Genomic characterization of growth differentiation factor 9 (<i>GDF9</i>): mutational and functional impact on reproductive traits of Cholistani Buchi sheep.

Growth Differentiation Factor 9 (GDF9) is a key candidate gene in sheep breeding. Various mutations within GDF9 influence reproductive performance by affecting ovarian function and fertility traits in sheep. As no study has been conducted so far to explore GDF9 mutations in infertile Cholistani Buchi sheep compared with fertile one, so the current study aimed to detect the GDF9 mutations and their possible structural and functional impact using comprehensive molecular and In Silico analyses. Blood samples were collected from 108 Cholistani Buchi sheep, and DNA was extracted. PCR and Sanger Sequencing analysis were performed to detect novel or already reported mutations in Cholistani Buchi sheep. The functional and structural impacts of detected mutations were examined using Dynmut2 and the ProtParam tool. 33 genetic mutations in infertile samples while 15 mutations in fertile samples were identified. Of them, V397I, T451N, G383V, A438V, V428G, I446M, S424R, I405L, C381F, C452R, E407G, T451P, R453S, E432Q, Y442N, D410Y, A388E, and P386T are the destabilizing mutations in infertile samples having negative impact on the encoded proteins. Protein-protein interaction (PPI) and gene ontology (GO) enrichment analysis indicated that the associated partners of GDF9 have diverse biological processes (BP), cellular components (CC), and molecular functions (MF). In conclusion, we have identified 18 novel genetic mutations associated with infertility in Cholistani Buchi sheep. The findings of this study will enhance the understanding of GDF9 mutations and their potential impact on growth and litter size traits in Cholistani Buchi sheep, thus, strengthening the genetic basis for future breeding strategies.