Analysis of diTPSs from Stevia.

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Analysis of diTPSs from Stevia. Analysis of diTPSs from Stevia. A, Phylogenetic analysis of SrCPS2 and SrKSL from Stevia. The maximum likelihood tree was constructed by the MEGA 6 program from an alignment of full-length SrCPS2 and SrKSL with other plant TPSs. Abbreviations and accession numbers of proteins are listed in Supplemental Table S2. B, Relative expression levels of Stevia diTPS (SrCPS, SrCPS2, SrKS1, and SrKSL) and P450 (SrKO1 and SrKAH) genes in leaf-trichomes (L-T) and trichomes (T). Transcript levels of genes expressed in L-T and T were measured by qRT-PCR. Amplification of Actin mRNA was used for normalization. C, Subcellular localization of Stevia diTPSs. YFP-fused SrCPS, SrCPS2, and SrKSL (SrCPS-YFP, SrCPS2-YFP, and SrKSL-YFP) were transiently expressed in N. benthamiana leaves by A. tumefaciens-mediated infiltration and visualized 3 d postinfiltration using the YFP channel of a confocal microscope. Auto, Chlorophyll autofluorescence; YFP, YFP channel image; Light, light microscope image; Merged, merged image between Auto and YFP. Bars = 10 µm (SrCPS-YFP) and 20 µm (SrCPS2-YFP and SrKSL-YFP). Mi Jung Kim et al. Plant Physiol. 2015;169:2462-2480 ©2015 by American Society of Plant Biologists