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小 RNA 谱分析鉴定参与菝葜皂苷生物合成调控的 miRNA。

Small RNA profiling for identification of miRNAs involved in regulation of saponins biosynthesis in Chlorophytum borivilianum.

机构信息

Department of Biotechnology, Panjab University, BMS Block-I, Sector 25, Chandigarh, 160014, India.

出版信息

BMC Plant Biol. 2017 Dec 28;17(1):265. doi: 10.1186/s12870-017-1214-0.

Abstract

BACKGROUND

MicroRNAs act as molecular regulator of cell signaling, plant growth and development, and regulate various primary and secondary plant metabolic processes. In the present study, deep sequencing of small RNAs was carried out to identify known and novel miRNAs from pharmaceutically important plant, Chlorophytum borivilianum.

RESULTS

Total 442 known miRNAs and 5 novel miRNAs were identified from young leaf small RNA library. Experimental validation with stem loop RT-PCR confirmed the in silico identification. Based on transcriptome data of root and leaf of C. borivilianum, Oryza sativa, and Arabidopsis thaliana target gene prediction was done using psRNAtarget and mirRanda. BLAST2GO helped in localization of predicted targets and KEGG (Kyoto Encyclopedia for Genes and Genomes) pathway analysis concluded that miR9662, miR894, miR172, and miR166 might be involved in regulating saponin biosynthetic pathway. The correlation between miRNA and its target gene was further validated by RT-qPCR analysis.

CONCLUSION

This study provides first elaborated glimpse of miRNA pool of C. borivilianum, which can help to understand the miRNA dependent regulation of saponin biosynthesis and to design further metabolic engineering experiment to enhance their contents in the plant.

摘要

背景

MicroRNAs 作为细胞信号、植物生长发育的分子调节剂,调节各种初级和次级植物代谢过程。本研究通过对药用植物吊兰的小 RNA 进行深度测序,从其中鉴定出已知和新的 miRNAs。

结果

从小叶小 RNA 文库中鉴定出 442 个已知 miRNAs 和 5 个新 miRNAs。通过茎环 RT-PCR 的实验验证,确认了这些 miRNAs 的存在。基于吊兰、水稻和拟南芥的根和叶的转录组数据,使用 psRNAtarget 和 mirRanda 对预测的靶基因进行预测。BLAST2GO 帮助预测靶基因的定位,KEGG(京都基因与基因组百科全书)通路分析表明,miR9662、miR894、miR172 和 miR166 可能参与调节皂苷生物合成途径。通过 RT-qPCR 分析进一步验证了 miRNA 与其靶基因之间的相关性。

结论

本研究首次详细描述了吊兰的 miRNA 组,这有助于理解 miRNA 对皂苷生物合成的调控,并设计进一步的代谢工程实验来提高植物中皂苷的含量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a834/5745966/74486c6cbc3c/12870_2017_1214_Fig1_HTML.jpg

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