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| Gene Synthesis | Gene expression of phenylalanine ammonialyase [BsPAL (Genbank accession no. KJ930039)], chalcone synthase [BsCHS (Genbank accession no. KJ930037)], flavanone 3-hydroxylas [BsF3H (Genbank accession no. KJ930038)], and anthocyanidin synthase [BsANS (Genbank accession no. KJ930039)] was examined by real-time quantitative RT-qPCR analysis. Total RNA was extracted from mixed leaf samples from three seedlings at each time point from each treatment using the CTAB method according to Liao et al. (2004) First-strand cDNA was synthetized using the Prime ScriptTM RT Reagent Kit with gDNA Eraser (TaKaRa Bio, Dalian, China). The gene-specific primers used for real-time RT-qPCR analysis were designed using Genescript software (GenScript Biotech Corp., Nanjing, China) (Supplemental Table 1). The 18S rRNA gene (Genbank accession no. KJ959633) from B. semperflorens was used as the internal reference gene to normalize any differences in the amount of each cDNA template. The RT-qPCR cycling conditions were as follows: 95 °C for 10 s, followed by 40 cycles of 95 °C for 10 s and 60 °C for 45 s. We employed the 2-△△CT method (Livak and Schmittgen, 2001) to analyze relative gene expression based on three biological replicates. The data and figures were generated using SPSS (version 16.0; IBM Corp., Armonk, NY) and Origin (version 8; OriginLab, Northampton, MA) software. | Get A Quote |
Leaves of Begonia semperflorens accumulate anthocyanins and turn red under low temperature (LT). In the present work, LT increased H2O2 content and superoxide anions production rate, causing significant increases in the activities of enzymes and contents of reduced components involved in the ascorbate-glutathione cycle (AsA-GSH cycle). As a result, LT-exposed seedlings increased the expression of genes involved in anthocyanin biosynthesis, and accumulated anthocyanin. Based on LT condition, application of N,N'-dimethylthiourea (DMTU) decreased reactive oxygen species (ROS) content, and unbalanced the AsA-GSH-controlled redox homeostasis. As a result, seedlings in the LT + DMTU group did not accumulate anthoc... More
