Functional Identification and Analysis of Ph4CL13 Gene in Petunia(Petunia hybrida)
MIAO Yun-Feng*, HU Xu-Hao*, LIN Bin-Ru, ZHONG Shi-Wei**
School of Landscape Architecture, Zhejiang Provincial Key Laboratory of Germplasm Innovation and Utilization for Garden Plants, Zhejiang A & F University, Hangzhou 311300, China
Abstract：The main pigment responsible for the coloration of plant petals is anthocyanin. Based on previous transcriptome data from petunias, the highly expressed 4-coumarate CoA ligase 13 (Ph4CL13) gene was identified during the bud coloring stage, suggesting its involvement in anthocyanin synthesis. Anthocyanin primarily determines petal color in plants. To investigate the role of Ph4CL13 in anthocyanin synthesis, this study used Petunia hybrida 'Ultra' as the material. The 1 653 bp cDNA fragment of Ph4CL13 (Peaxi162Scf00089g00045.1) was isolated from the petunia genome. Phylogenetic analysis revealed a close relationship between Ph4CL13 and Fa4CL-1 and Fa4CL-2. qPCR analysis revealed that Ph4CL13 was highly expressed during the coloring stages, suggested its involvement in anthocyanin synthesis and impact on petunia flower coloration. Silencing Ph4CL13 via virus-induced gene silencing (VIGS), the results showed that the corollas of petunia turned white, and the level of anthocyanin significantly decreased in Ph4CL13-silenced plant compared to control. qPCR analyses demonstrated that Ph4CL13 silencing transcriptionally repressed several anthocyanin biosynthesis genes, including chalconesynthase (CHS), chalcone isomerase (CHI), flavanone-3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), flavonoid synthase (FLS), suggested that Ph4CL13 played a critical role in the anthocyanin synthesis stage of flower coloring process, that might be a potential complex feedback mechanism among these genes related to anthocyanin synthesis. This study preliminarily revealed the important role of Ph4CL13 in anthocyanin synthesis, providing a reference for further exploring its molecular mechanisms in flower color development.
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