Zhao Wenlong, Xiao Weiyu, Sun Jinliang, Chen Mingxin, Ma Mingqing, Cao Yaqi, Cen Weijian, Li Rongbai, Luo Jijing
College of Life Science and Technology, State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, China.
Agriculture College, State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, China.
Plants (Basel). 2022 Apr 3;11(7):977. doi: 10.3390/plants11070977.
A chromosome single segment substitution line (CSSL) DC90, which was generated by introgressing , a locus derived from common wild rice ( Griff.), into the 9311 ( L. ssp. ) background, exhibits a chilling tolerance phenotype under chilling stress. Here, an integration of microRNA (miRNA) deep sequencing and transcriptomic sequencing analysis was performed to explore the expression profiles of miRNAs and their target genes mediated by under chilling stress, and to reveal the possible regulatory mechanisms of miRNAs that are involved in chilling tolerance. Integration analysis revealed that a number of differentially expressed miRNAs (DEMs) and putative target genes with different expression patterns and levels were identified in 9311 and DC90 under chilling stress. KEGG enrichment analysis revealed that the target genes that are regulated by chilling-induced miRNAs are involved in the regulation of various biological processes/pathways, including protein biosynthesis, redox process, photosynthetic process, and chloroplast development in two genotypes. CRISPR/Cas9 editing of the target genes of the key DEMs in a chilling tolerant rice variety Zhonghua 11 (ZH11) found that (), one of the putative target genes of osa-miR1846a/b-5p and encoding a wax synthesis protein, is correlated with a chilling stress tolerance phenotype, implying osa-miR1846a/b-5p/ plays an important role in -mediated chilling stress tolerance regulatory pathway(s). Therefore, we speculate that the may regulate the key miRNA target genes in response to chilling stress by differential regulation of miRNAs in wild rice, thereby resulting in the variation of chilling tolerance phenotype between 9311 and DC90.
一个染色体单片段代换系（CSSL）DC90，是通过将源自普通野生稻（Griff.）的一个位点渗入到9311（L. ssp.）背景中产生的，在冷胁迫下表现出耐冷表型。在此，进行了微小RNA（miRNA）深度测序和转录组测序分析的整合，以探索冷胁迫下miRNA及其靶基因的表达谱，并揭示参与耐冷性的miRNA的可能调控机制。整合分析表明，在冷胁迫下，在9311和DC90中鉴定出了许多具有不同表达模式和水平的差异表达miRNA（DEM）和推定靶基因。KEGG富集分析表明，受冷诱导的miRNA调控的靶基因参与了各种生物学过程/途径的调控，包括两种基因型中的蛋白质生物合成、氧化还原过程、光合作用过程和叶绿体发育。对耐冷水稻品种中华11（ZH11）中关键DEM的靶基因进行CRISPR/Cas9编辑发现，osa-miR1846a/b-5p的推定靶基因之一（）编码一种蜡合成蛋白，与耐冷胁迫表型相关，这意味着osa-miR1846a/b-5p/在介导的冷胁迫耐受调控途径中起重要作用。因此，我们推测可能通过对野生稻中miRNA的差异调控来调节关键的miRNA靶基因以响应冷胁迫，从而导致9311和DC90之间耐冷表型的差异。
