Key Laboratory of Horticultural Plant Biology (Ministry of Education), College of Horticulture and Forestry Science, Huazhong Agricultural University, Wuhan 430070, China.
Mol Biotechnol. 2012 May;51(1):44-57. doi: 10.1007/s12033-011-9439-x.
To identify novel as well as conserved miRNAs in citrus, deep sequencing of small RNA library combined with microarray was performed in precocious trifoliate orange (an early flowering mutant of trifoliate orange, Poncirus trifoliata L. Raf.), resulting in the obtainment of a total of 114 conserved miRNAs belonging to 38 families and 155 novel miRNAs. The miRNA star sequences of 39 conserved miRNAs and 27 novel miRNAs were also discovered among newly identified miRNAs, providing additional evidence for the existence of miRNAs. Through degradome sequencing, 172 and 149 genes were identified as targets of conserved miRNAs and novel miRNAs, respectively. GO and KEGG annotation revealed that high ranked miRNA-target genes were those implicated in biological and metabolic processes. To characterize those miRNAs expressed at the juvenile and adult development stages of citrus, further analysis on the expression profiles of these miRNAs through hybridizing the commercial microarray and real-time PCR was performed. The results revealed that some miRNAs were down-regulated at adult stage compared with juvenile stage. Detailed comparison of the expression patterns of some miRNAs and corresponding target genes revealed the negative correlation between them, while few of them are positively correlated.
为了鉴定柑橘中新的和保守的 microRNA，在早实枳（枳，芸香科柑橘属）中进行了小 RNA 文库的深度测序，并结合微阵列分析，总共获得了 114 个属于 38 个家族的保守 microRNA 和 155 个新的 microRNA。在新鉴定的 microRNA 中还发现了 39 个保守 microRNA 和 27 个新 microRNA 的 miRNA 星序列，为 microRNA 的存在提供了额外的证据。通过降解组测序，分别鉴定出 172 个和 149 个基因作为保守 microRNA 和新 microRNA 的靶基因。GO 和 KEGG 注释表明，高排名的 miRNA 靶基因与生物和代谢过程有关。为了研究这些在柑橘幼龄和成年发育阶段表达的 miRNA，通过杂交商业微阵列和实时 PCR 进一步分析了这些 miRNA 的表达谱。结果表明，与幼龄期相比，一些 miRNA 在成年期下调。对一些 miRNA 和相应靶基因的表达模式的详细比较表明，它们之间存在负相关，而少数则呈正相关。
