Journal:

Article Title: Truffles Regulate Plant Root Morphogenesis via the Production of Auxin and Ethylene 1 [C] [W] [OA]

doi: 10.1104/pp.109.141325

Figure Lengend Snippet: Changes in root morphology of Arabidopsis and C. incanus induced by truffle mycelia metabolites. Using the bioassay setup of Figure 1, two truffle species (T. melanosporum_1 = strain 1, T. melanosporum_2 = strain 2; and T. borchii_1 = strain 1, T_borchii_2 = strain 2) reduced primary root length of Arabidopsis (A and F) and C. incanus (B and F), and increased root branching (C, D, and F) and increased root hair length in Arabidopsis (E and F). F, Root morphology of 10-d-old seedlings (scale Arabidopsis 1.0 mm, scale Cistus 2.5 mm). Statistics: Asterisk (*) indicates statistically different results from control (P < 0.05, ANOVA on ranks with Dunn's posthoc test; for root length and branching n > 30 seedlings/treatment; for root hair length n = 300). Bars in E represent ses.

Article Snippet: Truffle mycelia of Tuber borchii (strains ATCC 96540 = strain 1, and 43BO = strain 2) were donated by Prof. Bonfante (University of Turin, Italy), while Dr. Chevalier (Institut National de la Recherche Agronomique, Clermont, France) provided Tuber melanosporum strains Bal1 (strain 1) and Rey_t (strain 2).

Techniques: Bioassay, Control

Journal:

Article Title: Truffles Regulate Plant Root Morphogenesis via the Production of Auxin and Ethylene 1 [C] [W] [OA]

doi: 10.1104/pp.109.141325

Figure Lengend Snippet: IAA concentrations in agar resulting from the exudation of truffle mycelia. After 10 d of growth in the dark, strains of T. borchii and T. melanosporum increased the IAA concentration from 3 to 10 times compared to the control petri dish containing no mycelium. IAA concentration after 10 more days in the growth chamber had dropped in all samples due to degradation by light (n = 3 or 4 petri dish/treatment, bars indicate sd; circles [○ for day 10 and • for day 10 + 10] indicate statistical differences compared to the respective controls, P < 0.05, Mann-Whitney).

Article Snippet: Truffle mycelia of Tuber borchii (strains ATCC 96540 = strain 1, and 43BO = strain 2) were donated by Prof. Bonfante (University of Turin, Italy), while Dr. Chevalier (Institut National de la Recherche Agronomique, Clermont, France) provided Tuber melanosporum strains Bal1 (strain 1) and Rey_t (strain 2).

Techniques: Concentration Assay, Control, MANN-WHITNEY

Journal:

Article Title: Truffles Regulate Plant Root Morphogenesis via the Production of Auxin and Ethylene 1 [C] [W] [OA]

doi: 10.1104/pp.109.141325

Figure Lengend Snippet: Ethylene biosynthesis in truffles. The occurrence of three ethylene biosynthesis pathways described in microorganisms was investigated in T. borchii using various ethylene precursors/inhibitor. Under 16-h photoperiods and without mycelium, addition of the ethylene precursor/inhibitors to malt extract did not significantly increase ethylene concentration (A). Under 16-h photoperiods, significantly higher ethylene levels were detected from mycelium grown on l-Met compared to the mycelium grown on unsupplemented medium (B). The other ethylene precursors (l-Gln, ACC, KMBA) did not increase the ethylene concentration compared to the mycelium grown on unsupplemented medium (B). Addition of AOA, an inhibitor of the ACC pathway, did not reduce ethylene concentration, confirming that l-Met was not transformed through the ACC pathway (B). The mycelium could only transform l-Met to ethylene under 16-h photoperiods, but not in the dark (C). In the contrary, KMBA was photodegraded to ethylene regardless of the presence/absence of the mycelium, while the molecule was not degraded in the dark (D). Taken together, these results demonstrate that ethylene is synthesized from l-Met probably through the KMBA pathway (marked in bold in E). Statistics: P < 0.05, Mann-Whitney. For A and B, asterisk (*) indicates difference from the unsupplemented malt extract; NS = nonsignificant. For C and D, different letters indicate statistically different results. For each treatment, n ≥ 3. Bars = ses (A–D).

Article Snippet: Truffle mycelia of Tuber borchii (strains ATCC 96540 = strain 1, and 43BO = strain 2) were donated by Prof. Bonfante (University of Turin, Italy), while Dr. Chevalier (Institut National de la Recherche Agronomique, Clermont, France) provided Tuber melanosporum strains Bal1 (strain 1) and Rey_t (strain 2).

Techniques: Concentration Assay, Transformation Assay, Synthesized, MANN-WHITNEY

Journal:

Article Title: Truffles Regulate Plant Root Morphogenesis via the Production of Auxin and Ethylene 1 [C] [W] [OA]

doi: 10.1104/pp.109.141325

Figure Lengend Snippet: Comparison of root morphology induced by truffles or IAA ± ACC. Root length (A and B), branching (C and D), and root hair length (E) of Arabidopsis (A, C, and E) or Cistus (B and D) subjected to mycelial metabolites of T. borchii strain 1 or T. melanosporum strain 1 (gray bars) or synthetic IAA alone (black bars), or IAA and ACC (1 μm; white bars). Error bars represent ses; n > 15 seedlings per treatment. The same symbols (*, ○) represent values that are statistically the same (P < 0.05, ANOVA on ranks with posthoc Dunn's test). Note that the root hair phenotype induced by the mycelium in Arabidopsis was only fully restored by supplying IAA and ACC together (E and F: root hair of 15-d-old seedlings, scale bars in F = 0.25 mm; ND = not determined).

Article Snippet: Truffle mycelia of Tuber borchii (strains ATCC 96540 = strain 1, and 43BO = strain 2) were donated by Prof. Bonfante (University of Turin, Italy), while Dr. Chevalier (Institut National de la Recherche Agronomique, Clermont, France) provided Tuber melanosporum strains Bal1 (strain 1) and Rey_t (strain 2).

Techniques: Comparison

Journal:

Article Title: Truffles Regulate Plant Root Morphogenesis via the Production of Auxin and Ethylene 1 [C] [W] [OA]

doi: 10.1104/pp.109.141325

Figure Lengend Snippet: Screening Arabidopsis mutants for resistance to truffle metabolites. Using the bioassay setup of Figure 1, the auxin aux1-7, ethylene ein2-LH, and double mutant aux1-7;ein2-LH were tested for their sensitivity to truffle (T. borchii strain 1) metabolites and compared to the wild type. A to C, Primary root length (A), root branching (B), and root hair length (C). Bars represent average values for 10-d-old seedlings, n > 20 seedlings/treatment; for root hair length n = 300, 15-d-old seedlings. Percentages represent changes versus respective controls (±ses). Different letters indicates statistically different values (P < 0.05, ANOVA on ranks and posthoc Dunn's test). D, Root tips of the Arabidopsis auxin reporter line DR5∷GFP. The mycelium increased the GFP signal intensity at the level of the first tier columnella cells (red arrows) and at the level of the epidermis (yellow arrows) compared to the mycelium control in DR5∷GFP roots. Hormones including IAA (0.1 μm) and ACC (1.0 μm) mimicked this increase in signal intensity fully when applied together (+IAA and ACC; D). For quantification of the GFP signal, refer to Supplemental Figure S7. MeOH control = control for the IAA, ACC, and IAA and ACC treatments. Scale bars in D = 25 μm.

Article Snippet: Truffle mycelia of Tuber borchii (strains ATCC 96540 = strain 1, and 43BO = strain 2) were donated by Prof. Bonfante (University of Turin, Italy), while Dr. Chevalier (Institut National de la Recherche Agronomique, Clermont, France) provided Tuber melanosporum strains Bal1 (strain 1) and Rey_t (strain 2).

Techniques: Bioassay, Mutagenesis, Control

Journal: Cell Death & Disease

Article Title: c-Jun N-terminal kinase has a key role in Alzheimer disease synaptic dysfunction in vivo

doi: 10.1038/cddis.2013.559

Figure Lengend Snippet: Synaptopathy occurred in TgCRND8 mice, already at an early stage of AD pathology. ( a – h ) Western blot and relative quantification performed on the TIF fraction of 2-months-old Wt and TgCRND8 mice. Tg mice showed a significant reduction in the PSD levels of GluN2A (31%) ( b ) and of GluN2B (54%) ( c ) subunits of NMDAr, as well as reduction of GluA1 (46%) ( d ) and of GluA2 (35%) ( e ) subunits of AMPAr, of PSD-95 (35%) ( f ) and of drebrin (60%) ( g ) if compared with age-matched Wt mice (Student's t -test, * P <0.05, ** P <0.01, *** P <0.001, n =6). Tubulin levels were not affected ( h ) (Student's t -test, P >0.05, n =6). ( i – p ) Western blot and relative quantification performed on the TIF fraction of 9-months-old Wt and TgCRND8 mice. Tg mice showed a severe reduction in the PSD levels of GluN2A (74%) ( j ) and of GluN2B (86%) ( k ) subunits of NMDAr, as well as reduction of GluA1 (66%) ( l ) and of GluA2 (71%) ( m ) subunits of AMPAr, of PSD-95 (80%) ( n ) and of drebrin (84%) ( o ) if compared with age-matched Wt mice (Student's t -test, * P <0.05, ** P <0.01, *** P <0.001, n =6). Tubulin levels were not affected ( p ) (Student's t -test, P >0.05, n =6)

Article Snippet: Primary antibodies were diluted in the same buffer (incubation overnight, 4 °C) using: anti GluN2A (1 : 2000, Gibco-Invitrogen, Paisley, Scotland, UK), anti GluN2B (1 : 2000, Gibco-Invitrogen), anti GluA1 (1 : 1000, Millipore, Billerica, MA, USA), anti GluA2 (1 : 1000, Millipore), anti PSD-95 (1 : 2000, Cayman Chemical Company, Ann Arbor, MI, USA), anti drebrin (1 : 2000, Assay Design, Ann Arbor, MI, USA), P-JNK (G-7) (1 : 1000, Cell Signaling Technology, Danvers, MA, USA), JNK (1 : 1000, Cell Signaling Technology), Cleaved caspase-3 (1 : 1000, Cell Signaling Technology).

Techniques: Western Blot, Quantitative Proteomics

Journal: Cell Death & Disease

Article Title: c-Jun N-terminal kinase has a key role in Alzheimer disease synaptic dysfunction in vivo

doi: 10.1038/cddis.2013.559

Figure Lengend Snippet: D-JNKI1 prevented A β -oligomers-induced loss of postsynaptic proteins from the PSD in TgCRND8 mice. ( a – h ) Western blot and relative quantification of TIF fraction obtained from hippocampus homogenate of 9-months-old Wt and TgCRND8 mice, chronically treated with vehicle (water) or D-JNKI1 (22 mg/kg for 5 months). TgCRND8 mice treated with vehicle showed a strong loss of GluN2A ( b ) and GluN2B ( c ) subunits of NMDAr, GluA1 ( d ) and GluA2 ( e ) subunits of AMPAr, PSD-95 ( f ) and drebrin ( g ) from the PSD. D-JNKI1 chronic treatment completely prevented these alterations in Tg mice, whereas it did not affect protein levels in Wt mice (two-way analysis of variance (ANOVA), Bonferroni post-hoc test, * P <0.05, ** P <0.01 Wt veh versus Tg veh; # P <0.05, # # P <0.01 Tg veh versus Tg D-JNKI1; § P <0.05 Wt D-JNKI1 versus Tg D-JNKI1, n =6). Tubulin levels ( h ) were not affected by the treatment or by mice genotype. ( i and j ) Western blot and relative quantification showing cleaved caspase-3 levels in the TIF fraction of 9-months-old Wt and TgCRND8 mice treated with vehicle (water) or D-JNKI1 (22 mg/kg for 5 months). D-JNKI1 chronic treatment completely prevented activation of caspase-3 in TgCRND8 mice (Two-way ANOVA, Bonferroni post-hoc test, ** P <0.01 Wt veh versus Tg veh; # # P <0.01 Tg veh versus Tg D-JNKI1, n =6)

Article Snippet: Primary antibodies were diluted in the same buffer (incubation overnight, 4 °C) using: anti GluN2A (1 : 2000, Gibco-Invitrogen, Paisley, Scotland, UK), anti GluN2B (1 : 2000, Gibco-Invitrogen), anti GluA1 (1 : 1000, Millipore, Billerica, MA, USA), anti GluA2 (1 : 1000, Millipore), anti PSD-95 (1 : 2000, Cayman Chemical Company, Ann Arbor, MI, USA), anti drebrin (1 : 2000, Assay Design, Ann Arbor, MI, USA), P-JNK (G-7) (1 : 1000, Cell Signaling Technology, Danvers, MA, USA), JNK (1 : 1000, Cell Signaling Technology), Cleaved caspase-3 (1 : 1000, Cell Signaling Technology).

Techniques: Western Blot, Quantitative Proteomics, Activation Assay