Identification And Function Analysis Of Long Noncoding RNA During The Development Of Sea Buckthorn Fruit

The eukaryotic genome is extremely complex,not only capable of producing coding m RNA but also a large number of noncoding RNAs,and about 98% of the regions in the eukaryotic genome are non-coding RNA regions.Research on noncoding RNA over the past decade has focused on short noncoding RNAs such as micro RNAs,small interfering RNAs,and Piwiassociated RNAs.With the development of high-throughput sequencing technology,a new class of noncoding RNA long noncoding RNA(lnc RNA)has been gradually discovered.Plant studies have found that lnc RNA can regulate important biological processes such as disease resistance,flowering,yield and fruit development of plants.The developmental process of sea buckthorn fruit is synergistically regulated by a variety of regulatory factors.Through further research on key regulatory genes and functional elements involved in the development of sea buckthorn fruit,it can help researchers better understand the molecular regulation during the development and development of seabuckthorn fruit.Mechanisms to better carry out the breeding and breeding of seabuckthorn.Therefore,in this study,the whole transcriptome was sequenced from three different developmental stages of the hybrid species 'hongji' sea buckthorn planted in Dengkou County,Inner Mongolia,and and the bioinformatics analysis process was established to identify sea buckthorn long noncoding RNA at the genome-wide level.Sequence characteristics,chromosome distribution characteristics and expression characteristics of the long noncoding RNA of sea buckthorn were obtained.Furthermore,differentially expressed genes and noncoding RNAs(miRNAs,lnc RNAs and circ RNAs),especially long noncoding RNAs,in fruits at different developmental stages were screened.Target gene prediction and functional prediction were used to further explore key long noncoding RNAs that play a regulatory role in the development of sea buckthorn fruit.The biological functions of the core-regulated lnc RNA in the development of sea buckthorn fruit were preliminarily verified by molecular biological experiments such as virus-mediated gene silencing.The main conclusions obtained are as follows:(1)A total of 68 miRNAs and 50 novel miRNAs were identified from small RNA sequencing data of sea buckthorn fruit.Furthermore,47 differentially expressed miRNAs were screened in the development sea buckthorn fruits.33 miRNAs were up-regulated,and 14 miRNAs were down-regulated.Bioinformatics enrichment analysis showed that these differentially expressed miRNAs are involved in the biosynthesis of various metabolites.(2)A total of 8907 reliable lnc RNAs,including 6676 linc RNA,1193 intronic lnc RNA and 1038 antisense lnc RNA,were identified from the whole transcriptome sequencing data of sea buckthorn fruit.The identified sea buckthorn lnc RNAs had short transcript length,low exon number and short ORF length,low sequence conservation than m RNA.(3)118 differentially expressed lnc RNAs(DElnc RNAs)were identified from three different developmental stages.The target genes predicted results showed that 84 lnc RNAs targeted 1061 genes by trans,and 67 lnc RNAs targeted 782 genes by cis.Predictive analysis of target gene function indicates that these DElnc RNAs are involved in the anabolic process of metabolites such as flavonoids and carotenoids.(4)22 DElnc RNAs were identified as target genes for 25 DEmiRNAs.Combined with miRNA-m RNA sequence information and targeted regulation,10 DElnc RNAs may regulate miRNA biological functions through endogenous targets mimics(e TM).In this way,a comprehensive regulatory network of lnc RNA-miRNA-m RNA was constructed.(5)We verified that two lnc RNAs(LNC1/LNC2)can regulate anthocyanin biosynthesis in the development fruit of sea buckthorn through the miRNA-lnc RNA-m RNA regulatory network by means of molecular localization analysis such as cell localization analysis and transient silencing,high performance liquid chromatography.In summary,this study is the first compre hensive disclosure of the various sequence characteristics of lnc RNA in sea buckthorn fruit,and identified a series of lnc RNA related to the development of sea buckthorn fruit,in order to further use molecular biology experimental methods to confirm the lnc RNA in sea buckthorn fruit.The specific regulatory functions and mechanisms of action during development provide the basis and data support for the research.At the same time,it promotes the in-depth study and application of transcriptional regulation in forest and fruit development research.