Root Samples Research Articles

Knockdown of microRNA390 Enhances Maize Brace Root Growth.

Brace root architecture is a critical determinant of maize's stalk anchorage and nutrition uptake, influencing root lodging resistance, stress tolerance, and plant growth. To identify the key microRNAs (miRNAs) in control of maize brace root growth, we performed small RNA sequencing using brace root samples at emergence and growth stages. We focused on the genetic modulation of brace root development in maize through manipulation of miR390 and its downstream regulated auxin response factors (ARFs). In the present study, miR167, miR166, miR172, and miR390 were identified to be involved in maize brace root growth in inbred line B73. Utilizing short tandem target mimic (STTM) technology, we further developed maize lines with reduced miR390 expression and analyzed their root architecture compared to wild-type controls. Our findings show that STTM390 maize lines exhibit enhanced brace root length and increased whorl numbers. Gene expression analyses revealed that the suppression of miR390 leads to upregulation of its downstream regulated ARF genes, specifically ZmARF11 and ZmARF26, which may significantly alter root architecture. Additionally, loss-of-function mutants for ZmARF11 and ZmARF26 were characterized to further confirm the role of these genes in brace root growth. These results demonstrate that miR390, ZmARF11, and ZmARF26 play crucial roles in regulating maize brace root growth; the involved complicated molecular mechanisms need to be further explored. This study provides a genetic basis for breeding maize varieties with improved lodging resistance and adaptability to diverse agricultural environments.

PRIMER REPORTE DE Macrophomina pseudophaseolina ASOCIADO A LA PUDRICIÓN DE RAÍZ EN CHILE APAXTLECO NATIVO (Capsicum annuum L.) EN GUERRERO, MÉXICO

<p><strong>Background.</strong> Chili is an important crop for Mexican culture. <strong>Objective. </strong>To identify the pathogen associated with the causal agent of root rot in a native chili pepper Apaxtleco in Guerrero, Mexico. <strong>Methodology.</strong> Root samples of diseased plants were taken from a chili plot in Apaxtla, Guerrero, and were taken to the Phytopathology Laboratory, at Colegio Superior Agropecuario del Estado de Guerrero (CSAEGRO), México. Subsequently, 1 cm root sections were made from healthy and diseased tissue, which were disinfected with 3% sodium hypochlorite for 1 min, and washed three times with sterile distilled water for 1 min, and dried with sterile absorbent paper, later, moistened sterilized paper was placed in the Petri dish and moistened with sterile distilled water and two slides were placed on it arranged in the shape of a cross and at each end of the slides and in the center of these, a piece of previously disinfected root was placed. The plates were kept at 28 °C ± 2 °C for 168 h. The purification of the pathogen was carried out using the hyphal tip and the identification was morpho-molecular. <strong>Results.</strong> The causal agent of root rot in Apaxtleco native chili plants was <em>Macrophomina pseudophaseolina</em>. <strong>Implications.</strong> Our result reveals the opportunity to improve the control of this pathogen at Guerrero, Mexico. <strong>Conclusion.</strong> <em>M. pseudophaseolina</em> causing root rot, in Apaxtleco native chili is reported for the first time.</p>

Evolution of vegetative morphoanatomical characters in Pabstiella (Pleurothallidinae: Orchidaceae)

The subtribe Pleurothallidinae encompasses approximately 48 genera, with Pabstiella standing out due to its 133 species, many of which are found in Brazil's Atlantic Forest. Molecular data strongly support the genus and its ten sections. However, only a few species have been anatomically described, and a comprehensive characterization of the genus for comparative purposes is still needed. Our study aimed to examine the main vegetative morphoanatomical characters of the root, stem, and leaves, and analyze their evolution based on the phylogeny of the genus. We analysed root, stem, and leaf samples from 57 species using light and scanning electron microscopy. We selected 35 characters and mapped them onto the molecular phylogeny. Pabstiella exhibits homogeneous leaf anatomy, with aquiferous tissue on both sides and chlorophyllous parenchyma in the middle, aligned with vascular bundles. The ramicaul anatomy displays a simple epidermis with primary or secondary walls, a simple to stratified hypodermis, a parenchymatous or aerenchymatous cortex, and a sheath of sclerenchyma surrounding the vascular bundles. The roots endovelamen cells are larger than those in the epivelamen, reticulated tilosomes of varying morphotypes, and giant cells in the cortex. Despite the common occurrence of homoplasy among vegetative traits, particularly in the leaves, our analyses revealed several synapomorphies for the genus. These include the presence of giant cells in the root cortex, endovelamen cells larger than those of the epivelamen, a specific distribution pattern of tilosomes, and 2–3 layers of aquifer tissue on the adaxial face of the leaf. Additionally, for the Pabstiella section, a lignified stem epidermis wall is a synapomorphic character state. Our findings demonstrate that vegetative morpho-anatomy is a useful diagnostic character for generic and infrageneric categories within the genus. When analysed from a phylogenetic perspective, it contributes to our comparative understanding of the genus.

Metataxonomic analysis of endophytic bacteria of blackberry (Rubus ulmifolius Schott) across tissues and environmental conditions

(1) Background: Endophytic bacteria represent an important component of plant wellness. They have been widely studied for their involvement in plant development and enhancement of stress tolerance. In this work, the endophytic communities of roots, stems, and leaves of blackberry (Rubus ulmifolius Schott) were studied in three different niches: natural, riverside, and human-impacted niches. (2) Results: The microbiome composition revealed that Sphingomonadaceae was the most abundant family in all samples, accounting for 9.4–45.8%. In contrast, other families seem to be linked to a specific tissue or niche. Families Microbacteriaceae and Hymenobacteraceae increased their presence in stem and leaf samples, while Burkholderiaceae abundance was important in riverside samples. Alpha and beta diversity analyses showed that root samples were the most diverse, and they gathered together in the same cluster, apart from the rest of the samples. (3) Conclusions: The analysis of the microbiome of R. ulmifolius plants revealed that the composition was essentially the same in different niches; the differences were primarily influenced by plant tissue factors with a core genome dominated by Sphingomonadaceae. Additionally, it was observed that R. ulmifolius can select its own microbiome, and this remains constant in all tissues evaluated regardless the niche of sampling.

Unlocking the growth-promoting and antagonistic power: A comprehensive whole genome study on Bacillus velezensis strains

Bacillus species are extensively documented as plant growth-promoting rhizobacteria, contributing significantly to the enhancement of soil fertility, nutrient recycling, and the control of phytopathogens. Utilizing them as biocontrol agents represents an environmentally friendly strategy, particularly within the rhizospheric community. This study presents the comprehensive genome sequences of three B. velezensis strains (LGMB12, LGMB319, and LGMB426) which were previously isolated from root samples of maize (Zea mays L.), along with a type strain FZB42. The research assesses the capability of the three strains for antagonizing fungi, specifically Fusarium graminearum, Fusarium verticillioides, Colletotrichum graminicola, and Stenocarpella sp. In paired cultures involving maize fungi, treatments containing bacteria B. velezensis exhibited statistically significant differences compared to both negative and positive treatments in terms of antagonism. Furthermore, genome mining techniques were employed to explore their inherent antagonistic potential. The assembly revealed that strains LGMB12, LGMB319, LGMB426, and FZB42 exhibit genome sizes of 4,187,541 bp, 4,244,954 bp, 3,976,537 bp, and 3,990,518 respectively. Their respective G + C content stands at 46.42 %, 46.50 %, 46.51 %, and 46.38 %. Moreover, the genomes present multiple gene clusters responsible for the synthesis of secondary metabolites and carbohydrate-active enzymes (CAZymes). These clusters highlight a diverse array of antibacterial and antifungal properties, complemented by numerous plant growth-promoting genes. These results highlight the potential of B. velezensis LGMB12, LGMB319, and LGMB426 strains as biocontrol and plant growth promotion agents, being promising candidates for further studies in agricultural production, including field trials.

Community assembly of ectomycorrhizal fungal communities in pure and mixed Pinus massoniana forests

Ectomycorrhizal (EcM) fungi play an important role in nutrient cycling and community ecological dynamics and are widely acknowledged as important components of forest ecosystems. However, little information is available regarding EcM fungal community structure or the possible relationship between EcM fungi, soil properties, and forestry activities in Pinus massoniana forests. In this study, we evaluated soil properties, extracellular enzyme activities, and fungal diversity and community composition in root and soil samples from pure Pinus massoniana natural forests, pure P. massoniana plantations, and P. massoniana and Liquidambar gracilipes mixed forests. The mixed forest showed the highest EcM fungal diversity in both root and bulk soil samples. Community composition and co-occurrence network structures differed significantly between forest types. Variation in the EcM fungal community was significantly correlated with the activities of β-glucuronidase and β−1,4-N-acetylglucosaminidase, whereas non-EcM fungal community characteristics were significantly correlated with β-1,4-glucosidase and β-glucuronidase activities. Furthermore, stochastic processes predominantly drove the assembly of both EcM and non-EcM fungal communities, while deterministic processes exerted greater influence on soil fungal communities in mixed forests compared to pure forests. Our findings may inform a deeper understanding of how the assembly processes and environmental roles of subterranean fungal communities differ between mixed and pure plantations and may provide insights for how to promote forest sustainability in subtropical areas.

Changes in the rhizosphere and root-associated bacteria community of white Guinea yam (Dioscorea rotundata Poir.) impacted by genotype and nitrogen fertilization

The bacterial diversity and composition of water yam (Dioscorea alata L. cv. A-19), which can grow without chemical fertilization, have recently been characterized with no significant differences compared with the use of chemical fertilization. However, the diversity and community structure of bacteria associated with the white Guinea yam (Dioscorea rotundata), the most cultivated and economically important yam in West Africa, have not yet been investigated. This study characterized the bacterial diversity and composition associated with bulk soil, rhizosphere, and plant roots in six white Guinea yam genotypes (S004, S020, S032, S042, S058, and S074) in field experiments in Ibadan, Nigeria under N-based chemical fertilizer application. The largest diversity of bacteria was found in the bulk soil, followed by the rhizosphere and roots. Based on the alpha diversity analysis, the bacterial diversity in both S020 and S042 increased with fertilizer application among the bulk soil samples. S058 grown under no-fertilizer conditions had the highest bacterial diversity among the rhizosphere samples. Beta diversity analysis highlighted the significant difference in the composition of bacteria associated with the genotypes and fertilizer treatments, and S032 had a unique bacterial composition compared to the other genotypes. The dominant phylum across all sample types was Proteobacteria. Actinobacteriota was the dominant phylum among bulk soil samples. At the genus level, Bacillus was the most abundant bacterial genus across both the control and treated samples. Pseudomonas was predominant across all rhizosphere samples. Chryseobacterium, Sphingobium, Delftia and Klebsiella associated with the rhizosphere were shown the altered relative abundance between the control and treated samples depending on genotypes. A genus related to symbiotic nitrogen-fixing bacteria, the Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium clade, showed higher relative abundance among all root samples, indicating that it is a core bacterial genus. Furthermore, the field application of chemical fertilizer had a significant impact on the relative abundances of two genera related to symbiotic nitrogen-fixers, Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium clade and Bradyrhizobium in the rhizosphere and root. These results suggest that N-based chemical fertilizers and plant genotypes would influence the compositional arrangement of associated bacterial communities, including symbiotic nitrogen-fixing bacteria.

Spatial variations impact the soil fungal communities of Larix gmelinii forests in Northeast China

Soil fungi play a critical role in the biogeochemical cycles of forest ecosystems. Larix gmelinii is a strong and important timber tree species, which forms close associations with a wide range of soil fungi. However, the temporal-spatial disparity effects on the assembly of soil fungal communities in L. gmelinii forests are poorly understood. To address these questions, a total of 120 samples, including 60 bulk soil and 60 root samples, were collected from Aershan and Genhe in July (summer) and October (autumn)2021. We obtained 7,788 operational taxonomic units (OTUs) after merging, filtering, and rarefying using high-throughput sequencing. The dominant phyla are Basidiomycota, Ascomycota, Mortierellomycota, and Mucoromycota. There were 13 dominant families, among which the families with average relative abundance more than 5% included Thelephoraceae, Mortierellaceae, Archaeorhizomycoaceae, and Inocybaceae. In the functional guilds, symbiotrophic fungi had a relative advantage in the identified functions, and the relative abundances of pathotrophic and saprotrophic fungi varied significantly between sites. There were 12 families differentially expressed across compartments, 10 families differentially expressed between seasons, and 69 families were differentially expressed between sites. The variation in alpha diversity in the bulk soil was greater than that in the rhizosphere soil. Among the three parts (compartment, season, and site), the site had a crucial effect on the beta diversity of the fungal community. Deterministic processes dominated fungal community assembly in Genhe, whereas stochastic processes dominated in Aershan. Soil physicochemical properties and climatic factors significantly affected fungal community structure, among which soil total nitrogen and pH had the greatest effect. This study highlights that spatial variations play a vital role in the structure and assembly of soil fungal communities in L. gmelinii forests, which is of great significance for us in maintaining the health of the forests.

Comparative Metabolome and Transcriptome Analyses Reveal Differential Enrichment of Metabolites with Age in Panax notoginseng Roots.

Panax notoginseng is a perennial plant well known for its versatile medicinal properties, including hepatoprotective, antioxidant, anti-inflammatory, anti-tumor, estrogen-like, and antidepressant characteristics. It has been reported that plant age affects the quality of P. notoginseng. This study aimed to explore the differential metabolome and transcriptome of 2-year (PN2) and 3-year-old (PN3) P. notoginseng plant root samples. Principal component analysis of metabolome and transcriptome data revealed major differences between the two groups (PN2 vs. PN3). A total of 1813 metabolites and 28,587 genes were detected in this study, of which 255 metabolites and 3141 genes were found to be differential (p < 0.05) between PN2 vs. PN3, respectively. Among differential metabolites and genes, 155 metabolites and 1217 genes were up-regulated, while 100 metabolites and 1924 genes were down-regulated. The KEGG pathway analysis revealed differentially enriched metabolites belonging to class lipids ("13S-hydroperoxy-9Z, 11E-octadecadionic acid", "9S-hydroxy-10E, 12Z-octadecadionic acid", "9S-oxo-10E, 12Z-octadecadionic acid", and "9,10,13-trihydroxy-11-octadecadionic acid"), nucleotides and derivatives (guanine and cytidine), and phenolic acids (chlorogenic acid) were found to be enriched (p < 0.05) in PN3 compared to PN2. Further, these differentially enriched metabolites were found to be significantly (p < 0.05) regulated via linoleic acid metabolism, nucleotide metabolism, plant hormone signal transduction, and arachidonic acid metabolism pathways. Furthermore, the transcriptome analysis showed the up-regulation of key genes MAT, DMAS, SDH, gallate 1-beta-glucosyltransferase, and beta-D-glucosidase in various plants' secondary metabolic pathways and SAUR, GID1, PP2C, ETR, CTR1, EBF1/2, and ERF1/2 genes observed in phytohormone signal transduction pathway that is involved in plant growth and development, and protection against the various stressors. This study concluded that the roots of a 3-year-old P. notoginseng plant have better metabolome and transcriptome profiles compared to a 2-year-old plant with importantly enriched metabolites and genes in pathways related to metabolism, plant hormone signal transduction, and various biological processes. These findings provide insights into the plant's dynamic biochemical and molecular changes during its growth that have several implications regarding its therapeutic use.

Presymptomatic Leaf Reflectance of Fusarium virguliforme-Infected Soybean Plants in Greenhouse Conditions

Sudden death syndrome of soybean is caused by a soilborne pathogen, Fusarium virguliforme. Prior to visible foliar symptoms, a destructive technique can be carried out to diagnose root infection. Hyperspectral sensors can be a presymptomatic and nondestructive alternative for plant disease diagnosis. This study was designed to relate leaf spectral reflectance to F. virguliforme root infection in the absence of foliar symptoms. Soybean plants were grown under controlled greenhouse conditions. The spectral reflectance of the plants was measured weekly beginning at 21 days after transplanting up until 42 days after transplanting using a swing hyperspectral imaging system fixed on a gantry. Destructive root sampling confirmed F. virguliforme root infection using real-time PCR. The most relevant wavelengths for discrimination were selected using the ReliefF algorithm. Three machine learning models (partial least squares discriminant analysis, support vector machine, and random forest) were evaluated for classification accuracy using the selected wavelengths. Relevant wavelengths for differentiating between the healthy and F. virguliforme-infected plants were found in the visible and red-edge region from 500 to 750 nm and the shortwave infrared region from 1,400 to 2,350 nm. In the absence of visible foliar symptoms, classification results showed over 79% mean F1 scores for all models. Partial least squares discriminant analysis was able to differentiate healthy and F. virguliforme-infected plants with a mean F1 score of 83.1 to 85.3% and a kappa statistic of 0.43 to 0.54. This work supports the use of hyperspectral remote sensing for early presymptomatic disease diagnosis under a controlled environment. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

The effect of endodontic instrumentation on fatigue, fracture load, and dentin crack formation in restored uniradicular teeth: in vitro study

Objective(s): The aim of this study was to evaluate the effect of manual, rotatory, and reciprocating endodontic instrumentation for uniradicular teeth on fatigue, load to failure, and dentin crack formation. Materials and Methods: Sixty-two human uniradicular teeth were selected. Root samples were standsdized at 14 mm (t0). Teeth were divided into three groups according to endodontic instrumentation: Manual (M), Rotatory (RT), or Reciprocating (RC) and filled with passive technique (t1). All teeth received the cementation of a glass fiber post and were restored with composite resin core (t2) and metallic crown. Samples were subjected to mechanical cycling (177 N, 2x106 cycles, 4 Hz) (t3), followed by a load to failure test. Failure analysis was performed and dentin crack analysis was conducted in two samples each time t0, t1, t2, and t3. Results: Load to failure was not different between groups (p=0.716). However, the M group presented more irreparable failures. M and RC groups presented a high percentage of root sections with defects, and t3 was the evaluation time which presented more dentin crack formation. Conclusion(s): Endodontic instrumentation system did not affect load to failure of uniradicular teeth. Still, it affected the failure mode, with manual and reciprocating instrumentations presenting more irreparable failures and high root dentin crack formation.

Revealing the diversity of Jojoba-associated fungi using amplicon metagenome approach and assessing the in vitro biocontrol activity of its cultivable community.

Jojoba shrubs are wild plants cultivated in arid and semiarid lands and characterized by tolerance to drought, salinity, and high temperatures. Fungi associated with such plants may be attributed to the tolerance of host plants against biotic stress in addition to the promotion of plant growth. Previous studies showed the importance of jojoba as jojoba oil in the agricultural field; however, no prior study discussed the role of jojoba-associated fungi (JAF) in reflecting plant health and the possibility of using JAF in biocontrol. Here, the culture-independent and culture-dependent approaches were performed to study the diversity of the jojoba-associated fungi. Then, the cultivable fungi were evaluated for in-vitro antagonistic activity and in vitro plant growth promotion assays. The metagenome analysis revealed the existence of four fungal phyla: Ascomycota, Aphelidiomycota, Basidiomycota, and Mortierellomycota. The phylum Ascomycota was the most common and had the highest relative abundance in soil, root, branch, and fruit samples (59.7%, 50.7%, 49.8%, and 52.4%, respectively). Alternaria was the most abundant genus in aboveground tissues: branch (43.7%) and fruit (32.1%), while the genus Discosia had the highest abundance in the underground samples: soil (24%) and root (30.7%). For the culture-dependent method, a total of 14 fungi were isolated, identified, and screened for their chitinolytic and antagonist activity against three phytopathogenic fungi (Fusarium oxysporum, Alternaria alternata and Rhizoctonia solani) as well as their in vitro plant growth promotion (PGP) activity. Based on ITS sequence analysis, the selected potent isolates were identified as AspergillusstellatusEJ-JFF3, Aspergillusflavus EJ-JFF4, Stilbocreasp. EJ-JLF1, Fusariumsolani EJ-JRF3, and AmesiaatrobrunneaEJ-JSF4. The endophyte strain A. flavus EJ-JFF4 exhibited the highest chitinolytic activity (9 Enzyme Index) and antagonistic potential against Fusarium oxysporum, Alternaria alternata, and Rhizoctonia solani phytopathogens with inhibitory percentages of 72, 70, and 80 respectively. Also, A. flavus EJ-JFF4 had significant multiple PGP properties, including siderophore production (69.3%), phosphate solubilization (95.4 µg ml-1). The greatest production of Indol-3-Acetic Acid was belonged to A.atrobrunnea EJ-JSF4 (114.5 µg ml-1). The analysis of FUNGuild revealed the abundance of symbiotrophs over other trophic modes, and the guild of endophytes was commonly assigned in all samples. For the first time, this study uncovered fungal diversity associated with jojoba plants using a culture-independent approach and in-vitro assessed the roles of cultivable fungal strains in promoting plant growth and biocontrol. The present study indicated the significance of jojoba shrubs as a potential source of diverse fungi with high biocontrol and PGP activities.

An Inhibitor-Monitorable Single-Tube Duplex Quantitative Real-Time PCR Assay for the Detection of 'Candidatus Liberibacter asiaticus'.

Huanglongbing (HLB) is a citrus infectious disease caused by 'Candidatus Liberibacter' spp. Recently, it has begun to spread rapidly worldwide, causing significant losses to the citrus industry. Early diagnosis of HLB relies on quantitative real-time PCR assays. However, the PCR inhibitors found in the nucleic acid extracted from plant materials pose challenges for PCR assays because they may result in false-negative results. Internal standard (IS) can be introduced to establish a single-tube duplex PCR for monitoring the influence of the PCR inhibitor, but it also brings the risk of false-negative results because the amplification of IS may compete with the target. To solve this problem, we proposed a mutation-enhanced single-tube duplex PCR (mSTD-PCR) containing IS with mutant-type primers. By introducing the 3'-terminal mutation in the primer of IS to weaken its amplification reaction and its inhibition of 'Candidatus Liberibacter asiaticus' (CLas) detection, the sensitivity and quantitative accuracy of CLas detection will not be affected by IS. In evaluating the sensitivity of CLas detection using simulation samples, the mSTD-PCR showed consistent sensitivity at 25 copies per test compared with the single-plex CLas assay. The detection result of 30 leaves and 30 root samples showed that the mSTD-PCR could recognize false-negative results caused by the PCR inhibitors and reduce workload by 48% compared with the single-plex CLas assay. Generally, the proposed mSTD-PCR provides a reliable, efficient, inhibitor-monitorable, quantitative screening method for accurately controlling HLB and a universal method for establishing a PCR assay for various pathogens.

First report of soybean root rot caused by Fusarium proliferatum in the Republic of Korea.

Fusarium species are widespread soilborne pathogens that can cause damping-off, root rot, and wilting in soybean [Glycine max (L.) Merrill], subsequently leading to significant yield suppression. Several Fusarium spp. have already been documented for their pathogenicity on soybean plants in the Republic of Korea. The nationwide monitoring of soybean diseases continues to identify new pathogenic Fusarium spp. In 2016, five plant samples at R3-R4 growth stages, showing symptoms of wilting in the upper parts and root rot, were collected in Suwon, Gyeonggi, Republic of Korea. Fungal colonies were obtained from the diseased root samples, with the surface sterilized in 1% sodium hypochlorite for 2 min, rinsed thrice with sterile distilled water, and placed on water agar at 25°C. Five isolates were collected and purified by single-spore isolation. The fungal mycelium was subsequently cultivated on potato dextrose agar for ten days. The isolates produced abundant, aerial, and white mycelium and became purple in old cultures. Macroconidia were slender, falcate to almost straight, usually 3 to 5 septated, and thin-walled. Microconidia were formed in chains from polyphalides, clavate or oval, usually single-celled with a flattened base. These characteristics of isolates were consistent with the description of F. proliferatum (Leslie and Summerrell 2006), and the representative isolate 16-19 was selected for molecular identification to confirm its identity as F. proliferatum. Two evolutionarily conserved genes, the translation elongation factor 1-alpha (EF-1α) and the second-largest subunit of RNA polymerase II (RPB2) genes, were partially amplified using the primers described by O'Donnell et al. (2008), resulting in nucleotide sequences of 680 and 382 base pairs, respectively. These two sequences (GenBank accession numbers: OQ992720 and OR060666) showed 100 and 99.5% identity to the EF-1α and RPB2 of F. proliferatum A40 (GenBank accession numbers: KP964907 and KP964842). For the Petri-dish pathogenicity assay (Broders et al. 2007), five surface-sterilized seeds were placed on water agar media with either sterile water or actively growing '16-19' culture. After 7 days of incubation in a growth chamber (25°C; 12-hour photoperiod), brown lesions were observed on the roots of the inoculated plants, while no symptoms were observed in the sterile water-treated controls. The experiment was conducted three times. For root-cut pathogenicity assay, conidial suspension (1×106 conidia/ml) of the isolate '16-19' was prepared with harvested mycelia cultured on PDA for 10 days with sterile water. The roots of 10-day-old soybean seedlings were partially cut and soaked in either the suspension or sterile water for 2 hours. The seedlings were transplanted into 12 cm plastic pots (11 cm in height) and grew in a greenhouse (26 ± 3°C, 13-h photoperiod). The experiment followed a completely randomized design with three replicates (i.e. three plants in a pot), and it was repeated twice. The inoculated plants began to wilt 7 days after inoculation, while the sterile water-treated controls remained healthy. Ten days after inoculation, all plants were collected, washed under running tap water, and evaluated for the presence and severity of root rot using a 0-4 scale (Chang et al. 2015). The inoculated plants exhibited reduced vigor and developed dark brown lesions on their roots. F. proliferatum was reisolated from symptomatic root tissues of the infected plants, while not from those of the controls. Its colony and spores were morphologically identical to those of the original isolate. F. proliferatum was previously reported as a causative agent of soybean root rot in the United States (Díaz Arias et al. 2011) and Canada (Chang et al. 2015). This is the first report of soybean root rot caused by F. proliferatum in the Republic of Korea. This finding implies that F. proliferatum may potentially threaten soybean production in the Republic of Korea and suggests that effective disease management strategies should be established for soybean protection against the disease, along with continuous surveillance.

Caracterización de Meloidogyne spp. y densidad poblacional de nematodos fitoparásitos asociados a cultivos de aliáceas en la región Arequipa, Perú

Alliaceae crops includes several species of agricultural importance for the Arequipa region, among them the onion is the most produced in the country, however, these crops are affected by phytoparasitic nematodes, including Meloidogyne spp. which causes significant losses in the yield. The aim of the present study was to characterise the root-knot nematode (Meloidogyne spp) and determine the population density of phytoparasitic nematodes in alliaceae crops in the main producing localities of the Arequipa region. Samples of roots and soil were collected in onion, chinese onion, leek and chives crops in the provinces of Arequipa, Castilla, Caylloma and Islay. The identification of nematode genus was found by morphological characteristics and the characterization of Meloidogyne were performed by perineal patterns and electrophoretic pattern of esterase (Est). In the 38 soil samples, different genus of plant-parasitic nematodes was found: Aphelenchus, Dorylaimus, Helicotylenchus, Hemicycliophora, Meloidogyne, Mesocriconema, Mononchus, OGMA, Pratylenchus, Rotylenchus, Tylenchus, Xiphinema and free-living nematodes, in root samples, eight populations of Meloidogyne spp. were found. distributed among three species: (1) M. incognita phenotype Est I1, Rm: 1.0 with 33.33% of frequency and Est I2, Rm: 1.03, 1.07 (50%); (2) M. javanica Est J3, Rm: 1.03, 1.24, 1.37 (16.67%) and an atypical species named Meloidogyne sp. Rm: 0.85 (33.33%). The identification of genus and species phytoparasitic nematodes in alliaceae will allow the adoption of more efficient integrated management measures, because have a distinct parasitism behavior.

Variation in Root Biomass and Distribution Based on the Topography, Soil Properties, and Tree Influence Index: The Case of Mt. Duryun in Republic of Korea.

Root biomass and distribution are influenced by abiotic factors, such as topography and soil physicochemical properties, determining belowground productivity. Hence, we investigated the variation in root biomass and vertical root distribution based on the topography, soil physicochemical properties, and tree influence index, and their relationships, across soil depths (0-10 cm, 10-20 cm, and 20-30 cm) and topographical gradients in a warm-temperate forest in Mt. Duryun, Republic of Korea. Two contrasting research sites were established: a lower slope oriented at ≤3° and an upper slope with a slope of 30°. Each site comprised eleven 400 m2 sampling plots from which root samples from various diameter classes (<2 mm, 2-5 mm, 5-10 mm, and >10 mm) were collected. While the bulk density increased with soil depth in the lower slope, the organic matter, available phosphorus, Ca2+, and Mg2+ showed a reversed pattern. Linear mixed-effects models generally revealed significant negative correlations between root biomass and soil pH, total nitrogen, and cation exchange capacity, particularly in small roots (βstd = -1.03 to -1.51) and coarse roots (βstd = -6.30). Root biomass exhibited a 10-15% increase in the upper slope compared to the lower slope, particularly in fine (median = 52.0 g m2-65.64 g m2) and medium roots (median = 56.04 g m2-69.52 g m2) at a 0-20 cm soil depth. While no significant correlation between root biomass and the tree influence index was found on the lower slope, a different pattern was found on the upper slope. Our results indicate that the variation in root biomass and distribution can also be explained by the differences in the soil environment and topographical positions.

Distribution and diversity studies of endophytic and rhizospheric fungi associated with the mangrove, Acanthus ilicifolius, using amplicon sequencing-based metagenomic approach

Fungal endophytes exhibit a broad diversity and are found universally associated with all plant species, playing a vital role in enhancing the host plant's resilience against both biotic and abiotic stressors. In this investigation conducted in Mumbai, India, the Illumina MiSeq sequencing platform was employed to study the endophytic fungal diversity of the common mangrove, Acanthus ilicifolius and its rhizosphere utilizing the amplification of the fungal ribosomal ITS2 region. The results revealed the creation of 307 Operational Taxonomic Units (OTUs) from a total of 663,600 sequencing reads, clustered at a 97 percent similarity level. Evaluation using Simpson and Shannon indices indicated that the highest community diversity was observed in stem tissue compared to root and leaf samples. The total OTUs were categorized into five phyla, 41 orders, 89 families, and 133 genera, with Ascomycota being the predominant phylum at 76.9% relative abundance, followed by Basidiomycota at 22.87%. The most abundant genera in the soil were Malassezia (35%) and Aspergillus (29.5%), while Vishniacozyma (61%) dominated in leaf samples. Unlike stem and root, no single genus dominated these tissues. The findings demonstrated significant distinctions between the fungal endophytic communities in plant tissues and the fungal microbiome in the soil. Notably, the endophyte fungi community in roots exhibited a closer resemblance to stem samples than to leaf samples. FunGuild analysis also revealed characteristic communities in different samples indicating specific functions of these genera. Overall, this study provides novel insights into the intricate composition of endophytic fungi within the tissues of A. ilicifolius and its rhizosphere.

Chitosan nanoemulsion incorporated with Carum carvi essential oil as ecofriendly alternative for mitigation of aflatoxin B1 contamination in stored herbal raw materials

The present investigation entails the first report on entrapment of Carum carvi essential oil (CCEO) into chitosan polymer matrix for protection of stored herbal raw materials against fungal inhabitation and aflatoxin B1 (AFB1) production. Physico-chemical characterization of nanoencapsulated CCEO was performed through Fourier transform infrared spectroscopy, dynamic light scattering, X-ray diffractometry, and scanning electron microscopy. The nanoencapsulated CCEO displayed improved antifungal and AFB1 suppressing potentiality along with controlled delivery over unencapsulated CCEO. The encapsulated CCEO nanoemulsion obstructed the ergosterol production and escalated the efflux of cellular ions, thereby suggesting plasma membrane as prime target of antifungal action in Aspergillus flavus cells. The impairment in methyglyoxal production and modeling based carvone interaction with Afl-R protein validated the antiaflatoxigenic mechanism of action. In addition, CCEO displayed augmentation in antioxidant potentiality after encapsulation into chitosan nanomatrix. Moreover, the in-situ study demonstrated the effective protection of Withania somnifera root samples (model herbal raw material) against fungal infestation and AFB1 contamination along with prevention of lipid peroxidation. The acceptable organoleptic qualities of W. somnifera root samples and favorable safety profile in mice (animal model) strengthen the application of nanoencapsulated CCEO emulsion as nano-fungitoxicant for preservation of herbal raw materials against fungi and AFB1 mediated biodeterioration.

Comparative transcriptome analysis between two different cadmium-accumulating genotypes of soybean (Glycine max) in response to cadmium stress

BackgroundCadmium (Cd) is extremely toxic and non-essential for plants. Different soybean varieties differ greatly in their Cd accumulation ability, but little is known about the underlying molecular mechanisms.ResultsHere, we performed transcriptomic analysis using Illumina pair-end sequencing on root tissues from two soybean varieties (su8, high-Cd-accumulating (HAS) and su7, low Cd-accumulating (LAS)) grown with 0 or 50 μM CdSO4. A total of 18.76 million clean reads from the soybean root samples were obtained after quality assessment and data filtering. After Cd treatment, 739 differentially expressed genes (DEGs; 265 up and 474 down) were found in HAS; however, only 259 DEGs (88 up and 171 down) were found in LAS, and 64 genes were same between the two varieties. Pathway enrichment analysis suggested that after cadmium treatment, the DEGs between LAS and HAS were mainly enriched in glutathione metabolism and plant-pathogen interaction pathways. KEGG analysis showed that phenylalanine metabolism responding to cadmium stress in LAS, while ABC transporters responding to cadmium stress in HAS. Besides we found more differential expressed heavy metal transporters such as ABC transporters and zinc transporters in HAS than LAS, and there were more transcription factors differently expressed in HAS than LAS after cadmium treatment in two soybean varieties, eg. bHLH transcription factor, WRKY transcription factor and ZIP transcription factor.ConclusionsFindings from this study will shed new insights on the underlying molecular mechanisms behind the Cd accumulation in soybean.

Deciphering mycobiota and its functional dynamics in root hairs of Rhododendron campanulatum D. Don through Next-gen sequencing

The Himalayas provide unique opportunities for the extension of shrubs beyond the upper limit of the tree. However, little is known about the limitation of the biotic factors belowground of shrub growth at these cruising altitudes. To fill this gap, the present study deals with the documentation of root-associated microbiota with their predicted functional profiles and interactions in the host Rhododendron campanulatum, a krummholz species. While processing 12 root samples of R. campanulatum from the sites using Omics we could identify 134 root-associated fungal species belonging to 104 genera, 74 families, 39 orders, 17 classes, and 5 phyla. The root-associated microbiota members of Ascomycota were unambiguously dominant followed by Basidiomycota. Using FUNGuild, we reported that symbiotroph and pathotroph as abundant trophic modes. Furthermore, FUNGuild revealed the dominant prevalence of the saptroptroph guild followed by plant pathogens and wood saprotrophs. Alpha diversity was significantly different at the sites. The heatmap dendrogram showed the correlation between various soil nutrients and some fungal species. The study paves the way for a more in-depth exploration of unidentified root fungal symbionts, their interactions and their probable functional roles, which may serve as an important factor for the growth and conservation of these high-altitude ericaceous plants.