serendipita indica strain  (ATCC)

Journal: Molecular cell

Article Title: Modulation of fungal phosphate homeostasis by the plant hormone strigolactone.

doi: 10.1016/j.molcel.2024.09.004

Figure Lengend Snippet: Figure 6. Pho84 contains a transmembrane binding pocket required for SL response (A) Left, EY917 expressing either a pho84-G256D or a pho84-A262P mutation. Right, BY4742 strains expressing either a pho84-G256D or a pho84-A262P mutation stained for AP activity. (B) Phosphate uptake in wild-type PHO84, pho84-G256D, and pho84-A262P strains in the presence and absence of 50 mM rac-GR24. Phosphate uptake is measured by the ratio of optical density at 700 nm versus 600 nm. (C) Structure of the yeast Pho84 protein modeled on an occluded Serendipita indica high-affinity phosphate transporter (PDB: 7SP5). Enlargement, pocket domain with two mutant amino acid substitutions (pho84-G256D and pho84-A262P). The Gly256Asp substitution clashes with a docked (+)-(20R)-GR24 isomer (cyan sticks). (D) Left, interactions between Pho84 binding pocket residues (green sticks) and a (+)-(20R)-GR24 isomer (blue sticks). Right, red fans indicate potential hydro- phobic contacts. (E) Analysis of mutations in SL-binding pocket in Pho84. Positions of Pho84 amino acid residues, AA74, 212, and 259 are represented by yellow spheres. AA256 is represented by a red sphere. Cyan sticks represent docked (+)-(20R)-GR24 isomer. Growth of EY917 expressing either pho84-L74F, pho84-S212F, or pho84- L259F on 100 mM rac-GR24. (F) Pho84 localization in response to SL. A line carrying the wild-type PHO84-GFP or the pho84-L259F-GFP allele is imaged before and after a 2-h 50 mM rac- GR24 incubation. Arrows, white: plasma membrane, yellow: vacuole. Scale bar, 5 mm.

Article Snippet: The Serendipita indica strain was purchased from ATCC with the strain designation END1 [DSM 11827] (product code: 204458).

Techniques: Binding Assay, Expressing, Mutagenesis, Staining, Activity Assay, Incubation, Clinical Proteomics, Membrane

Journal: Molecular cell

Article Title: Modulation of fungal phosphate homeostasis by the plant hormone strigolactone.

doi: 10.1016/j.molcel.2024.09.004

Figure Lengend Snippet: Figure 7. Plant-associated fungi respond to SL (A) Manganese (Mn2+) sensitivity of Fusarium graminearum (Fusarium) on synthetic media containing 10 mM MnCl2 in the presence or absence of 10 mM rac- GR24. Plates were photographed from above (top) and below (bottom). (B) Mn2+ sensitivity of Serendipita indica (Serendipita) on synthetic media containing 0.75 mM MnCl2 in the presence or absence of 5 mM rac-GR24. Growth was quantified for MnCl2-treated cultures (n = 8) by measuring fungal area (cm2). Cultures grown on 5 mM rac-GR24 had significantly greater colony areas, *** p value < 0.001. (C) Growth of Fusarium on synthetic media in the presence or absence of 5 mM rac-GR24 on increasing levels of Pi. (D) Acid phosphatase activity of homogenized Serendipita cultures spotted onto synthetic media containing 1 mM Pi and supplemented with 5 mM rac-GR24 or 1 mM of a naturally occurring SL, 5-deoxystrigol. Increased red staining indicates higher AP activity. Intensities of red staining on SL relative to mock (DMSO) treatment were quantified in the graph below. (E) SL works through Pho84 to regulate hyphal branching and spore formation in Fusarium. Left, a representative image of Fusarium hyphae in the presence or absence of 15 mM rac-GR24 3 h post-inoculation. Right, quantification of spore production of a wild-type PHO84 and mutant pho84 knockout line in the presence or absence of rac-GR24. ** p value < 0.01, *** p value < 0.001, N.S., not significant. (F) Structural models of Serendipita (SiPho84) and Fusarium (FgPho84) Pho84. Red, key amino acids involved in SL recognition. (G) Proportion of amino acids in the SL-binding motif at each position in Pho84 sequences from the fungal kingdom. Arrow, Saccharomyces cerevisiae laboratory strain (S288c). Black lines indicate that gene could not be identified.

Article Snippet: The Serendipita indica strain was purchased from ATCC with the strain designation END1 [DSM 11827] (product code: 204458).

Techniques: Activity Assay, Staining, Mutagenesis, Knock-Out, Binding Assay