Analysis On The Differences Of MRNA And LncRNA In Adult Testis Tissues Among Three Cattle Breeds

Yellow cattle are the major contribution to the beef cattle industry in my country.The breeding of bulls can speed up the process of improving the genetic performance of beef cattle.The development of testicular tissue is the basis for the excellent reproductive performance of bulls,as well as the prerequisite for testicular development and sperm production.Testicular development and sperm production depend on the precise regulation of transcription and post-transcriptional levels of related genes,and long non-coding RNA(lncRNA)has been proven to play a very important role in the regulation of cell differentiation,organ development,transcription and epigenetic genes.There are three paternal origins Y1,Y2,and Y3 for cattle in the world.The Y1 and Y2 haplotype groups are of Bos taurus populations origin,and the Y3 haplotype group is of Bos indicus populations origin.Chinese cattle mainly have Y2 and Y3 haplotypes.In order to explore the transcriptome characteristics of lncRNA and mRNA in the testicular tissue of Chinese yellow cattle(Bos taurus and Bos indicus)after sexual maturity,this study selected Hainan cattle(Y3 haplotype group)and Mongolian cattle(Y2 haplotype group)as representatives of Chinese taurus and indicus,respectively,with Black Angus cattle(Y1haplotype group)as the representative of European taurus cattle.Each breed were selected 3testicular tissue samples after the sexual maturity,total constructed 9 specific libraries by using the technology of RNA-seq,then screened its mRNAs and candidate lncRNAs through bioinformatics analysis,analyzed coding protein genes and the target genes of idifferential lncRNAs,and performed annotation,functional enrichment analysis and RT-q PCR verification.The main results are as follows:(1)Analysis of differentially expressed mRNA in adult testis tissues among three cattle breedsThere are 1468 differentially expressed genes between Hainan cattle and Mongolian cattle.Among them,AR,ZFP42,BCL2,LRP2,1EIF2S2 were related to gonadal development,and GSTA3 and HPGD were related to prostaglandin metabolism.In Hainan cattle and Black Angus cattle populations,there were 2353 differentially expressed genes,PATZ1,HSPA2,AXL1,SPATA9,CYLC1 were enriched in the spermatogenesis process.MAMLD1,LRP2,GATA6,NR5A1,OSR1 were related to male gonadal development,and CNTFR,DMRT1,DMRT3 were related to sex differentiation.There are 550 differentially expressed genes between Mongolian cattle and Black Angus cattle breeds,BMP7 and RXRA were associated with prostaglandin morphology.Analyzing the unique gene functions of each of the three breeds,the specific genes of Black Angus cattle ware enriched in African trypanosomiasis,malaria,amoebiasis and other autoimmune functions,while the specific genes of Mongolian cattle were enriched in Melanin production and partly related to immune response,Hainan cattle-specific genes ware more biased towards the regulation of hormones in animals,the formation and development of arteries,and the regulation of functions related to male reproduction.22 differentially expressed genes were randomly selected for RT-q PCR verification,and the results of quantitative analysis were basically consistent with the results of sequencing analysis.(2)Analysis of the differentially expressed lncRNA in adult testis tissues among three cattle breedsThere are 3967 lncRNAs between Hainan cattle and Mongolian cattle breeds,of which406 were differentially expressed lncRNAs.The target genes DMRT1,FSHR and NUP210 L were related to the development of Sertoli cells.There are 4635 lncRNAs between Hainan cattle and Black Angus cattle breeds,of which 541 were differential lncRNAs.The target genes DMRT1,FSHR,NANOS2 and DMRT3 were related to the development testicular sertoli and sex differentiation.There are 5584 lncRNAs between Mongolian and Black Angus breeds,of which 116 differentially express lncRNAs.TTLL5 and SPEF2 were related to the process of sperm cell metamorphosis.Randomly select 22 differentially expressed lncRNAs for RT-q PCR verification,and found that the quantitative analysis results are consistent with the sequencing analysis results.In summary,this study conducted RNA-seq analysis on the testis tissues of three cattle breeds after sexual maturation,and found that a large number of differentially expressed genes and lncRNAs in the testis tissues of the three cattle breeds after sexual maturity were related to testicular development and sperm production.It provides a theoretical basis for the selection of excellent breeding bulls using molecular-assisted breeding technology.