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Cloning and Expression Analysis of Anthocyanin Synthetase Gene ApANS in Acer palmatum
Zhong Huaiqin1,2,3, Chen Yude4, Lin Rongyan1,2,3, Luo Yuanhua1,2,3, Lin Bing1,2,3, Chen Shilin3,5*
1Institute of Crop Sciences, Fujian Academy of Agricultural Science, Fuzhou 350013, China; 2Flowers Research Center, Fujian Academy of Agricultural Science, Fuzhou 350013, China; 3Fujian Engineering Research Center for Characteristic Floriculture, Fuzhou 350013 China; 4Fujian Academy of Agricultural Science, Fuzhou 350003, China; 5Biotechnology Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China
Abstract: Anthocyanin synthetase (ANS) is one of the key enzymes， involved in the end of biosynthesis pathway that converts lcucoanthocyanidin into colored anthocyanidin in plants. In this study， the full-length cDNA sequence of ANS gene was obtained from crimson leaves of Acer palmatum ‘De-shojo’ by using RACE techniques and designated as ApANS. The full-length of ApANS was 1 371 bp， and it included a complete open reading frame of 1 083 bp， which encoded a 360-amino-acid protein. The protein was belonging to the 2-oxoglutarate dependent dioxygenase superfamily， which possessed a typical conserved structural domain of the 2-oxoglutarate and Fe2+-dependent oxygenase， containing 2-oxoglutarate and iron ion combination sites. Sequence alignment and phylogenetic tree analyses showed that ApANS was the most closely related to Dimocarpus longan which belonged to the same sapindales and shared up to 90% homology. The fluorescent quantitative PCR analysis indicated that ApANS was high expression in red leaves， trace expression in yellow and green leaves with slightly red and no expression in green leaves. The transcript level was highest in germinal-leaf stage， unfold-leaf stage and colorationleaf stage. Then with turning green in leaves， the transcript level sharply declined. It’s description that ApANS plays an important role in anthocyanin metabolism and the formation of color in leaves of Acer palmatum.
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