Identification and characterization of microRNAs from citrus expressed sequence tags
Identification and characterization of microRNAs from citrus expressed sequence tags
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DOI：
10.1007/s11295-010-0319-5
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发表时间：
2011-02
影响因子：
2.4
通讯作者：
Xiao-Meng Wu;Mei-Ya Liu;Qiang Xu;Wenwu Guo
中科院分区：
文献类型：
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作者：
Xiao-Meng Wu;Mei-Ya Liu;Qiang Xu;Wenwu Guo
MicroRNA (miRNA) 是一组一般大小为 21-24 个核苷酸的单链非编码 RNA，它们通过抑制基因翻译或降解目标 mRNA 来负向调节转录后基因表达。尽管 miRNA 在植物中具有重要功能，但人们对柑橘中的 miRNA 知之甚少。在这里，我们通过将已知植物 miRNA 与 560,271 个柑橘 EST 进行比较，提出了柑橘中 microRNA 前体的生物信息学鉴定研究。来自 10 个柑橘品种和 3 个杂交品种的 78 个 EST 被鉴定为推定的 miRNA 前体，编码 51 个独特的 miRNA 序列，代表 28 个 miRNA 家族。对柑橘单基因和 EST 的推定 miRNA 进行 Blastn 搜索，以鉴定目标基因。结果，36 个 unigene 和 77 个 EST 被确定为 25 个柑橘 miRNA 家族的推定靶标。假定的靶标主要是转录因子，在发育、代谢和抗逆性中发挥重要作用。为了验证预测的 miRNA，应用 qRT-PCR 检测柑橘中 9 个假定 miRNA 的组织特异性表达。巴伦西亚。通过 5'RACE 绘制了 3 个 miRNA 家族靶向的 5 个 mRNA 上的切割位点。本研究提供了柑橘 miRNA 前体、成熟 miRNA 和 miRNA 靶标的信息，有助于未来柑橘 miRNA 功能的研究。
MicroRNAs (miRNAs) are a group of single-stranded noncoding RNAs with general size of 21–24 nucleotides, which negatively regulate gene expression posttranscriptionally by repressing gene translation or degrading targeted mRNAs. Despite the important functions of miRNAs in plants, little is known about miRNAs in citrus. Here we present a study of bioinformatics identification of microRNA precursors in citrus by comparing known plant miRNAs against 560,271 citrus ESTs. Seventy-eight ESTs from ten citrus species and three hybrids were identified as putative miRNA precursors, encoding 51 unique miRNA sequences, representing 28 miRNA families. Blastn search of the putative miRNAs against citrus unigenes and ESTs was performed to identify the target genes. As a result, 36 unigenes and 77 ESTs were identified as putative targets of 25 citrus miRNA families. The putative targets are mainly transcription factors and play important roles in development, metabolism, and stress resistance. To validate the predicted miRNAs, qRT-PCR was applied to detect the tissue-specific expression of nine putative miRNAs inCitrus sinensiscv. Valencia. The cleavage sites on five mRNAs targeted by three miRNA families were mapped by 5′RACE. This study provided information on citrus miRNA precursors, mature miRNAs, and miRNA targets and is helpful for future research of miRNA function in citrus.