关键词: 西方蜜蜂, 长链非编码RNA, 反式作用, 内源性竞争RNA, 时空表达谱

Abstract:  【Aim】This study aims to offer a scientific basis for investigating the regulatory function and mechanism of lncRNA13922 in Apis mellifera by determining its expression profiles in different developmental stages and adult tissues of workers and analyzing its manipulation modes and function. 【Methods】The expression levels of lncRNA13922 in different tissues (antennae, venom gland, brain, midgut, fat body, cuticle and hypopharyngeal gland) of adult workers of A. mellifera were validated using RT-PCR. The expression levels of lncRNA13922 in different developmental stages (egg, larva, prepupa, pupa and adult) and tissues (antennae, venom gland, brain, midgut, fat body, cuticle and hypopharyngeal gland) of adult workers of A. mellifera were determined using RT-qPCR. The Pearson correlation analysis was used to predict the co-expressed mRNAs of lncRNA13922 of A. mellifera. The nucleotide sequence of lncRNA13922 was aligned to the miRBase database using miRDeep2 software, and miRNA precursors were predicted through miRPara software. Miranda, RNAhybrid, and TargetScan software was respectively utilized to predict miRNAs targeted by lncRNA13922 and mRNAs targeted by miRNAs, the intersection was adopted as the high-confidence target candidate set, and then the competitive endogenous RNA (ceRNA) regulatory network was constructed based on the targeting relationships. GO and KEGG database functional annotation of target mRNAs was performed by using related software. 【Results】The target fragment with expected size (about 127 bp) of lncRNA13922 was amplified in the antennae, brain, midgut, fat body, hypopharyngeal gland, cuticle and venom gland of adult workers of A. mellifera. lncRNA13922 was differentially expressed in the eggs, 3-day-old larvae, 1- and 2-day-old prepupae and 4-day-old pupae of A. mellifera, and its expression level in the eggs was the highest and significantly higher than those in the 3-day-old larvae and 1- and 2-day-old prepupae. lncRNA13922 was expressed in the 1-, 2-, 6-, 12-, 15- and 17-day-old adults, with differential expression levels, and the expression level of lncRNA13922 in the 2-day-old adult was the highest and significantly higher than those in the 1- and 17-day-old adults. lncRNA13922 was differentially expressed in the above-mentioned seven tissues of adult workers of A. mellifera, and its expression level in the venom gland was the highest and significantly higher than those in the brain, midgut, fat body and hypopharyngeal gland. lncRNA13922 was potentially co-expressed with 29 mRNAs, with their expression levels showing positive correlation relationships, while lncRNA13922 was potentially co-expressed with 12 mRNAs, with their expression levels showing negative correlation relationships. lncRNA13922 was predicted to be the precursors for two miRNAs. lncRNA13922 could target 75 miRNAs, further targeting 1 833 mRNAs. These target mRNAs were involved in 603 GO terms such as cell part and cell, as well as 56 KEGG pathways such as endocytosis and purine metabolism. 【Conclusion】lncRNA13922 is dynamically and differentially expressed in different developmental stages and adult tissues of A. mellifera workers, and potentially participates in the regulation of developmental process through trans-acting, miRNA precursor, and ceRNA network.

Key words: Apis mellifera, long non-coding RNA; trans-action, endogenous competitive RNA, spatiotemporal expression profile