medicago nsp2 mutant  (Medicago)

Journal: Advanced Science

Article Title: Nodulation Signaling Pathway 1 and 2 Modulate Vanadium Accumulation and Tolerance of Legumes

doi: 10.1002/advs.202306389

Figure Lengend Snippet: NSP1 and NSP2 mutations increase vanadium (V) sensitivity. a,b) Shoot (a) and root (b) phenotypes of wild‐type (R108) and mutant nsp1 and nsp2 Medicago truncatula plants under different V conditions. Four‐week‐old seedlings were treated with the indicated concentrations of V for one week. Bars, 2 cm. c) Biomass of R108, nsp1 , and nsp2 plants under different V conditions. Plants were treated as described in (a) and (b). The treatment concentrations of V were 0 mg L −1 (V0), 20 mg L −1 (V20), 200 mg L −1 (V200), and 2000 mg L −1 (V2000). Data are expressed as mean ± SD values ( n = 6). Different letters above the bars indicate significant differences at P < 0.05 (Duncan's test). d,e) Intracellular ROS in the leaves (d) and roots (e) of R108, nsp1 , and nsp2 plants detected using 2′−7′ dichlorodihydrofluorescein diacetate (H 2 DCFDA). Three‐week‐old seedlings were cultured with ½‐strength Hoagland nutrient solution prepared with soil extract. After nodulation of R108, plants were treated with (+V) or without (‐V) 100 mg L −1 V for 2 days. Then, leaves and roots were obtained for ROS detection. Arrows indicate where ROS accumulated in tissues.

Article Snippet: Here, it is found that nsp1 and nsp2 , Rhizobium symbiosis defective mutants of Medicago truncatula , are sensitive to V. Concentrations of phosphorus (P), iron (Fe), and sulfur (S) with V are negatively correlated in the shoots of wild‐type R108, but not in mutant nsp1 and nsp2 shoots.

Techniques: Mutagenesis, Cell Culture

Journal: Advanced Science

Article Title: Nodulation Signaling Pathway 1 and 2 Modulate Vanadium Accumulation and Tolerance of Legumes

doi: 10.1002/advs.202306389

Figure Lengend Snippet: NSP1 and NSP2 mutations alter the accumulation of vanadium (V) and the correlations of V concentration with phosphorus (P), iron (Fe), sulfur (S), and calcium (Ca) concentrations. a) The V concentration in the shoots and roots of 5‐week‐old wild‐type (R108), nsp1 , and nsp2 Medicago truncatula plants treated with different concentrations of V for one week. The treatment concentrations of V were 0 mg L −1 (V0), 20 mg L −1 (V20), and 200 mg L −1 (V200). b) The translocation factor of V from roots to shoots of R108, nsp1 , and nsp2 plants. Data were calculated from shoot‐to‐root ratios of V concentrations in (a). c–f) Concentrations of P, Fe, S, and Ca in shoots and roots of wild‐type (R108) and mutant nsp1 and nsp2 plants described in (a). Data in (a–f) are expressed as mean ± SD values ( n = 4). Statistical significance is denoted by asterisks based on independent sample t ‐tests (* P < 0.05, ** P < 0.01, *** P < 0.001). g–i) Correlation analysis of the V concentration with P, Fe, S, and Ca in shoots of wild‐type (R108) (g), nsp1 (h), and nsp2 (i) and roots of j) R108, k) nsp1 , and l) nsp2 using the data in (a) and (c–f).

Article Snippet: Here, it is found that nsp1 and nsp2 , Rhizobium symbiosis defective mutants of Medicago truncatula , are sensitive to V. Concentrations of phosphorus (P), iron (Fe), and sulfur (S) with V are negatively correlated in the shoots of wild‐type R108, but not in mutant nsp1 and nsp2 shoots.

Techniques: Concentration Assay, Translocation Assay, Mutagenesis

Journal: Advanced Science

Article Title: Nodulation Signaling Pathway 1 and 2 Modulate Vanadium Accumulation and Tolerance of Legumes

doi: 10.1002/advs.202306389

Figure Lengend Snippet: The deficiency of phosphorus (P), iron (Fe), or sulfur (S) increases the sensitivity of plants to vanadium (V) stress. a–c) Images of F0 (a), Fm (b), and Fv/Fm ratios (c) in the leaves of wild type (R108) and mutant nsp1 and nsp2 Medicago truncatula plants. Three‐week‐old seedlings were cultured with ½‐strength Hoagland nutrient solution prepared with soil extract. After nodulation of R108, plants were treated with 100 mg L −1 V (+V) under low P (LP, 1 µ m PO 4 3− ), low Fe (L‐Fe, 1 µ m Fe 2+ ), or low S (LS, 1 µ m SO 4 2− ) conditions for 16 h. Then, leaves were obtained for chlorophyll fluorescence imaging. d–f) Statistical analysis of F0 (d), Fm (e), and Fv/Fm ratios (f) in (a‐c). Data are expressed as mean ± SD values ( n = 4). Different letters above the bars indicate significant differences at P < 0.05 (Duncan's test).

Article Snippet: Here, it is found that nsp1 and nsp2 , Rhizobium symbiosis defective mutants of Medicago truncatula , are sensitive to V. Concentrations of phosphorus (P), iron (Fe), and sulfur (S) with V are negatively correlated in the shoots of wild‐type R108, but not in mutant nsp1 and nsp2 shoots.

Techniques: Mutagenesis, Cell Culture, Fluorescence, Imaging

Journal: Advanced Science

Article Title: Nodulation Signaling Pathway 1 and 2 Modulate Vanadium Accumulation and Tolerance of Legumes

doi: 10.1002/advs.202306389

Figure Lengend Snippet: Mutation or overexpression of phosphate (P), iron (Fe) and sulfate (S) transporter genes can enhance plant tolerance to vanadium (V). a) Growth phenotypes of Medicago truncatula wild‐type (R108) plants and the vacuolar phosphate transporter mutant mtvpt3‐1 plants under V stress. Five‐week‐old seedlings were treated with 1000 mg L −1 V for five days. b) Growth phenotypes of Arabidopsis thaliana wild‐type (Col‐0), P transporter mutant ( pht1;1 , pht1;9 , vpt1 , and pho1 ), and VPT1 ‐overexpressing plants ( VPT1‐ OE) under V stress. Three‐week‐old seedlings were treated with 1000 mg L −1 V for 10 days. c) Growth phenotypes of wild‐type (Col‐0) and MtVPT3 ‐overexpressing plants ( MtVPT3‐ OE) under V stress. Three‐week‐old seedlings were treated with 1000 mg L −1 V for 10 days. d) Growth phenotypes of wild‐type (Col‐0) and Fe transporter mutant irt1 plants under V stress. Three‐week‐old seedlings were treated with 1000 mg L −1 V for 10 days. e) Growth phenotypes of wild‐type (Col‐0) and S transporter mutant ( sultr1;1 , sultr1;2 , sultr3;3/3;4/3;5 , sultr3;1/3;2/3;3/3;5 , sultr3;1/3;2/3;3/3;4/3;5 , sultr4;1 , and sultr4;2 ) plants under V stress. Three‐week‐old seedlings were treated with 1000 mg L −1 V for one week.

Article Snippet: Here, it is found that nsp1 and nsp2 , Rhizobium symbiosis defective mutants of Medicago truncatula , are sensitive to V. Concentrations of phosphorus (P), iron (Fe), and sulfur (S) with V are negatively correlated in the shoots of wild‐type R108, but not in mutant nsp1 and nsp2 shoots.

Techniques: Mutagenesis, Over Expression

Journal: Advanced Science

Article Title: Nodulation Signaling Pathway 1 and 2 Modulate Vanadium Accumulation and Tolerance of Legumes

doi: 10.1002/advs.202306389

Figure Lengend Snippet: NSP1 and NSP2 are required for both plant tolerance to vanadium (V) and the response of phosphate (P), iron (Fe), and sulfate (S) transporter genes and are also dependent on the existence of soil microbes. a) Growth phenotypes of wild‐type (R108) and nsp1 and nsp2 mutant Medicago truncatula plants under V stress in the presence and absence of soil microorganisms. Three‐week‐old seedlings were cultured with ½‐strength Hoagland nutrient solution prepared with non‐sterile (Natural) or sterile soil extract (Sterilized Soil Extract Nutrient Solution). After nodulation of R108, plants were treated with (+V) or without (‐V) 30 mg L −1 V for 1 week. Bars, 2 cm. b,c) Heatmap of relative expression levels of P, Fe, and S transporter genes in the shoots and roots of wild‐type (R108), nsp1 , and nsp2 plants under V stress in the presence (b) and absence (c) of soil microorganisms. Plants were treated as in (a). The color codes indicate the fold changes in gene expression after V treatment, expressed as log 2 [(expression with V (+V))/(expression without V (‐V))]. Data were calculated based on the values in Figures (Supporting Information). Asterisks indicate statistically significant differences based on independent sample t ‐tests (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: Here, it is found that nsp1 and nsp2 , Rhizobium symbiosis defective mutants of Medicago truncatula , are sensitive to V. Concentrations of phosphorus (P), iron (Fe), and sulfur (S) with V are negatively correlated in the shoots of wild‐type R108, but not in mutant nsp1 and nsp2 shoots.

Techniques: Mutagenesis, Cell Culture, Sterility, Expressing, Gene Expression

Journal: Advanced Science

Article Title: Nodulation Signaling Pathway 1 and 2 Modulate Vanadium Accumulation and Tolerance of Legumes

doi: 10.1002/advs.202306389

Figure Lengend Snippet: NSP1 and NSP2 mutations alter the β‐diversity and relative abundance of rhizobacteria under vanadium (V) stress. a–c) Principal coordinate analysis (PCoA) depicting the β‐diversity of rhizobacteria in wild‐type (R108) (a), nsp1 (b), and nsp2 (c) Medicago truncatula plants treated with different V concentrations. In the legend, R, P1, and P2 represent R108, nsp1 , and nsp2 , respectively, while 0, 20, 200, and 2000 represent V treatment concentrations in mg L −1 . The table presents the corresponding P ‐values for comparisons between different concentrations within the same plant material. d–g) Principal coordinate analysis (PCoA) of β‐diversity of rhizobacteria among wild‐type (R108), nsp1 , and nsp2 treated with 0 (d), 20 (e), 200 (f), or 2000 (g) mg L −1 V. The meaning of the legend and the content shown in the table are consistent with (a–c). Permutational multivariate analysis of variance (PERMANOVA) (a–g) was conducted based on Bray–Curtis distances with a 90% confidence interval. h) Relative abundance of the top ten rhizobacterial families in wild‐type (R108), nsp1 , and nsp2 plants. R, P1, and P2 represent R108, nsp1 , and nsp2 , respectively. The numbers following the points of R, P1, and P2 indicate the V treatment concentration in mg L −1 .

Article Snippet: Here, it is found that nsp1 and nsp2 , Rhizobium symbiosis defective mutants of Medicago truncatula , are sensitive to V. Concentrations of phosphorus (P), iron (Fe), and sulfur (S) with V are negatively correlated in the shoots of wild‐type R108, but not in mutant nsp1 and nsp2 shoots.

Techniques: Concentration Assay

Journal: Advanced Science

Article Title: Nodulation Signaling Pathway 1 and 2 Modulate Vanadium Accumulation and Tolerance of Legumes

doi: 10.1002/advs.202306389

Figure Lengend Snippet: NSP1 and NSP2 mutations alter rhizobacterial co‐occurrence networks. a–c) Co‐occurrence networks of rhizobacteria in wild‐type (R108) (a), nsp1 (b), and nsp2 (c) Medicago truncatula plants. Low frequency operational taxonomic units (OTUs) with frequencies less than 92% were removed from all samples. V‐responsive OTUs (VrOTUs) are highlighted and color‐coded. The edges of the co‐occurrence network were selected based on criteria of Spearman's rho > 0.7 and P ‐value < 0.001. d–f) Mantel test of plant element concentrations and VrOTU modules in R108 (d), nsp1 (e), and nsp2 (f). Plant element concentration data were sourced from Figure . Lines with significant correlations are indicated in color.

Article Snippet: Here, it is found that nsp1 and nsp2 , Rhizobium symbiosis defective mutants of Medicago truncatula , are sensitive to V. Concentrations of phosphorus (P), iron (Fe), and sulfur (S) with V are negatively correlated in the shoots of wild‐type R108, but not in mutant nsp1 and nsp2 shoots.

Techniques: Concentration Assay

Journal: Advanced Science

Article Title: Nodulation Signaling Pathway 1 and 2 Modulate Vanadium Accumulation and Tolerance of Legumes

doi: 10.1002/advs.202306389

Figure Lengend Snippet: NSP1 and NSP2 mutations alter the rhizobacterial biomarkers and association between biomarkers and plant element concentrations. a–c) Abundance and importance assessment of rhizobacterial biomarkers in R108 (a), nsp1 (b), and nsp2 (c) Medicago truncatula plants, along with Spearman's correlation with plant element concentrations. The bubble chart displays the frequency of the top 20 biomarkers. The bar chart shows the importance of the top 20 biomarkers. The correlation matrix illustrates the relationships between the top 20 biomarkers and plant element concentrations. Plant element concentration data were sourced from Figure . Spearman's correlation coefficients were calculated with statistical significance indicated as follows: * P < 0.05, ** P < 0.01, and *** P < 0.001. d,e) A proposed model illustrating the response mechanism of Medicago truncatula to vanadium (V) stress and the effect of NSP1 and NSP2 mutations on it. In (d), V may share transporters with phosphate (P) and sulfate (S). Under V stress, the wild‐type (R108) plant downregulates the expression of P and S transporter genes ( MtPT1 and MtSULTR1;1 ) in roots to reduce V absorption. Furthermore, the upregulation of MtVPT2 , MtVPT3 , and MtPHO1;2 accelerate the vacuolar compartmentalization of V and its loading into other tissues to reduce the toxicity of V to roots. The downregulation of the Fe transporter genes MtZIP6 and MtZIP9 ensures the response of P transporter genes to V through Fe‐P balance regulation. The upregulated expression of MtZIP6 , MtPT1 , MtPT2 , MtSULTR1;1 , and MtSULTR1;2 in the shoots may increase the ability of Fe, P, and S to resist V toxicity. However, the tolerance of R108 to V and the expression response of genes in roots largely depend on the presence of soil microbes. R108 can maintain the diversity of the rhizobacterial community under V stress and actively regulate rhizobacteria to coordinate the response to V stress. In (e), both nsp1 and nsp2 mutants cannot form nodules. Subsequently, P, S, and Fe transporter genes are unable to respond positively or appropriately to V stress. Additionally, the rhizobacteria become fragile and disordered. Therefore, nsp1 and nsp2 exhibit a more sensitive V stress phenotype compared to R108.

Article Snippet: Here, it is found that nsp1 and nsp2 , Rhizobium symbiosis defective mutants of Medicago truncatula , are sensitive to V. Concentrations of phosphorus (P), iron (Fe), and sulfur (S) with V are negatively correlated in the shoots of wild‐type R108, but not in mutant nsp1 and nsp2 shoots.

Techniques: Concentration Assay, Expressing