Fig. 2

Properties of LiMLDP. a Subcellular localization of LiMLDP-mVenus in N. tabacum pollen tubes. 6 h after germination, cells were stained for LBs using Nile Red and fluorescence was documented by confocal laser scanning microscopy. From top to bottom: Nile Red, mVenus, merged image. Scale bar = 10 μm. 9 out of 9 pollen tubes analyzed showed comparable results. The characteristic punctate pattern of the mVenus signal was also observed in 10 out of 10 unstained pollen tubes. b Hydrophobicity of LiMLDP compared to HpOGP and A. thaliana OLEOSIN1 according to the Kyte & Doolittle amino acid scale. c Changes in LiMLDP expression in response to nitrogen starvation as determined by Illumina RNA sequencing. Samples from two L. incisa cultures were sequenced in 4 technical replicates each. The fold change is given after 3 days of nitrogen starvation compared to nitrogen replete conditions and the significance of this change is given as the P value adjusted for multiple testing at a false discovery rate of 0.05. d Changes in LiMLDP expression in response to varying nitrogen supply as determined by quantitative real-time PCR (qRT-PCR). Transcript levels were normalized to RIBOSOMAL PROTEIN S21. Expression is shown relative to time point 0 and error bars represent the standard error of the mean for three batches cultivated in parallel. The dotted line indicates the onset of nitrogen repletion and total fatty acid (TFA) levels are shown for comparison. e Phylogenetic tree of MLDPs determined by the maximum likelihood method. Proteins that have previously been studied are marked with an asterisk and LiMLDP is highlighted in bold. LDP = LIPID DROPLET PROTEIN, LDSP = LIPID DROPLET SURFACE PROTEIN