Abstract：Putian black duck (Anas anas domesticus) is the sole native variety in black-feather laying duck, which is rare waterfowl resource in China. The microphthalmia-associated transcription factor (MITF), the most important transcription factor of melanocytes, could be induced by α-MSH (melanocyte-stimulating hormone) and bind to the cAMP responsive element binding protein (CREB) to activate MITF gene expression in melanocytes which plays a key role in the synthesis of melanin. To investigate the relationship between the expression of the MITF gene and regularity of eumelanin deposition in different tissues of Putian black duck, the MITF gene was firstly cloned and analyzed from the skin of Putian black duck with nested PCR and sequenced (GenBank No. MG516571). MITF gene expression was detected in various tissues of Putian black ducks via real-time quantitative PCR, and deposition of eumelanin in the tissues was measured by UV spectrophotometry. The results showed that the length of MITF gene from Putian black duck was 1 341 bp, which contained the complete CDS sequence (12~1 334 bp) and encoded 440 amino acids. The amino acid homology compared with Anas platyrhynchos was 100%, and it was above 95% compared with chicken (Gallus gallus) and quail (Coturnix japonica). The genetic relationship of MITF between Putian black duck and Anas platyrhynchos was the closest, followed with Anser cygnoides, Silky flow, Gallus gallus, and relationship compared with pig (Sus scrofa), sheep (Ovis aries), and other mammals was the farthest. The mRNA expression of MITF gene in Putian black duck skin tissue was the highest in the tissues (skin＞kidney＞gizzard＞liver＞muscle) (P＜0.01). The deposition of eumelanin was basically consistent with the expression of MITF gene in each tissue of Putian black duck. The expression of MITF gene had significant positive correlation with the production of eumelanin in the liver tissues of Putian black duck (P＜0.05). In summary, the expression of MITF gene had tissue specificity in Putian black duck, which might have positive regulation on the deposition of eumelanin and could be used as a candidate gene for the regulation of melanin deposition. The study could provide basic material for the selection of melanin trait in Putian black duck.
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