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Keywords = arbuscular mycorrhizal fungi

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15 pages, 3171 KiB  
Article
Genome-Wide Identification, Expression, and Protein Interaction of GRAS Family Genes During Arbuscular Mycorrhizal Symbiosis in Poncirus trifoliata
by Fang Song, Chuanya Ji, Tingting Wang, Zelu Zhang, Yaoyuan Duan, Miao Yu, Xin Song, Yingchun Jiang, Ligang He, Zhijing Wang, Xiaofang Ma, Yu Zhang, Zhiyong Pan and Liming Wu
Int. J. Mol. Sci. 2025, 26(5), 2082; https://doi.org/10.3390/ijms26052082 - 27 Feb 2025
Viewed by 132
Abstract
Arbuscular mycorrhizal (AM) fungi establish mutualistic symbiosis with most land plants, facilitating mineral nutrient uptake in exchange for photosynthates. As one of the most commercially used rootstocks in citrus, Poncirus trifoliata heavily depends on AM fungi for nutrient absorption. The GRAS gene family [...] Read more.
Arbuscular mycorrhizal (AM) fungi establish mutualistic symbiosis with most land plants, facilitating mineral nutrient uptake in exchange for photosynthates. As one of the most commercially used rootstocks in citrus, Poncirus trifoliata heavily depends on AM fungi for nutrient absorption. The GRAS gene family plays essential roles in plant growth and development, signaling transduction, and responses to biotic and abiotic stresses. However, the identification and functional characterization of GRAS family genes in P. trifoliata remains largely unexplored. In this study, a comprehensive genome-wide analysis of PtGRAS family genes was conducted, including their identification, physicochemical properties, phylogenetic relationships, gene structures, conserved domains, chromosome localization, and collinear relationships. Additionally, the expression profiles and protein interaction of these genes under AM symbiosis were systematically investigated. As a result, 41 GRAS genes were identified in the P. trifoliata genome, and classified into nine distinct clades. Collinearity analysis revealed seven segmental duplications but no tandem duplications, suggesting that segmental duplication played a more important role in the expansion of the PtGRAS gene family compared to tandem duplication. Additionally, 18 PtGRAS genes were differentially expressed in response to AM symbiosis, including orthologs of RAD1, RAM1, and DELLA3 in P. trifoliata. Yeast two-hybrid (Y2H) screening further revealed that PtGRAS6 and PtGRAS20 interacted with both PtGRAS12 and PtGRAS18, respectively. The interactions were subsequently validated through bimolecular fluorescence complementation (BiFC) assays. These findings underscored the crucial role of GRAS genes in AM symbiosis in P. trifoliata, and provided valuable candidate genes for improving nutrient uptake and stress resistance in citrus rootstocks through molecular breeding approaches. Full article
(This article belongs to the Special Issue Molecular Research of Tropical Fruit (2nd Edition))
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Figure 1

Figure 1
<p>Chromosome localization of <span class="html-italic">PtGRAS</span> family genes. The text on the left represents the number of chromosomes, and the scale on the left represents the chromosome size.</p>
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<p>Phylogenetic relationships of GRAS proteins between <span class="html-italic">Poncirus trifoliata</span> (Pt), <span class="html-italic">Arabidopsis thaliana</span> (At), and <span class="html-italic">Medicago trunctula</span> (Mt). Red squares represent PtGRAS proteins, blue stars represent AtGRAS proteins, and yellow circles represent MtGRAS proteins. The phylogenetic tree was constructed by MEGA X using the Maximum Likelihood Method (1000 bootstrap). The different colors of backgrounds indicated eight clades of <span class="html-italic">GRAS</span> family genes.</p>
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<p>Conserved motifs and gene structures of <span class="html-italic">PtGRAS</span> family genes. (<b>A</b>) Conserved motifs of <span class="html-italic">PtGRAS</span> family genes using MEME algorithm. The different colors indicated 12 identified motifs. (<b>B</b>) The gene structures are based on the sequences of <span class="html-italic">PtGRAS</span> family genes. The yellow color and green color indicated CDS and UTR, and the lines indicated Intron.</p>
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<p>Collinearity analysis of <span class="html-italic">PtGRAS</span> genes. (<b>A</b>) Gray lines indicated all duplicated genes, dark lines indicated segmentally duplicated genes, and the heatmap and line graph were gene densities. (<b>B</b>) Collinearity analysis of <span class="html-italic">PtGRAS</span> genes with <span class="html-italic">A. thaliana</span> and <span class="html-italic">M. truncatula</span>. The gray lines represented all collinear pairs of <span class="html-italic">P. trifoliata</span> with <span class="html-italic">A. thaliana</span> and <span class="html-italic">M. truncatula</span> at the genome level. The black lines represented collinearity gene pairs.</p>
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<p>The expression profiles of <span class="html-italic">PtGRAS</span> genes in response to arbuscular mycorrhizal symbiosis. (<b>A</b>). Heatmap analysis of RNA-seq data. (<b>B</b>). Relative expression of qRT-PCR analysis. AM, arbuscular mycorrhizal inoculated <span class="html-italic">P. trifoliata</span> roots; NM, non-mycorrhizal control roots. The asterisks indicated significant differences of student’s <span class="html-italic">t</span>-test (** <span class="html-italic">p</span> &lt; 0.01).</p>
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<p>Identification of the interaction of AM symbiosis-related PtGRAS proteins. (<b>A</b>) Y2H analyses screening the interaction among PtGRAS proteins using SD/–LW and SD/–LWHA selective medium. AD-RecT + BD-53 and AD + BD were utilized as positive control and negative control, respectively. The experimental controls (positive and negative controls) are shown in <a href="#app1-ijms-26-02082" class="html-app">Figure S3</a>. (<b>B</b>) BiFC assay validation of the interaction between PtGRAS6, PtGRAS12, and PtGRAS18 proteins in <span class="html-italic">N. benthamiana</span> leaves. Scale bars, 30 µm. The overlapping of GFP and mCherry fluorescence is marked with red arrow. FIB2:mCherry was utilized as a nucleus marker, YFPn + YFPc was provided as the negative control.</p>
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27 pages, 4855 KiB  
Article
Metagenomic Characterization of the Maerua crassifolia Soil Rhizosphere: Uncovering Microbial Networks for Nutrient Acquisition and Plant Resilience in Arid Ecosystems
by Sumayah M. Alharbi, Nadiah Al-Sulami, Hadba Al-Amrah, Yasir Anwar, Ola A. Gadah, Lina Ahmed Bahamdain, Mohammed Al-Matary, Amnah M. Alamri and Ahmed Bahieldin
Genes 2025, 16(3), 285; https://doi.org/10.3390/genes16030285 - 26 Feb 2025
Viewed by 170
Abstract
Background/Objectives:Maerua crassifolia, a threatened medicinal species endemic to drylands, exhibits a pronounced drought sensitivity. Despite the critical role of microorganisms, particularly bacteria and fungi, the microbial consortia in M. crassifolia’s rhizosphere remain underexplored. Methods: Metagenomic whole genome shotgun sequencing (WGS) was [...] Read more.
Background/Objectives:Maerua crassifolia, a threatened medicinal species endemic to drylands, exhibits a pronounced drought sensitivity. Despite the critical role of microorganisms, particularly bacteria and fungi, the microbial consortia in M. crassifolia’s rhizosphere remain underexplored. Methods: Metagenomic whole genome shotgun sequencing (WGS) was employed to elucidate the taxonomic composition of bacterial and fungal communities inhabiting the soil rhizosphere of M. crassifolia. Results: The data revealed a marked predominance of bacterial genomes relative to fungal communities, as evidenced by non-redundant gene analysis. Notably, arbuscular mycorrhizal fungi (AMF), specifically Rhizophagus clarus, Rhizophagus irregularis and Funneliformis geosporum, are key rhizosphere colonizers. This study confirmed the presence of phosphate-solubilizing bacteria (PSB), such as Sphingomonas spp., Cyanobacteria and Pseudomonadota, underscoring the critical role of these microorganisms in the phosphorus cycle. Additionally, the study uncovered the presence of previously uncharacterized species within the phylum Actinobacteria, as well as unidentified taxa from the Betaproteobacteria, Gemmatimonadota and Chloroflexota phyla, which may represent novel microbial taxa with potential plant growth-promoting properties. Conclusions: Findings suggest a complex, symbiotic network where AMF facilitate phosphorus uptake through plant–root interactions. In a tripartite symbiosis, PSB enhance inorganic phosphorus solubilization, increasing bioavailability, which AMF assimilate and deliver to plant roots, optimizing nutrition. This bacterial–fungal interplay is essential for plant resilience in arid environments. Future investigations should prioritize the isolation and characterization of underexplored microbial taxa residing in the rhizosphere of M. crassifolia, with particular emphasis on members of the Actinobacteria, Betaproteobacteria, Gemmatimonadota and Chloroflexota phyla to uncover their roles in nutrient acquisition and sustainability. Full article
(This article belongs to the Section Genes & Environments)
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<p>The figure depicts the location where the samples were collected.</p>
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<p>ANOSIM results of soil metagenomes associated with <span class="html-italic">M. crassifolia</span> plants. ANOSIM was used to compare microbial community structure between groups A and B at the (<b>A</b>) phylum, (<b>B</b>) genus and (<b>C</b>) species levels.</p>
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<p>PCA results of soil metagenomes associated with <span class="html-italic">M. crassifolia</span> plants. PCA was performed based on gene abundances at the (<b>A</b>) phylum, (<b>B</b>) genus and (<b>C</b>) species levels. The <span class="html-italic">x</span>-axis and <span class="html-italic">y</span>-axis represent the first and second principal components (PC1 and PC2), respectively. The percentage of variation explained by each principal component is indicated in parentheses.</p>
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<p>Microbial abundance at the domain level, representing bacteria, Eukaryota and others. The abundance values are based on nRG identified in metagenomic data of <span class="html-italic">M. crassifolia</span> plant across soil types. “Others” include archaea, viruses and unidentified taxa.</p>
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<p>The microbial abundance of the top ten bacterial (purple columns) and eukaryotic (pink columns) phyla, based on nRG identified in the metagenomic data of the <span class="html-italic">M. crassifolia</span> plant across different soil types, including the NOVO_MIX samples.</p>
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<p>The microbial abundance of the top ten bacterial (purple columns) and eukaryotic (pink columns) genera, based on nRG identified in the metagenomic data of the <span class="html-italic">M. crassifolia</span> plant across different soil types, including the NOVO_MIX samples.</p>
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<p>The microbial abundance of the top ten bacterial (purple columns) and eukaryotic (pink columns) species, based on nRG identified in the metagenomic data of the <span class="html-italic">M. crassifolia</span> plant across different soil types, including the NOVO_MIX samples.</p>
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<p>The relative microbial abundance of the top ten bacterial phyla, based on nRG identified in the metagenomic data of the <span class="html-italic">M. crassifolia</span> plant across different soil types (e.g., rhizosphere and bulk soil).</p>
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<p>The relative microbial abundance of the top ten bacterial genera, based on nRG identified in the metagenomic data of the <span class="html-italic">M. crassifolia</span> plant across different soil types (e.g., rhizosphere and bulk soil).</p>
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<p>The relative microbial abundance of the top ten bacterial species, based on nRG identified in the metagenomic data of the <span class="html-italic">M. crassifolia</span> plant across different soil types (e.g., rhizosphere and bulk soil).</p>
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<p>The relative microbial abundance of the top ten eukaryotic phyla, based on nRG identified in the metagenomic data of the <span class="html-italic">M. crassifolia</span> plant across different soil types (e.g., rhizosphere and bulk soil).</p>
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<p>The relative microbial abundance of the top ten eukaryotic genera, based on nRG identified in the metagenomic data of the <span class="html-italic">M. crassifolia</span> plant across different soil types (e.g., rhizosphere and bulk soil).</p>
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<p>The relative microbial abundance of the top ten eukaryotic species, based on nRG identified in the metagenomic data of the <span class="html-italic">M. crassifolia</span> plant across different soil types (e.g., rhizosphere and bulk soil).</p>
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<p>(<b>A</b>) The LEFSe line graph represents species that show a significant difference in abundance, with an LDA threshold of 4.0. The length of each bar in the graph is proportional to the size of the effect (LDA score), with longer bars indicating greater differences in abundance. The <span class="html-italic">x</span>-axis represents the LDA score for the bulk soil ‘B’ group. (<b>B</b>) The phylogenetic tree illustrates the relative abundance and evolutionary links between taxa. The nodes on the tree denote taxonomic classifications, while the branching patterns indicate putative ancestral relationships. The size of each node correlates with the observed abundance of that taxon. The highlighted red node labeled “a” identifies “Unclassified Acidimicrobiales” as a biomarker taxon distinct from the indistinguishable yellow nodes. The red marker “B” means that this biomarker taxon is associated with the bulk soil habitat.</p>
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<p>(<b>A</b>) The LEFSe line graph represents species that show a significant difference in abundance, with an LDA threshold of 4.0. The length of each bar in the graph is proportional to the size of the effect (LDA score), with longer bars indicating greater differences in abundance. The <span class="html-italic">x</span>-axis represents the LDA score for the bulk soil ‘B’ group. (<b>B</b>) The phylogenetic tree illustrates the relative abundance and evolutionary links between taxa. The nodes on the tree denote taxonomic classifications, while the branching patterns indicate putative ancestral relationships. The size of each node correlates with the observed abundance of that taxon. The highlighted red node labeled “a” identifies “Unclassified Acidimicrobiales” as a biomarker taxon distinct from the indistinguishable yellow nodes. The red marker “B” means that this biomarker taxon is associated with the bulk soil habitat.</p>
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<p>The microbiome of the rhizosphere is mainly structured by chemotaxis, i.e., the controlled movement of microbes in response to chemicals. This behavior can be described as either negative chemotaxis, in which microbes move away from a chemical repellent, or positive chemotaxis, in which they move toward a chemical attractant. Under the guidance of chemical cues from root exudates, PGPMs show positive chemotaxis towards plant roots. These microbes can colonize the rhizosphere more easily thanks to this controlled migration, which improves plant development. On the other hand, certain chemicals contained in root exudates or released by PGPMs, such as <span class="html-italic">Nonomuraea</span> spp. in the rhizosphere, can induce negative chemotaxis on phytopathogens, including <span class="html-italic">Aspergillus</span> spp., <span class="html-italic">Fusarium</span> spp. and <span class="html-italic">R. arrhizus</span>. This repellent effect restricts their access to the rhizosphere, confining them to the bulk soil and protecting the plant.</p>
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13 pages, 1829 KiB  
Article
Identifying AMF-Rich Tir Wheat Rhizospheres to Foster Microbial Inoculants Useful in Sustainable Agriculture: Evidence from the Van Lake Basin
by Solmaz Najafi, Mehmet Ülker, Younes Rezaee Danesh, Semra Demir, Erol Oral, Fevzi Altuner, Siyami Karaca, Meriç Balci, Burak Özdemir, Bulut Sargin, Aynur Dilsiz, Çağlar Sagun, Ezelhan Selem, Sana Jamal Salih, Mina Najafi, Beatrice Farda and Marika Pellegrini
Sustainability 2025, 17(4), 1676; https://doi.org/10.3390/su17041676 - 18 Feb 2025
Viewed by 356
Abstract
Arbuscular mycorrhizal fungi (AMF) play a pivotal role in sustainable agriculture by enhancing nutrient efficiency and reducing the dependence on synthetic fertilizers. Developing these sustainable, effective products requires knowledge of the target plant and its associated microbial communities in the production landscape of [...] Read more.
Arbuscular mycorrhizal fungi (AMF) play a pivotal role in sustainable agriculture by enhancing nutrient efficiency and reducing the dependence on synthetic fertilizers. Developing these sustainable, effective products requires knowledge of the target plant and its associated microbial communities in the production landscape of interest. This study focused on AMF populations associated with Tir wheat in six main locations of Türkiye’s Van Lake Basin. The Erçek-Özalp-Saray region exhibited the highest organic matter values. Higher available phosphorous contents were found for Erciş-Patnos and Muradiye. The Erciş-Patnos region exhibited the highest AMF density (120 spores/10 g soil) and frequency (75%), while the lowest AMF density (45 spores/10 g soil) was recorded in Muradiye. Sand contents correlated positively with spore number and mycorrhizal frequency and negatively with silt and clay. Based on these results, Erciş-Patnos was elected as the best location for the isolation of AMF spores suitable for the development of microbial-based tools for Tir wheat cultivation. These results are very important in the current context of climate change, which mandates the use of low-impact environmental strategies. Further research should explore the interactions of AMFs with other microorganisms to optimize their ecological benefits. However, the results of this study provide a valuable basis for future investigations of AMF-based products for use in sustainable Tir wheat cultivation. Full article
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Figure 1
<p>Results of soil physicochemical parameters (i.e., electrical conductivity, organic matter, available phosphorous, calcium carbonate contents, and USDA (United States Department of Agriculture) soil texture classification) organized based on the six sampling locations: Bitlis-Ahlat, Erciş-Patnos, Erçek-Özalp-Saray, Muradiye, Muş, and Van-Gevaş. For organic matter and phosphorous, results followed by different case letters are significantly different according to the Conover–Iman test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>AMF parameters (spore number, mycorrhizal frequency, and mycorrhizal density) determined in soil samples collected from different locations. Means followed by the same case letter (a–c) are not significantly different according to the Conover–Iman test (α = 0.05).</p>
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<p>Correlations among physicochemical and AMF soil parameters.</p>
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<p>Linear regression plots showing relationships between soil texture components (sand, silt, and clay) and AMF parameters. Significant differences were recorded for all correlations investigated (<span class="html-italic">p</span> &lt; 0.05). Coefficients of determination (R<sup>2</sup>) and <span class="html-italic">p</span>-values: SN vs. sand (R<sup>2</sup> = 0.074, <span class="html-italic">p</span> = 0.003), SN vs. silt (R<sup>2</sup> = 0.089, <span class="html-italic">p</span> = 0.001), SN vs. clay (R<sup>2</sup> = 0.036, <span class="html-italic">p</span> = 0.043), F% vs. sand (R<sup>2</sup> = 0.105, <span class="html-italic">p</span> = 0.0004), F% vs. silt (R<sup>2</sup> = 0.124, <span class="html-italic">p</span> = 0.0001) (spore number—SN, mycorrhizal frequency—F%).</p>
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17 pages, 1662 KiB  
Article
Enhancing Sorghum Growth: Influence of Arbuscular Mycorrhizal Fungi and Sorgoleone
by Isabela Figueiredo de Oliveira, Maria Lúcia Ferreira Simeone, Ubiraci Gomes de Paula Lana, Cristiane de Carvalho Guimarães and Sylvia Morais de Sousa Tinôco
Microorganisms 2025, 13(2), 423; https://doi.org/10.3390/microorganisms13020423 - 15 Feb 2025
Viewed by 375
Abstract
The low availability of phosphorus (P) in soil is one of the main constraints on crop production. Plants have developed several strategies to increase P use efficiency, including modifications in root morphology, the exudation of different compounds, and associations with microorganisms such as [...] Read more.
The low availability of phosphorus (P) in soil is one of the main constraints on crop production. Plants have developed several strategies to increase P use efficiency, including modifications in root morphology, the exudation of different compounds, and associations with microorganisms such as arbuscular mycorrhizal fungi (AMF). This study aimed to investigate the effect of sorgoleone compound on AMF colonization and its subsequent impact on P uptake, rhizosphere microbiota, and sorghum growth. The experiment was conducted in a greenhouse using the sorghum genotype P9401, known for low sorgoleone production. Three doses of purified sorgoleone (20 μM, 40 μM, and 80 μM) were added to low-P soil and plants were harvested after 45 days. Treatments included inoculation with the arbuscular mycorrhizal fungi Rhizophagus clarus and a negative control without inoculum. The addition of 40 and 80 μM of sorgoleone did not significantly increase mycorrhization. However, treatment with 20 μM sorgoleone combined with R. clarus inoculation significantly increased total sorghum biomass by 1.6-fold (p ≤ 0.05) compared to the non-inoculated treatment. AMF inoculation influenced only AMF colonization and the fungal microbiota, without affecting the bacterial community, whereas sorgoleone showed no effect on either. The activities of acid and alkaline phosphatases in the rhizospheric soil did not differ significantly among the treatments. Furthermore, the sorghum genes CYP71AM1, associated with sorgoleone biosynthesis, and Sb02g009880, Sb06g002560, Sb06g002540, and Sb03g029970 (related to phosphate transport induced by mycorrhiza) were significantly upregulated (p ≤ 0.05) in fine roots under these conditions. The 20 μM concentration of sorgoleone can enhance AMF colonization in sorghum and promote plant growth under low-P conditions, without significantly altering the microbiota. Full article
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<p>Chromatogram obtained using UPLC-QToF equipment for the sorgoleone standard at 500 ng mL<sup>−1</sup> (<b>A</b>) and extracted from sorghum genotype BR007B (<b>B</b>) analyzed on a Waters Acquity UPLC BEH column in positive mode, with extracted ion <span class="html-italic">m</span>/<span class="html-italic">z</span> 359.2222 [M+H]+. The analysis was performed at Embrapa Agroindústria Tropical in Fortaleza, CE. The arrow indicates the retention time of the sorgoleone standard (11.23 min) and BR007B (11.25 min), followed by its relative area (71—standard and 142—BR007B).</p>
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<p>Principal component analysis (PCA) for sorghum plants grown in a greenhouse under low-phosphorus soil supplemented with 0, 20, 40, and 80 µM of sorgoleone, and with (M+) and without (M−) inoculation of <span class="html-italic">Rhizophagus clarus</span>. SA: total surface area; TDW: total dry weight; P: total P content; and MYC: mycorrhization.</p>
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<p>Relative expression of genes of <span class="html-italic">CYP71AM1</span> (<span class="html-italic">21G12</span>), <span class="html-italic">RiEF</span>, <span class="html-italic">AM3</span> (<span class="html-italic">SbAM3</span>), <span class="html-italic">Sb02g009880</span> (<span class="html-italic">SbPT8</span>), <span class="html-italic">Sb06g002560</span> (<span class="html-italic">SbPT9</span>), <span class="html-italic">Sb06g002540</span> (<span class="html-italic">SbPT10</span>), and <span class="html-italic">Sb03g029970</span> (<span class="html-italic">SbPT11</span>) measured in the roots of sorghum plants grown in a greenhouse under low-P conditions with 0 (0SGL) and 20 µM (20SGL) of sorgoleone, with (M+) and without inoculation (M−) of <span class="html-italic">Rhizophagus clarus</span> after 45 days. Error bars are standard error of the mean (SEM) of three biological replications. The bars with the same letter do not differ by the Tukey test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Profile based on non-metric multidimensional scaling (NMDS) using the Bray–Curtis distance matrix to analyze the genetic diversity of the bacterial community (<b>A</b>) and arbuscular mycorrhizal fungi (<b>B</b>), relative abundance of the phyla of the bacterial community (<b>C</b>) and alkaline and acid phosphatase activities (<b>D</b>) from rhizosphere soil of sorghum grown in a greenhouse under low P with and without inoculation of <span class="html-italic">Rhizophagus clarus</span> and supplemented with 0 and 20 µM of sorgoleone. Error bars are standard error of the mean (SEM) of three biological replications. The bars with the same letter do not differ by the Tukey test (<span class="html-italic">p</span> &lt; 0.05).</p>
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22 pages, 2987 KiB  
Article
Responses of Arbuscular Mycorrhizal Fungi and Plant Communities to Long-Term Mining and Passive Restoration
by Sofía Yasmín Utge Perri, María Victoria Valerga Fernández, Adalgisa Scotti, Roxana Paula Colombo, Florencia González, Lautaro Valenzuela, Alicia Margarita Godeas and Vanesa Analía Silvani
Plants 2025, 14(4), 580; https://doi.org/10.3390/plants14040580 - 14 Feb 2025
Viewed by 349
Abstract
Mining activities cause strong soil alterations, such as heavy metal (HM) pollution, which decreases the diversity of plant communities and rhizospheric microorganisms, including arbuscular mycorrhizal (AM) fungi. The polymetallic Paramillos de Uspallata mine in the Andes Mountains, the first mining exploitation in Argentina, [...] Read more.
Mining activities cause strong soil alterations, such as heavy metal (HM) pollution, which decreases the diversity of plant communities and rhizospheric microorganisms, including arbuscular mycorrhizal (AM) fungi. The polymetallic Paramillos de Uspallata mine in the Andes Mountains, the first mining exploitation in Argentina, provides a unique scenario to study AM fungal resilience after long-term disturbance following over 40 years of inactivity. This study aimed to analyze mycorrhizal status and AM fungal communities in the mine and a nearby unexploited area and to evaluate their associations with soil parameters to elucidate life history strategies. Long-term exposure to elevated Fe, Pb, Zn, and Ag concentrations and high electrical conductivity (EC, 5.46 mS/cm) led to the dominance of Entrophospora infrequens in association with Pappostipa speciosa, demonstrating that this AM species is a stress-tolerant strategist in symbiosis with a pioneer perennial plant, resilient in the most impacted mine areas. In contrast, the unexploited area, with an EC of 0.48 mS/cm and low HM contents, supported competitive and ruderal species, revealing distinct ecological strategies of AM fungi in disturbed versus undisturbed environments. These findings highlight the potential of E. infrequens for bioremediation and ecological restoration in post-mining landscapes. Full article
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<p>Number of plants of each species and level of AM root colonization in different areas within Paramillos de Uspallata mine and outside the mine (Off-mine). AM colonization level: high = H; medium = M; low = L; No: No AM root colonization. Urban Ruins: Urban Ruins area; Exp 1: Exploitation area 1; Exp 2: Exploitation area 2; Off-mine: Off-mine area.</p>
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<p>Spore density (number of AM spores per 10 g dry soil) for each area in Paramillos de Uspallata mine and an area without mining exploitation. The mean ± standard error values (bars) are reported. Urban Ruins: Urban Ruins area; Exp 1: Exploitation area 1; Exp 2: Exploitation area 2; Off-mine: Off-mine area. The ends of the whiskers represent the minima and maxima, the bottom and top of the box are the first and third quartiles, respectively, and the line inside the box is the median.</p>
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<p>Relative abundance (%) of AM fungal species based on morphological taxonomy of spores for each area in Paramillos de Uspallata. The mean ± standard error values (bars) are reported. Urban Ruins (<b>a</b>); Exploitation area 1 (<b>b</b>); Exploitation area 2 (<b>c</b>); Off-mine area (<b>d</b>).</p>
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<p>AM fungal families (%) found in each area. Urban Ruins: Urban Ruins area; Exp 1: Exploitation area 1; Exp 2: Exploitation area 2; Off-mine: Off-mine area.</p>
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<p>Principal component analysis for the environmental variables analyzed in each area within Paramillos de Uspallata mine (Urban Ruins; Exploitation area 1: Exp 1; Exploitation area 2: Exp 2) and outside mine (Off-mine area). The color code and the angles of the vectors indicate the contribution of each variable to the PC 1 and PC 2 axes. AMF spores: abundance of AM spores; AMF richness: richness of AM fungal species; TDS: total dissolved solutes; EC: electrical conductivity; Total C: total carbon; Total P: total phosphorus.</p>
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<p>A non-metric multidimensional scaling (NMDS) plot performed using the Bray–Curtis dissimilarity matrix for the AM fungal communities and environmental parameters. Each green diamond represents AM fungal species and the color circles the areas of Urban Ruins; Exploitation area 1: Exp 1; Exploitation area 2: Exp 2, and Off-mine area. AMF spores: abundance of AM spores; AMF richness: richness of AM fungal species; TDS: total dissolved solutes; EC: electrical conductivity; Total C: total carbon; Total P: total phosphorus.</p>
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<p>Proportionality (%) of AM functional groups based on the life history strategies C-S-R in each area within Paramillos de Uspallata mine (Urban Ruins; Exploitation area 1: Exp 1; Exploitation area 2: Exp 2) and outside the mine (Off-mine area).</p>
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<p>Geographical location of Paramillos de Uspallata mine (<b>a</b>) in Mendoza (Argentina) (<b>left</b>) and the sampling areas (<b>right</b>): Urban Ruins area (<b>b</b>), Exploitation 1 area (<b>c</b>), Exploitation 2 area (<b>d</b>,<b>e</b>), and Off-mine area (<b>f</b>).</p>
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21 pages, 4477 KiB  
Article
Diversity of Arbuscular Mycorrhiza Fungi in the Arid Desert Ecosystems of Kuwait: Detection and Identification from Perennial Native Grass Roots
by Tareq A. Madouh, Majda K. Suleiman, Ali. M. Quoreshi and Merlin K. Davidson
Diversity 2025, 17(2), 130; https://doi.org/10.3390/d17020130 - 14 Feb 2025
Viewed by 332
Abstract
Arbuscular Mycorrhizal Fungi (AMF) play a key role in enduring stresses in desert ecosystems, as they enhance the moisture and nutrient supply to desert plants. An investigation was carried out to detect the existence of AMF in the root regions of five perennial [...] Read more.
Arbuscular Mycorrhizal Fungi (AMF) play a key role in enduring stresses in desert ecosystems, as they enhance the moisture and nutrient supply to desert plants. An investigation was carried out to detect the existence of AMF in the root regions of five perennial native desert grasses of Kuwait (Cenchrus ciliaris L., Cenchrus setigerus Vahl, Lasiurus scindicus Henrard, Pennisetum divisum (Forssk.) Henr., and Panicum turgidum Forssk.) in comparison with a non-native grass (Panicum virgatum L.). The native plants, C. setigerus and P. divisum had the highest colonization (100%) with vesicles, followed by P. turgidum (90%). The colonization of arbuscules was highest in the non-native grass, P. virgatum (60%), followed by C. setigerus (50%). Phylogenetic analysis for molecular identification to determine the genetic diversity of the AMF community in association with the native plant roots of the two dominant desert species, i.e., L. scindicus and P. turgidum, against P. virgatum, revealed a rich diversity. The AMF, Claroideoglomus lamellosum, and Rhizophagus sp. were identified from L. scindicus roots, Rhizophagus iranicus from P. turgidum roots, and Claroideoglomus lamellosum, from the non-native grass, P. virgatum, with almost 98–100% sequence similarity, indicating a significant difference between the mycorrhizal species in the nativity of grasses. This research confirms the diversity of AMF associated with native desert plants and emphasizes their symbiosis and host specificity. Thus, this study provides insight into AMF community structures, functions, and profiling, allowing us to understand their ecological and economic impacts, and ultimately implement strategies for sustainable biodiversity, productivity, and ecosystem management. Full article
(This article belongs to the Special Issue Biodiversity and Ecology of Soil Fungal Communities)
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<p>Map showing the location of the sampling area from the KISR’s Station for Research and Innovation—East of the State of Kuwait.</p>
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<p>Schematic representation of root staining procedure to determine the rate of AM fungal colonization.</p>
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<p>Percentage of AMF colonization in various native perennial desert grassroots; the common letters are not significantly different by DMRT (<span class="html-italic">p</span> = 0.05), (<span class="html-italic">n</span> = 5).</p>
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<p>AM Fungi structures observed in the root system of various native perennial desert grasses of Kuwait. Assessment of AM colonization revealed the presence of well-developed arbucules, vesicles, and hyphae with the roots of various desert grasses. Arrow indicates arbuscules in the roots of <span class="html-italic">Pennisetum divisum</span> (<b>A</b>), arbuscules in the roots of <span class="html-italic">Panicum turgidum</span> (<b>B</b>), vesicles and hyphae in the roots of <span class="html-italic">Cenchrus setigerus</span> (<b>C</b>), vesicles in the roots of <span class="html-italic">Lasiurus scindicus</span> (<b>D</b>), vesicles and hyphae in the roots of <span class="html-italic">Pennisetum divisum</span> (<b>E</b>), and vesicles and hyphae in the roots of <span class="html-italic">Panicum virgatum</span> (<b>F</b>), and confirmed that all above plants examined colonized by AM fungi with varied degree of colonization.</p>
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14 pages, 7540 KiB  
Article
The Effect of Rhizophagus intraradices on Cadmium Uptake and OsNRAMP5 Gene Expression in Rice
by Xiaoqi Bao, Junli Liu, Gaoyang Qiu, Xiaodong Chen, Junbo Zhang, Hua Wang, Quan Zhang and Bin Guo
Int. J. Mol. Sci. 2025, 26(4), 1464; https://doi.org/10.3390/ijms26041464 - 10 Feb 2025
Viewed by 520
Abstract
The molecular mechanism of arbuscular mycorrhizal fungi (AMF) in reducing cadmium (Cd) accumulation in plants remains unclear. In this respect, the effects of Rhizophagus intraradices (Ri) inoculation under Cd stress on rice growth, the uptake of Cd along with other elements, and the [...] Read more.
The molecular mechanism of arbuscular mycorrhizal fungi (AMF) in reducing cadmium (Cd) accumulation in plants remains unclear. In this respect, the effects of Rhizophagus intraradices (Ri) inoculation under Cd stress on rice growth, the uptake of Cd along with other elements, and the expression of Cd transport genes, including OsNRAMP1/5, were studied using wild-type (WT) and osnramp5 mutant rice. The results showed that Ri inoculation did not affect rice growth. The uptake of Cd of the osnramp5 mutant was much lower than the WT, as 27.6%, 17.5%, and 39.9% of Cd were noted in the grains, shoots, and roots, respectively. For the WT, Cd alone significantly promoted the OsNRAMP5 expression in shoots, but Ri inoculation significantly suppressed OsNRAMP5 expression and significantly reduced its grain and shoot Cd by 44.4% and 62.3%, respectively, compared to the Cd alone treatment. In contrast, for the osnramp5 mutant, Ri inoculation did not influence OsNRAMP5 expression or the grain and shoot Cd. Furthermore, the expression of other Cd transporters (OsIRT1, OsZIP3/7, OsCAX1a) in both varieties were not changed under the treatments. In conclusion, Ri inoculating significantly reduced Cd uptake by rice, with the molecular mechanism by negative regulation of expression of the OsNRAMP5 gene. Full article
(This article belongs to the Collection Advances in Molecular Plant Sciences)
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<p>The colonization picture (<b>a</b>) and total colonization rate (<b>b</b>) of the WT and <span class="html-italic">P</span> plant roots under different treatments (bar = 50 μm). “WT” indicates wild-type rice grown under normal conditions; “<span class="html-italic">P</span>” indicates the <span class="html-italic">osnramp5</span> mutant grown under normal conditions; “WT+Ri” indicates wild-type rice inoculated with Ri; “WT+Cd” indicates wild-type rice subjected to Cd stress; and “WT+Ri+Cd” indicates wild-type rice inoculated with Ri under Cd exposure. The “<span class="html-italic">P</span>+Ri” <span class="html-italic">osnramp5</span> mutant was inoculated with Ri; the “<span class="html-italic">P+</span>Cd” <span class="html-italic">osnramp5</span> mutant was subjected to Cd stress; and the “<span class="html-italic">P</span>+Ri+Cd” <span class="html-italic">osnramp5</span> mutant was inoculated with Ri under Cd exposure. The red arrow indicates the arbuscules.</p>
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<p>Comparison of the growth status of WT and <span class="html-italic">P</span> at maturity under different treatments (bar = 14.3 cm). The abbreviations used in <a href="#ijms-26-01464-f002" class="html-fig">Figure 2</a> were same as that of <a href="#ijms-26-01464-f001" class="html-fig">Figure 1</a>.</p>
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<p>The Cd concentration in seeds, shoots, and roots of the WT (<b>a</b>,<b>b</b>) and <span class="html-italic">P</span> (<b>c</b>,<b>d</b>) plants under the Ri and Cd treatments. For the data (means ± SEs, <span class="html-italic">n</span> = 5), different letters (a, b, c, d) indicate significant differences (<span class="html-italic">p</span> &lt; 0.05, Duncan’s test). A two-way ANOVA was used to test the interaction between the Ri treatment and Cd pollution. N: not significant, * <span class="html-italic">p</span> &lt; 0.05. The abbreviations used in <a href="#ijms-26-01464-f003" class="html-fig">Figure 3</a> were same as that of <a href="#ijms-26-01464-t001" class="html-table">Table 1</a>.</p>
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<p>The impact of the Ri and Cd treatments on the relative expression level of Cd transporters in the WT (<b>a</b>–<b>f</b>) and <span class="html-italic">P</span> (<b>g</b>–<b>l</b>) plants. Leaves were harvested separately for qRT-PCR analysis. Fold changes in the gene expression were normalized against the reference gene <span class="html-italic">OsACTIN</span>. For the data (means ± SEs, <span class="html-italic">n</span> = 5), different letters (a, b, c, d) indicate significant differences (<span class="html-italic">p</span> &lt; 0.05, Duncan’s test). A two-way ANOVA was used to test the interaction between the Ri treatment and Cd pollution. N: not significant, * <span class="html-italic">p</span> &lt; 0.05. The abbreviations used in <a href="#ijms-26-01464-f004" class="html-fig">Figure 4</a> were same as that of <a href="#ijms-26-01464-t001" class="html-table">Table 1</a>.</p>
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28 pages, 9460 KiB  
Article
Biocontrol of Verticillium Wilt of Olive Using Rhizophagus irregularis: Insights into Oxidative Stress and Phenolic Pathways
by Hanane Boutaj, Abdelilah Meddich, Said Wahbi, Zainab El Alaoui-Talibi, Allal Douira, Abdelkarim Filali-Maltouf and Cherkaoui El Modafar
Int. J. Plant Biol. 2025, 16(1), 21; https://doi.org/10.3390/ijpb16010021 - 8 Feb 2025
Viewed by 608
Abstract
Verticillium wilt of olive (VWO), caused by Verticillium dahliae (Vd), represents a significant threat to olive cultivation, particularly to the Moroccan cultivar “Picholine Marocaine”, by reducing yield, tree survival, and overall productivity. Current chemical and cultural control strategies provide limited efficacy, highlighting the [...] Read more.
Verticillium wilt of olive (VWO), caused by Verticillium dahliae (Vd), represents a significant threat to olive cultivation, particularly to the Moroccan cultivar “Picholine Marocaine”, by reducing yield, tree survival, and overall productivity. Current chemical and cultural control strategies provide limited efficacy, highlighting the need for alternative approaches such as biological control. This study investigated the potential of Rhizophagus irregularis (RI) as a biological agent to mitigate VWO over a 10-month period following V. dahliae inoculation. Disease severity, incidence, and defoliation rate as well as oxidant stress markers, antioxidant enzyme activities, and the phenolic pathway were evaluated in a greenhouse experiment. Our results showed that R. irregularis significantly reduced (p < 0.05) disease severity and incidence by about 31% and 26%, respectively, and decreased defoliation rates by 35% in RI-infected plants (RIV). The presence of R. irregularis triggered enhanced antioxidant enzyme activities (e.g., superoxide dismutase, catalase, and polyphenoloxidase), increased lignin deposition, elevated phenylalanine ammonia-lyase activity, and increased total phenol content in olive tissues. These biochemical and structural changes reduced lipid peroxidation, oxidative stress, and physiological damage, resulting in delayed disease progression. Enhanced accumulation of lignin and total phenolic compounds, particularly in aboveground tissues, was observed. This likely reinforced cell walls and enhanced resistance to pathogen infection. This study demonstrates that R. irregularis offers a promising biocontrol agent for combating soil-borne diseases in olives and other susceptible crops by mitigating VWO, boosting antioxidant defenses, and modulating the phenolic pathway with tissue-specific responses. Full article
(This article belongs to the Section Plant–Microorganisms Interactions)
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<p>Kinetic of disease severity of the “Picholine Marocaine” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and then post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means (<span class="html-italic">n</span> = 62).</p>
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<p>Kinetic of disease incidence of the “Picholine Marocaine” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and then post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means (<span class="html-italic">n</span> = 62).</p>
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<p>Kinetic of defoliation rate of the “Picholine Marocaine” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and then post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means (<span class="html-italic">n</span> = 10) and SDs are compared using Tukey’s HSD test at 5% level. Different letters indicate significant differences.</p>
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<p>Kinetic of hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) content in the leaves (<b>a</b>), stems (<b>b</b>), and roots (<b>c</b>) of the “Picholine Marocaine” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and subsequently post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means ± SDs were compared within each month. Bars with the same lowercase letter do not differ significantly over time (0–10 months) within each treatment, according to Tukey’s HSD test (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters (A, B, C, and D) indicate significant differences between treatments.</p>
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<p>Kinetic of hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) content in the leaves (<b>a</b>), stems (<b>b</b>), and roots (<b>c</b>) of the “Picholine Marocaine” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and subsequently post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means ± SDs were compared within each month. Bars with the same lowercase letter do not differ significantly over time (0–10 months) within each treatment, according to Tukey’s HSD test (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters (A, B, C, and D) indicate significant differences between treatments.</p>
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<p>Kinetic of malonyldialdehyde (MDA) content in the leaves (<b>a</b>), stems (<b>b</b>), and roots (<b>c</b>) of the “<span class="html-italic">Picholine Marocaine</span>” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and subsequently post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means ± SDs were compared within each month. Bars with the same lowercase letter do not differ significantly over time (0–10 months) within each treatment, according to Tukey’s HSD test (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters (A, B, C, and D) indicate significant differences between treatments.</p>
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<p>Kinetic of malonyldialdehyde (MDA) content in the leaves (<b>a</b>), stems (<b>b</b>), and roots (<b>c</b>) of the “<span class="html-italic">Picholine Marocaine</span>” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and subsequently post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means ± SDs were compared within each month. Bars with the same lowercase letter do not differ significantly over time (0–10 months) within each treatment, according to Tukey’s HSD test (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters (A, B, C, and D) indicate significant differences between treatments.</p>
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<p>Kinetic of superoxide dismutase (SOD) activity in the leaves (<b>a</b>), stems (<b>b</b>), and roots (<b>c</b>) of the “Picholine Marocaine” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and subsequently post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means ± SDs were compared within each month. Bars with the same lowercase letter do not differ significantly over time (0–10 months) within each treatment, according to Tukey’s HSD test (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters (A, B, C, and D) indicate significant differences between treatments.</p>
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<p>Kinetic of polyphenol oxidase (PPO) activity in the leaves (<b>a</b>), stems (<b>b</b>), and roots (<b>c</b>) of the “<span class="html-italic">Picholine Marocaine</span>” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and subsequently post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means ± SDs were compared within each month. Bars with the same lowercase letter do not differ significantly over time (0–10 months) within each treatment, according to Tukey’s HSD test (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters (A, B, C, and D) indicate significant differences between treatments.</p>
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<p>Kinetic of peroxidase (POD) activity in the leaves (<b>a</b>), stems (<b>b</b>), and roots (<b>c</b>) of the “Picholine Marocaine” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and subsequently post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means ± SDs were compared within each month. Bars with the same lowercase letter do not differ significantly over time (0–10 months) within each treatment, according to Tukey’s HSD test (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters (A, B, C, and D) indicate significant differences between treatments.</p>
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<p>Kinetic of catalase (CAT) activity in the leaves (<b>a</b>), stems (<b>b</b>), and roots (<b>c</b>) of the “<span class="html-italic">Picholine Marocaine</span>” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and subsequently post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means ± SDs were compared within each month. Bars with the same lowercase letter do not differ significantly over time (0–10 months) within each treatment, according to Tukey’s HSD test (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters (A, B, C, and D) indicate significant differences between treatments.</p>
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<p>Kinetic of lignin content in the leaves (<b>a</b>), stems (<b>b</b>), and roots (<b>c</b>) of the “<span class="html-italic">Picholine Marocaine</span>” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and subsequently post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means ± SDs were compared within each month. Bars with the same lowercase letter do not differ significantly over time (0–10 months) within each treatment, according to Tukey’s HSD test (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters (A, B, C, and D) indicate significant differences between treatments.</p>
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<p>Kinetic of phenylalanine ammonia-lyase (PAL) activity in the leaves (<b>a</b>), stems (<b>b</b>), and roots (<b>c</b>) of the “<span class="html-italic">Picholine Marocaine</span>” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and subsequently post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means ± SDs were compared within each month. Bars with the same lowercase letter do not differ significantly over time (0–10 months) within each treatment, according to Tukey’s HSD test (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters (A, B, C, and D) indicate significant differences between treatments.</p>
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<p>Kinetic of total phenol content in the leaves (<b>a</b>), stems (<b>b</b>), and roots (<b>c</b>) of the “<span class="html-italic">Picholine Marocaine</span>” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and subsequently post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Means ± SDs were compared within each month. Bars with the same lowercase letter do not differ significantly over time (0–10 months) within each treatment, according to Tukey’s HSD test (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters (A, B, C, and D) indicate significant differences between treatments.</p>
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<p>Principal component analysis (PCA) of different plant parts of the “Picholine Marocaine” olive cultivar under different treatments. Olive plants were pre-inoculated with <span class="html-italic">R. irregularis</span> for three months and subsequently post-inoculated with <span class="html-italic">V. dahliae</span> for 10 months. Oxidative stress, antioxidant defense, and phenolic enzyme pathway variables are represented in red, while the four treatments are shown in blue. Abbreviations: SOD: superoxide dismutase, H2O2: hydrogen peroxide, PPO: polyphenol oxidase, POD: peroxidase, CAT: catalase, and PAL: phenylalanine ammonia-lyase.</p>
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21 pages, 5901 KiB  
Article
Arbuscular Mycorrhizal Fungi Alleviate Cadmium Phytotoxicity by Regulating Cadmium Mobility, Physiological Responses, and Gene Expression Patterns in Malus hupehensis Rehd
by Xiaolei Zhuang, Siyu Liu, Shengzhe Xu, Sijun Qin, Deguo Lyu, Jiali He and Jiangtao Zhou
Int. J. Mol. Sci. 2025, 26(4), 1418; https://doi.org/10.3390/ijms26041418 - 7 Feb 2025
Viewed by 444
Abstract
Arbuscular mycorrhizal fungi (AMF) affect cadmium (Cd) accumulation and tolerance in host plants. However, the effects of AMF on Cd accumulation and phytotoxicity and their underlying mechanism in apples remain uncharacterized. In this study, the comprehensive physiological and molecular responses of uninoculated and [...] Read more.
Arbuscular mycorrhizal fungi (AMF) affect cadmium (Cd) accumulation and tolerance in host plants. However, the effects of AMF on Cd accumulation and phytotoxicity and their underlying mechanism in apples remain uncharacterized. In this study, the comprehensive physiological and molecular responses of uninoculated and Rhizophagus intraradices-inoculated Malus hupehensis Rehd. rootstocks exposed to 0 or 300 μM Cd were investigated. AMF inoculation mitigated Cd-induced growth and photosynthesis inhibition and nutrient ion disorders. It also lowered the concentrations of Cd in all tissues and reduced Cd transport to the shoots. Compared to uninoculated apple plants, those inoculated with mycorrhizal fungi reduced the mobility and toxicity of Cd by altering its form and binding it to the cell walls of the roots and leaves. AMF inoculation ameliorated Cd stress by altering endogenous phytohormone levels and triggering enzymatic and non-enzymatic antioxidant systems. Transcriptome analysis revealed that the differentially expressed genes (DEGs) associated with AMF under Cd stress regulated carbohydrate and amino acid biosynthesis and metabolism, as well as phytohormone biosynthesis and signal transduction. Furthermore, AMF inoculation downregulated certain genes involved in Cd uptake and transport while upregulating other genes involved in detoxification. These results suggest that AMF alleviate Cd phytotoxicity by orchestrated physiological and transcriptomic regulation in M. hupehensis Rehd., providing valuable insights into the efficacy of AMF inoculation in improving the heavy metal resistance of fruit trees. Full article
(This article belongs to the Special Issue Advances in Arbuscular Mycorrhizal Symbiosis)
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Figure 1
<p>Dry mass (<b>a</b>) and nutrient element content (<b>b</b>) of roots and leaves of <span class="html-italic">M. hupehensis</span> Rehd. without <span class="html-italic">R. intraradices</span> inoculation (NM) or with <span class="html-italic">R. intraradices</span> inoculation (M) exposed to 0 µM CdCl<sub>2</sub> or 300 µM CdCl<sub>2</sub> (+Cd) for 60 d. Data are means ± standard error of the mean (SE; <span class="html-italic">n</span> = 3). Different letters on bars indicate significant differences between treatments. <span class="html-italic">p</span>-values for ANOVA of CdCl<sub>2</sub> (Cd), AMF (M), and their interactions are shown. **: <span class="html-italic">p</span> ≤ 0.01; ****: <span class="html-italic">p</span> ≤ 0.0001; ns: not significant.</p>
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<p>Cd localization (<b>a</b>) in the roots and leaves; Cd concentrations (<b>b</b>) in the roots, stems, and leaves; and bioconcentration factors (BCFs) of the roots and aerial tissues and translocation factors (T<span class="html-italic"><sub>f</sub></span>) (<b>c</b>) for <span class="html-italic">M. hupehensis</span> Rehd. without <span class="html-italic">R. intraradices</span> inoculation (NM) or with <span class="html-italic">R. intraradices</span> inoculation (M) exposed to 0 µM CdCl<sub>2</sub> or 300 µM CdCl<sub>2</sub> (+Cd) for 60 d. Data are means ± standard error of the mean (SE; <span class="html-italic">n</span> = 3). Different letters on bars indicate significant differences between treatments. <span class="html-italic">p</span>-values for ANOVA of CdCl<sub>2</sub> (Cd), AMF (M), and their interactions are shown. *: <span class="html-italic">p</span> ≤ 0.05; **: <span class="html-italic">p</span> ≤ 0.01; ****: <span class="html-italic">p</span> ≤ 0.0001.</p>
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<p>Strigolactone (SL), indoleacetic acid (IAA), jasmonic acid (JA), and abscisic acid (ABA) content in the roots and leaves of <span class="html-italic">M. hupehensis</span> Rehd. without <span class="html-italic">R. intraradices</span> inoculation (NM) or with <span class="html-italic">R. intraradices</span> inoculation (M) exposed to 0 µM CdCl<sub>2</sub> or 300 µM CdCl<sub>2</sub> (+Cd) for 60 d. Data are means ± standard error of the mean (SE; <span class="html-italic">n</span> = 3). Different letters on bars indicate significant differences between treatments.</p>
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<p>Histochemical detection (<b>a</b>) of O<sub>2</sub><sup>•−</sup> and H<sub>2</sub>O<sub>2</sub>; O<sub>2</sub><sup>•−</sup> and H<sub>2</sub>O<sub>2</sub> concentrations (<b>b</b>); and principal component analysis (PCA) (<b>c</b>) of reactive oxygen species (ROS), non-enzymatic antioxidants, and antioxidant enzymes in the roots and leaves of <span class="html-italic">M. hupehensis</span> Rehd. without <span class="html-italic">R. intraradices</span> inoculation (NM) or with <span class="html-italic">R. intraradices</span> inoculation (M) exposed to 0 µM CdCl<sub>2</sub> or 300 µM CdCl<sub>2</sub> (+Cd) for 60 d. Data are means ± standard error of the mean (SE; <span class="html-italic">n</span> = 3). Values were normalized. Different letters indicate significant differences between treatments.</p>
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<p>Numbers of upregulated and downregulated differentially expressed genes (DEGs) (<b>a</b>) and Venn diagram of DEGs (<b>b</b>) in the roots of <span class="html-italic">M. hupehensis</span> Rehd. without <span class="html-italic">R. intraradices</span> inoculation (NM) or with <span class="html-italic">R. intraradices</span> inoculation (M) exposed to 0 µM CdCl<sub>2</sub> or 300 µM CdCl<sub>2</sub> (+Cd) for 60 d.</p>
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<p>Significantly differentially expressed genes involved in carbohydrate metabolism, amino acid metabolism, and lignin biosynthesis (<b>a</b>); and phytohormone signal transduction (<b>b</b>) in the roots of <span class="html-italic">M. hupehensis</span> Rehd. without <span class="html-italic">R. intraradices</span> inoculation (NM) or with <span class="html-italic">R. intraradices</span> inoculation (M) exposed to 0 µM CdCl<sub>2</sub> or 300 µM CdCl<sub>2</sub> (+Cd) for 60 d. Gradient color barcode represents normalized FPKM values. TPP: trehalose-6-phosphate phosphatase; GPI: glucose-6-phosphate isomerase; SUS: sucrose synthase; ATP-PFK: ATP-dependent 6-phosphofructokinase; TAT: tyrosine aminotransferase; CAD: cinnamyl-alcohol dehydrogenase; POD: peroxidase; AOS: allene oxide synthase; OPR: 12-oxophytodienoate reductase; OPCL: 4-coumarate-CoA ligase-like; MYC: myelocytomatosis; GH: Gretchen Hagen; BHLH: basic helix–loop–helix; IAA: indole-3-acetic acid inducible; ARF: auxin response factor.</p>
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<p>Schematic model of AMF-enhanced Cd tolerance in <span class="html-italic">M. hupehensis</span> Rehd. exposed to 0 µM CdCl<sub>2</sub> or 300 µM CdCl<sub>2</sub> (+Cd) for 60 d. Cd<sup>2+</sup> uptake, accumulation and detoxification in non-mycorrhizal seedling (left) and mycorrhizal seedling (right). Mycorrhizal seedlings exhibited lower root Cd accumulation and root-to-shoot translocation capacity but higher antioxidant defense activity, resulting in lower growth reduction than non-mycorrhizal seedlings. ZIP1.1, ZIP1.2, and ZIP5: zinc/iron-regulated transporter-related proteins 1.1, 1.2, and 5; YSL3.1 and YSL3.2: yellow stripe-like transporter 3.1 and 3.2; MT2: metallothionein 2.</p>
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19 pages, 2318 KiB  
Article
The Role of Vitekang Soil Conditioner and Arbuscular Mycorrhizae Fungi in Mitigating Cadmium Stress in Solanum lycopersicum Plants
by Qianqian Wang, Yue Liu, Guangxin Chen, Xing Liu, Mohsin Tanveer, Yongjun Guo, Peng Zeng and Liping Huang
Horticulturae 2025, 11(2), 179; https://doi.org/10.3390/horticulturae11020179 - 7 Feb 2025
Viewed by 426
Abstract
Solanum lycopersicum, a widely cultivated vegetable crop globally, faces soil cadmium (Cd) contamination issues due to Cd’s high mobility, posing potential threats to Solanum lycopersicum growth and human health. In light of this, this study selected three representative Solanum lycopersicum varieties: Micro [...] Read more.
Solanum lycopersicum, a widely cultivated vegetable crop globally, faces soil cadmium (Cd) contamination issues due to Cd’s high mobility, posing potential threats to Solanum lycopersicum growth and human health. In light of this, this study selected three representative Solanum lycopersicum varieties: Micro Tom, Red Guanyin, and Taiwan Pink King, and designed a series of experiments to investigate their growth performance under Cd stress. Experimental treatments included the sole application of different concentrations of Vitekang soil conditioner (VT), as well as the individual and combined application of VT and arbuscular mycorrhizal fungi (AMF). By thoroughly analyzing agronomic traits, cellular membrane lipid peroxidation levels, the activities of antioxidant enzymes (Catalase (CAT), Superoxide Dismutase (SOD), and Peroxidase (POD)), and the expression levels of genes related to Cd transport and detoxification (SLNRAMP6 and SlHMA3), this study comprehensively evaluated the effectiveness of different treatments in mitigating Cd stress in the three Solanum lycopersicum varieties. The results indicated that when VT was applied at a concentration of 2.4 g/kg in combination with AMF, it significantly reduced the detrimental effects of Cd on Micro Tom, Red Guanyin, and Taiwan Pink King. The specific experimental outcomes were as follows: (i) significantly decreased Cd accumulation in Solanum lycopersicum roots and leaves; (ii) effectively mitigated cellular membrane lipid peroxidation; (iii) significantly increased antioxidant enzyme activities; and (iv) influenced expression patterns of genes related to Cd transport and detoxification. This study further confirms that, compared to the sole application of VT or AMF, the combined application of these two treatments serves as a more effective practical method, exhibiting significant advantages in alleviating soil Cd contamination, promoting Solanum lycopersicum growth, and improving agronomic traits. This study not only advances research progress on VT and AMF in Solanum lycopersicumes, providing a solid theoretical and experimental foundation for cultivating high-quality Solanum lycopersicumes, but also holds significant importance for improving and optimizing the “VIP+N” technology, achieving farmland soil protection, and enhancing agricultural product quality. Full article
(This article belongs to the Section Vegetable Production Systems)
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<p>The diagram of agronomic traits.</p>
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<p>The effects of different treatments on MDA content (<b>a</b>), CAT activity (<b>b</b>), SOD activity (<b>c</b>), and POD activity (<b>d</b>) in <span class="html-italic">Solanum lycopersicum</span> leaves under cadmium stress, different letters in the figure represents significant difference.</p>
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<p>Different treatments of DAB (<b>a</b>) and NBT (<b>b</b>) staining of <span class="html-italic">Solanum lycopersicum</span> leaves under cadmium stress.</p>
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<p>Effects of different concentration treatments on the expression of <span class="html-italic">NRAMP6</span> (<b>a</b>) and <span class="html-italic">HMA3</span> (<b>b</b>) genes in <span class="html-italic">Solanum lycopersicum</span> (<b>c</b>) seedlings under cadmium stress. Different letters in the figure represents significant difference.</p>
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<p>The effect of different treatments on cadmium accumulation in <span class="html-italic">Solanum lycopersicum</span> roots (<b>a</b>) and leaves (<b>b</b>) under cadmium stress. Different letters in the figure represents significant difference.</p>
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<p>The effects of different treatments on MDA content (<b>a</b>), CAT activity (<b>b</b>), SOD activity (<b>c</b>), and POD activity (<b>d</b>) in <span class="html-italic">Solanum lycopersicum</span> leaves under cadmium stress, different letters in the figure represents significant difference.</p>
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19 pages, 3083 KiB  
Article
Effects of Nitrogen Addition on Soil Aggregate Stability and Mycorrhizal Morphological Characteristics: Differential Responses of Arbuscular Mycorrhizal and Ectomycorrhizal Fungi
by Mingxin Zhou and Yibo Li
Forests 2025, 16(2), 282; https://doi.org/10.3390/f16020282 - 6 Feb 2025
Viewed by 598
Abstract
Soil aggregate stability plays a pivotal role in ecosystem functioning and carbon sequestration. Nitrogen deposition influences aggregate stability and drives differential responses in AM and ECM fungi, yet the underlying mechanisms remain unclear. This study aimed to determine how N addition influences soil [...] Read more.
Soil aggregate stability plays a pivotal role in ecosystem functioning and carbon sequestration. Nitrogen deposition influences aggregate stability and drives differential responses in AM and ECM fungi, yet the underlying mechanisms remain unclear. This study aimed to determine how N addition influences soil aggregation, mycorrhizal morphological characteristics, and soil organic carbon (SOC) across two mycorrhizal types. A temperate forest experiment was conducted in northeastern China using 12 plots subjected to four N treatments (control, low, medium, high). Soil properties, soil aggregate traits, mycorrhizal morphological characteristics, and aggregate distributions were quantified. Relationships were examined via correlation analyses, random forest models, and structural equation modeling. N enrichment substantially increased SOC and soil water content, enhancing the proportion of large soil aggregates (4–8 mm) and elevating mean weight diameter (MWD) and geometric mean diameter (GMD), two critical indicators of soil aggregate stability. AM fungi exhibited a stronger response to N addition than ECM, reflected in greater hyphal development and carbon accumulation. SOC and water content correlated positively with aggregate stability, whereas soil pH showed a negative association. N inputs enhance soil structural stability by promoting SOC and water retention, with AM fungi demonstrating heightened sensitivity to N addition. These findings emphasize the integral role of mycorrhizal dynamics in shaping soil carbon stabilization under increasing N deposition. Full article
(This article belongs to the Section Forest Soil)
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<p>Sample plot and diagram of the experimental design in Wuying National Nature Reserve, Yichun City, Heilongjiang Province, northeastern China.</p>
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<p>Changes in mean weight diameter (<b>A</b>), geometric mean diameter (<b>B</b>), total hyphae length (<b>C</b>), total hyphae surface area (<b>D</b>), mean hyphae diameter (<b>E</b>), and mycorrhizal root colonization (<b>F</b>) across different treatments. CK represents the control, TL represents the low-N treatment (25 kg N·ha<sup>−1</sup>·yr<sup>−1</sup>), TM represents the medium-N treatment (50 kg N·ha<sup>−1</sup>·yr<sup>−1</sup>), and TH represents the high-N treatment (75 kg N·ha<sup>−1</sup>·yr<sup>−1</sup>). Different lowercase letters indicate significant differences between different treatments, while different capital letters denote statistically significant differences within the same treatment across different mycorrhizal types (ECM and AM). A significant difference is indicated when <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Heatmap of environmental factors with MWD and GMD (<b>A</b>); heatmap of environmental factors and mycorrhizal morphological characteristics (<b>B</b>). All data were log-transformed to meet the normality assumption. The color intensity in the heatmap represents the strength of the correlation, with the legend on the right side displaying the corresponding correlation coefficient values. Strong positive correlations are shown in shades of red, while weaker correlations are shown in shades of blue. The closer the value is to 1 or −1, the stronger the linear relationship between the variables. Symbols and acronyms of traits: SOC represents soil organic carbon, SWC represents soil water content, TN represents total nitrogen, TP represents total phosphorus, MWD represents mean weight diameter, GMD represents geometric mean diameter, THL represents total hyphae length, THS represents total hyphae surface area, MHD represents mean hyphae diameter, and MRC represents mycorrhizal root colonization.</p>
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<p>Relationships between soil organic carbon and mean weight diameter (<b>A</b>), geometric mean diameter (<b>B</b>), total hyphae length (<b>C</b>), total hyphae surface area (<b>D</b>), mean hyphae diameter (<b>E</b>), and mycorrhizal root colonization (<b>F</b>).</p>
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<p>Soil aggregate distribution under different treatments and different mycorrhizal types. CK represents the control, TL represents the low-N treatment (25 kg N·ha<sup>−1</sup>·yr<sup>−1</sup>), TM represents the medium-N treatment (50 kg N·ha<sup>−1</sup>·yr<sup>−1</sup>), and TH represents the high-N treatment (75 kg N·ha<sup>−1</sup>·yr<sup>−1</sup>). Different lowercase letters indicate significant differences between aggregate sizes in different treatments in the same aggregate size at the level of 0.05. Different capital letters indicate significant differences between different soil aggregate sizes in the same treatment at the level of 0.05.</p>
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<p>Random forest variable importance plot. The variables are ranked in order of relevance in predicting soil aggregate stability ((<b>A</b>) MWD, (<b>B</b>) GMD, (<b>C</b>) THL, (<b>D</b>) THS, (<b>E</b>) MHD, (<b>F</b>) MRC); colors in the graph represent the relative importance of each variable, dynamically assigned based on correlation strength. The importance measure considered for the analysis is the mean decrease in accuracy computed via the random forest classification algorithm. SOC represents soil organic carbon, SWC represents soil water content, TN represents total nitrogen, TP represents total phosphorus, MWD represents mean weight diameter, GMD represents geometric mean diameter, THL represents total hyphae length, THS represents total hyphae surface area, MHD represents mean hyphae diameter, and MRC represents mycorrhizal root colonization. A single asterisk (*) represents <span class="html-italic">p</span> &lt; 0.05; double asterisks (**) represent <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Structural equation model of soil aggregates traits and mycorrhizal morphological characteristics. The red arrow indicates positive correlation; the grey arrow indicates no significant correlation. SOC represents soil organic carbon, MWD represents mean weight diameter, GMD represents geometric mean diameter, THL represents total hyphae length, THS represents total hyphae surface area, MHD represents mean hyphae diameter, and MRC represents mycorrhizal root colonization. *** <span class="html-italic">p</span> &lt; 0.001.</p>
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14 pages, 3474 KiB  
Article
Indigenous Arbuscular Mycorrhizal Fungi Consortium Enhances Growth and Protects Boufeggous Gharas Date Palm Against Fusarium oxysporum f. sp. albedinis Infection in Figuig Oasis (Morocco)
by Elmostafa Gagou, Halima Bouchentouf, Khadija Chakroune, Mahmoud Abbas, Touria Lamkami, Mondher El Jaziri and Abdelkader Hakkou
Int. J. Plant Biol. 2025, 16(1), 20; https://doi.org/10.3390/ijpb16010020 - 6 Feb 2025
Viewed by 621
Abstract
An indigenous consortium of arbuscular mycorrhizal fungi (AMF) from the Figuig oasis in southern Morocco, comprising Rhizophagus sp., Funneliformis sp., Acaulospora sp., Sclerocystis sp., and Scutellospora sp., was evaluated for its effects on the growth and sensitivity of Phoenix dactylifera L. variety (cv. [...] Read more.
An indigenous consortium of arbuscular mycorrhizal fungi (AMF) from the Figuig oasis in southern Morocco, comprising Rhizophagus sp., Funneliformis sp., Acaulospora sp., Sclerocystis sp., and Scutellospora sp., was evaluated for its effects on the growth and sensitivity of Phoenix dactylifera L. variety (cv. Boufeggous Gharas) to Fusarium oxysporum f. sp. Albedinis (Foa), the causal agent of Bayoud disease. Despite its high fruit quality and local appreciation, the Boufeggous Gharas variety is highly susceptible to Foa and is currently at risk of extinction, underscoring the urgent need for its sustainable management. The results demonstrated that Boufeggous Gharas seedlings inoculated with a consortium of indigenous AMF showed significantly improved shoot and root length, leaf number, and biomass as compared to non-mycorrhized seedlings. In contrast, Foa-infected seedlings showed significantly reduced growth, with a 46.6% decrease in shoot height and a 50.4% reduction in root length compared to non-infected seedlings. Interestingly, AMF inoculation mitigated this sensitivity to Foa, significantly restoring growth parameters. Seedlings treated with AMF + Foa showed a 51% increase in shoot height and a 61% improvement in root length, along with over 100% gains in shoot and root biomass compared to seedlings infected solely with Foa. This study provides the first evidence of integrating AMF into sustainable date palm cultivation practices to mitigate the impacts of biotic stresses, thereby promoting the preservation and valorization of vulnerable date palm varieties. The protective effects of AMF are attributed to improved nutrient uptake, enhanced root architecture, and systemic resistance induced by AMF colonization. Full article
(This article belongs to the Section Plant–Microorganisms Interactions)
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<p>Experimental protocol developed for this research.</p>
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<p>Ten-week-old date palm seedlings (cv. Boufeggous Gharas) grown under control conditions.</p>
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<p>Representative photomicrographs of AMF spores from the consortium isolated in the Figuig oasis. (<b>A</b>) Intra- and extraradical spores of <span class="html-italic">Rhizophagus</span> sp. (<b>B</b>) Loose clusters of <span class="html-italic">Funneliformis</span> sp. spores. (<b>C</b>) Compact and dense clusters of <span class="html-italic">Rhizophagus</span> sp. spores. (<b>D</b>) Aggregated (fascicle) spores of <span class="html-italic">Sclerocystis</span> sp. (<b>E</b>) Spores of <span class="html-italic">Scutellospora</span> sp. mounted in Melzer’s reagent. (<b>F</b>) <span class="html-italic">Acaulospora</span> sp. spores showing hyphal connections. Scale bars: A = 1 mm; B = 2 mm; C = 500 µm; D = 2 mm; E = 100 µm; F = 100 μm.</p>
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<p>Morphological characteristics of <span class="html-italic">Fusarium oxysporum</span> f. sp. <span class="html-italic">albedinis</span> (Foa) isolated from Boufeggous Gharas rachis fragments from trees suffering from vascular fusariosis. Cottony white-to-pink mycelium on PDA after 7 days of incubation at 27 °C. Scale bar: 2 cm.</p>
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<p>Mycorrhization of Boufeggous Gharas seedling roots with the indigenous consortium of AMF after 16 weeks of culture. (<b>A</b>) Intraradical spores (S) observed under optical microscopy. Scale bar = 500 µm. (<b>B</b>) Extraradical mycelium (ME) and spores (S) colonizing the roots of date palm seedlings. Scale bar = 500 µm. (<b>C</b>) Frequency and intensity of mycorrhization (%). NM: Non-mycorrhized seedlings. M: Mycorrhized seedlings. Error bars represent standard deviation, and different letters indicate significant differences according to Student’s <span class="html-italic">t</span>-test at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Growth responses in Boufeggous Gharas seedlings after 16 weeks of culture in presence of AMF consortium. (<b>A</b>) Shoot height (cm) and root height (cm). (<b>B</b>) Number of leaves per plant. (<b>C</b>) Shoot fresh weight (g) and root fresh weight (g). (<b>D</b>) Shoot dry weight (g) and root dry weight (g). NM: Non-mycorrhized seedlings. M: Mycorrhized seedlings. Different letters above the bars indicate significant differences according to Student’s <span class="html-italic">t</span>-test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Growth responses in 10-week-old Boufeggous Gharas seedlings. Control condition for 48 weeks (c). Infected with Foa for 32 weeks (Foa). First inoculated with AMF for 16 weeks followed by infection with Foa for additional 32 weeks (AMF + Foa). (<b>A</b>) Shoot height (cm) and root height (cm). (<b>B</b>) Shoot fresh weight (g) and root fresh weight (g). (<b>C</b>) Shoot dry weight (g) and root dry weight (g). (<b>D</b>) Number of leaves per plant. Different letters above the bars indicate significant differences according to one way ANOVA followed by Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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19 pages, 6500 KiB  
Article
Half-Century Scientometric Analysis: Unveiling the Excellence of Fungi as Biocontrol Agents and Biofertilisers
by Ziqi Yuan, Qi Shen, Kefei Yu, Yan Liu, Huabao Zheng, Yanlai Yao and Baolei Jia
J. Fungi 2025, 11(2), 117; https://doi.org/10.3390/jof11020117 - 4 Feb 2025
Viewed by 851
Abstract
Reducing the use of chemical inputs is becoming a major challenge in developing sustainable agriculture. Fungi, known as biocontrol agents (BCAs) and biofertilisers, are crucial in scientific research and are celebrated for their efficacy, eco-friendliness, and multifaceted roles. In this study, a bibliometric [...] Read more.
Reducing the use of chemical inputs is becoming a major challenge in developing sustainable agriculture. Fungi, known as biocontrol agents (BCAs) and biofertilisers, are crucial in scientific research and are celebrated for their efficacy, eco-friendliness, and multifaceted roles. In this study, a bibliometric analysis was conducted on 5349 articles related to fungi as BCAs and biofertilisers over the past half-century using the Web of Science Core Collection (WoSCC) database. The publications on fungi, such as BCAs and biofertilisers, have increased significantly over the last 20 years, with a maximum growth rate of 33.7%. The USA and China lead in this field. Keyword clustering analysis revealed that entomopathogenic fungi, including Hemiptera, Coleoptera, and Lepidoptera, can be used to manage plant pests. It also showed that fungi can be used as biofertilisers to promote plant growth. The analysis of research trends shows that Beauveria bassiana in biological control is highly significant. This study also showed that entomopathogenic fungi control plant pests by infiltrating the insect cuticles. Trichoderma spp. exert biocontrol effects by producing antibiotics. Arbuscular mycorrhizal fungi can trigger plant defence mechanisms by modulating secondary metabolite synthesis. This study contributes to the current knowledge of fungi as BCAs and biofertilisers and can guide future research. Full article
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<p>Comprehensive overview and flowchart of the study design.</p>
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<p>Literature publication and citations of fungi as BCAs and biofertilisers. (<b>A</b>) Number of publications on fungi as BCAs and biofertilisers and total citations per year, 1976–2024. (<b>B</b>) Total citations and average annual citations of the top ten most cited literatures on fungi as BCAs and biofertilisers [<a href="#B38-jof-11-00117" class="html-bibr">38</a>,<a href="#B39-jof-11-00117" class="html-bibr">39</a>,<a href="#B40-jof-11-00117" class="html-bibr">40</a>,<a href="#B41-jof-11-00117" class="html-bibr">41</a>,<a href="#B42-jof-11-00117" class="html-bibr">42</a>,<a href="#B43-jof-11-00117" class="html-bibr">43</a>,<a href="#B44-jof-11-00117" class="html-bibr">44</a>,<a href="#B45-jof-11-00117" class="html-bibr">45</a>,<a href="#B46-jof-11-00117" class="html-bibr">46</a>,<a href="#B47-jof-11-00117" class="html-bibr">47</a>].</p>
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<p>Number of publications on fungi as BCAs and biofertilisers in different countries and international cooperation. (<b>A</b>) The annual growth curve of article frequency for the top six countries on fungi as BCAs and biofertilisers from 1976 to 2024. (<b>B</b>) The top ten countries by total citation count. Numbers represent separately the total citations and the average article citations. (<b>C</b>) Global cooperation network in the field of fungal BCAs and biofertilisers. The intensity of the colour shades corresponds to the scientific productivity levels, with deeper colours indicating higher productivity. The numbers indicate the number of publications from each country.</p>
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<p>Sankey diagram showing the proportion of fungi as research subjects for BCAs and biofertilisers by affiliation and journal (<b>A</b>) and by country and journal (<b>B</b>) The sankey diagram consists of a series of “Nodes” and “Arcs”, that are read from left to right, with the thickness of each line proportional to the value it is representing.</p>
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<p>Keywords co-occurrence map on fungi as BCAs and biofertilisers. (<b>A</b>) Keywords co-occurrence network map. The blue sections illustrate the role of fungi as BCAs and biofertilisers in the comprehensive management of plant pests. The green section provides the types of pests that fungi can eliminate. The red sections of the analysis highlight the application of fungi as BCAs and biofertilisers in the effective control of plant diseases. (<b>B</b>) Co-occurrence network map of the year of occurrence (Frequency 100). Different colours correspond to different years. The closer the colour is to yellow, the more recent the year it represents. (<b>C</b>) Word cloud of keywords. The size of the font indicates the frequency of keyword occurrences. The larger the font, the higher the frequency. The circle indicates the frequency of keyword occurrence.</p>
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<p>Trends on topics related to fungi as BCAs and biofertilisers. (<b>A</b>) The development trend of keywords with a frequency exceeding 100 occurrences. The size and colour intensity of the circles represent changes in frequency. Larger circles and darker colours indicate higher occurrence rates. (<b>B</b>) Thematic map of keywords. The horizontal axis represents the level of importance, while the vertical axis indicates the level of development. The first quadrant represents niche topics, characterised by low importance but high development. The second quadrant includes popular topics, which are highly important and well developed. The third quadrant represents declining topics with low importance and low development, while the fourth quadrant contains basic topics, which are of high importance but still in the early stages of development. (<b>C</b>) The thematic evolution of keywords in three stages. The length of the stripes represents the frequency of keyword occurrence. Longer stripes signify higher keyword frequencies during that period. The thickness of each line is directly proportional to the value it represents.</p>
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<p>Mechanism of fungi as BCAs and biofertilisers. (<b>A</b>) Mechanism and effect of EPF as BCAs (insecticides). (<b>B</b>) The biocontrol mechanism and effect of <span class="html-italic">Trichoderma</span> in controlling plant pathogens and promoting crop health. (<b>C</b>) Mechanism and effect of AMF in controlling plant diseases and promoting plant growth. Figures created with BioRender.com.</p>
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21 pages, 3439 KiB  
Article
Labile Carbon Input Mitigates the Negative Legacy Effects of Nitrogen Addition on Arbuscular Mycorrhizal Symbiosis in a Temperate Grassland
by Sitong Liu, Yuxiao Zhang, Xiaoqian Yu, Meng Cui, Liangchao Jiang, Tao Zhang and Yingzhi Gao
Plants 2025, 14(3), 456; https://doi.org/10.3390/plants14030456 - 4 Feb 2025
Viewed by 422
Abstract
Nitrogen (N) deposition and carbon (C) addition significantly influence the dynamics of plant–microbe interactions, particularly altering the symbiotic relationship between plants and arbuscular mycorrhizal fungi (AMF). However, the effects and underlying mechanisms of labile C input on the relationship between AMF and various [...] Read more.
Nitrogen (N) deposition and carbon (C) addition significantly influence the dynamics of plant–microbe interactions, particularly altering the symbiotic relationship between plants and arbuscular mycorrhizal fungi (AMF). However, the effects and underlying mechanisms of labile C input on the relationship between AMF and various plant species in a nitrogen-enriched environment remain a knowledge gap. A seven-year field experiment was conducted to examine how six levels of N and three levels of labile C addition impact AMF colonization in four key plant species: Leymus chinensis (Trin. ex Bunge) Tzvelev, Stipa baicalensis Roshev., Thermopsis lanceolata R. Br. and Potentilla bifurca Linn. Our results showed that N and C additions exert significantly different effects on the relationship between AMF and various plant species. Labile C addition mitigated historical N negative effects, particularly for S. baicalensis, enhancing AMF infection and promoting nutrient exchange under high-N and low-C conditions. The relationship between AMF and both L. chinensis and T. lanceolata changed to weak mutualism under low-N and high-C conditions, with significant decreases in vesicular and arbuscular abundance. Plant root stoichiometry plays a critical role in modulating AMF symbiosis, particularly under high-N and -C conditions, as reflected in the increased AMF infection observed in T. lanceolata and P. bifurca. Our findings emphasize the species-specific and nutrient-dependent AMF symbiosis, revealing that targeted C input can mitigate the legacy effects of N enrichment. Effective nutrient management is of crucial importance for ecological restoration efforts in temperate grasslands affected by long-term N enrichment. Full article
(This article belongs to the Special Issue Plant-Soil Microbe Interactions in Ecosystems)
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<p>Effects of historical N enrichment and subsequent labile C addition on arbuscular mycorrhizal fungi (AMF) infection density of plant roots across different plant species. (<b>a</b>) <span class="html-italic">L. chinensis</span> (Trin. ex Bunge) Tzvelev (perennial rhizomatous grass); (<b>b</b>) <span class="html-italic">S. baicalensis</span> Roshev. (perennial bunchgrass); (<b>c</b>) <span class="html-italic">T. lanceolata</span> R. Br. (legume); (<b>d</b>) <span class="html-italic">P. bifurca</span> Linn. (forb). CK represents control treatment with only water added, while Low C and High C correspond to C addition levels of 200 and 2000 g C m<sup>−2</sup> yr<sup>−1</sup>, respectively. Error bars represent ±SE (n = 8). Data were fitted using linear, polynomial or nonlinear curve fitting methods, selecting the most appropriate curve type for each C level.</p>
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<p>Effects of historical N enrichment and subsequent labile C addition on arbuscular mycorrhizal fungi (AMF) structural abundance in different plant species. (<b>a</b>–<b>d</b>) Vesicular abundance; (<b>e</b>–<b>h</b>) Hyphal abundance; (<b>i</b>–<b>l</b>) Arbuscular abundance. CK denotes control plots with only water added, while Low C and High C correspond to C addition levels of 200 and 2000 g C m<sup>−2</sup> yr<sup>−1</sup>, respectively. Error bars represent ±SE (n = 8). Data were fitted using linear, polynomial or nonlinear curve fitting methods, with the most suitable curve type chosen for each C level.</p>
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<p>Effects of historical N enrichment and subsequent labile C addition on surface soil (0–10 cm) hyphal length density of arbuscular mycorrhizal fungi. CK represents control treatment with only water added, while Low C and High C correspond to carbon addition levels of 200 and 2000 g C m<sup>−2</sup> yr<sup>−1</sup>, respectively. Error bars represent ±SE (n = 8). Data were fitted using linear, polynomial or nonlinear curve fitting methods, selecting the most appropriate curve type for each C level.</p>
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<p>Effects of historical N enrichment and subsequent labile C addition on root C, N and P stoichiometric ratios in different plant species. (<b>a</b>–<b>d</b>) Root C/P ratio; (<b>e</b>–<b>h</b>) Root N/P ratio; (<b>i</b>–<b>l</b>) Root C/N ratio. CK denotes control with only water added; Low C and High C correspond to carbon addition levels of 200 and 2000 g C m<sup>−2</sup> yr<sup>−1</sup>, respectively. Error bars represent ±SE (n = 8). Data were fitted using linear, polynomial or nonlinear curve fitting methods, selecting the most suitable curve type for each C level.</p>
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<p>Importance of historical N enrichment and subsequent labile C addition in explaining variance in the first principal component (PC1) of AMF infection metrics across different plant species based on random forest analysis. The plant species represented are (<b>a</b>) <span class="html-italic">L. chinensis</span> (Trin. ex Bunge) Tzvelev, explaining 66.47% of the variance; (<b>b</b>) <span class="html-italic">S. baicalensis</span> Roshev., explaining 77.40% of the variance; (<b>c</b>) <span class="html-italic">T. lanceolata</span> R. Br., explaining 69.51% of the variance; and (<b>d</b>) <span class="html-italic">P. bifurca</span> Linn., explaining 74.48% of the variance. The importance of each variable is measured as the percentage increase in mean squared error (%IncMSE) when the variable is randomly permuted. NH<sub>4</sub><sup>+</sup> indicates soil NH<sub>4</sub><sup>+</sup>-N; NO<sub>3</sub><sup>−</sup> indicates soil NO<sub>3</sub><sup>−</sup>-N; RC, RN, RP, RC/N, RC/P and RN/P represent root carbon, nitrogen, phosphorus content and their ratios; SIN refers to soil inorganic N; SOC indicates soil organic C; SWC indicates soil water content; DOC indicates soil dissolved organic C; DN indicates soil dissolved N; C/N represents the ratio of DOC to DN; AP indicates soil available P; TP indicates soil total P. Significant variables are marked with asterisks: * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Structural equation models (SEMs) illustrating the relationships between N addition rates, C addition rates, key variables and the first principal component (PC1) of AMF infection metrics across different plant species. The models illustrate the direct and indirect effects of N and C addition on AMF infection metrics through selected variables. (<b>a</b>) <span class="html-italic">L. chinensis</span> (Trin. ex Bunge) Tzvelev: PC1 explains 66.47% of the variance; (<b>b</b>) <span class="html-italic">S. baicalensis</span> Roshev.: PC1 explains 77.40% of the variance; (<b>c</b>) <span class="html-italic">T. lanceolata</span> R. Br.: PC1 explains 69.51% of the variance; (<b>d</b>) <span class="html-italic">P. bifurca</span> Linn.: PC1 explains 74.48% of the variance. Path coefficients are indicated on each arrow. Green solid lines represent significant positive correlations, while red solid lines represent significant negative correlations (<span class="html-italic">p</span> &lt; 0.05). Dotted lines represent non-significant relationships (<span class="html-italic">p</span> &gt; 0.05). The thickness of the lines corresponds to the strength of the relationship. Model fit indices at the bottom of each panel indicate a good fit for each SEM (e.g., χ<sup>2</sup>/df, RMSEA, CFI, GFI, TLI). NH<sub>4</sub><sup>+</sup>-N indicates soil NH<sub>4</sub><sup>+</sup>-N; NO<sub>3</sub><sup>−</sup>-N indicates soil NO<sub>3</sub><sup>−</sup>-N; Root C, Root C/N and Root C/P represent root C, the ratio of root C and N and the ratio of root C and P; DOC indicates soil dissolved organic C. Significance levels are indicated as follows: * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Diagram illustrating the effects of historical N enrichment and C addition on AMF symbiosis. (<b>a</b>) Low historical N and low C: A mutualistic symbiosis between host plants and AMF is established through balanced nutrient exchange. Host plants provide C to AMF in exchange for essential minerals. (<b>b</b>) High N and low C: Phosphorus becomes limited due to N accumulation. Plants increasingly rely on AMF for P, thereby strengthening the mutualistic relationship. (<b>c</b>) The addition of sucrose allowed the plants to increase the amount of carbon transported to the AMF through the mycorrhizal interface without being constrained by the resource of soil nutrients and the relationship became a weak mutualism. (<b>d</b>) High N and high C: While nitrogen is abundant, increased C input reduces N availability and exacerbates P limitation. This fosters a strong mutualistic relationship, with AMF playing a crucial role in plant nutrient acquisition. Figure adapted from Ma et al. [<a href="#B13-plants-14-00456" class="html-bibr">13</a>].</p>
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20 pages, 4652 KiB  
Article
Optimizing Tomato Seedling Production in the Tropics: Effects of Trichoderma, Arbuscular Mycorrhizal Fungi, and Key Agronomical Factors
by Teresa Leuratti, Lorenzo Fellin, Nicola Michelon, Juan Bosco Palacios Tario, Jaime Ernesto Santamaria Gutiérrez, Giorgio Gianquinto, Francesco Orsini and Giampaolo Zanin
Agronomy 2025, 15(2), 392; https://doi.org/10.3390/agronomy15020392 - 31 Jan 2025
Viewed by 516
Abstract
Agriculture remains a key contributor to Central America’s economy, despite climate change posing a significant threat to the sector. In the Trifinio region, already afflicted by arid summers, temperatures are expected to rise in the near future, potentially exacerbating the vulnerability of smallholder [...] Read more.
Agriculture remains a key contributor to Central America’s economy, despite climate change posing a significant threat to the sector. In the Trifinio region, already afflicted by arid summers, temperatures are expected to rise in the near future, potentially exacerbating the vulnerability of smallholder farmers. This study investigates the effects of two fungal symbionts, Trichoderma asperellum (TR) and the Arbuscular mycorrhiza fungi (AMF) Glomus cubense, and agronomic choices and practices such as cultivar selection, substrate type, and fertigation management on tomato (Solanum lycopersicum L.) seedling growth and quality. Results showed that nutrient solution and the adoption of forest topsoil as substrate significantly enhanced morphological, physiological, and quality parameters. Modifying the nutrient solution to allow for an increase in plant height of 170% and a dry weight of 163% and enhancing Dickson’s quality index (DQI) by 64.5%, while the use of forest topsoil resulted in plants 58.6% higher, with an increase of 101% in dry weight and of 90.1% in the DQI. Both T. asperellum and G. cubense had positive effects on specific growth parameters; for instance, TR increased leaf number (+6.95%), while AMF increased stem diameter (+3.56%) and root length (+19.1%), although they did not, overall, significantly increase the seedling’s biomass and quality. These findings underscore the importance of agronomic practices in mitigating the impacts of climate change on tomato production, offering valuable insights for farmers in semi-arid regions. Full article
(This article belongs to the Section Horticultural and Floricultural Crops)
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Figure 1

Figure 1
<p>Climatic parameters recorded from the start (S) to the end (E) of the experiment. Black lines represent the daily temperatures (T), while the grey line indicates the relative humidity (RH).</p>
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<p>Effect of the five treatment variables on: plant height (<b>A</b>), leaf number (<b>B</b>), stem diameter (<b>C</b>), and root length (<b>D</b>). Histograms represent the average values, and error bars represent the standard error mean (SEM). Only significant differences for <span class="html-italic">p</span> ≤ 0.01 are reported, resulting from the Tukey’s honestly significant difference test (HSD). **: <span class="html-italic">p</span> ≤ 0.01. ***: <span class="html-italic">p</span> ≤ 0.001. US = forest soil, PEAT = peat substrate; NM = no mycorrhization; M = no mycorrhization; NTR = no Trichoderma; TR = with Trichoderma; WA = only water; NS = nutrient solution; PON = Pony; VAL = Valiente.</p>
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<p>Effect of the five treatment variables on total dry weight (<b>A</b>), dry matter partitioning (<b>B</b>), percentage of dry matter (<b>C</b>), and shoot/root ratio (<b>D</b>). Histograms represent the average values, and error bars represent the standard error mean (SEM). Only significant differences for <span class="html-italic">p</span> ≤ 0.01 are reported, resulting from the Tukey’s honestly significant difference test (HSD). **: <span class="html-italic">p</span> ≤ 0.01. ***: <span class="html-italic">p</span> ≤ 0.001. See the caption of <a href="#agronomy-15-00392-f002" class="html-fig">Figure 2</a> for abbreviations.</p>
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<p>Leaf temperature parameter recorded for the five treatment variables. Histograms represent the average values, and error bars represent the standard error mean (SEM). Only significant differences for <span class="html-italic">p</span> ≤ 0.01 are reported, resulting from the Tukey’s honestly significant difference test (HSD). **: <span class="html-italic">p</span> ≤ 0.01. ***: <span class="html-italic">p</span> ≤ 0.001. See the caption of <a href="#agronomy-15-00392-f002" class="html-fig">Figure 2</a> for abbreviations.</p>
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<p>Effect of the five treatment variables on stem diameter ratio (<b>A</b>), sturdiness quotient (<b>B</b>), root dry weight/root length ratio (<b>C</b>), and Dickson Quality Index (<b>D</b>). Histograms represent the average values, and error bars represent the standard error mean (SEM). Only significant differences for <span class="html-italic">p</span> ≤ 0.01 are reported, resulting from the Tukey’s honestly significant difference test (HSD). **: <span class="html-italic">p</span> ≤ 0.01. ***: <span class="html-italic">p</span> ≤ 0.001. See the caption of <a href="#agronomy-15-00392-f002" class="html-fig">Figure 2</a> for abbreviations.</p>
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<p>Effect of the substrate × irrigation interaction on plant height (<b>A</b>), stem diameter ratios (<b>B</b>), leaf temperature (<b>C</b>), and shoot/root ratio (<b>D</b>), percentage of dry matter partitioning (<b>E</b>). Histograms represent the average value, and error bars indicate the standard error mean (SEM). Only significant differences for <span class="html-italic">p</span> ≤ 0.01 are reported, resulting from the Tukey’s honestly significant difference test (HSD) for <span class="html-italic">p</span> ≤ 0.01. Different lowercase letters indicate significant differences. See the caption of <a href="#agronomy-15-00392-f002" class="html-fig">Figure 2</a> for abbreviations.</p>
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<p>Effect of the substrate × AMF interaction on plant height (<b>A</b>), leaf temperature (<b>B</b>), and percentage of dry matter (<b>C</b>). Histograms represent the average value, and error bars indicate the standard error mean (SEM). Only significant differences for <span class="html-italic">p</span> ≤ 0.01 are reported, resulting from the Tukey’s honestly significant difference test (HSD) for <span class="html-italic">p</span> ≤ 0.01. Different lowercase letters indicate significant differences. See the caption of <a href="#agronomy-15-00392-f002" class="html-fig">Figure 2</a> for abbreviations.</p>
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<p>Effect of the substrate × <span class="html-italic">Trichoderma</span> interaction on leaf number (<b>A</b>), root length (<b>B</b>), root dry weight/root length (<b>C</b>), and Dickson Quality Index (<b>D</b>). Histograms represent the average value, and error bars indicate the standard error mean (SEM). Only significant differences for <span class="html-italic">p</span> ≤ 0.01 are reported, resulting from the Tukey’s honestly significant difference test (HSD) for <span class="html-italic">p</span> ≤ 0.01. Different lowercase letters indicate significant differences. See the caption of <a href="#agronomy-15-00392-f002" class="html-fig">Figure 2</a> for abbreviations.</p>
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<p>Effect of the <span class="html-italic">Trichoderma</span> × fertigation interaction on plant height (<b>A</b>), leaf number (<b>B</b>) and stem diameter (<b>C</b>). Histograms represent the average value, and error bars indicate the standard error mean (SEM). Only significant differences for <span class="html-italic">p</span> ≤ 0.01 are reported, resulting from the Tukey’s honestly significant difference test (HSD) for <span class="html-italic">p</span> ≤ 0.01. Different lowercase letters indicate significant differences. See the caption of <a href="#agronomy-15-00392-f002" class="html-fig">Figure 2</a> for abbreviations.</p>
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<p>Effect of the AMF × fertigation interaction on root length (<b>A</b>), and leaf temperature (<b>B</b>). Histograms represent the average value, and error bars indicate the standard error mean (SEM). Only significant differences for <span class="html-italic">p</span> ≤ 0.01 are reported, resulting from the Tukey’s honestly significant difference test (HSD) for <span class="html-italic">p</span> ≤ 0.01. Different lowercase letters indicate significant differences. See the caption of <a href="#agronomy-15-00392-f002" class="html-fig">Figure 2</a> for abbreviations.</p>
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<p>Effect of Substrate × variety interaction on root dry weight / root length ration (<b>A</b>), shoot/root ratio (<b>B</b>), dry matter partitioning (<b>C</b>), and Dickson Quality Index (<b>D</b>). Histograms represent the average value, and error bars indicate the standard error mean (SEM). Only significant differences for <span class="html-italic">p</span> ≤ 0.01 are reported, resulting from the Tukey’s honestly significant difference test (HSD) for <span class="html-italic">p</span> ≤ 0.01. Different lowercase letters indicate significant differences. See the caption of <a href="#agronomy-15-00392-f002" class="html-fig">Figure 2</a> for abbreviations.</p>
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<p>Variety × AMF interaction on percentage of dry matter Histograms represent the average value, and error bars indicate the standard error mean (SEM). Only significant differences for <span class="html-italic">p</span> ≤ 0.01 are reported, resulting from the Tukey’s honestly significant difference test (HSD) for <span class="html-italic">p</span> ≤ 0.01. Different lowercase letters indicate significant differences. See the caption of <a href="#agronomy-15-00392-f002" class="html-fig">Figure 2</a> for abbreviations.</p>
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