Molecular cloning of full-length Dmrt1 cDNA of Reeves turtle (Chinemys reevesii)

We cloned and sequenced a full-length open reading frame turtle dmrt1 cDNA (Crdmrt1) that was 1,504 bp in length and encoded 371 amino acid residues. RT-PCR analysis in different tissues of adult male turtle showed that the Crdmrt1 cDNA fragment was only detected in the testis. The amino acid sequence derived from Crdmrt1 demonstrated high homology to sequences from dmrt1 of Pelodiscus sinensis (92% identities and 93% positives) and Elaphe quadrivirgata (75% identities and 83% positives). The deduced amino acid from Crdmrt1 contained a conserved DM domain, a male-specific motif, and a P/S-rich region. In DMRT1 from reptiles, birds, mammals, amphibians, and fish, the amino acid identities and positives for DM domains were 85-100% and 88-100%, respectively, those for male-specific motifs were 47-100% and 60-100%, and those for P/S-rich regions were 24-100% and 35-100%. The module consisted of intertwined CCHC and HCCC Zn(2+)-binding sites in the DM domain and was conserved in all 11 species analyzed in this study. Amino acid sequences of Crdmrt1 and previously reported DMRT1s, DMRT2s, and DMRT3s were subjected to phylogenetic analysis. The resulting tree showed that CrDMRT1 belongs to DMRT1, and turtles are a sister group to a cluster of birds and snakes. This is the first study of the cloning of full-length dmrt1 from a reptile.     

Identification of a novel gene (Apq1) from the indica rice cultivar ‘Habataki’ that improves the quality of grains produced under high temperature stress

The appearance of brown rice grown under high temperature conditions is an important characteristic for improvement in Japanese rice breeding programs. We performed a QTL analysis of the appearance quality of brown rice using chromosome segment substitution lines of the indica cultivar ‘Habataki’ in the ‘Koshihikari’ genetic background. A line carrying a ‘Habataki’ segment on chromosome 7 showed a high percentage of perfect grains produced under high temperature conditions during the ripening period. To verify the role of this segment, and to narrow down the region containing the useful allele, substitution mapping was performed using multiple paired lines. As a result, the chromosomal location of a gene that we named Appearance quality of brown rice 1 (Apq1) was delimited to a 48-kb region. In addition, we developed an Apq1-near isogenic line (NIL) to evaluate the effect of Apq1 on various agronomic traits. Under high temperature conditions during the ripening period, the Apq1-NIL produced significantly higher percentages of perfect grains than ‘Koshihikari’. Other agronomic traits, including yield and palatability, were similar between the Apq1-NIL and ‘Koshihikari’. Therefore, the ‘Habataki’ allele of Apq1 will be useful in breeding programs aimed at improving the quality of grains ripened under high temperature conditions.     

Preliminary study of the effects of condensed barley distillers soluble on rumen fermentation and plasma metabolites in Japanese Black cows

In Japan, condensed barley distillers soluble (CBDS) is a widely known liquor byproduct that contains a high level of protein and is used as a supplementary protein feed for cattle. The present study evaluated the effects of CBDS feed on rumen fermentation and plasma metabolites in Japanese Black cows. Applying a replicated 3 × 3 Latin square design, nine cows were offered CBDS and hay (CBDS‐t), soy bean meal and hay (Soybean‐t) and only hay (Hay‐t) over 35 days. We collected ruminal fluid and plasma just before feeding and at 3 h after feeding. The concentrations of propionate and butyrate in the rumen before feeding were lower in the CBDS‐t than in the Soybean‐t group (P < 0.05). However, after 3 h, the concentrations were higher in the CBDS‐t than in the Soybean‐t and Hay‐t groups (P < 0.05). Although, there were no differences in the compositions (% mol) of propionate and butyrate in the rumen and the concentration of plasma β‐hydroxybutyric acid before feeding between treatments, after 3 h they were significantly higher in the CBDS‐t than in the Soybean‐t and Hay‐t groups (P < 0.05). These results indicate that feeding CBDS promotes rumen fermentation and butyrate metabolism.