水稻育性转换相关lncRNA(MSTRG.100908.3)及其介导的ceRNA调控网络解析OA

Long non-coding RNAs(lncRNAs)are a class of regulatory non-coding RNAs with a length of more than 200 nt.They mainly play the role of competing endogenous RNAs(ceRNAs)by binding to specific microRNAs(miRNAs),thereby indirectly regulating the expression of target genes.In the whole transcriptome sequencing for the young panicles of the fertile line and the sterile line of the photo-thermo-sensitive genic male sterile(PTGMS)line Wuxiang S(WXS)in the critical period of fertility transition,a lncRNA(MSTRG.100908.3)with significantly high expression in the fertile line had been screened out and the ceRNA regulatory network of lncRNA(MSTRG.100908.3)-miRNA(miR160a)-messenger RNA(mRNA)(ARF10/ARF22)interaction was predicted.In order to further reveal the regulatory role of lncRNA(MSTRG.100908.3)in the fertility transition of WXS,an expression analysis was performed.The results showed that,during the three different stages of the development of young panicles in the WXS fertile line,the expression level of MSTRG.100908.3 was positively correlated with those of ARF10(LOC_Os04g43910)and ARF22(LOC_Os10g33940),both of which were negatively correlated with the expression level of miR160a.Based on the obtained core sequence,the full-length sequence of MSTRG.100908.3 was successfully obtained by using the technology of rapid amplification of cDNA ends(RACE)through 5'RACE and 3'RACE.The bioinformatics analysis showed that MSTRG.100908.3 conformed to the characteristics of lncRNA,and it was hypothesized that the full-length sequence belonged to lncRNA.Then,the core sequence of MSTRG.100908.3 and the precursor sequence of miR160a were constructed into the overexpression vector pCXUN,and the coding sequences(CDS)of ARF10 and ARF22 were constructed into the luciferase reporter vector pGreenⅡ 0800-LUC.Then,for the purpose of verification,the two recombinant vectors were co-transformed into the protoplasts of WXS.The results showed that miR160a down-regulated the expression of ARF10 and ARF22.MSTRG.100908.3 acted as a ceRNA to bind to miR160a,thereby indirectly up-regulating the expression of ARF10 and ARF22.