High-Tech Research Center, Shandong Academy of Agricultural Sciences, Key Laboratory of Crop Genetic Improvement and Biotechnology, Huanghuaihai, Ministry of Agriculture, Jinan 250100, PR China.
BMC Plant Biol. 2010 Jan 5;10:3. doi: 10.1186/1471-2229-10-3.
MicroRNAs (miRNAs) are a new class of small, endogenous RNAs that play a regulatory role in the cell by negatively affecting gene expression at the post-transcriptional level. miRNAs have been shown to control numerous genes involved in various biological and metabolic processes. There have been extensive studies on discovering miRNAs and analyzing their functions in model species, such as Arabidopsis and rice. Increasing investigations have been performed on important agricultural crops including soybean, conifers, and Phaselous vulgaris but no studies have been reported on discovering peanut miRNAs using a cloning strategy.
In this study, we employed the next generation high through-put Solexa sequencing technology to clone and identify both conserved and species-specific miRNAs in peanuts. Next generation high through-put Solexa sequencing showed that peanuts have a complex small RNA population and the length of small RNAs varied, 24-nt being the predominant length for a majority of the small RNAs. Combining the deep sequencing and bioinformatics, we discovered 14 novel miRNA families as well as 75 conserved miRNAs in peanuts. All 14 novel peanut miRNAs are considered to be species-specific because no homologs have been found in other plant species except ahy-miRn1, which has a homolog in soybean. qRT-PCR analysis demonstrated that both conserved and peanut-specific miRNAs are expressed in peanuts.
This study led to the discovery of 14 novel and 22 conserved miRNA families from peanut. These results show that regulatory miRNAs exist in agronomically important peanuts and may play an important role in peanut growth, development, and response to environmental stress.
MicroRNAs (miRNAs) 是一类新的内源性小分子 RNA,通过负调控转录后水平的基因表达来发挥调控作用。miRNAs 被证明可以控制许多参与各种生物和代谢过程的基因。在模式物种(如拟南芥和水稻)中已经进行了广泛的研究来发现 miRNAs 并分析它们的功能。越来越多的研究已经在包括大豆、针叶树和菜豆在内的重要农业作物上进行,但尚未有研究报道使用克隆策略在花生中发现 miRNAs。
在本研究中,我们采用下一代高通量 Solexa 测序技术克隆和鉴定花生中的保守和种特异性 miRNAs。下一代高通量 Solexa 测序表明,花生具有复杂的小 RNA 群体,小 RNA 的长度不同,大多数小 RNA 的长度为 24nt。结合深度测序和生物信息学,我们在花生中发现了 14 个新的 miRNA 家族和 75 个保守 miRNA。除了在大豆中有同源物的 ahy-miRn1 外,所有 14 个新的花生 miRNA 都被认为是种特异性的,因为在其他植物物种中没有发现同源物。qRT-PCR 分析表明,保守和花生特异性 miRNAs 都在花生中表达。
本研究从花生中发现了 14 个新的和 22 个保守的 miRNA 家族。这些结果表明,调节性 miRNAs 存在于具有重要农艺学意义的花生中,可能在花生的生长、发育和对环境胁迫的反应中发挥重要作用。