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马铃薯保守 microRNAs 的预测和验证

Prediction and validation of conservative microRNAs of Solanum tuberosum L.

机构信息

Institute of Bioengineering, Zhejiang Sci-Tech University, No. 2 Road, Xiasha, Hangzhou, 310018, People's Republic of China.

出版信息

Mol Biol Rep. 2010 Oct;37(7):3081-7. doi: 10.1007/s11033-009-9881-z. Epub 2009 Oct 31.

Abstract

Potato (Solanum tuberosum) is an important crop around the world, and accounts for a significant amount of the food consumed by humans. However, little information is available about potato miRNAs which play important regulatory roles in plant growth and development. In the present study, computational prediction of potential miRNAs from potato revealed 71 miRNAs belonging to 48 families. Amongst these 71 mRNAs, 65 were predicted for the first time. Most potato miRNA families have one to three members, and sequence analysis showed that the candidate pre-miRNA sequences varied from 48 to 224 bp in length. To verify the predicted miRNAs, specific stem-loop RT primers were designed and real-time PCR assays were used to profile the expression levels of seven miRNAs from different tissues of potato. The results showed that all the selected miRNAs were successfully amplified. Most of them had their highest expression levels in leaves, and the lowest levels in the stem, while miR159 and miR164 presented a different expression pattern. The specific expression levels of each miRNAs in the tested tissues may be related to their particular functions in regulating potato vegetative growth and organ development.

摘要

马铃薯(Solanum tuberosum)是一种在世界各地都很重要的作物,占人类食用食物的很大一部分。然而,关于在植物生长和发育中起重要调节作用的马铃薯 miRNA 的信息却很少。在本研究中,对马铃薯潜在 miRNA 的计算预测揭示了 48 个家族中的 71 个 miRNA。在这 71 个 miRNA 中,有 65 个是首次预测的。大多数马铃薯 miRNA 家族有一到三个成员,序列分析表明候选前体 miRNA 序列的长度从 48 到 224bp 不等。为了验证预测的 miRNA,设计了特定的茎环 RT 引物,并使用实时 PCR 分析来分析马铃薯不同组织中七种 miRNA 的表达水平。结果表明,所有选定的 miRNA 都成功扩增。它们大多数在叶片中的表达水平最高,在茎中的表达水平最低,而 miR159 和 miR164 则表现出不同的表达模式。在测试的组织中每个 miRNA 的特定表达水平可能与其在调节马铃薯营养生长和器官发育中的特定功能有关。

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