Fig. S1 Phylogenetic tree of Cer synthases from plants showing the branches leading to LOH1/LOH3 and LOH2.

Fig. S2 High-pressure liquid chromatography chromatograms of sphingoid bases liberated from whole Saccharomyces cerevisiae cells and from freeze-dried leaves of 5-wk-old Arabidopsis thaliana plants by strong alkaline hydrolysis.

Fig. S3 T-DNA insertion lines for LOH1, LOH2, and LOH3.

Fig. S4 The levels of the phytohormones salicylic acid (SA), abscisic acid (ABA), indole-3-acetic acid (IAA), and jasmonic acid (JA) in the T-DNA insertion lines for LOH1, LOH2 and LOH3.

Fig. S5 The proportions of Cer and GlcCer species having zero, one, or two double bonds for Col-0, loh1, loh2, and loh3 Arabidopsis thaliana plants.

Table S1 Polymerase chain reaction primers

Table S2 QuantiTect Primer Assays (Qiagen) for real-time PCR

Table S3 Molecular species of Cer and GlcCer detected by ultra performance liquid chromatography/electrospray ionization time-of-flight mass spectrometry (UPLC/ESI-TOF-MS) in the lipid extracts from Arabidopsis thaliana and Saccharomyces cerevisiae

Methods S1 Additional details for cloning of Arabidopsis thaliana LOH1, LOH2 and LOH3, in vitro Cer synthase assay, lipid extraction from yeast cells and A.  thaliana leaves, mild alkaline hydrolysis, fractionation of the lipid extract, analysis by UPLC/MS-TOF, analysis of phytohormones and real-time PCR.

Notes S1 The two groups of Cer and GlcCer species distinguished by their level of desaturation.

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