Circular RNA ame_circ_000115 regulates expression of genes in larval gusts of <i>Apis mellifera ligustica</i> stressed by <i>Ascosphaera apis</i>.

Yaping YE; Jie Wang; Jiaxin Zhang; Kaiyao Zhang; Xiaoyu GU; Yutong Yao; Zhongmin Ren; Yang Zhang; Dafu Chen; Rui Guo

Return

Circular RNA ame_circ_000115 regulates expression of genes in larval gusts of Apis mellifera ligustica stressed by Ascosphaera apis.

Author: Yaping YE ¹ ; Jie WANG ¹ ; Jiaxin ZHANG ¹ ; Kaiyao ZHANG ¹ ; Xiaoyu GU ¹ ; Yutong YAO ¹ ; Zhongmin REN ¹ ; Yang ZHANG ¹ ; Dafu CHEN ¹ ; Rui GUO ¹
Author Information

1. College of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China.
Publication Type:Journal Article
Keywords: Apis mellifera ligustica; Ascosphaera apis; Apis mellifera; ame-miR-13b; ame_circ_000115; competing endogenous RNA (ceRNA); immune response
MeSH: Bees/genetics*; Animals; Larva/metabolism*; RNA, Circular/genetics*; RNA, Small Interfering/genetics*; MicroRNAs/genetics*
From: Chinese Journal of Biotechnology 2023;39(1):217-230
CountryChina
Language:Chinese
Abstract: Circular RNAs (circRNAs) are a new class of non-coding RNAs, which have been confirmed to regulate insect gene expression and immune response through multiple manners such as competing endogenous RNA (ceRNA) regulatory network. Currently, function of circRNA in honey bee immune response remains unclear. In this study, PCR and Sanger sequencing were performed to validate the back splicing (BS) site of ame_circ_000115 (in short ac115). RT-qPCR was used to detect the expression profile of ac115 in larval guts of Apis mellifera ligustica stressed by Ascosphaera apis. Dual-luciferase reporter gene assay was conducted to verify the binding relationship between ac115 and ame-miR-13b. Interference of ac115 in larval guts was carried out by feeding specific siRNA, followed by determination of the effect of ac115 interference on expression of six genes relevant to host immune response. The results confirmed the existence of BS site within ac115. Compared with the un-inoculated group, the expression of ac115 in 4-day-old larval gut of the A. apis-inoculated group was up-regulated with extreme significance (P < 0.000 1), while that in 5- and 6-day-old larval guts were significantly up-regulated (P < 0.05). The brightness of specific band for ac115 in 4-, 5- and 6-day-old larval guts of the siRNA-circ_000115-fed group gradually became weak, whereas that of the siRNA-scrambl-fed group was pretty high without obvious variation. Compared with that of the siRNA-scramble-fed group, the expression of ac115 in 4-day-old larval gut of the siRNA-circ_000115-fed group was significantly down-regulated (P < 0.05), whereas that of the 5- and 6-day-old larval guts were down-regulated with extreme significance (P < 0.001). Ame-miR-13b was truly existed and expressed in A. m. ligustica larval guts, and there was true binding relationship between ac115 and ame-miR-13b. Compared with that of the siRNA-scramble-fed group, the expression of antimicrobial peptide genes hymenoptaecin and abaecin in 6-day-old larval gut of the siRNA-circ_000115-fed group was significantly up-regulated (P < 0.05), while that of ecdysone receptor (Ecr) was down-regulated with extreme significance (P < 0.01). These results indicate that ac115 is truly expressed in A. m. ligustica larval guts, BS site truly exists within ac115, and effective interference of ac115 in A. m. ligustica larval guts can be achieved via feeding siRNA. Moreover, ac115 potentially regulates Ecr expression through adsorption of ame-miR-13b and expression of hymenoptaecin and abaecin using a non-ceRNA manner, further participating in host stress-response.