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Pest Management Science Mar 2024The biocontrol potential of soil microbes can reduce the extensive use of hazardous synthetic fungicides. This study was designed to find a strain of rhizobacteria...
BACKGROUND
The biocontrol potential of soil microbes can reduce the extensive use of hazardous synthetic fungicides. This study was designed to find a strain of rhizobacteria indigenous to Pakistan with potential biocontrol against early blight of tomato caused by Alternaria solani and to characterize its biocontrol mechanisms.
RESULTS
Among 88 strains tested for antagonism against A. solani on agar media, S27, Dt10 and 423, identified by 16S rRNA sequencing as strains of Bacillus amyloliquefaciens, B. cereus and Stenotrophomonas rhizophila, respectively, were the most inhibitory. When applied to detached tomato leaflets in Petri dish assays, the strains reduced lesion development by over 30% compared to the control. In greenhouse pot trials, the bacterial strains reduced early blight severity by over 50%. In three field trials, all three strains applied to tomato foliage slowed early blight disease progress and reduced disease severity, with B. amyloliquefaciens S27 reducing the area under the disease progress curve by up to 70%. All three strains showed protease, catalase and oxidase activities in vitro, but none produced β-1,3-glucanase and only B. cereus Dt10 showed slight chitinase activity. In a greenhouse experiment in which the bacteria were applied to tomato foliage prior to pathogen inoculation, bacteria-treated leaves had higher β-1,3-glucanase and chitinase levels than leaves inoculated only with the pathogen, indicating priming induction of response.
CONCLUSION
Three rhizobacteria strains have the potential to control early blight of tomato under Pakistan's growing conditions, with B. amyloliquefaciens S27 being the most promising candidate for commercial development. Antagonism and induction of the priming response may be mechanisms of biocontrol by the bacterial strains. © 2023 Society of Chemical Industry.
Topics: Solanum lycopersicum; Pakistan; RNA, Ribosomal, 16S; Chitinases; Plant Diseases
PubMed: 37939120
DOI: 10.1002/ps.7872 -
Langmuir : the ACS Journal of Surfaces... Nov 2023Hydrophilicity is a requisite attribute for the 2D cell culture substrate's surface, facilitating cell adhesion and spreading. Conventional poly(dimethylsiloxane) (PDMS)...
Hydrophilicity is a requisite attribute for the 2D cell culture substrate's surface, facilitating cell adhesion and spreading. Conventional poly(dimethylsiloxane) (PDMS) microfluidic chips necessitate protein coatings to enhance hydrophilicity; however, this approach is afflicted by issues of transient efficacy, interference with cell analysis, and high costs. This paper presents a protein-free microfluidic chip, termed a "microfluidic Petri dish-chip (MPD-chip)", integrating PDMS as the cover and a tissue culture-treated (TC-treated) Petri dish as the substrate. Microstructures are hot-embossed onto the Petri dish substrate using a silicon mold. This meticulous replication process serves to establish stable flow field dynamics within the chip. A simplified method for irreversible bonding, utilizing plasma activation and silylation, is proposed for affixing the PDMS cover onto the microstructured Petri dish substrate. The prepared composite chip exhibits remarkable tightness, boasting a notable bond strength of 2825 kPa. Furthermore, the composite microfluidic chip demonstrates the capability to withstand flow velocities of at least 200 μL/min, effectively meeting the required injection standards for both cell suspension and culture medium. SH-SY5Y and HeLa cells are cultured dynamically in the MPD-chip and control groups. Outcomes encompassing normalized cell density, cell adhesion area, and cell viability metrics unequivocally highlight the superiority of the MPD-chip in facilitating long-term two-dimensional (2D) cell cultures.
Topics: Humans; Microfluidics; Microfluidic Analytical Techniques; HeLa Cells; Neuroblastoma; Cell Culture Techniques; Proteins
PubMed: 37906157
DOI: 10.1021/acs.langmuir.3c01982 -
Stem Cell Research & Therapy Oct 2023Thymic epithelial cells (TECs) are responsible for shaping the repertoires of T cells, where their postnatal regeneration depends on a subset of clonogenic TECs. Despite...
BACKGROUND
Thymic epithelial cells (TECs) are responsible for shaping the repertoires of T cells, where their postnatal regeneration depends on a subset of clonogenic TECs. Despite the implications for regenerative medicine, their cultivation and expansion remain challenging. Primary explant cell culture is a technique that allows the seeding and expansion of difficult-to-culture cells. Here, we report a reliable and simple culture system to obtain functional TECs and thymic interstitial cells (TICs).
METHODS
To establish primary thymic explants, we harvested 1 mm cleaned fragments of thymus from 5-week-old C57/BL6 mice. Tissue fragments of a complete thymic lobe were placed in the center of a Petri dish with 1 mL of DMEM/F-12 medium supplemented with 20% fetal bovine serum (FBS) and 1% penicillin‒streptomycin. To compare, thymic explants were also cultivated by using serum-free DMEM/F-12 medium supplemented with 10% KnockOut™.
RESULTS
We obtained high numbers of functional clonogenic TECs and TICs from primary thymic explants cultivated with DMEM/F-12 with 20% FBS. These cells exhibited a highly proliferative and migration profile and were able to constitute thymospheres. Furthermore, all the subtypes of medullary TECs were identified in this system. They express functional markers to shape T-cell and type 2 innate lymphoid cells repertoires, such as Aire, IL25, CCL21 and CD80. Finally, we also found that ≥ 70% of lineage negative TICs expressed high amounts of Aire and IL25.
CONCLUSION
Thymic explants are an efficient method to obtain functional clonogenic TECs, all mTEC subsets and different TICs AireIL25 with high regenerative capacity.
Topics: Mice; Animals; Immunity, Innate; Lymphocytes; Thymus Gland; Epithelial Cells; T-Lymphocytes; Cell Differentiation
PubMed: 37904232
DOI: 10.1186/s13287-023-03529-8 -
Ying Yong Sheng Tai Xue Bao = the... Sep 2023The use of artificial cyanobacteria crusts is one of the effective methods to prevention and control of desertification. Soil fine substance is one of the important...
The use of artificial cyanobacteria crusts is one of the effective methods to prevention and control of desertification. Soil fine substance is one of the important factors limiting the colonization and growth of artificial cyanobacteria crusts. We compared the growth of artificial cyanobacterial crusts with different fine substance contents by setting the volume ratios of fine substance to quicksand as 0:1, 1:1, 2:1, 4:1 and 1:0. The results showed that the cover of artificial cyanobacteria crusts increased gradually with the increases of fine substance contents, while the contents of chlorophyll a and extracellular polysaccharide firstly increased and then decreased slightly. The optimum growth of artificial cyanobacterial crusts was achieved under the treatment of 4:1 ratio. Under such treatment after 60 days of incubation, artificial cyanobacteria crusts cover was 70%, and the contents of chlorophyll a, loosely bound exopolysaccharide (LB-EPS), tightly bound exopolysaccharide (TB-EPS), and glycocalyx exopolysaccharide (G-EPS) were 17.5, 70.0, 175.0, and 200.0 μg·cm, respectively. Increasing the amount of cyanobacteria under the condition of low fine substance content could promote the formation and growth of artificial cyanobacterial crusts (0.5 g of cyanobacteria per petri dish was the optimal). It could provide a new idea for the large-scale culture of artificial cyanobacterial crusts inoculum.
Topics: Chlorophyll A; Soil; Soil Microbiology; Cyanobacteria
PubMed: 37899105
DOI: 10.13287/j.1001-9332.202309.014 -
Journal of the American Chemical Society Nov 2023We report the use of acid-diffusion to assemble core-shell supramolecular gel beads with different low-molecular-weight gelators (LMWGs) in the core and shell. These gel...
We report the use of acid-diffusion to assemble core-shell supramolecular gel beads with different low-molecular-weight gelators (LMWGs) in the core and shell. These gel beads grow a shell of dibenzylidenesorbitol-based DBS-COOH onto a core comprising DBS-CONHNH and agarose that has been loaded with acetic acid. Diffusion of the acid from the core triggers shell assembly. The presence of DBS-CONHNH enables the gel core to be loaded with metal nanoparticles (NPs) as acyl hydrazide reduces metal salts . The pH-responsiveness of DBS-COOH allows responsive assembly of the shell with both temporal and spatial control. By fixing multiple gel beads in a Petri dish, the cores become linked to one another by the assembled DBS-COOH gel shell─a process we describe as diffusion-adhesion assembly. By controlling the geometry of the beads with respect to one another, it is possible to pattern the structures, and using a layer-by-layer approach, 3D objects can be fabricated. If some of the beads are loaded with basic DBS-carboxylate instead of CHCOOH, they act as a "sink" for diffusing protons, preventing DBS-COOH shell assembly in the close proximity. Those beads do not adhere to the remainder of the growing gel object and can be simply removed once diffusion-assembly is complete, acting as templates, and enabling the fabrication of 3D "imprinted" multigel architectures. Preloading the gel beads with AuNPs or AgNPs suspends these functional units within the cores at precisely defined locations within a wider gel object. In summary, this approach enables the dynamic fabrication of shaped and patterned gels with embedded metal NPs─such objects have potential next-generation applications in areas including soft nanoelectronics and regenerative medicine.
PubMed: 37885219
DOI: 10.1021/jacs.3c07376 -
Plant Disease Oct 2023The state of Puebla is the main producer of cabbage ( var. ) in Mexico, with an area of approximately 1,858 ha (SIAP 2023). In April 2023, a field sampling was conducted...
The state of Puebla is the main producer of cabbage ( var. ) in Mexico, with an area of approximately 1,858 ha (SIAP 2023). In April 2023, a field sampling was conducted in the San Luis Ajajalpan, Tecali de Herrera (18°55.57'N, 97°55.607'W), Puebla, Mexico. The average temperature was 24°C and the relative humidity was 95% for five consecutive days. Cabbage plants cv. 'American Taki San Juan' close to harvest, with head rot symptoms were found in a commercial area of approximately 3 ha, at an estimated incidence of 35 to 45%. More than 70% of the leaves were symptomatic on severely affected plants. Typical symptoms included chlorosis of older foliage, soft rot with abundant white to gray mycelium, and abundant production of large and irregularly-shaped sclerotia. The fungus was isolated from 30 symptomatic plants. Sclerotia were collected from symptomatic heads, surface sterilized in 3% NaOCl, rinsed twice with sterile distilled water, and plated on Potato Dextrose Agar (PDA) with sterile forceps. Subsequently, a dissecting needle was used to place fragments of mycelium directly on PDA. Plates were placed in an incubator at 25°C in the dark. A total of 30 representative isolates were obtained by the hyphal-tip method, one from each diseased plant (15 isolates from sclerotia and 15 from mycelial fragments). After 8 days, colonies had fast-growing, dense, cottony-white aerial mycelium forming irregular sclerotia of 3.75 ± 0.8 mm (mean ± standard deviation, n=100). Each Petri dish produced 14-25 sclerotia (mean = 18, n = 50), after 10 days. The sclerotia were initially white and gradually turned black. The isolates were identified as based on morphological characteristics (Saharan and Mehta 2008). Two representative isolates were chosen for molecular identification, and genomic DNA was extracted by a CTAB protocol. The ITS region and the glyceraldehyde 3-phosphate dehydrogenase (G3PDH) gene were sequenced for two isolates (White et al. 1990; Staats et al. 2005). The ITS and G3PDH sequences of a representative isolate (SsC.1) were deposited in the GenBank (ITS- OR286628; G3PDH- OR333495). BLAST analysis of the partial sequences ITS (509 bp) and G3PDH (915 bp) showed 100% similarity to S. sclerotiorum isolates (GenBank: MT436756.1 and OQ790148). Pathogenicity was confirmed by inoculating 10 detached cabbage heads of 'American Taki San Juan', using the SsC.1 isolate, according to Sanogo et al. (2015). Heads were placed on the rim of a plastic container and inserted in a moisture box with 2 cm of water on its bottom. The box was covered with a plastic sheet to maintain humidity. The control plants were inoculated with a plug of noncolonized PDA. The inoculated cabbages were covered with white to gray mycelia and abundant sclerotia within 10 days, whereas no symptoms were observed on non-inoculated controls. The fungus was re-isolated from the inoculated cabbages as described above, fulfilling Koch's postulates. The pathogenicity tests were repeated three times. White mold caused by on Brussels sprouts was recently reported in Mexico (Ayvar-Serna et al. 2023). In 2015, . was reported on cabbage in New Mexico, causing head rot (Sanogo et al. 2015). To our knowledge, this is the first report of . causing white mold on cabbage in Mexico. This research is essential for designing management strategies and preventing spread to other production areas.
PubMed: 37884482
DOI: 10.1094/PDIS-08-23-1534-PDN -
Journal of the Mechanical Behavior of... Dec 2023Alginate gel scaffolds are biocompatible and biodegradable materials that have been used in a variety of tissue engineering applications. The porosity of alginate gel...
Alginate gel scaffolds are biocompatible and biodegradable materials that have been used in a variety of tissue engineering applications. The porosity of alginate gel scaffolds is an important property that affects their performance. However, it is difficult to predict the porosity of alginate gel scaffolds accurately. In this study, a GA-coupled ANN model was developed to predict the porosity of alginate gel scaffolds. The model was trained on a dataset of 107 scaffolds with known porosities. The model was able to achieve a mean absolute error of 0.13, which suggests that it is able to accurately predict the porosity of alginate gel scaffolds. The alginate scaffold was fabricated by a microfluidic technique using a syringe pump and a flow device. The crosslinker solution was poured into the Petri dish to crosslink the polymer to the gel structure. The Archimedes method was used to determine the scaffold's apparent porosity. The artificial neural network has been used to model the porosity of the gel scaffold using the input parameters such as alginate-pluronic viscosity, surface tension, and contact angle etc. The maximum porosity was modelled to be 96.4 % using GA whereas the experimental value for the same was measured to be 92.8 ± 2 %. A 3.7% variation in the porosity was found from modelled value. To the best of our knowledge, this study is the first to develop an integrated ANN-coupled GA model to predict the maximum porosity of the gel scaffold. The result indicates that artificial intelligence has great potential for optimizing the parameters to fabricate the gel scaffold that can be used for tissue engineering applications.
Topics: Tissue Scaffolds; Porosity; Alginates; Artificial Intelligence; Tissue Engineering; Biocompatible Materials
PubMed: 37883894
DOI: 10.1016/j.jmbbm.2023.106204 -
Plant Disease Oct 2023Pseudostellaria heterophylla is one of the traditional medicines in China. From 2020 to 2022, postharvest wet root rot disease was observed with an incidence of 2~5% on...
Pseudostellaria heterophylla is one of the traditional medicines in China. From 2020 to 2022, postharvest wet root rot disease was observed with an incidence of 2~5% on the tuberous roots of the harvested P. heterophylla in Zherong county, Fujian province, China, which usually occurs under damp and unventilated conditions. The symptoms of the disease were as follows: white mycelia grew on the surface of tuberous root initially and gradually wrapped around the roots, the internal root tissue turned yellow and became wet decay finally. To identify the causal agent, a total of 20 samples with symptomatic tuberous roots were collected. Small pieces (3 mm2) were treated by surface disinfection with 75% ethanol and 1% NaOCl, then rinsed 3 times with sterile water. These treated pieces were transferred onto potato dextrose agar (PDA) and incubated at 25°C in the dark for 7 d. Ten pure cultures were obtained using single-spore isolation method. The fungus colonies initially produced white aerial mycelium, subsequently exhibited yellow pigmentation. Mycelia were consisted of smooth, hyaline, branched, and septate hyphae. The conidia were solitary or clustered, brown or dark brown, smooth, ellipsoidal to spherical, 6.66 (5.50-7.81)×5.65 (4.17-7.22) µm (n=50) in size. The conidiophores were hyaline or pale brown and produced conidiogenous cells, which were pale brown, smooth, ampulliform, and 10.14 (8.82-15.30) um long (n=50). Based on these morphological characteristics, the fungus was identified as the genus Apiospora (Arthrinium). The rDNA-ITS region and partial β-tubulin gene (BenA) were amplified using the primers ITS1/ITS4 (White et al. 1990) and Bt2a/Bt2b (Glass and Donaldson 1995), respectively. The sequences of isolates FJAT-32563 and FJAT-32564 were deposited in GenBank (ITS, OM920984 and OM920985; BenA, OM953823 and OM953824). All sequences had more than 99% similarity with those of A. arundinis strain CBS:106.12 (ITS, KF144883; BenA, KF144973). In the multilocus phylogenetic analysis (ITS + BenA), the two isolates clustered together with other strains of A. arundinis with 100% bootstrap support. The isolates were therefore identified as A. arundinis based on both morphological and molecular characteristics. To confirm the pathogenicity, fresh tuberous roots were selected and surface disinfected, then the roots were immersed with a quarter length in the conidial suspension (106/mL) for 30 min, whereas the control roots were immersed with sterile water (n=30). They were placed in petri dish with wet filter paper at 25±2℃, maintaining 80% relative humidity in the dark. The white aerial mycelium appeared at 5 days after inoculation, and wet root rot decaying occurred after inoculation for 21 days. The symptoms were similar to those described above, whereas the control roots were asymptomatic. The same fungus was re-isolated from the infected roots, showing similar morphological characteristics and molecular traits. Koch's postulates were completed and the pathogenicity test for the isolates has been repeated thrice. Previously, A. arundinis was reported to infect peach and sugarcane (Ji et al. 2020; Liao et al. 2022). To our knowledge, this is the first report of A. arundinis causing wet root rot of P. heterophylla in China. The disease would be a potentially new threat to this medicinal plant.
PubMed: 37883638
DOI: 10.1094/PDIS-05-23-0911-PDN -
Environmental Science and Pollution... Nov 2023The possibility of using the non-nitrogen-fixing cyanobacterium (Chroococcus sp.) for the reduction of soil nitrate contamination was tested through Petri dish...
The possibility of using the non-nitrogen-fixing cyanobacterium (Chroococcus sp.) for the reduction of soil nitrate contamination was tested through Petri dish experiments. The application of 0.03, 0.05 and 0.08 mg/cm Chroococcus sp. efficiently removed NO-N from the soil through assimilation of nitrate nutrient and promotion of soil denitrification. At the optimal application dose of 0.05 mg/cm, 44.06%, 36.89% and 36.17% of NO-N were removed at initial NO-N concentrations of 60, 90 and 120 mg/kg, respectively. The polysaccharides released by Chroococcus sp. acted as carbon sources for bacterial denitrification and facilitated the reduction of soil salinity, which significantly (p < 0.05) stimulated the growth of denitrifying bacteria (Hyphomicrobium denitrificans and Hyphomicrobium sp.) as well as significantly (p < 0.05) elevated the activities of nitrate reductase and nitrite reductase by 1.07-1.23 and 1.15-1.22 times, respectively. The application of Chroococcus sp. promoted the dominance of Nocardioides maradonensis in soil microbial community, which resulted in elevated phosphatase activity and increased available phosphorus content. The application of Chroococcus sp. positively regulated the growth of soil bacteria belonging to the genera Chitinophaga, Prevotella and Tumebacillus, which may contribute to increased soil fertility through the production of beneficial enzymes such as invertase, urease and catalase. To date, this is the first study verifying the remediation effect of non-nitrogen-fixing cyanobacteria on nitrate-contaminated soil.
Topics: Nitrates; Cyanobacteria; Nitrate Reductase; Nitrite Reductases; Soil; Denitrification
PubMed: 37870669
DOI: 10.1007/s11356-023-30383-1 -
Computational and Structural... 2023Performing lifespan assays with () nematodes manually is a time consuming and laborious task. Therefore, automation is necessary to increase productivity. In this...
Performing lifespan assays with () nematodes manually is a time consuming and laborious task. Therefore, automation is necessary to increase productivity. In this paper, we propose a method to automate the counting of live using deep learning. The survival curves of the experiment are obtained using a sequence formed by an image taken on each day of the assay. Solving this problem would require a very large labeled dataset; thus, to facilitate its generation, we propose a simplified image-based strategy. This simplification consists of transforming the real images of the nematodes in the Petri dish to a synthetic image, in which circular blobs are drawn on a constant background to mark the position of the . To apply this simplification method, it is divided into two steps. First, a Faster R-CNN network detects the , allowing its transformation into a synthetic image. Second, using the simplified image sequence as input, a regression neural network is in charge of predicting the count of live nematodes on each day of the experiment. In this way, the counting network was trained using a simple simulator, avoiding labeling a very large real dataset or developing a realistic simulator. Results showed that the differences between the curves obtained by the proposed method and the manual curves are not statistically significant for either short-lived N2 (p-value log rank test 0.45) or long-lived (p-value log rank test 0.83) strains.
PubMed: 37867965
DOI: 10.1016/j.csbj.2023.10.007