Fig 6. PP-InsP catabolic enzymes contribute to Pi and nitrate homeostasis in Marchantia.

(A) Growth phenotypes of 3-week-old Tak-1, Mppfa-dsp1^ge, Mpnudt1^ge, Mpvip1^ge, MpvipΔpd^ge and Mpvip1^ge MpvipΔpd^ge plants grown from single gemmae on ^½B5 plates in continuous light at 22°C. Scale bar = 1 cm. Representative single gemmae cups are shown alongside, scale bar = 0.1 cm. (B) Quantification of projected thallus surface areas of 3-week-old Tak-1, Mppfa-dsp1^ge, Mpnudt1^ge, Mpvip1^ge, MpvipΔpd^ge and Mpvip1^ge MpvipΔpd^ge plants. Tukey-type all-pairs comparisons between the genotypes [107] were performed in the R package multcomp [106]. (C) Rhizoid mass normalized to thallus mass of 4-week-old Tak-1, Mppfa-dsp1^ge, Mpnudt1^ge, Mpvip1^ge, MpvipΔpd^ge and Mpvip1^ge MpvipΔpd^ge plants. Rhizoids were manually peeled with forceps. The weight of the rhizoids was normalized by the thallus weight of the same plant. Statistical significance was assessed with a Dunnett test with Tak-1 as reference (**** p < 0.001, *** p < 0.005, ** p < 0.01, * p < 0.05). (D) Number of gemmae cups of 4-week-old Tak-1, Mppfa-dsp1^ge, Mpnudt1^ge, Mpvip1^ge, MpvipΔpd^ge and Mpvip1^ge MpvipΔpd^ge plants. Statistical significance was assessed with a Dunnett test with Tak-1 as reference (**** p < 0.001, *** p < 0.005, ** p < 0.01, * p < 0.05). (E) Nitrate quantification of 2-week-old Tak-1, Mppfa-dsp1^ge, Mpnudt1^ge, Mpvip1^ge, MpvipΔpd^ge and Mpvip1^ge MpvipΔpd^ge plant lines grown under nitrate starvation or control conditions. 8 plants were used per genotype. Nitrate was quantified using the Miranda spectrophotometric method [104]. Technical triplicates were done for the standards and duplicates for all samples. Statistical significance was assessed with a Dunnett test with Tak-1 as reference (**** p < 0.001, *** p < 0.005, ** p < 0.01, * p < 0.05). (F) Total Pi levels of 2-week-old Tak-1, Mppfa-dsp1^ge, Mpnudt1^ge, Mpvip1^ge, MpvipΔpd^ge and Mpvip1^ge MpvipΔpd^ge plants grown in Pi-starvation or Pi-sufficient (0.5 mM K₂HPO₄/KH₂PO₄) conditions. Technical triplicates were done for the standards and duplicates for all samples. Statistical significance was assessed with a Dunnett test with Tak-1 as reference (**** p < 0.001, *** p < 0.005, ** p < 0.01, * p < 0.05).