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Expression and tissue-specific localization of nitrate-responsive miRNAs in roots of maize seedlings

机译：硝酸盐反应性miRNA在玉米幼苗根系中的表达及组织特异性定位

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摘要

Nitrogen availability seriously affects crop productivity and environment. The knowledge of post-transcriptional regulation of plant response to nutrients is important to improve nitrogen use efficiency of crop. This research was aimed at understanding the role of miRNAs in the molecular control of plant response to nitrate. The expression profiles of six mature miRNAs were deeply studied by quantitative real time polymerase chain reaction and in situ hybridization (ISH). To this aim, a novel optimized protocol was set up for the use of digoxygenin-labelled Zip Nucleic Acid-modified oligonucleotides as probes for ISH. Significant differences in miRNAs' transcripts accumulation were evidenced between nitrate-supplied and nitrate-depleted roots. Real-time PCR analyses and in situ detection of miRNA confirmed the array data and allowed us to evidence distinct miRNAs spatio-temporal expression patterns in maize roots. Our results suggest that a prolonged nitrate depletion may induce post-transcriptionally the expression of target genes by repressing the transcription of specific miRNAs. In particular, the repression of the transcription of miR528a/b, miR528a*/b*, miR169i/j/k, miR169i*/j*/k*, miR166j/k and miR408/b upon nitrate shortage could represent a crucial step integrating nitrate signals into developmental changes in maize roots.
机译：氮的供应严重影响作物的生产力和环境。转录后调控植物对养分的响应的知识对于提高作物的氮利用效率非常重要。这项研究旨在了解miRNA在植物对硝酸盐反应的分子控制中的作用。通过定量实时聚合酶链反应和原位杂交（ISH）深入研究了六个成熟miRNA的表达谱。为此，建立了一种新的优化方案，以使用地高辛配基标记的Zip核酸修饰的寡核苷酸作为ISH探针。硝酸根补充和硝酸根耗尽的根之间在miRNA转录物积累上存在显着差异。实时PCR分析和miRNA的原位检测证实了阵列数据，使我们能够证明玉米根中不同的miRNA时空表达模式。我们的结果表明，延长的硝酸盐消耗可能通过抑制特定miRNA的转录而在转录后诱导靶基因的表达。特别是在硝酸盐缺乏时对miR528a / b，miR528a * / b *，miR169i / j / k，miR169i * / j * / k *，miR166j / k / n和miR408 / b转录的抑制可能是至关重要的将硝酸盐信号整合到玉米根系发育变化中的步骤。

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