Screening and Functional Prediction of Differential miRNAs Associated with Broodiness in Black Muscovy Duck (Cairna moschata)
LI Li1,2, ZHANG Lin-Li2, Nemat O. KEYHANI3, XIN Qing-Wu2, MIAO Zhong-Wei2, ZHU Zhi-Ming2, QIU Jun-Zhi1,*, ZHENG Nen-Zhu2,*
1 College of Life Science, Fujian Agriculture and Forestry University, Fuzhou 363000, China; 2 Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China; 3 Institute of Food and Agricultural Sciences, University of Florida, Gainesville, FL 32611, USA
Abstract The black Muscovy duck (Cairna moschata) is excellent lean meat ducks that is drought-tolerant and rough-fed, which has important value and special status in the poultry industry. However, the black Muscovy duck has strong broodiness and low fecundity, which restricts rapid development of the industry. The ovarian tissues of 3 black Muscovy ducks at the laying and broody stages were collected, respectively. The TRIzol method was used to extract total RNA from the ovarian tissues, the library was constructed and high-throughput sequencing technology was used to analyze the differential expression of miRNA between the 2 groups, then the known, novel miRNAs were predicted for target genes, and the functions of the target genes were enriched and analyzed by GO and KEGG, at last the high-throughput sequencing results were verified by qPCR. The results showed that the raw reads produced by sequencing exceeded 11 099 443 pieces in both groups, and the proportion of clean reads after filtering was all higher than 96.15%, which suggested the data could be used for subsequent analysis. A total of 344 miRNAs were identified, including 275 known miRNAs and 69 newly discovered miRNAs. Sixteen significantly differently expressed miRNAs were screened, including 5 down-regulated and 11 down-regulated in broody ducks with 440 predicted target genes, in which growth hormone secretagogue receptor (GHSR), follistatin (FST) and other target genes were related to reproduction and ovarian development. Thus, it was speculated that the corresponding gga-mir-16c-5p, gga-mir-146a-3p and novel_ 247、novel_ 325 and other miRNAs might be related to broodiness of black Muscovy duck. GO enrichment analysis showed that chromosome organization, nuclear chromosome, spindle microtubule, DNA binding, and nucleotide kinase activity were related to reproduction and development. KEGG pathway annotation placed 234 target genes into 101 signaling pathways, including Wnt and insulin signaling pathways which might be related to germ cell differentiation and development. qPCR confirmed that relative expression levels of up-regulated and down-regulated miRNAs were consistent with the high-throughput sequencing results. This study screened key miRNAs related to broodiness in Muscovy ducks, providing basic material for analyzing broodiness mechanism of Muscovy duck from transcriptional or post transcriptional regulatory level and accelerating the breeding of new high-yield strains.
LI Li,ZHANG Lin-Li,Nemat O. KEYHANI, et al. Screening and Functional Prediction of Differential miRNAs Associated with Broodiness in Black Muscovy Duck (Cairna moschata)[J]. 农业生物技术学报, 2022, 30(4): 726-738.
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