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16 pages, 4957 KiB  
Article
Growth-Promoting Effects of Grass Root-Derived Fungi Cadophora fastigiata, Paraphoma fimeti and Plectosphaerella cucumerina on Spring Barley (Hordeum vulgare) and Italian Ryegrass (Lolium multiflorum)
by Izolda Pašakinskienė, Violeta Stakelienė, Saulė Matijošiūtė, Justas Martūnas, Marius Rimkevičius, Jurga Būdienė, Algis Aučina and Audrius Skridaila
Microorganisms 2025, 13(1), 25; https://doi.org/10.3390/microorganisms13010025 - 26 Dec 2024
Viewed by 199
Abstract
Many endophytic fungi are approved as plant growth stimulants, and several commercial biostimulants have already been introduced in agricultural practice. However, there are still many species of fungi whose plant growth-promoting properties have been understudied or not studied at all. We examined the [...] Read more.
Many endophytic fungi are approved as plant growth stimulants, and several commercial biostimulants have already been introduced in agricultural practice. However, there are still many species of fungi whose plant growth-promoting properties have been understudied or not studied at all. We examined the growth-promoting effect in spring barley (Hordeum vulgare) and Italian ryegrass (Lolium multiflorum) induced by three endophytic fungi previously obtained from the roots of Festuca/Lolium grasses. Surface-sterilized seeds were inoculated with a spore suspension of Cadophora fastigiata (isolate BSG003), Paraphoma fimeti (BSG010), Plectosphaerella cucumerina (BSG006), and their spore mixture. Before harvesting, the inoculated plants were grown in a greenhouse, with the barley being in multi-cavity trays for 30 days and ryegrass being placed in an original cylindric element system for 63 days. All three newly tested fungi had a positive effect on the growth of the barley and ryegrass plants, with the most pronounced impact observed in their root size. The fungal inoculations increased the dry shoot biomass between 11% and 26% in Italian ryegrass, but no such impact was observed in barley. The highest root increment was observed in barley. Herein, P. cucumerina and C. fastigiata inoculations were superior to other treatments, showing an increase in root dry weight of 50% compared to 20%, respectively. All fungal inoculations significantly promoted root growth in Italian ryegrass, resulting in a 20–30% increase in dry weight compared to non-inoculated plants. Moreover, a strong stimulatory effect of the fungi-emitted VOCs on the root development was observed in plate-in-plate arrays. In the presence of C. fastigiata and P. cucumerina cultures, the number of roots and root hairs in barley seedlings doubled compared to control plants. Thus, in our study, we demonstrated the potential of the grass root-derived endophytes C. fastigiata, P. fimeti, and P. cucumerina as growth promoters for spring barley and Italian ryegrass. These studies can be extended to other major crops and grasses by evaluating different fungal isolates. Full article
(This article belongs to the Special Issue Rhizosphere Bacteria and Fungi That Promote Plant Growth)
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<p>Spring barley plants affected by fungal inoculations after 30 days of growth in multi-cavity trays. (<b>A</b>) Control plants, no fungal inoculation; (<b>B</b>–<b>E</b>) inoculations as follows: <span class="html-italic">Cadophora fastigiata</span> BSG003 (<b>B</b>), <span class="html-italic">Paraphoma fimeti</span> BSG010 (<b>C</b>); <span class="html-italic">Plectosphaerella cucumerina</span> BSG006 (<b>D</b>); the three fungi mix (<b>E</b>); (<b>F</b>,<b>G</b>) representative views of the plants from <span class="html-italic">P. cucumerina</span> treatment in a multi-cavity tray (<b>G</b>), and a root display from the bottom (<b>F</b>). Scale bar 3 cm.</p>
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<p>Spring barley growth parameters affected by fungal inoculations after 30 days of growth in multi-cavity trays. (<b>A</b>) green shoot biomass; (<b>B</b>) dry root biomass. Different letters (a, b, c and d) above the bars indicate significant differences between the treatments (<span class="html-italic">p</span> ≤ 0.05) based on Tukey’s HSD post hoc test.</p>
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<p>A view of the experimental panel of Italian ryegrass plants affected by fungal inoculations after 63 days of growth in the cylinder elements system. (<b>A</b>) A representative view of five blocks of plants from <span class="html-italic">Cadophora fastigiata</span> BSG003, <span class="html-italic">Paraphoma fimeti</span> BSG010, <span class="html-italic">Plectosphaerella cucumerina</span> BSG006, the three fungi mix inoculations and the control; (<b>B</b>–<b>D</b>) representative views of the plants from <span class="html-italic">C. fastigiata</span> treatment: roots in the soil at the moment of the opening of geotextile bags (<b>B</b>), control plant roots measured (<b>C</b>) next to the plant roots from <span class="html-italic">C. fastigiata</span> inoculation (<b>D</b>).</p>
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<p>Italian ryegrass growth parameters affected by fungal inoculations after 63 days of growth in the cylinder elements system. (<b>A</b>) Shoot and root height; (<b>B</b>) shoot green and dry biomass; (<b>C</b>) shoot number; (<b>D</b>) dry root biomass. Different letters (a, b, c and d) above the bars indicate significant differences between the treatments (<span class="html-italic">p</span> ≤ 0.05) based on Tukey’s HSD post hoc test. In (<b>A</b>), regular font for root length, bold—for shoot height; in (<b>B</b>), regular—for green shoot weight, bold—for dry shoot weight; in (<b>C</b>), regular—for total stem no., bold—for nodding stem no.</p>
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<p>The effect on of barley root development exposed to VOCs of endophytic fungi <span class="html-italic">Cadophora fastigiata</span> BSG003 and <span class="html-italic">Plectosphaerella cucumerina</span> BSG006. (<b>A</b>) The effect on the number of main and lateral roots after 7 days of growth; (<b>B</b>) the effect on the number of root hairs on the 5th, 6th and 7th days in a 0.5 mm<sup>2</sup> area. (<b>C</b>–<b>E</b>) Spring barley plants grown in plate-in plate assays (top images) and microscopical images of the roots (bottom images) under the exposure of fungal cultures: (<b>C</b>) control, no fungal culture in small plates, (<b>D</b>) barley with <span class="html-italic">C. fastigiata</span> BSG003, (<b>E</b>) barley with <span class="html-italic">P. cucumerina</span> BSG010. Different letters (a, b, c and d) above the bars indicate significant differences between the treatments (<span class="html-italic">p</span> ≤ 0.05) based on Tukey’s HSD post hoc test. In (<b>A</b>), regular font for main root no., bold—for lateral root no.; in (<b>B</b>), regular—for control, bold—for <span class="html-italic">C. fastigiata</span>, italicized—for <span class="html-italic">P. cucumerina</span>.</p>
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21 pages, 20591 KiB  
Article
New Species of Diaporthales (Ascomycota) from Diseased Leaves in Fujian Province, China
by Xiayu Guan, Taichang Mu, Nemat O. Keyhani, Junya Shang, Yuchen Mao, Jiao Yang, Minhai Zheng, Lixia Yang, Huili Pu, Yongsheng Lin, Mengjia Zhu, Huajun Lv, Zhiang Heng, Huiling Liang, Longfei Fan, Xiaoli Ma, Haixia Ma, Zhenxing Qiu and Junzhi Qiu
J. Fungi 2025, 11(1), 8; https://doi.org/10.3390/jof11010008 - 26 Dec 2024
Viewed by 163
Abstract
Fungal biota represents important constituents of phyllosphere microorganisms. It is taxonomically highly diverse and influences plant physiology, metabolism and health. Members of the order Diaporthales are distributed worldwide and include devastating plant pathogens as well as endophytes and saprophytes. However, many phyllosphere Diaporthales [...] Read more.
Fungal biota represents important constituents of phyllosphere microorganisms. It is taxonomically highly diverse and influences plant physiology, metabolism and health. Members of the order Diaporthales are distributed worldwide and include devastating plant pathogens as well as endophytes and saprophytes. However, many phyllosphere Diaporthales species remain uncharacterized, with studies examining their diversity needed. Here, we report on the identification of several diaporthalean taxa samples collected from diseased leaves of Cinnamomum camphora (Lauraceae), Castanopsis fordii (Fagaceae) and Schima superba (Theaceae) in Fujian province, China. Based on morphological features coupled to multigene phylogenetic analyses of the internal transcribed spacer (ITS) region, the large subunit of nuclear ribosomal RNA (LSU), the partial beta-tubulin (tub2), histone H3 (his3), DNA-directed RNA polymerase II subunit (rpb2), translation elongation factor 1-α (tef1) and calmodulin (cal) genes, three new species of Diaporthales are introduced, namely, Diaporthe wuyishanensis, Gnomoniopsis wuyishanensis and Paratubakia schimae. This study contributes to our understanding on the biodiversity of diaporthalean fungi that are inhabitants of the phyllosphere of trees native to Asia. Full article
(This article belongs to the Special Issue Diversity of Microscopic Fungi)
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<p>Consensus tree of <span class="html-italic">Diaporthe virgiliae</span> species complex inferred from Bayesian inference analyses based on the combined ITS, <span class="html-italic">cal, his3, tef1</span> and <span class="html-italic">tub2</span> sequence dataset, with <span class="html-italic">Diaporthe shennongjiaensis</span> (CNUCC 201905) as the outgroup. The Maximum likelihood (ML) bootstrap support values and Bayesian posterior probabilities (BPPs) above 80% and 0.90 are shown at the nodes. Strains marked with “T” are ex-type, ex-epitype and ex-neotype. The isolates from this study are indicated in red.</p>
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<p>Consensus tree of <span class="html-italic">Gnomoniopsis</span> inferred from Bayesian inference analyses based on the combined ITS, <span class="html-italic">tef1</span> and <span class="html-italic">tub2</span> sequence dataset, with <span class="html-italic">Melanconis stilbostoma</span> (CBS 109778) as the outgroup. The Maximum likelihood (ML) bootstrap support values and Bayesian posterior probabilities (BPPs) above 80% and 0.90 were shown at the nodes. Strains marked with “T” are ex-type, ex-epitype and ex-neotype. The isolates from this study are indicated in red.</p>
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<p>Consensus tree of <span class="html-italic">Tubakiaceae</span> inferred from Bayesian inference analyses based on the combined ITS, LSU, <span class="html-italic">rpb2</span>, <span class="html-italic">tef1</span> and <span class="html-italic">tub2</span> sequence dataset, with <span class="html-italic">Greeneria uvicola</span> (FI12007) as the outgroup. The Maximum likelihood (ML) bootstrap support values and Bayesian posterior probabilities (BPPs) above 80% and 0.90 were shown at the nodes. Strains marked with “T” are ex-type, ex-epitype and ex-neotype. The isolates from this study are indicated in red.</p>
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<p><span class="html-italic">Diaporthe wuyishanensis</span> (HMAS 352949). (<b>a</b>) Diseased leaves of <span class="html-italic">Cinnamomum camphora</span>; (<b>b</b>,<b>c</b>) surface and reverse sides of colony after 7 days on PDA (<b>d</b>,<b>e</b>) and 14 days; (<b>f</b>,<b>g</b>) conidiomata; (<b>h</b>) conidiogenous cells and conidia; and (<b>i</b>,<b>j</b>) alpha conidia. Scale bars: (<b>h</b>–<b>j</b>) 10 µm.</p>
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<p><span class="html-italic">Gnomoniopsis wuyishanensis</span> (HMAS 353149). (<b>a</b>) Diseased leaves of <span class="html-italic">Castanopsis fordii</span>; (<b>b</b>) surface and reverse sides of colony after 7 days on PDA (<b>c</b>) and 14 days; (<b>d</b>,<b>e</b>) conidiomata; (<b>f</b>–<b>k</b>) conidiogenous cells and conidia; and (<b>l</b>,<b>m</b>) conidia. Scale bars: (<b>f</b>–<b>m</b>) 10 µm.</p>
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<p><span class="html-italic">Paratubakia schimae</span> (HMAS 353150). (<b>a</b>) Diseased leaves of <span class="html-italic">Schima superba</span>; (<b>b</b>) surface and reverse sides of colony after 7 days on PDA (<b>c</b>) and 14 days; (<b>d</b>,<b>e</b>) conidiomata; (<b>f</b>–<b>j</b>) conidiogenous cells and conidia; and (<b>k</b>,<b>l</b>) conidia. Scale bars: (<b>f</b>–<b>l</b>) 10 µm.</p>
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17 pages, 1151 KiB  
Article
Fungal Biodiversity of Apple Bark, Leaves, Stems, and Fruit Under Rain Shelters with Reduced Fungicide Schedule
by Claudia Maria Oliveira Longa, Lidia Nicola, Massimo Pindo, Elisa Bozza, Carmela Sicher, Daniel Bondesan, Ilaria Pertot and Michele Perazzolli
Agriculture 2025, 15(1), 17; https://doi.org/10.3390/agriculture15010017 - 25 Dec 2024
Viewed by 26
Abstract
The use of rain shelters is a promising agronomic practice to protect crops from rainfall, reducing the need for fungicides to control certain pathogens that take advantage of leaf wetness. However, the combined condition of absence of rain and reduced fungicide schedule can [...] Read more.
The use of rain shelters is a promising agronomic practice to protect crops from rainfall, reducing the need for fungicides to control certain pathogens that take advantage of leaf wetness. However, the combined condition of absence of rain and reduced fungicide schedule can affect the fungal populations, possibly favoring biocontrol agents and/or other pathogens. In this study, the effects this practice on epiphytic and endophytic fungal communities associated with barks, leaves, flowers, and fruits of two apple cultivars (Fuji and Golden Delicious) were evaluated across two seasons. Apple plants were grown under two conditions in a commercial-like orchard: (1) covered by rain shelters with reduced fungicide schedule and (2) uncovered with standard integrated pest management (IPM) schedule. The use of rain shelters combined with reduced fungicide applications affects the overall fungal community structure and their abundance of specific taxa. Leaf epiphytes were the most impacted community, and fungal communities also differed between the two apple cultivars. The use of rain shelters helped reduce fungicide input in the orchard, but it increased the abundance of potential pathogens compared to the IPM in open field conditions, such as powdery mildew and apple scab. Understanding how the plant microbiome responds to new practices that help in reducing fungicides can help developing strategies that avoid the build-up of potentially new pathogens. Full article
(This article belongs to the Section Crop Protection, Diseases, Pests and Weeds)
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<p>Relative abundances of fungal communities at phylum level for each apple plant tissue of Golden and Fuji cultivars, sampled inside (IN) and outside (OUT) the rain shelter. IN and OUT conditions were treated according to integrated pest management guidelines, with IN condition not receiving fungicide treatments against apple scab.</p>
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<p>Composition of the most abundant fungal families of endophytes and epiphytes of different apple tissues. Taxa composition is reported for each plant cultivar (Golden or Fuji), time of sampling (T0, T1, T2, T3, T4; and T5), and growth condition, such as inside (IN) or outside (OUT) the rain shelter.</p>
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<p>Richness (Observed Species) and diversity indices (Shannon and Simpson) of endophytes and epiphytes of different apple tissues. Taxa composition is reported for each time of sampling (T0, T1, T2, T3, T4, and T5) and growth condition in the IN condition (rain shelter + reduced fungicide applications) and in the OUT condition (open field + standard IPM program).</p>
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<p>Principal Coordinates Analysis (PCoA) based on the Bray–Curtis dissimilarity distance matrix of Illumina sequencing data of fungal communities in apple tree tissues (Fuji cultivar in round symbol and Golden cultivar in triangle symbol) in the IN condition (rain shelter + reduced fungicide applications; green color) and in the OUT condition (open field + standard IPM program; orange color).</p>
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20 pages, 3582 KiB  
Article
Diversity and Plant Growth-Promoting Properties of Rhodiola rosea Root Endophytic Bacteria
by Inga Tamošiūnė, Muhammad Fahad Hakim, Odeta Buzaitė, Vidmantas Stanys, Jurgita Vinskienė, Elena Andriūnaitė and Danas Baniulis
Microorganisms 2025, 13(1), 13; https://doi.org/10.3390/microorganisms13010013 - 25 Dec 2024
Viewed by 36
Abstract
Plants inhabiting environments with suboptimal growth conditions often have a more pronounced capacity to attract and sustain microbial communities that improve nutrient absorption and expand abiotic stress tolerance. Rhodiola rosea L. is a succulent plant of the Crassulaceae family adapted to survive in [...] Read more.
Plants inhabiting environments with suboptimal growth conditions often have a more pronounced capacity to attract and sustain microbial communities that improve nutrient absorption and expand abiotic stress tolerance. Rhodiola rosea L. is a succulent plant of the Crassulaceae family adapted to survive in sandy or rocky soils or dry tundra. The aim of the present study was to investigate the diversity and plant growth-stimulating potential of R. rosea endophytic microbiota. Metataxonomic analysis of the bacterial diversity in the rhizome of R. rosea revealed 108 families. Among these, three families were found exclusively in the core microbiome of 1-year-old plants, while nine families were unique to the core microbiome of mature plants grown in the field for more than 4 years. Seventy-three endophytic bacteria isolates were obtained from the rhizome of R. rosea plants and were assigned into 14 distinct bacterial genera of Firmicutes (26%) or Proteobacteria (74%) phyla. Screening for functional genes related to the nitrogen cycle, phosphorus mineralisation or dissolution, and traits associated with nitrogen fixation (56% of isolates), siderophore production (40%), inorganic phosphorus solubilisation (30%), and production of indole-related compounds (51%) led to the classification of the isolates into 16 distinct clusters. Co-cultivation of 45 selected isolates with germinating Arabidopsis seedlings revealed 18 and 5 isolates that resulted in more than a 20% increase in root or shoot growth, respectively. The study results established the complexity of the succulent R. rosea endophytic microbiome and identified isolates for potential plant growth-stimulating applications. Full article
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<p>Composition and relative abundance of family-level endophytic bacterial taxa in 1-year-old and &gt;4-year-old <span class="html-italic">R. rosea</span> rhizome samples assessed using metataxonomic sequencing of <span class="html-italic">16S rRNA</span> variable region V4 and V5–7 amplicons.</p>
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<p>Bacterial diversity variation in 1-year-old and &gt;4-year-old <span class="html-italic">R. rosea</span> rhizome samples. The nonmetric multidimensional scaling (NMDS) analysis of the ASV datasets generated using metataxonomic sequencing of <span class="html-italic">16S rRNA</span> variable region V4 and V5–7 amplicons was carried out using the Bray–Curtis dissimilarity matrix.</p>
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<p>Core microbiome of <span class="html-italic">R. rosea</span> 1-year-old (<b>upper panel</b>) and &gt;4-year-old (<b>lower panel</b>) rhizome samples estimated based on <span class="html-italic">16S rRNA</span> variable region V4 and V5–7 amplicon metataxonomic sequencing. The threshold of &gt;5% counts and &gt;30% samples was used for calculation using the MicrobiomeAnalyst server [<a href="#B57-microorganisms-13-00013" class="html-bibr">57</a>]. Bacterial classes are represented by different colours.</p>
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<p>Phylogenetic tree reconstruction for endophytic isolates obtained from the rhizome of <span class="html-italic">R. rosea</span>. A phylogenetic tree was built using the iTOL server v.7 [<a href="#B56-microorganisms-13-00013" class="html-bibr">56</a>] and representative sequence data. The numbers represent the count of isolates.</p>
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<p>Plant growth-promoting traits of endophytic bacteria isolated from the rhizome of <span class="html-italic">R. rosea</span>. The numbers in red font indicate representative clusters. The isolate numbers shown in blue or green colour font indicate isolates selected for growth regulation analysis using Arabidopsis seedlings, and those marked in green colour font showed the most prominent growth-enhancing effect. Positive test results are indicated by “+” sign. Abbreviations: NF—nitrogen fixation; PS—phosphate solubilisation; SP—siderophore production; IRC—indole-related compound production (mg mL<sup>−1</sup>); RMA—cellular redox-modulating activity in tobacco cell culture (% of control).</p>
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<p>Arabidopsis seedling radicle (blue fill colour)- and hypocotyl (green fill colour)-growth-modulating effect of selected endophytic bacterial isolates obtained from <span class="html-italic">R. rosea</span> rhizome. The data are shown as boxplots representing the median, minimum and maximum scores, and lower and upper quartiles; the red dashed line indicates the mean value of control; numbers indicate clusters as shown in <a href="#microorganisms-13-00013-f005" class="html-fig">Figure 5</a>; different letters denote significant differences between the analysed groups (<span class="html-italic">p</span> ≤ 0.05).</p>
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24 pages, 1704 KiB  
Article
In Vitro Antioxidant and Antibacterial Activities of Ethyl Acetate Extracts of Ziziphus lotus Leaves and Five Associated Endophytic Fungi
by Amel Ghazi-Yaker, Bart Kraak, Jos Houbraken, El-hafid Nabti, Cristina Cruz, Noria Saadoun and Karim Houali
Microorganisms 2024, 12(12), 2671; https://doi.org/10.3390/microorganisms12122671 - 23 Dec 2024
Viewed by 373
Abstract
The exploration of new pharmacological compounds from endophytic fungi offers infinite possibilities. The aim of this study was to evaluate the antibacterial and antioxidant activities of extracts from the leaves of Ziziphus lotus and five of its endophytic fungi and investigate the chemical [...] Read more.
The exploration of new pharmacological compounds from endophytic fungi offers infinite possibilities. The aim of this study was to evaluate the antibacterial and antioxidant activities of extracts from the leaves of Ziziphus lotus and five of its endophytic fungi and investigate the chemical diversity of the secondary metabolites produced. Isolated, purified, and molecularly identified endophytes and plant leaves were subjected to ethyl acetate extraction. The antibacterial potential of the extracts was assessed by the disc diffusion method against five bacterial strains: Staphylococcus aureus ATCC 25923; Staphylococcus aureus MU50; Enterococcus faecalis WDCM00009; Escherichia coli ATCC 25922; and Pseudomonas aeruginosa ATCC 27853. DPPH and reducing power tests were performed to assess antioxidant potential. GC–MS analysis was used to identify volatile compounds in extracts. Fungal endophytes were identified as Aspergillus cavernicola, Aspergillus persii, Alternaria alternata, Cladosporium asperlatum, and Fusarium incarnatum–equiseti complex, with respective accession numbers DTO 412-G6, DTO 412-I5, DTO 413-E7, DTO 412-G4, and DTO 414-I2. GC–MS analysis revealed a large number of bioactive compounds. All extracts showed antibacterial activity against at least two of the bacteria tested, and most showed antioxidant activity. The Aspergillus cavernicola extract stood out for its higher phenolic content and higher antioxidant and antibacterial activities in all tests. Full article
(This article belongs to the Special Issue Endophytic Fungus as Producers of New and/or Bioactive Substances)
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<p>Antibacterial activity of ethyl acetate extracts (<b>A</b>) <span class="html-italic">S. aureus</span> ATCC 25923, (<b>B</b>) <span class="html-italic">S. aureus</span> MU50, (<b>C</b>) <span class="html-italic">E. faecalis</span> WDCM00009 (<b>D</b>) <span class="html-italic">E. coli</span> ATCC 25922, and (<b>E</b>) <span class="html-italic">P. aerugenosa</span> ATCC 27853 (F.iec: <span class="html-italic">Fusarium incarnatum–equiseti</span> complex; <span class="html-italic">A.c: Aspergillus cavernicola</span>; <span class="html-italic">A.p: Aspergillus persii</span>; C.a: <span class="html-italic">Cladosporium asperlatum</span>; A.a: <span class="html-italic">Alternaria alternata</span>; Z.l: <span class="html-italic">Ziziphus lotus</span>; DMSO: Dimethylsulfoxide; AT: antibiotic Chloramphenicol).</p>
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<p>DPPH radical scavenging activities of endophytic fungi and <span class="html-italic">Z. lotus</span> leaves ethyl acetate extracts in comparison with ascorbic acid.</p>
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<p>Reducing power activity of samples and ascorbic acid at 1 mg/mL.</p>
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<p>Total phenolic contents in <span class="html-italic">Z. lotus</span> and its endophytic fungi.</p>
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13 pages, 2165 KiB  
Article
Phaseolus vulgaris (L.) Growth Promotion by Cladosporium halotolerans Inoculation Under Salinity Stress
by Samah Alzahrani, Azhar Najjar, Samah Noor, Nidal M. Zabermawi, Shaza Y. Qattan, Shahira A. Hassoubah, Rania M. Makki and Manal El-Zohri
Soil Syst. 2024, 8(4), 135; https://doi.org/10.3390/soilsystems8040135 - 23 Dec 2024
Viewed by 220
Abstract
Salinization induced by salt stress is a critical environmental factor, and limits the expansion of agricultural areas and population distribution in continental regions, including Saudi Arabia. Common beans, a vital source of protein, energy, and dietary fibers, are negatively affected by salt stress. [...] Read more.
Salinization induced by salt stress is a critical environmental factor, and limits the expansion of agricultural areas and population distribution in continental regions, including Saudi Arabia. Common beans, a vital source of protein, energy, and dietary fibers, are negatively affected by salt stress. In this research, the endophytic fungus Cladosporium halotolerans was utilized to remediate saline soils and enhance common bean growth and resilience. The results of this study demonstrated that soil treatment with C. halotolerans enhances the soil properties by decreasing soil pH and increasing soil organic matter content under saline conditions. Inoculation by C. halotolerans also significantly improved plant growth parameters, induced systemic resistance to salinity, and increased the levels of chlorophyll b and carotenoids. Fungal inoculation also causes stress relief as indicated by reducing malondialdehyde concentration by 27.4% lower than stressed plants. Microscopic images revealed the active association and colonization of C. halotolerans within the roots of the Phaseolus vulgaris both under control and saline conditions. Therefore, utilizing endophytic fungi C. halotolerans for saline soil remediation appears to be a promising alternative in plant treatments, highlighting their potential as valuable resources for both research and commercial applications under salinity stress. Full article
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<p>Fresh weight (FW) of (<b>a</b>) shoot, (<b>b</b>) root and dry weight (DW) of (<b>c</b>) shoot, and (<b>d</b>) root of non-salinized and salinized <span class="html-italic">P. vulgaris</span> plant as affected by <span class="html-italic">C. halotolerans</span> inoculation. Each column represents a mean value of three replicates (<span class="html-italic">n</span> = 3) with vertical bars representing the standard error of the mean. Bars with different letters indicate a significant difference (<span class="html-italic">p</span> &lt; 0.05) between SNP treatments at all studied water levels as determined by two-way ANOVA and Bonferroni multiple comparison test.</p>
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<p>(<b>a</b>) Shoot length, (<b>b</b>) root length, and (<b>c</b>) leaf area of non-salinized and salinized <span class="html-italic">P. vulgaris</span> plant as affected by <span class="html-italic">C. halotolerans</span> inoculation. Each column represents a mean value of three replicates (<span class="html-italic">n</span> = 3) with vertical bars representing the standard error of the mean. Bars with different letters indicate a significant difference (<span class="html-italic">p</span> &lt; 0.05) between SNP treatments at all studied water levels as determined by two-way ANOVA and Bonferroni multiple comparison test.</p>
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<p>Concentrations of (<b>a</b>) chlorophyll a, (<b>b</b>) chlorophyll b, and (<b>c</b>) carotenoids in leaves of non-salinized and salinized <span class="html-italic">P. vulgaris</span> plant as affected by <span class="html-italic">C. halotolerans</span> inoculation. Each column represents a mean value of three replicates (<span class="html-italic">n</span> = 3) with vertical bars representing the standard error of the mean. Bars with different letters indicate a significant difference (<span class="html-italic">p</span> &lt; 0.05) between SNP treatments at all studied water levels as determined by two-way ANOVA and Bonferroni multiple comparison test.</p>
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<p>Malondialdehyde (MDA) concentration in roots of non-salinized and salinized <span class="html-italic">P. vulgaris</span> plant as affected by <span class="html-italic">C. halotolerans</span> inoculation. Each column represents a mean value of three replicates (<span class="html-italic">n</span> = 3) with vertical bars representing the standard error of the mean. Bars with different letters indicate a significant difference (<span class="html-italic">p</span> &lt; 0.05) between SNP treatments at all studied water levels as determined by two-way ANOVA and Bonferroni multiple comparison test.</p>
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<p>(<b>a</b>) Root sectioning of uninoculated plants. (<b>b</b>,<b>c</b>) Intracellular colonization by endophytic <span class="html-italic">C. halotolerans</span>, dark septate hyphae within a healthy root tissue of the treated plant under salinity. The red-colored arrow head indicates the presence of fungal mycelia as observed under a Vickers compound microscope stained with trypan blue staining.</p>
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12 pages, 2112 KiB  
Article
Foliar Spraying with Endophytic Trichoderma Biostimulant Increases Drought Resilience of Maize and Sunflower
by András Csótó, György Tóth, Péter Riczu, Andrea Zabiák, Vera Tarjányi, Erzsébet Fekete, Levente Karaffa and Erzsébet Sándor
Agriculture 2024, 14(12), 2360; https://doi.org/10.3390/agriculture14122360 - 22 Dec 2024
Viewed by 452
Abstract
Microbial biostimulants that promote plant growth and abiotic stress tolerance are promising alternatives to chemical fertilizers and pesticides. Although Trichoderma fungi are known biocontrol agents, their biostimulatory potential has been scarcely studied in field conditions. Here, the mixture of two endophytic Trichoderma strains [...] Read more.
Microbial biostimulants that promote plant growth and abiotic stress tolerance are promising alternatives to chemical fertilizers and pesticides. Although Trichoderma fungi are known biocontrol agents, their biostimulatory potential has been scarcely studied in field conditions. Here, the mixture of two endophytic Trichoderma strains (Trichoderma afroharzianum TR04 and Trichoderma simmonsii TR05) was tested as biostimulant in the form of foliar spray on young (BBCH 15-16) maize (5.7 ha) and sunflower (5.7 and 11.3 ha) fields in Hungary. The stimulatory effect was characterized by changes in plant height, the number of viable leaves, and the chlorophyll content, combined with yield sensor collected harvest data. In all trials, the foliar treatment with Trichoderma spores increased photosynthetic potential: the number of viable leaves increased by up to 6.7% and the SPAD index by up to 19.1% relative to the control. In extreme drought conditions, maize yield was doubled (from 0.587 to 1.62 t/ha, p < 0.001). The moisture content of the harvested seeds, as well as sunflower height, consistently increased post-treatment. We concluded that foliar spraying of young plants with well-selected endophytic Trichoderma strains can stimulate growth, photosynthesis, and drought tolerance in both monocot maize and dicots sunflower crops in field conditions. Full article
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<p>Experimental design to study the effect of the endophytic <span class="html-italic">Trichoderma</span> formulation. Aerial view of Experimental Site 1 (<b>a</b>) and Experimental Site 2 (<b>b</b>). Blue lines indicate the borders of the Experimental Sites, red lines indicate the borders of the plots. (<b>c</b>) Experimental designs C1–4 indicate control plots; T1–4 indicate plots treated with <span class="html-italic">Trichoderma</span>.</p>
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<p>Average NDVI mean values with natural breaks coloring method, using 80% overlapping (Experimental Site I). Light blue polygon outline color shows the lower average NDVI values (below 0.8). C1–4 indicate control plots; T1–4 indicate Trichoderma-treated plots.</p>
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<p>Monthly weather data of the experimental years 2022 and 2023. (<b>a</b>) Average, minimal and maximal temperatures. (<b>b</b>) Monthly average of solar radiation and precipitation.</p>
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<p>Changes in average number of viable leaves (<b>a</b>,<b>c</b>) and chlorophyll content (<b>b</b>,<b>d</b>, measured as SPAD values) of maize (<b>a</b>,<b>b</b>) and sunflower (<b>c</b>,<b>d</b>) plants following treatment with <span class="html-italic">Trichoderma</span> spp. compared to untreated control plants. Data points represent mean values, and error bars indicate standard error. *: <span class="html-italic">t</span>-test <span class="html-italic">p</span> &lt; 0.01, **: <span class="html-italic">t</span>-test <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Average yield (<b>a</b>) and average moisture content of harvested seeds (<b>b</b>) of maize and sunflower plants treated with <span class="html-italic">Trichoderma</span> spp. compared to untreated controls. Error bars represent standard error. **: <span class="html-italic">t</span>-test <span class="html-italic">p</span> &lt; 0.001.</p>
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23 pages, 7582 KiB  
Article
Endophytic Bacteria from the Desiccation-Tolerant Plant Selaginella lepidophylla and Their Potential as Plant Growth-Promoting Microorganisms
by Maria Guadalupe Castillo-Texta, José Augusto Ramírez-Trujillo, Edgar Dantán-González, Mario Ramírez-Yáñez and Ramón Suárez-Rodríguez
Microorganisms 2024, 12(12), 2654; https://doi.org/10.3390/microorganisms12122654 - 21 Dec 2024
Viewed by 458
Abstract
Bacteria associated with plants, whether rhizospheric, epiphytic, or endophytic, play a crucial role in plant productivity and health by promoting growth through complex mechanisms known as plant growth promoters. This study aimed to isolate, characterize, identify, and evaluate the potential of endophytic bacteria [...] Read more.
Bacteria associated with plants, whether rhizospheric, epiphytic, or endophytic, play a crucial role in plant productivity and health by promoting growth through complex mechanisms known as plant growth promoters. This study aimed to isolate, characterize, identify, and evaluate the potential of endophytic bacteria from the resurrection plant Selaginella lepidophylla in enhancing plant growth, using Arabidopsis thaliana ecotype Col. 0 as a model system. Plant growth-promotion parameters were assessed on the bacterial isolates; this assessment included the quantification of indole-3-acetic acid, phosphate solubilization, and biological nitrogen fixation, a trehalose quantification, and the siderophore production from 163 endophytic bacteria isolated from S. lepidophylla. The bacterial genera identified included Agrobacterium, Burkholderia, Curtobacterium, Enterobacter, Erwinia, Pantoea, Pseudomonas, and Rhizobium. The plant growth promotion in A. thaliana was evaluated both in Murashige Skoog medium, agar-water, and direct seed inoculation. The results showed that the bacterial isolates enhanced primary root elongation and lateral root and root hair development, and increased the fresh and dry biomass. Notably, three isolates promoted early flowering in A. thaliana. Based on these findings, we propose the S. lepidophylla bacterial isolates as ideal candidates for promoting growth in other agriculturally important plants. Full article
(This article belongs to the Section Plant Microbe Interactions)
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<p>Scheme of inoculation of <span class="html-italic">Arabidopsis thaliana</span> plants in MS 50% and AW culture medium with endophytic bacteria.</p>
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<p>Representative image of the in vitro plant growth-promotion parameters. (<b>A</b>) Solubilization of phosphates. AB, <span class="html-italic">A. brasilense</span> Cd; <span class="html-italic">Pseudomonas</span> sp. SlL121, higher PS for rainy season; <span class="html-italic">Pantoea</span> sp. SlL46, lower PS for rainy season; <span class="html-italic">Erwinia</span> sp. SlS1, higher PS for drought season; <span class="html-italic">Pseudomonas</span> sp. SlS14, lower PS for drought season. (<b>B</b>) Production of siderophores. AC, <span class="html-italic">A. chlorophenolicus</span> 30.16; AB, <span class="html-italic">A. brasilense</span> Cd; <span class="html-italic">Pseudomonas</span> sp. SlL121 is a major producer of rainy season hydroxamate-type siderophores; <span class="html-italic">Burkholderia</span> sp. SlL91 is the largest producer of rainy season catechol-type siderophores; <span class="html-italic">Pseudomonas</span> sp. SlS14 is a major producer of hydroxamate-type siderophores for drought season; <span class="html-italic">Pseudomonas</span> sp. SlS1 is a major producer of catechol-type siderophores for drought season.</p>
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<p>Promotion of plant growth of <span class="html-italic">A. thaliana</span> in MS 50% culture medium. (<b>A</b>) Length of the plant, rainy season. (<b>B</b>) Biomass in fresh and dry weight, rainy season. (<b>C</b>) Length of the plant, dry season. (<b>D</b>) Biomass in fresh and dry weight, dry season. The bars represent the mean value. Differences statistically significant with respect to the control bacteria were determined with a completely randomized ANOVA (Pr &gt; F =&lt; 0.0001), followed by a Tukey analysis (α = 0.05); the means with the same letter are not significantly different, and the error bars indicate the standard deviation of three repetitions. The biomass only represents the value of a replicate with 30 plants for each bacterial isolate.</p>
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<p>Appearance of <span class="html-italic">A. thaliana</span> plants inoculated with the different bacterial isolates in MS 50% culture medium.</p>
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<p>Promotion of plant growth of <span class="html-italic">A. thaliana</span> in AW culture medium. (<b>A</b>) Length of the plant, rainy season. (<b>B</b>) Biomass in fresh and dry weight, rainy season. (<b>C</b>) Length of the plant, dry season. (<b>D</b>) Biomass in fresh and dry weight, dry season. The bars represent the mean value. Differences that were statistically significant with respect to the control bacteria were determined with a completely randomized ANOVA (Pr &gt; F = &lt; 0.0001), followed by a Tukey analysis (α = 0.05); the means with the same letter are not significantly different, and the error bars indicate the standard deviation of three repetitions. The biomass represents the value of a replicate with 30 plants for each bacterial isolate.</p>
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<p>Appearance of <span class="html-italic">A. thaliana</span> plants inoculated with different bacterial isolates in AW culture medium.</p>
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<p>Promotion of plant growth of <span class="html-italic">A. thaliana</span> in AW culture medium. (<b>A</b>) Length of the plant, both seasons. (<b>B</b>) Biomass in fresh and dry weight, both seasons. (<b>C</b>) The germination percentage on days 1, 4, 7 and 10. (<b>D</b>) The germination speed on day 10. (<b>E</b>) The germination speed of <span class="html-italic">A. thaliana</span> seeds. The bars represent the mean value, and the statistically significant differences with respect to the control bacteria were determined with a completely randomized ANOVA (Pr &gt; F = &lt; 0.0001), followed by a Tukey analysis (α = 0.05); the means with the same letter are not significantly different, and the error bars indicate the standard deviation of three repetitions.</p>
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<p>Appearance of <span class="html-italic">A. thaliana</span> plants inoculated directly on the seeds with different bacterial isolates in AW culture medium.</p>
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14 pages, 1316 KiB  
Article
Genomic Insights and Plant Growth-Promoting Potential of Rhizobial Strains from Agave americana
by Luis Galdino García-Pérez, Clara Ivette Rincón-Molina, Esperanza Martínez-Romero, Marco Antonio Rogel, Yunuen Tapia-Torres, Luis Alberto Manzano-Gómez, Julio César Maldonado-Gómez, Francisco Alexander Rincón-Molina and Reiner Rincón-Rosales
Horticulturae 2024, 10(12), 1370; https://doi.org/10.3390/horticulturae10121370 - 20 Dec 2024
Viewed by 339
Abstract
Symbiotic relationships between plants and bacteria play a pivotal role in both natural and agricultural ecosystems, particularly through endophytic colonization or rhizospheric interactions. This study evaluated the plant growth-promoting potential (PGP) of native rhizobial strains Rhizobium sp. ACO-34A and Mesorhizobium sp. 28A (now [...] Read more.
Symbiotic relationships between plants and bacteria play a pivotal role in both natural and agricultural ecosystems, particularly through endophytic colonization or rhizospheric interactions. This study evaluated the plant growth-promoting potential (PGP) of native rhizobial strains Rhizobium sp. ACO-34A and Mesorhizobium sp. 28A (now reclassified within the genus Kumtagia), isolated from Agave americana. Through phenotypic characterization, PGP evaluation, and comparative genomic analysis, both strains demonstrated the ability to thrive under diverse salinity levels and pH conditions, reflecting their adaptability to challenging environments. Rhizobium sp. ACO-34A exhibited superior resistance to antibiotics and heavy metals, coupled with robust PGP traits, such as phosphate solubilization and indole-3-acetic acid (IAA) production, which are crucial for enhancing nutrient availability and root development. Similarly, Mesorhizobium sp. 28A showed exceptional phosphate solubilization efficiency and contributed to improved seedling performance. These findings highlight the capacity of rhizobia associated with agave species to improve plant growth, reduce dependence on chemical fertilizers, and support sustainable agriculture, particularly in nutrient-depleted or semi-arid soils. Genomic analyses revealed the presence of genes linked to stress resilience and nutrient acquisition, underlining the functional versatility of these strains. By leveraging these native rhizobial strains, agricultural practices can achieve higher productivity and sustainability, making them valuable tools for enhancing the agronomic yield and ecological resilience of agave crops for agro-industries. Full article
(This article belongs to the Section Genetics, Genomics, Breeding, and Biotechnology (G2B2))
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<p>Distribution and coverage of functional subsystem categories in the genomes of <span class="html-italic">Rhizobium</span> sp. ACO-34A (<b>A</b>) and, <span class="html-italic">Mesorhizobium</span> sp. 28A (<b>B</b>).</p>
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<p>Effect of inoculation with native rhizobial bacteria on <span class="html-italic">Agave americana</span> seeds. (<b>A</b>) Germination percentage, (<b>B</b>) germination velocity, (<b>C</b>) plumule length, and (<b>D</b>) radicle length.</p>
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21 pages, 3321 KiB  
Article
Microbiome Structures and Beneficial Bacteria in Soybean Roots Under Field Conditions of Prolonged High Temperatures and Drought Stress
by Sandeep Gouli, Aqsa Majeed, Jinbao Liu, David Moseley, M. Shahid Mukhtar and Jong Hyun Ham
Microorganisms 2024, 12(12), 2630; https://doi.org/10.3390/microorganisms12122630 - 19 Dec 2024
Viewed by 457
Abstract
Drought stress has a significant impact on agricultural productivity, affecting key crops such as soybeans, the second most widely cultivated crop in the United States. Endophytic and rhizospheric microbial diversity analyses were conducted with soybean plants cultivated during the 2023 growing season amid [...] Read more.
Drought stress has a significant impact on agricultural productivity, affecting key crops such as soybeans, the second most widely cultivated crop in the United States. Endophytic and rhizospheric microbial diversity analyses were conducted with soybean plants cultivated during the 2023 growing season amid extreme weather conditions of prolonged high temperatures and drought in Louisiana. Specifically, surviving and non-surviving soybean plants were collected from two plots of a Louisiana soybean field severely damaged by extreme heat and drought conditions in 2023. Although no significant difference was observed between surviving and non-surviving plants in microbial diversity of the rhizosphere, obvious differences were found in the structure of the endophytic microbial community in root tissues between the two plant conditions. In particular, the bacterial genera belonging to Proteobacteria, Pseudomonas and Pantoea, were predominant in the surviving root tissues, while the bacterial genus Streptomyces was conspicuously dominant in the non-surviving (dead) root tissues. Co-occurrence patterns and network centrality analyses enabled us to discern the intricate characteristics of operational taxonomic units (OTUs) within endophytic and rhizospheric networks. Additionally, we isolated and identified bacterial strains that enhanced soybean tolerance to drought stresses, which were sourced from soybean plants under a drought field condition. The 16S rDNA sequence analysis revealed that the beneficial bacterial strains belong to the genera Acinetobacter, Pseudomonas, Enterobacter, and Stenotrophomonas. Specific bacterial strains, particularly those identified as Acinetobacter pittii and Pseudomonas sp., significantly enhanced plant growth metrics and reduced drought stress indices in soybean plants through seed treatment. Overall, this study advances our understanding of the soybean-associated microbiome structure under drought stress, paving the way for future research to develop innovative strategies and biological tools for enhancing soybean resilience to drought. Full article
(This article belongs to the Section Plant Microbe Interactions)
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<p>The drought-damaged soybean field where the plant samples were collected (Alexandria, Louisiana). Plot A and Plot B neighbored each other.</p>
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<p>The records of rainfalls (<b>A</b>) and temperatures (<b>B</b>) in Alexandria, Louisiana, during the 2022 and 2023 growing seasons. These graphs were obtained from Weather Spark (<a href="https://weatherspark.com" target="_blank">https://weatherspark.com</a>) (data source: The Alexandria International Airport, accessed on 4 January 2024).</p>
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<p>Taxa bar plot showing seven abundant bacterial genera either in surviving or dry/dead soybean root tissues (root endosphere).</p>
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<p>Relative abundance of bacterial genera dominant in the root endosphere of drought-damaged soybean plants.</p>
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<p>The co-occurrence networks analyses. Four groups each for endophytic (<b>A</b>) and rhizospheric networks (<b>B</b>) are illustrated. Individual types of networks within each category are indicated. Nodes, edges, and clusters for each network are specified. The color of nodes signifies OTUs from the same module in each network, while line color indicates positive (orange) and negative (blue) correlation coefficients. Network construction employed Spearman’s correlation coefficient, with R &gt; 0.9 and <span class="html-italic">p</span> &lt; 0.05 as criteria.</p>
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13 pages, 3351 KiB  
Article
Identification and Characterization of Endophytic Fungus DJE2023 Isolated from Banana (Musa sp. cv. Dajiao) with Potential for Biocontrol of Banana Fusarium Wilt
by Longqi Jin, Rong Huang, Jia Zhang, Zifeng Li, Ruicheng Li, Yunfeng Li, Guanghui Kong, Pinggen Xi, Zide Jiang and Minhui Li
J. Fungi 2024, 10(12), 877; https://doi.org/10.3390/jof10120877 - 17 Dec 2024
Viewed by 387
Abstract
This study characterized an endophytic fungus, DJE2023, isolated from healthy banana sucker of the cultivar (cv.) Dajiao. Its potential as a biocontrol agent against banana Fusarium wilt was assessed, aiming to provide a novel candidate strain for the biological control of the devastating [...] Read more.
This study characterized an endophytic fungus, DJE2023, isolated from healthy banana sucker of the cultivar (cv.) Dajiao. Its potential as a biocontrol agent against banana Fusarium wilt was assessed, aiming to provide a novel candidate strain for the biological control of the devastating disease. The fungus was isolated using standard plant tissue separation techniques and fungal culture methods, followed by identification through morphological comparisons, multi-gene phylogenetic analyses, and molecular detection targeting Fusarium oxysporum f. sp. cubense (Foc) race 1 and race 4. Furthermore, assessments of its characteristics and antagonistic effects were conducted through pathogenicity tests, biological trait investigations, and dual-culture experiments. The results confirmed isolate DJE2023 to be a member of the Fusarium oxysporum species complex but distinct from Foc race 1 or race 4, exhibiting no pathogenicity to banana plantlets of cv. Fenza No.1 or tomato seedlings cv. money maker. Only minute and brown necrotic spots were observed at the rhizomes of banana plantlets of ‘Dajiao’ and ‘Baxijiao’ upon inoculation, contrasting markedly with the extensive necrosis induced by Foc tropical race 4 strain XJZ2 at those of banana cv Baxijiao. Notably, co-inoculation with DJE2023 and XJZ2 revealed a significantly reduced disease severity compared to inoculation with XJZ2 alone. An in vitro plate confrontation assay showed no significant antagonistic effects against Foc, indicating a suppressive effect rather than direct antagonism of DJE2023. Research on the biological characteristics of DJE2023 indicated lactose as the optimal carbon source for its growth, while maltose favored sporulation. The optimal growth temperature for this strain is 28 °C, and its spores can germinate effectively within the range of 25–45 °C and pH 4–10, demonstrating a strong alkali tolerance. Collectively, our findings suggest that DJE2023 exhibits weak or non-pathogenic properties and lacks direct antagonism against Foc, yet imparts a degree of resistance against banana Fusarium wilt. The detailed information provides valuable insight into the potential role of DJE2023 in integrated banana disease control, presenting a promising candidate for biocontrol against banana Fusarium wilt. Full article
(This article belongs to the Special Issue Fusarium spp.: A Trans-Kingdom Fungus)
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<p>Identification of endophytic fungus DJE2023 from banana cv. Dajiao. (<b>a</b>) The colony morphology of DJE2023. (<b>b</b>) Conidia. (<b>c</b>) Chlamydospores. (<b>d</b>) Multigene system phylogenetic tree. (<b>e</b>) Molecular identification using specific primers for <span class="html-italic">Fusarium oxysporum</span> f. sp. <span class="html-italic">cubense</span> (<span class="html-italic">Foc</span>) races 1 and 4. Among them, W106F/W106R are the universal primers for <span class="html-italic">F. oxysporum</span>, W1805F/W1805R are the specific primers for race 1of <span class="html-italic">Foc</span>, and W2987F/W2987R are the specific primers for race 4 of <span class="html-italic">Foc</span>. M: DL2000 Marker; 1,2: DJE-2023; 3: FOC Race 1; 4: FOC Race 4; 5: Water control.</p>
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<p>The pathogenicity test results for the banana endophytic strain DJE2023 and the <span class="html-italic">Fusarium oxysporum</span> f. sp. <span class="html-italic">cubense</span> (<span class="html-italic">Foc</span>) race 4 strain XJZ2. (<b>a</b>) The typical symptoms exhibited in the rhizomes of various banana cultivars. (<b>b</b>) The disease incidence statistics resulting from inoculating strain DJE2023 onto various banana cultivars. (<b>c</b>) The disease incidence statistics resulting from inoculating DJE2023 and XJZ2 onto banana cv. Baxijiao. The inoculation data of 30 plantlets were randomly divided into three groups for statistical analysis and the mean ± S.D. (n = 3). The significant difference (Duncan test, <span class="html-italic">p</span> &lt; 0.05) between two groups is represented by the superscript letters above the error bars.</p>
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<p>The biocontrol effect of DJE2023 on the banana plantlets cv. Baxijiao. (<b>a</b>) XJZ2, DJE2023, XJZ2, and DJE2023 co-infect the roots of banana plantlets; CK is water control. The disease conditions of leaves and roots are observed 45 days later. (<b>b</b>) Disease severity levels of banana roots; each treatment was replicated with 30 banana plantlets. (<b>c</b>) Disease index analyzed by diseased plantlet number of different disease grades. The inoculation data of 30 plantlets were randomly divided into three groups for statistical analysis and the mean ± S.D. (n = 3). The significant difference (Duncan test, <span class="html-italic">p</span> &lt; 0.05) between two groups is represented by the superscript letters above the error bars.</p>
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<p>Effects of carbon source on colony morphology, growth diameter, and sporulation of DJE2023. (<b>a</b>) The effect of carbon sources on the colony morphology of DJE2023. (<b>b</b>) The effect of carbon sources on the growth diameter of DJE2023. (<b>c</b>) The effect of carbon sources on the spore production of DJE2023. All data are presented as the mean ± standard deviation (n = 3). The significant difference (Duncan test, <span class="html-italic">p</span> &lt; 0.05) between two groups is represented by the superscript letters above the error bars.</p>
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<p>Effect of temperature on the morphology and growth diameter of DJE2023 colonies. (<b>a</b>) Colony morphology of DJE2023 under different temperatures. (<b>b</b>) Effect of temperature on the growth diameter of DJE2023. All data are presented as the mean ± standard deviation (n = 3). The significant difference (Duncan test, <span class="html-italic">p</span> &lt; 0.05) between two groups is represented by the superscript letters above the error bars.</p>
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<p>Effect of pH on colony morphology and growth diameter of DJE2023. (<b>a</b>) Colony morphology of DJE2023 under different pH levels. (<b>b</b>) Effect of pH on the growth diameter of DJE2023. All data are presented as the mean ± standard deviation (n = 3). The significant difference (Duncan test, <span class="html-italic">p</span> &lt; 0.05) between two groups is represented by the superscript letters above the error bars.</p>
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19 pages, 2832 KiB  
Article
Isolation and Characterization of Bacillus Strains from Egyptian Mangroves: Exploring Their Endophytic Potential in Maize for Biological Control of Spodoptera frugiperda
by Hayam M. Fathy, Mona Awad, Nawal A. Alfuhaid, El-Desoky S. Ibrahim, Moataz A. M. Moustafa and Ayatollah S. El-Zayat
Biology 2024, 13(12), 1057; https://doi.org/10.3390/biology13121057 - 17 Dec 2024
Viewed by 458
Abstract
The widespread use of pesticides to manage Spodoptera frugiperda has led to significant challenges. This insect has developed resistance to 47 active insecticide ingredients. Therefore, endophytic entomopathogenic bacteria have been explored as an alternative pest management strategy, offering the potential to reduce reliance [...] Read more.
The widespread use of pesticides to manage Spodoptera frugiperda has led to significant challenges. This insect has developed resistance to 47 active insecticide ingredients. Therefore, endophytic entomopathogenic bacteria have been explored as an alternative pest management strategy, offering the potential to reduce reliance on chemical pesticides. The current study aims to evaluate the colonization potential of indigenous marine Bacillus strains as endophytes in maize plants and to assess their insecticidal activity against S. frugiperda. Four inoculation methods—foliar application, seed treatment, soil drenching, and a combination of all three—were used to establish the Bacillus strains as endophytes in maize plants. Our results showed that the promising native Bacillus strains exhibited both antibacterial and insecticidal effects against S. frugiperda neonates under laboratory conditions. Foliar application of Bacillus sp. Esh39 caused the highest mortality rate (65%), followed by Bacillus tequilensis R39 (60%). However, this method did not significantly enhance plant height or chlorophyll content. The potential of these native Bacillus strains warrants further investigation to improve biological control via endophytic mediation. Our findings provide valuable insights into the bacterial diversity and functionality of mangrove ecosystems and pave the way for innovative, sustainable insect management strategies. Full article
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<p>Total isolates from different mangrove spheres and sediments using two isolation media.</p>
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<p>Percentage mortality of <span class="html-italic">S. frugiperda</span> neonates by thirty-three bacterial isolates.</p>
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<p>Percentage mortality of <span class="html-italic">S. frugiperda</span> neonates by thirty-three bacterial isolates.</p>
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<p>Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences of three bacterial isolates (highlighted with red circles) and closest matches from the NCBI database.</p>
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25 pages, 5413 KiB  
Article
Whole-Genome Profiling of Endophytic Strain B.L.Ns.14 from Nigella sativa Reveals Potential for Agricultural Bioenhancement
by Dimitra Douka, Tasos-Nektarios Spantidos, Polina C. Tsalgatidou, Panagiotis Katinakis and Anastasia Venieraki
Microorganisms 2024, 12(12), 2604; https://doi.org/10.3390/microorganisms12122604 - 16 Dec 2024
Viewed by 705
Abstract
Endophytic microbes in medicinal plants often possess beneficial traits for plant health. This study focuses on the bacterial endophyte strain B.L.Ns.14, isolated from Nigella sativa leaves, which demonstrated multiple plant growth-promoting properties. In vitro tests showed that B.L.Ns.14 supports plant growth, colonization, and [...] Read more.
Endophytic microbes in medicinal plants often possess beneficial traits for plant health. This study focuses on the bacterial endophyte strain B.L.Ns.14, isolated from Nigella sativa leaves, which demonstrated multiple plant growth-promoting properties. In vitro tests showed that B.L.Ns.14 supports plant growth, colonization, and tolerance to abiotic stress. The strain also exhibited antifungal activity against phytopathogens such as Rhizoctonia solani, Colletotrichum acutatum, Verticillium dahliae, and Fusarium oxysporum f. sp. radicis-lycopersici. Whole-genome analysis, supported by ANI and dDDH values, identified B.L.Ns.14 as Bacillus halotolerans. Genome mining revealed 128 active carbohydrate enzymes (Cazymes) related to endophytism and biocontrol functions, along with genes involved in phosphate solubilization, siderophore and IAA production, biofilm formation, and motility. Furthermore, genes for osmolyte metabolism, Na+/H+ antiporters, and stress response proteins were also identified. The genome harbors 12 secondary metabolite biosynthetic gene clusters, including those for surfactin, plipastatin mojavensin, rhizocticin A, and bacilysin, known for their antagonistic effects against fungi. Additionally, B.L.Ns.14 promoted Arabidopsis thaliana growth under both normal and saline conditions, and enhanced Solanum lycopersicum growth via seed biopriming and root irrigation. These findings suggest that Bacillus halotolerans B.L.Ns.14 holds potential as a biocontrol and plant productivity agent, warranting further field testing. Full article
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<p>Illustration of antifungal activity of B.L.Ns.14 in vitro by using dual culture assay.</p>
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<p>Plant growth-promoting traits of B.L.Ns.14; (<b>a</b>) siderophore production, (<b>b</b>) phosphate solubilization, (<b>c</b>) protease secretion, (<b>d</b>) cellulase secretion, (<b>e</b>) urease production, and (<b>f</b>) acetoin production.</p>
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<p>Colonization-related traits of B.L.Ns.14; (<b>a</b>) swarming motility, (<b>b</b>) swimming motility, and (<b>c</b>) biofilm formation ability.</p>
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<p>Survival ability of B.L.Ns.14 under drought and salt stress conditions, as well as under different temperatures.</p>
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<p>Antibiotic susceptibility test of B.L.Ns.14 by using six commercial antibiotics of three fixed concentrations (μg/mL); (A) ampicillin, (T) tetracycline, (S) streptomycin, (R) rifampicin, (K) kanamycin, and (C) chloramphenicol. The absence or presence of the clear zone around the soaked paper disks shows resistance or susceptibility of the strain. Red arrows indicate the susceptibility of the strain defined by the halo formed around the paper disks.</p>
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<p>Sequence-based phylogenomic tree constructed on TYGS (<a href="https://tygs.dsmz.de/" target="_blank">https://tygs.dsmz.de/</a>) depicting the position of bacterial strain B.L.Ns.14 relative to other phylogenetically close species. The tree was generated with FastME from Genome BLAST Distance Phylogeny (GBDP) distances [<a href="#B49-microorganisms-12-02604" class="html-bibr">49</a>]. The numbers above the branches are GBDP pseudo-bootstrap support values &gt;60% from 100 replications. The branch lengths are scaled in terms of GBDP distance formula d5 and the tree was rooted at the midpoint. The accession numbers of genome sequences are listed in parentheses.</p>
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<p>Results of antiSMASH analysis of the <span class="html-italic">B. halotolerans</span> B.L.Ns.14 genome. Illustration of detecting genomic regions where biosynthetic gene clusters of secondary metabolites are located. The closest core biosynthetic gene clusters of known and unknown BGCs according to the MIBiG database are depicted along with some of the best hits in ClusterBlast. Core biosynthetic genes of the cyclic lipopeptides and the aminoacid sequences are reported, as well as the gene similarity percentage given by antiSMASH.</p>
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<p>The beneficial activity of Β.L.Ns.14 on <span class="html-italic">A. thaliana</span> Col-0 seedlings in vitro, under normal and saline conditions (100 mM NaCl). (<b>A</b>) Representative images of the seedlings for the two treatments under normal (NC) and saline (SC) conditions; (<b>B</b>) shoot fresh weight of the seedlings (mg) (<span class="html-italic">n</span> = 12); (<b>C</b>) root fresh weight of the seedlings (mg) (n = 12); (<b>D</b>) primary root length (cm) (n = 12); and (<b>E</b>) total lateral root number (n = 12). Data represent the mean (SD) of seedlings from one representative experiment. Asterisks indicate statistically significant differences after <span class="html-italic">t</span>-test analysis (**, <span class="html-italic">p</span> &lt; 0.01, ***, <span class="html-italic">p</span> &lt; 0.001, ****, and <span class="html-italic">p</span> &lt; 0.0001).</p>
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<p>The beneficial activity of Β.L.Ns.14 on <span class="html-italic">A. thaliana</span> Col-0 seedlings in vitro via volatile emission, under normal and saline conditions (100 mM NaCl). (<b>A</b>) Representative images of the seedlings for the two treatments under normal (NC) and saline (SC) conditions; (<b>B</b>) shoot fresh weight of the seedlings (mg) (n = 12); and (<b>C</b>) rosette diameter (cm) (n = 12). Data represent the mean (SD) of seedlings from one representative experiment. Asterisks indicate statistically significant differences after <span class="html-italic">t</span>-test analysis (*, <span class="html-italic">p</span> &lt; 0.1, **, <span class="html-italic">p</span> &lt; 0.01, and ****, <span class="html-italic">p</span> &lt; 0.0001).</p>
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<p>Plant growth-promoting effect of Β.L.Ns.14 on <span class="html-italic">S. lycopersicum</span> var. Chondrokatsari Messinias; depiction of tomato seedlings (<b>A</b>) treated by seed biopriming method (scale bar = 1 cm) after 5 days; (<b>B</b>) treated by seed biopriming and root irrigation after 4 weeks; (<b>C</b>) radical growth (cm) emerged from bioprimed seeds from 3 replicates after using inoculants with bacterial suspensions (10<sup>6</sup> CFU/mL and 10<sup>8</sup> CFU/mL), each containing 15 seeds (n = 3); (<b>D</b>) germination (%) of bioprimed seeds with bacterial suspensions (10<sup>6</sup> CFU/mL and 10<sup>8</sup> CFU/mL), each containing 15 seeds (n = 3); and (<b>E</b>) data of shoot length (cm), shoot fresh weight (gr), and shoot dry weight (gr) of tomato seedlings (n = 30) after 4 weeks emerged from bioprimed seeds and root irrigation with bacterial suspension (10<sup>8</sup> CFU/mL) in pots. Data represent the mean (SD) of seedlings and asterisks indicate statistically significant differences after <span class="html-italic">t</span>-test analysis (ns, non-significant; ****, <span class="html-italic">p</span> &lt; 0.0001).</p>
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24 pages, 9449 KiB  
Article
Intercropping Shapes the Metabolome and Microbiome of Medicinal Giant Lily (Cardiocrinum giganteum) in Bamboo, Chinese Fir, and Mixed Forests
by Jie Zhang, Yilin Ning, Haoyu Wu, Guibin Gao, Zhizhuang Wu, Yuwen Peng, Zhiyuan Huang and Xiaoping Zhang
Forests 2024, 15(12), 2201; https://doi.org/10.3390/f15122201 - 13 Dec 2024
Viewed by 431
Abstract
Intercropping is a promising strategy for sustainable medicinal plant cultivation, but its impact on plant–microbe interactions remains poorly understood. This study investigated the influence that intercropping giant lily (Cardiocrinum giganteum) with bamboo (BG), Chinese fir (FG), and mixed forests (MG) had [...] Read more.
Intercropping is a promising strategy for sustainable medicinal plant cultivation, but its impact on plant–microbe interactions remains poorly understood. This study investigated the influence that intercropping giant lily (Cardiocrinum giganteum) with bamboo (BG), Chinese fir (FG), and mixed forests (MG) had on the giant lily metabolome and microbiome compared to a monoculture control (GG). Metabolomic analysis revealed that BG significantly increased the accumulation of terpenoids (e.g., yucalexin B22, 19.39-fold), alkaloids (e.g., anabasine, 2.97-fold), and steroids (e.g., O-acetyl-lariciresinol, 4.49-fold), while MG induced the production of stress-related metabolites (e.g., aflatoxin G2, 128.62-fold), and FG enhanced nitrogen metabolism (e.g., putrescine, 2.47-fold). Intercropping altered the rhizosphere and endophytic microbial communities, with BG enriching beneficial bacteria (e.g., Acidobacteria and Alphaproteobacteria) and FG promoting symbiotic fungi (e.g., Serendipita and Xylariales). Network analysis revealed strong correlations between specific microbial taxa (e.g., Bacillus and Ceratobasidiaceae) and key metabolites (e.g., norpandamarilactonine A, methylgingerol), indicating their potential roles in shaping the metabolic profiles of giant lily. These findings highlight the complex interplay between intercropping systems, microbial communities, and medicinal plant metabolism and provide a basis for developing targeted cultivation strategies to enhance the production of bioactive compounds in giant lily and other medicinal plants. Full article
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<p>Effects of different intercropping systems on the secondary metabolism of giant lily bulbs. (<b>a</b>) Partial least squares discriminant analysis (PLS-DA) score plot showing the metabolic profiles of giant lily bulbs under different intercropping systems: bamboo–giant lily (BG), Chinese fir–giant lily (FG), mixed forest-giant lily (MG), and forest gap–giant lily (GG). (<b>b</b>) Permutation test for PLS-DA model validation. (<b>c</b>) Heatmap displaying the relative abundance of differentially accumulated metabolites among the four intercropping systems.</p>
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<p>Effects of different intercropping systems on the secondary metabolism of giant lily bulbs. (<b>a</b>–<b>c</b>) Volcano plots illustrating the differentially abundant metabolites. (<b>d</b>) KEGG pathway enrichment analysis of the differentially abundant metabolites.</p>
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<p>Microbial diversity analysis. (<b>a</b>–<b>d</b>) Alpha diversity (Shannon index) comparisons of rhizosphere bacteria (<b>a</b>), rhizosphere fungi (<b>b</b>), endophytic bacteria (<b>c</b>), and endophytic fungi (<b>d</b>) among the four intercropping systems. (<b>e</b>,<b>f</b>) Principal coordinates analysis plots depicting the beta diversity of bacterial (<b>e</b>) and fungal (<b>f</b>) communities in the rhizosphere and endosphere of giant lily. “*” shows at <span class="html-italic">p</span> ≤ 0.05, “**” shows at <span class="html-italic">p</span> ≤ 0.01.</p>
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<p>Microbiota of rhizosphere and endophytes. (<b>a</b>) Relative abundance of bacterial phyla in the rhizosphere and endosphere. (<b>b</b>) Relative abundance of bacterial genera in the rhizosphere and endosphere. (<b>c</b>) Relative abundance of fungal phyla in the rhizosphere and endosphere. (<b>d</b>) Relative abundance of fungal genera in the rhizosphere and endosphere.</p>
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<p>Linear discriminant analysis effect size (LEfSe) results of microbial groups in the rhizosphere and endophytic bacteria of different intercropping systems. (<b>a</b>) Rhizosphere bacteria; (<b>b</b>) rhizosphere fungi; (<b>c</b>) endophytic bacteria; (<b>d</b>) endophytic fungi.</p>
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<p>Procrustes analysis of microbial community composition and metabolic profiles in giant lily across intercropping systems. (<b>a</b>) Rhizosphere bacteria; (<b>b</b>) rhizosphere fungi; (<b>c</b>) endophytic bacteria; (<b>d</b>) endophytic fungi. The connecting line represents the vector residuals of the two sorted configurations, which can be used to evaluate the variation between the two. A shorter connecting line indicates a higher level of consistency between the two datasets.</p>
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<p>Interactions between differentially abundant microorganisms and differentially abundant metabolites. (<b>a</b>) Co-expression network of differentially abundant microorganisms and differentially abundant metabolites (|rho| &gt; 0.90); (<b>b</b>) Redundancy analysis results of typical differentially abundant microorganisms and differentially abundant metabolites.</p>
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18 pages, 4899 KiB  
Review
Endophytic Entomopathogenic Fungi: Their Role in Enhancing Plant Resistance, Managing Insect Pests, and Synergy with Management Routines
by Krishnamoorthy Aravinthraju, Mookiah Shanthi, Marimuthu Murugan, Ramasamy Srinivasan, Lourena Arone Maxwell, Narayanan Manikanda Boopathi and Rangasamy Anandham
J. Fungi 2024, 10(12), 865; https://doi.org/10.3390/jof10120865 - 13 Dec 2024
Viewed by 742
Abstract
The interaction between plants and microorganisms plays a major role in plant growth promotion and disease management. While most microorganisms directly influence plant health, some indirectly support growth through pest and disease suppression. Endophytic entomopathogenic fungi are diverse, easily localized, and have long-lasting [...] Read more.
The interaction between plants and microorganisms plays a major role in plant growth promotion and disease management. While most microorganisms directly influence plant health, some indirectly support growth through pest and disease suppression. Endophytic entomopathogenic fungi are diverse, easily localized, and have long-lasting effects on insect pests. When inhabiting plants, these fungi alter secondary metabolites, volatile organic compounds, and microbiomes, enhancing plant resistance to pests and diseases and sometimes improving growth. However, their persistence in plant systems may be challenged by the plant’s defense mechanisms or by human interventions such as insecticides, fungicides, herbicides, and phyto-insecticides, which are common in agriculture. As effective biocontrol agents, endophytic entomopathogenic fungi can also be integrated with other pest management strategies like predators, parasitoids, and chemicals. This review will explore the impact of endophytic entomopathogens on plant systems and their compatibility with other management practices. Full article
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<p>Endophytic entomopathogenic fungi and their associated host against insect pests.</p>
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<p>Metabolic changes in host plants due to endophytic entomopathogenic fungi. The endophytic entomopathogenic fungus (1) enters the plant system through the roots and colonizes within the plant tissues (2). This colonization may alter plant volatiles (3), secondary metabolites, and hormones, which can result in repelling insect pests (4) and attracting natural enemies, such as predators (5) and parasitoids (6).</p>
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<p>Compatibility of IPM components for its success. Microbes have a two-way interaction: they can be antagonistic to natural enemies while also being affected by chemical pesticides. Therefore, their compatibility with other integrated pest management (IPM) practices is crucial for successful pest management.</p>
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