Clostridium butyricum MIYAIRI is effective for both the treatment and the prophylaxis of AAD in children, as it normalizes the intestinal flora disturbed by antibiotics.

Flower color intensity is largely determined by the amount of accumulated anthocyanins. Delphinium flowers show a wide range of colors from pale pink to deep orange to red to dark blue. Here, we demonstrated that the level of anthocyanin accumulation in dark blue, orange and red varieties was higher than in pale blue and pale pink varieties. Since dihydroflavonol 4-reductase (DFR) is a key enzyme in anthocyanin biosynthesis and accumulation in plants, we investigated the relationship between flower color intensity and the level of DFR gene expression. Six delphinium varieties with different flower colors were analyzed. Varieties that accumulated relatively high levels of anthocyanin also had high levels of DFR expression and enzyme activity in crude protein extracts. By contrast, DFR expression and activity was low in varieties with low anthocyanin accumulation. Alignment of DFR amino acid sequences in the six varieties showed the presence of two types, termed DgDFR and DnDFR. Recombinant DgDFR and DnDFR proteins had similar substrate specificities, but the kinetic turnover rate of the DnDFR enzyme was higher than that of DgDFR. We conclude that DFR expression level is closely correlated with flower color intensity and that DFR is an important factor that determines anthocyanin accumulation and delphinium flower color intensity.

Although delphiniums are famed for their blue flowers, a few varieties display reddish flowers, such the pale-pink garden varieties 'Ehime Kou 9 (Kou 9)' and 'F 1 Super Happy Pink (SHP), ' and the orange-red flowered species Delphinium nudicaule (NDC). Blue delphinium flowers contain a delphinidin-derived anthocyanin, whereas the three varieties mentioned above have anthocyanins derived from the aglycone pelargonidin in their sepals. As flavonoid 3′,5′-hydroxylase (F3′5′H) is known to be the key enzyme in biosynthesis of delphinidin, we compared the structure and function of the F3′5′H gene in three varieties of delphiniums with reddish flowers to one that has blue flowers. We found that the F3′5′H gene of 'Kou 9' had a point mutation that generated a stop codon in the first exon. Genomic PCR analysis indicated that the 'SHP' variety lacked F3′5′H. Although the nucleotide sequence of the F3′5′H open reading frame was identical in 'NDC' to that of the wild type, it lacked an intron and no F3′5′H transcripts could be detected in this variety. We conclude that the red flower phenotypes of these delphiniums derive from independent mutations of the F3′5′H gene. This is the first report on the delphinium F3′5′H gene. At a practical level, these mutations should be of value for breeding new pink and red flower varieties.