Daweishan Mini chicken is a valuable chicken breed in China. In this study, the complete mitochondrial genome sequence of Daweishan Mini chicken using PCR amplification, sequencing and assembling has been obtained for the first time. The total length of the mitochondrial genome was 16,785 bp, with the base composition of 30.26% A, 23.73% T, 32.51% C, 13.51% G. It contained 37 genes (2 ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA genes) and a major non-coding control region (D-loop region). The protein start codons are ATG, except for COX1 that begins with GTG. The complete mitochondrial genome sequence of Daweishan Mini chicken provides an important data set for further investigation on the phylogenetic relationships within Gallus gallus.
SUMMARYThe molecular mechanism underlying the yellow seed trait has been a subject of quality breeding in Brassica. Thus, uncovering the biosynthetic pathway of proanthocyanidin (PA) accumulation in the Brassica seed coat is a promising research programme. Arabidopsis thaliana BANYULS (BAN) encodes anthocyanidin reductase, which is involved in seed coat pigmentation. In the current study, 2 and 4 BAN homologues were isolated using one pair of primers from Brassica nigra and Brassica juncea, respectively. Reverse transcription polymerase chain reaction (PCR) analysis showed that BAN was expressed abundantly in the seed coat of black seeds and in the embryos of all lines, but not in the seed coat of yellow seeds. Primers incorporating B genome-specific nucleotide variations were designed according to previously published BAN gene sequences of Brassica species to discern the BAN sequence located in B genome origin of Brassica using allele-specific PCR amplification. Proanthocyanidins were also detected by p-dimethylaminocinnamaldehyde staining and a butanol–hydrochloric acid (BuOH–HCl) colorimetric assay in the seed coat of black seeds, but not in the seed coat of yellow seeds. Anthocyanins were not also detected in the seed coat of Brassica species by the BuOH–HCl assay. Both transcriptional and chemical analyses suggested that BAN genes could be involved in both the biosynthesis of PAs and colour formation in the seed coat of Brassica species, whereas no expression of the BAN gene could block biosynthesis of PAs in the yellow seed coat.
Current technologies that are used for genome-wide microRNA (miRNA) prediction are mainly based on BLAST tool. They often produce a large number of false positives. Here, we describe an effective approach for identifying orthologous pre-miRNAs in several primates based on syntenic information. Some of them have been validated by small RNA high throughput sequencing data. This approach uses the synteny information and experimentally validated miRNAs of human, and incorporates currently available algorithms and tools to identify the pre-miRNAs in five other primates. First, we identified 929 potential pre-miRNAs in the marmoset in which miRNAs have not yet been reported. Then, we predicted the miRNAs in other primates, and we successfully re-identified most of the published miRNAs and found 721, 979, 650 and 639 new potential pre-miRNAs in chimpanzee, gorilla, orangutan and rhesus macaque, respectively. Furthermore, the miRNA transcriptome in the four primates have been re-analyzed and some novel predicted miRNAs have been supported by the small RNA sequencing data. Finally, we analyzed the potential functions of those validated miRNAs and explored the regulatory elements and transcription factors of some validated miRNA genes of interest. The results show that our approach can effectively identify novel miRNAs and some miRNAs that supported by small RNA sequencing data maybe play roles in the nervous system.
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