目的 microRNAs（miRNA）是一類非編碼的單鏈小分子RNA，在植物的生長發(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)通過生物信息學手段預(yù)測新的地黃miRNA，為今后地黃miRNA的生物學研究奠定基礎(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)庫中的已知植物miRNA與通過高通量測序獲得的93172條地黃EST序列進行同源比對，按照miRNA前體應(yīng)具備的標準進行篩選。結(jié)果 預(yù)測到分屬于8個家族的8條潛在地黃miRNA序列，并通過實時熒光定量PCR對8條預(yù)測的miRNA進行了檢測，證實8條miRNA在地黃中真實存在。隨后利用軟件對8條地黃miRNA的靶基因進行預(yù)測，發(fā)現(xiàn)其靶基因主要編碼與地黃生長發(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)測的地黃miRNA及其靶基因為今后研究它們在地黃中的生物學功能奠定了基礎(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.

國家自然科學基金資助項目（30901969）