目的 microRNAs（miRNA）是一類非編碼的單鏈小分子RNA，在植物的生長(zhǎng)發(fā)育、逆境適應(yīng)和代謝調(diào)控等過程中發(fā)揮著至關(guān)重要的作用。本研究擬利用地黃EST數(shù)據(jù)通過生物信息學(xué)手段預(yù)測(cè)新的地黃miRNA，為今后地黃miRNA的生物學(xué)研究奠定基礎(chǔ)。方法 本研究根據(jù)miRNA 家族在不同物種中的保守性，將miRBase數(shù)據(jù)庫(kù)中的已知植物miRNA與通過高通量測(cè)序獲得的93172條地黃EST序列進(jìn)行同源比對(duì)，按照miRNA前體應(yīng)具備的標(biāo)準(zhǔn)進(jìn)行篩選。結(jié)果 預(yù)測(cè)到分屬于8個(gè)家族的8條潛在地黃miRNA序列，并通過實(shí)時(shí)熒光定量PCR對(duì)8條預(yù)測(cè)的miRNA進(jìn)行了檢測(cè)，證實(shí)8條miRNA在地黃中真實(shí)存在。隨后利用軟件對(duì)8條地黃miRNA的靶基因進(jìn)行預(yù)測(cè)，發(fā)現(xiàn)其靶基因主要編碼與地黃生長(zhǎng)發(fā)育、代謝以及脅迫響應(yīng)等過程相關(guān)的蛋白。結(jié)論 新預(yù)測(cè)的地黃miRNA及其靶基因?yàn)榻窈笱芯克鼈冊(cè)诘攸S中的生物學(xué)功能奠定了基礎(chǔ)。

Objective MicroRNAs (miRNA) are a class of small non-coding RNA molecules, playing vital roles in plant growth and development, stress adaption, and metabolic pathway regulation. In order to establish the material basis for future studies on the biological function of miRNAs, we aimed to predict novel miRNAs from EST sequences of Rehmannia glutinosa by using bioinformatic strategies. Methods Since most of the plant miRNAs were conserved in plant species, all plant miRNAs deposited in miRNase were aligned to the 93 172 EST sequences generated by next generation high-throughput RNA sequencing technology and the putative miRNA precursors were screened according to serious criteria. Results Eight novel rehmannia miRNAs were identified that belonged to eight different families, which were further validated by real-time PCR analysis. Then the eight rehmannia miRNAs were subjected to target prediction analysis, and the results showed that the target genes encoded the proteins related to root growth, metabolism, stress responses, and other processes. Conclusion The miRNAs and their target genes identified in this study will provide clues to their biological functions in R. glutinosa.

國(guó)家自然科學(xué)基金資助項(xiàng)目（30901969）