Mouse fat storage-inducing transmembrance protein 2 (FIT2) promotes lipid droplet accumulation in plants Page: 826
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0- . * *Figure 1 FIT2 expression in Nicotiana benthamiana leaves elaborates a neutral lipid compartment. (a) Representative confocal images of
N. benthamiana leaves expressing FIT2 or mutated FIT2 proteins (i.e. FIT2-N 801A or FIT2-FLL157-1591AAA) in the presence or absence of LEC2. Cells were
(co)transformed as indicated by the panel label, and LDs (green) were visualized by staining with BODIPY 493/503. Red colour corresponds to
chloroplast autofluorescence. Images are 2D projections of Z-stacks of 50 optical sections. Bar = 20 pm. (b) Number of total LDs and LDs in different
size categories per image area. The numbers and sizes of LDs were quantified by Imagei as BODIPY-stained lipid area in 2D projections of Z-stacks (see
Experimental procedures for details). Small LDs: BODIPY-stained lipid area <3 pm2. Medium LDs: 3-6 pm2. Large LDs: 6-10 pm2. Super-sized LDs:
>10 pm2. Averages and SDs were calculated from three biological replicates (three Z-stacks from each replicate). Asterisks indicate significant difference
relative to the mock control at P s 0.05 determined by Student's t-test. (c) and (d) TAG and SE content in infiltrated N. benthamiana leaves. Averages
and SDs were plotted from four biological replicates. Asterisks indicate significant difference in comparison with the P19 control at P < 0.05 determined
by Student's t-test.(Figure 1 c and d). Detailed analysis of the TAG and SE molecular
species by conventional, direct-infusion ESI-MS/MS (electrospray
ionization tandem mass spectrometry) (Figure S1) indicated that,
generally, the TAGs and SEs that are normally present in leaves
were all increased upon FIT2 expression. These same molecularspecies of TAG and SE were exaggerated further by co-expression
of FIT2 and LEC2, making the overall increase in TAGs and SE
even more obvious (Figure S1). Notably, there was no further
increase in TAG and SE in leaves co-expressing LEC2 and the
enhanced FIT2 variant (FLLI157-1591AAA) in comparison with2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd., 15, 824-836
826 Yingqi Cai et al.
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Cai, Yingqi; McClinchie, Elizabeth; Price, Ann; Nguyen, Thuy N.; Gidda, Satinder K.; Watt, Samantha C. et al. Mouse fat storage-inducing transmembrance protein 2 (FIT2) promotes lipid droplet accumulation in plants, article, December 2, 2016; Hoboken, New Jersey. (https://digital.library.unt.edu/ark:/67531/metadc984098/m1/3/: accessed April 24, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; crediting UNT College of Arts and Sciences.