Study on bioinformatics and expression pattern of nce-miR-11248 and target gene SGT1 in Nosema ceranae.

ZHANG Kaiyao; ZHAO Haodong; ZHANG Yiqiong; ZHAO Xiao; GAO Xuze; ZOU Peiyuan; ZHANG Kuihao; CHEN Dafu; GUO Rui; NIU Qingsheng

[Objective] In this study, previously identified nce-miR-11248 in Nosema ceranae was expressed and verified, its target gene was projected and analyzed, and their expression patterns during the Nosema ceranae infection to Apis mellifera ligustica workers were assessed, aiming to provide references or further exploration of the mechanism underlying N. ceranae infection regulated by nce-miR-11248. [Method] Stem-loop RT-PCR and Sanger sequencing were used to prove the authenticity of nce-miR-1248. Related bioinformatic software were employed to predict its target gene and analyze molecular chaacteristics of the encoded protein. MEME and TBtools were utilized to predict conserved motifs and strutural domains in the encode protein. Multiple alignment of amino acid sequences was conducted by Mega 1.0 software, followed by construction of phylogenetic tree with the neighbor-joining method. The midgut issues of A.m. ligustica workers at 1, 2, 4, 6 and 8 d post N. ceranae infestation were collected. RT-qPCR was used to determine expression profiles of nce-miR-11248 and the target gene. [Result] It was proved hat nce-miR-11248 was truly existed in N. ceranae spores, and the sequence length was 25 bp. The target gene of nce-miR-11248 was SGT1. The molecular formula of SGT1 was C765 H1213N195 O241 S4 with molecular weight of approximately 17.10 ku, its lipolysis coefficient, isoelectric point and hydrophilic coefficient were 82.67, 5.50 and -0.709, respectively. There was no classical signal peptide and transmembrane domain in SGT1, and it simultaneously located in nucleus, mitochondrion and peroxisome. Additionally, SGT1 proeins in N. ceranae, Nosema bombycis, Pancytospora, Encephalitozoon intestinalis and Encephalitozoon romaleae contained four conserved motifs of Motif 1, Motif 2, Motif 3 and Motif 4 and one structural domain of SGT1 superfamily. Furthermore, SGT1 proteins in N. ceranae, N. bombycis, E. intestinalis and E. romaleae were grouped into one cladcand N. ceranae SGT1 and N. bombycis SGT1 had the closest evolutionary distance. As compared to 1 day post N. ceranae inoculation, the expression level of nce-miR-11248 was significantly down-regulated at 2, 4, 6 and 8 days post inoculation (P<0.05), the expression level of SGT1 was down-regulated at 2, 4, 6 and 8 days post inoculation, and the differences in expression level at 4, 6 and 8 days post inoculation reached significant levels (P<0.05). [Conclusion] This study showed that nce-miR-11248 was truly existed in N. ceranae spores. N. ceranae SGT1 and N. bombycis SGT1 had the closest genetic distance. Both nce-miR-11248 and target gene SGT1 presented continuous decreases during the N. ceranae infection to A. m. ligustica workers, and nce-miR-11 248 and SGT1 were poential factors involved in modulation of the microsporidian infection process.

GENE expression; NOSEMA ceranae; TRANSMEMBRANE domains; CHEMICAL formulas; AMINO acid sequence

Journal of Northwest A & F University - Natural Science Edition, 2024, Vol 52, Issue 2, p32

1671-9387

Academic Journal

10.13207/j.cnki.jnwafu.2024.02.004