Fig. 5

Properties of LiLBP36. a Subcellular localization of LiLBP36-mVenus in N. tabacum pollen tubes. Following 6 h of pollen germination, cells were stained for LBs using Nile Red and fluorescence was documented by laser scanning microscopy. From left to right: Nile Red fluorescence, mVenus fluorescence, merged image. Scale bar = 10 μm. 5 out of 5 pollen tubes analyzed showed comparable results. The characteristic punctate pattern of the mVenus signal was also observed in 13 out of 13 unstained pollen tubes. b Changes in expression of LiLBP36 in response to varying nitrogen supply as determined by Illumina RNA sequencing. Samples from two L. incisa cultures were sequenced in 4 technical replicates each. The fold change of gene expression 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. c Changes in expression of LiLBP36 in response to varying nitrogen supply as determined by qRT-PCR. Transcript levels were normalized to RIBOSOMAL PROTEIN S21 transcripts. Expression is shown relative to time point 0 and error bars represent the standard error of the mean for 3 batches cultivated in parallel. The dotted line indicates the onset of nitrogen repletion and total fatty acid (TFA) levels are shown for comparison. d Schematic representation of LiLBP36 features with numbers indicating amino acid positions. The full line illustrates the section of the amino acid sequence that bears strong resemblance to fungal TAG-, DAG- and MAG- lipases according to secondary structure modelling using Phyre2. The section includes the conserved GXSXG motif as well as a second catalytic residue (D403)