Peucedanum praeruptorum, Dunn, Dunn

Sui, Ziwei, Luo, Jun, Yao, Ruolan, Huang, Chuanlong, Zhao, Yucheng & Kong, Lingyi, 2019, Functional characterization and correlation analysis of phenylalanine ammonia-lyase (PAL) in coumarin biosynthesis from Peucedanum praeruptorum Dunn, Phytochemistry 158, pp. 35-45 : 36

2.1. Identification of the cDNA encoding PAL in P. praeruptorum

Using PAL sequences reported previously and our transcriptome database of P. praeruptorum (accession no. SRX997427) ( Zhao et al., 2015), a BLAST search was conducted. According to the E -value of alignments, only one PAL was identified and cloned. This cDNA, which encodes a 719 amino acid residue protein (Table S1), was designated PpPAL. According to bioinformatics, PpPAL is most closely related to PAL of Angelica sinensis (AsPAL) , which also belongs to Apiaceae ( Fig. S2 ). The amino acid sequences of PpPAL also contain a highly conserved MIO group, which is formed by Ala-Ser-Gly residues ( Fig. S3 ). The high sequence similarity revealed that they might display the same enzymatic functions such as substrate specificity, optimal pH and catalytic mechanism.

Figures

Fig. 2. The content changes of Praeruptorin A, Praeruptorin B and Imperatorin. A. The content of three coumarin compounds in different tissues (Root, Stem and Leaf). B. The content changes of three coumarin compounds under MeJA elicitation. C. The content changes of three coumarin compounds under H2O2 treatment. D. The content changes of three coumarin compounds under UV elicitation. E. The content changes of three coumarin compounds under cold treatment. F. The content changes of three coumarin compounds under heat treatment. Values represent means ± SD (n = 3).

Fig. 3. Subcellular localization of PpPAL in Arabidopsis protoplasts. Green panels show GFP fluorescence, red panels show mCherry fluorescence and chloroplast autofluorescence is shown in blue panels. Merged panels represent combined fluorescence from GFP, mCherry and chloroplasts. Arabidopsis protoplasts containing GFP were used as a blank control and AtGAPC1-mCherry was used as a positive control for cytosol localization. Bars = 20 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)