Cloning and Expression Analysis of Light Signal Transduction Regulatory Factor LhSPA1 Gene in Lily
HUANG Li1, YANG Jie2, LYU Bo2, SONG Ju-Rong2, CHEN Feng2, GUO Cong2, YANG Yuan-Yuan2, XIANG Fa-Yun2,*, XU Ben-Bo1,*
1 Engineering Research Center of Ecology and Agricultural Use of Wetland, Ministry of Education, Yangtze University, Jingzhou 434025, China; 2 Industrial Crops Institute, Hubei Academy of Agricultural Sciences/Hubei Research Center of Flower, Wuhan 430064, China
Abstract：Anthocyanin is one of the key substances determining the color of lily (Lilium brownii var. viridulum) flowers, and its biosynthesis is susceptible to various environmental factors. SPA1 (suppressor of phyA-105) transcription factor plays a crucial role in regulating plant light signal transduction. To further understand the role of LhSPA1 in the regulation of anthocyanin induction in oriental hybrid lily (Lilium oriental hybrid), this study utilized 'Sunny Martinique' cultivar as material. The differences of anthocyanin content in lily buds treated with different light treatments were compared, and ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to detect the perianth slices of flower buds in the complete color development period. Combined with the previous transcriptome data, LhSPA1 was cloned and then subjected to bioinformatics analysis; and the expression levels of genes related to anthocyanin biosynthesis under different light conditions were analyzed using qRT-PCR. The results showed that the main color components in the tepals of 'Sunny Martinique' were cyanidin and pelargonidin. Shading treatment reduced the accumulation of anthocyanin in lily buds. The results of gene expression analysis showed that the expression of anthocyanin biosynthesis-related genes was down-regulated under shading treatment, but the expression of LhSPA1 was significantly higher than that under normal light conditions. The coding sequence length of LhSPA1 (GenBank No. PQ499086) was 2 931 bp, encoding 976 amino acids. Phylogenetic tree analysis showed that LhSPA1 protein was closely related to ZmSPA1 protein in monocotyledonous plant. Sequence analysis revealed that LhSPA1 protein shared conserved domains with SPA1 protein from other species. Subcellular localization results revealed that the fluorescence signal of LhSPA1 protein was located in the nucleus of Nicotiana tabacum. Transient overexpression of LhSPA1 and constitutive photomorphogenic 1 (LhCOP1) in lily petals resulted in light lily flower color. In conclusion, it was speculated that LhSPA1 played an important role in light-induced anthocyanin biosynthesis in lily. This study lays a foundation for analyzing the molecular mechanism of light signal regulating lily anthocyanin biosynthesis pathway.
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