Project description:Whole genome sequencing of Red Chittagong Cattle (RCC) cattle and insight into candidate gene mutations associated with disease resistance
Project description:Malnad Gidda is one among the 43 registered cattle breeds of India with unique traits and spread over Western Ghats and coastal regions of Karnataka state in India. Selection of highly elite fertile bulls for the breeding purpose is a critical control point in animal breeding programmes. Therefore, to characterize the semen proteome and to understand the semen biology of this breed, a comprehensive proteomic analysis of spermatozoa and seminal plasma has been carried out by employing SCX and bRPLC fractionation strategies in a mass-spectrometry platform. The semen samples from three Malnad Gidda bulls maintained at Southern Regional Station of ICAR-NDRI under standard managemental conditions were used in the study. The proteomic characterisation of semen from Malnad Gidda breed resulted in the identification of 5, 84,520 PSMs, from 24,467 peptides from 2,815 proteins in spermatozoa and identification of 2, 77,583 PSMs from 12,047 peptides, which resulted in 1,974 proteins from seminal plasma. Out of 2,815 proteins in spermatozoa and 1,974 proteins from seminal plasma, 969 proteins were common to both seminal plasma and spermatozoa. The biological processes and cellular localization of spermatozoa proteins were studied using DAVID tool and were further enriched the identical GO terms using REVIGO online tool. The functional enrichment analysis of identified proteins indicated their roles in the biological processes like sperm motility, spermatid development, spermatogenesis, and so on. GO studies showed the commonalities and differences in the molecular functions of the proteins exclusively identified in spermatozoa, seminal plasma and common proteins. This is the first proteomic investigation conducted on the semen samples of an Indian indigenous breed; therefore, we believe that our preliminary data should significantly advance our understanding of semen proteome of Indian cattle.
Project description:Background: The Malnad Gidda are unique dwarf Bos indicus cattle native to heavy rainfall Malnad and coastal areas of Karnataka in India. These cattle are highly adapted to harsh climatic conditions and are more resistant to Foot and Mouth disease as compared to other breeds of B.indicus. Since the first genome reference became available from B.taurus Hereford breed, only a few other breeds have been genotyped using high-throughput platforms. Also despite the known reports on high diversity within indicine breeds as compared to taurine breeds, only one draft genome of Nellore and horn transcriptome of Kankrej breed were sequenced at base level resolution. Because of the special characteristics Malnad Gidda possess, it becomes the choice of breed among many indicine cows to study at molecular level and genotyping. Results: Sequencing mRNA from the PBMCs isolated from blood of one selected Malnad Gidda bull resulted in generation of 55 million paired-end reads of 100bp length. Raw sequencing data is processed to trim the adaptor and low quality bases, and are aligned against the whole genome and transcript assemblies of Bos taurus UMD 3.1 and Bos indicus (Nellore breed) respectively. About 72% of the sequenced reads from our study could be mapped against the B.taurus genome where as only 41% of reads could be mapped against the Bos indicus transcript assembly. Transcript assembly from the alignment carried out against the annotated B.taurus UMD 3.1 genome resulted in identification of ~10,000 genes with significant expression (FPKM>1). In a similar analysis against the B.indicus Kankrej assembled transcripts we could identify only ~6,000 transcripts. From the variant analysis of the sequencing data we found ~10,000 SNPs in coding regions among which ~9,000 are novel and ~6,400 are amino acid changing. Conclusions: For the first time we have genotyped and explored the transcriptome of B.indicus Malnad Gidda breed. A comparative analysis of mapping the RNA-Seq data against the available reference genome and transcript sequences is demonstrated. An enhanced utility of transcript sequencing could be achieved by improving or completing the sequence assembly of any B.indicus breed to better characterize the indicine breeds for productivity features and selective breeding.
Project description:Profiling of miRNA in serum of FMDV infected cattle, to understand disease pathogenesis and to identify FMD specific miRNA biomarkers to aid early pre-clinical diagnosis