Laboratory for Molecular Plant Biology, VKR Center Proactive Plant, Department of Plant Biology and Biotechnology, Faculty of Life Sciences, University of Copenhagen, DK-1871 Frederiksberg C, Copenhagen, Denmark.
Physiol Plant. 2010 Sep 1;140(1):57-68. doi: 10.1111/j.1399-3054.2010.01384.x. Epub 2010 May 8.
MicroRNAs (miRNAs) are short RNA chains (20-24 bp) which are emerging as important regulators of gene expression. miRNAs are encoded by specific genes, and in Arabidopsis, 190 genes have presently been identified. It has been shown that miR399 is essential for the phosphate starvation response, and recent studies have shown transcriptional changes in a number of additional miRNAs in response to a shortage of phosphate. In this study, global profiles of the miRNA in shoots of Arabidopsis plants grown on limited phosphate or full nutrient in combination with sucrose feed were analysed using the miRCURY LNA microRNA Array system. Furthermore, changes in miRNA transcript were compared between a mutant lacking the transcription factor phosphate starvation responses 1 (PHR1) and wild-type plants. The global analysis identified miRNAs belonging to nine families to respond to P deprivation, sucrose or PHR1. Among these, miR399d, miR827, miR866, miR391 and miR163 were most prominently induced upon P starvation, whereas miR169b/c was strongly induced in previously starved plants when provided with sufficient P and more so when combined with an addition of sucrose. This study shows that array analysis is in general agreement with data obtained by other high-throughput technologies. The array data were confirmed by real-time reverse transcriptase-polymerase chain reaction analyses of selected pri-miRNAs. Our data corroborate the implication that several miRNAs are involved in the P-starvation response and further identify miR866 and miR163 as new candidates of miRNAs associated with the regulation of the P-starvation response.
MicroRNAs (miRNAs) 是短链 RNA（20-24bp），作为基因表达的重要调控因子正在兴起。miRNAs 由特定基因编码，在拟南芥中，目前已经鉴定出 190 个基因。已经证明 miR399 对于磷饥饿响应是必需的，最近的研究表明在响应磷缺乏时，许多其他 miRNAs 的转录发生变化。在这项研究中，使用 miRCURY LNA microRNA Array 系统分析了在有限磷或全营养与蔗糖喂养条件下生长的拟南芥植物的 shoot 中的 miRNA 的全局图谱。此外，还比较了缺乏转录因子磷饥饿响应 1 (PHR1) 的突变体和野生型植物之间 miRNA 转录的变化。全局分析确定了属于九个家族的 miRNA 响应 P 剥夺、蔗糖或 PHR1。在这些 miRNA 中，miR399d、miR827、miR866、miR391 和 miR163 在 P 饥饿时最显著诱导，而 miR169b/c 在先前饥饿的植物中在提供足够的 P 时强烈诱导，当与添加蔗糖结合时更强烈诱导。这项研究表明，阵列分析通常与其他高通量技术获得的数据一致。通过对选定的 pri-miRNAs 的实时逆转录聚合酶链反应分析证实了阵列数据。我们的数据证实了几种 miRNAs 参与 P 饥饿响应的含义，并进一步确定 miR866 和 miR163 作为与 P 饥饿响应调控相关的新 miRNA 候选物。
