进一步研究月季植物生长素调控机制提供了理论基础。
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Analysis of the effect of exogenous auxin on the regulatory relationship between miRNAs and their target genes in rose using high-throughput sequencing
ABSTRACT Rose (Rosa hybrida L.) is one of the major cut flowers in the world. Studying the molecular mechanism of auxin regulation in growth is of great significance for enhancing the understanding of the growth and development processes in rose and informing accurate exogenous auxin application in rose production. However, the response mechanism of rose to miRNA-mediated auxin signal transduction is unclear. In this study, rose plants were treated with IAA, and 75 known miRNAs and 168 novel miRNAs were identified by small RNA sequencing. Among them, 19 known miRNAs and 42 miRNAs were differentially expressed. Many differential miRNAs demonstrated staged responses to auxin treatment. The targeted relationship between miRNA and key transcription factors regulated by auxin in rose was analysed, and the target genes in the ARF family and AUX/IAA family were screened. By using quantitative real-time PCR(qRT-PCR) to verify the expression patterns of the miRNA regulating the auxin signal transduction pathway and its target gene, we found that miR156a, miR160a, miR164a, miR167d, miR396b-3p, novel_miR_189, novel_miR_74, novel_miR_8, and novel_miR_207 were anti-correlated with the ARF family, and miR390a-3p and novel_miR_101 were anti-correlated with the AUX/IAA family. These results provide a theoretical basis for further studies on the auxin regulatory mechanisms in rose.
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