Polymorphism Analysis and Functional Study of Anthocyanidin Synthase Gene (LiANS) Among Different Flower-color Varieties of Lagerstroemia indica
LU Di1, YUAN Tian-Yi1, HOU Xiao-Yu1, YU Can-Nan1, ZHAO Kai-Zuo1, LIN Jing-Jing1, GUO Zheng-Hao1, ZHANG Jian1,2, YU Chun-Mei1,2,*
1 School of Life Science, Nantong University, Nantong 226019, China; 2 Nantong Key Laboratory of Landscape Plant Genetics and Breeding, Nantong 226019, China
Abstract：Anthocyanidin synthase (ANS) catalyzes the conversion of colorless proanthocyanidins into colored anthocyanidins. This study used 4 crape myrtle (Lagerstroemia indica) cultivars with white, pink, purple, and red petals to investigate the characteristics of the LiANS gene. Through multiple sequence alignment, phylogenetic tree construction, homology modeling, detection of the activity of prokaryotically expressed fusion protein LiANS-His6, qPCR, and promoter cloning, the characteristics of LiANS in different cultivars were analyzed. The results showed that the ORF of the LiANS gene was 1 068 bp, encoding 355 amino acids. According to the differences among LiANS, it was found that there were 4 LiANS allelic variants in 4 cultivars. Homology modeling and enzyme activity assays of the 3 purified LiANS-His6 variants showed that these variations had no effect on enzyme activity. qPCR analysis revealed that the LiANS gene was expressed constitutively in leaves from upper, middle, and lower part of shoots, and the flower at 5 developmental stages, but the expression patterns were different. The overall anthocyanin content peaked at the S3 stage of petal development. Among the samples, the red-flowered cultivar 'Dynamite' maintained relatively high expression levels during the full bloom and senescence stages (S3 and S4). The expression levels of the LiANS gene were not always positively correlated among the 4 varieties. Promoter sequence analysis indicated that 3 allelic variants of the promoter existed among the 4 cultivars, featuring different G-box elements and MYB binding sites. Notably, the white-flowered cultivar 'Pure white' exhibited a 195 bp deletion in the proANS region. This study provides genetic resources for molecular breeding of crape myrtle cultivars with different flower colors and lays a foundation for further elucidation of its regulatory mechanisms.
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