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Abstract
The Chinese swamp buffalo (Bubalis bubalis) is vital to the lives of small farmers and has tremendous economic importance. However, a lack of genomic information has hampered research on augmenting marker assisted breeding programs in this species. Thus, a high-throughput transcriptomic sequencing of B. bubalis was conducted to generate transcriptomic sequence dataset for gene discovery and molecular marker development. Illumina paired-end sequencing generated a total of 54,109,173 raw reads. After trimming, de novo assembly was performed, which yielded 86,017 unigenes, with an average length of 972.41 bp, an N50 of 1,505 bp, and an average GC content of 49.92%. A total of 62,337 unigenes were successfully annotated. Among the annotated unigenes, 27,025 (43.35%) and 23,232 (37.27%) unigenes showed significant similarity to known proteins in NCBI non-redundant protein and Swiss-Prot databases (E-value < 1.0E-5), respectively. Of these annotated unigenes, 14,439 and 15,813 unigenes were assigned to the Gene Ontology (GO) categories and EuKaryotic Ortholog Group (KOG) cluster, respectively. In addition, a total of 14,167 unigenes were assigned to 331 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Furthermore, 17,401 simple sequence repeats (SSRs) were identified as potential molecular markers. One hundred and fifteen primer pairs were randomly selected for amplification to detect polymorphisms. The results revealed that 110 primer pairs (95.65%) yielded PCR amplicons and 69 primer pairs (60.00%) presented polymorphisms in 35 individual buffaloes. A phylogenetic analysis showed that the five swamp buffalo populations were clustered together, whereas two river buffalo breeds clustered separately. In the present study, the Illumina RNA-seq technology was utilized to perform transcriptome analysis and SSR marker discovery in the swamp buffalo without using a reference genome. Our findings will enrich the current SSR markers resources and help spearhead molecular genetic research studies on the swamp buffalo.
Highlights
The water buffalo (Bubalus bubalis), which belongs to the Bubalus genus of the Bovidae family, is an economically significant livestock that has been used as dairy, meat, and source of draught power [1]
We obtained a total of 54,109,173 raw reads, which after removal of redundant reads, trimming of adaptors and filtering for low-quality sequences resulted in 52,979,055 high-quality clean reads with 10,595,811,000 bp of sequence data (Table 1)
The results of FastQC v0.11.3 analysis showed that the Q20 percentage and GC percentage were 97.91% and
Summary
The water buffalo (Bubalus bubalis), which belongs to the Bubalus genus of the Bovidae family, is an economically significant livestock that has been used as dairy, meat, and source of draught power [1]. These animals are typically found in tropical and subtropical regions, wet grasslands, marshes and swamps. The milk yield of crossbreeds Nili-Ravi F1 and F2 reached 2,041.2 kg and 2,351.3 kg respectively, which were 86.8% and 115.2% higher than that of selected buffaloes (P < 0.01) [5]. One of the main long-term hindrances in the buffalo industry in China is the lack of breeds with high milk and reproductive performance
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