Colony Growth Rate Research Articles

Identification and characterization of Botrytis cinerea causing gray mold on tomato in Malaysia

Botrytis cinerea, commonly known as gray mold, is a pervasive fungal pathogen that affects a wide range of plant species, leading to significant agricultural losses. The identification of Botrytis cinerea in Malaysia is crucial for protecting the agricultural sector, minimizing economic losses, ensuring food security, maintaining export quality, addressing environmental concerns, and advancing scientific research. In the present research, tomato fruits collected from Cameron Highlands, Pahang, Malaysia showed gray mold disease symptoms of B. cinerea. The fungal isolates were examined morphologically for colony colour, growth rate, conidiophores, conidia shape, and sclerotia on PDA and V8 agar. According to the results, conidiophores appeared in grape shape and length was range of 21.26-32.52 μm, ovoid conidial dimensions were in the range of 10.03-16.08 × 7.37-11.15 μm and sclerotia size was range 1.91-4.50 × 1.70-4.00 mm. All isolates were attributed to the morphospecies Botrytis cinerea on account of these characteristics. The resulting sequences deposited in GenBank were accessions MT012053 to MT012062, respectively. A BLAST analysis of the resulting 550-bp nucleotide sequences showed 99-100% identity closest matched to B. cinerea. The pathogenicity experiments showed P6 isolates of B. cinerea were highly pathogenic and caused gray mold development on tomato fruits that led to severe symptoms in five days. Meanwhile, the least pathogenic isolate was P9. In terms of temperature, B. cinerea grew faster on PDA at 20ºC, slower grew below 20ºC and did not grow at 25ºC. Identification and characterization of B. cinerea on tomato could potentially provide information to assist disease management strategies for B. cinerea.

The antagonistic activity of Streptomyces spiroverticillatus (No. HS1) against of poplar canker pathogen Botryosphaeria dothidea

BackgroundPoplar canker caused by Botryosphaeria dothidea is one of the most severe plant disease of poplars worldwide. In our study, we aimed to investigate the modes of antagonism by fermentation broth supernatant (FBS) of Streptomyces spiroverticillatus HS1 against B. dothidea.ResultsIn vitro, the strain and FBS of S. spiroverticillatus HS1 significantly inhibited mycelial growth and biomass accumulation, and also disrupted the mycelium morphology of B. dothidea. On the 3rd day after treatment, the inhibition rates of colony growth and dry weight were 80.72% and 52.53%, respectively. In addition, FBS treatment damaged the plasma membrane of B. dothidea based on increased electrical conductivity in the culture medium, and malondialdehyde content of B. dothidea mycelia. Notably, the analysis of key enzymes in glycolysis pathway showed that the activity of hexokinase (HK), phosphofructokinase (PFK), and pyruvate kinase (PK), Ca2+Mg2+-ATPase were significantly increased after FBS treatment. But the glucose contents were significantly reduced, and pyruvate contents were significantly increased in B. dothidea after treatment with FBS.ConclusionsThe inhibitory mechanism of S. spiroverticillatus HS1 against B. dothidea was a complex process, which was associated with multiple levels of mycelial growth, cell membrane structure, material and energy metabolism. The FBS of S. spiroverticillatus HS1 could provide an alternative approach to biological control strategies against B. dothidea.

Discovery and Characterization of Epichloë Fungal Endophytes from Elymus spp. in Northwest China.

Epichloë fungal endophytes hold promise in sustainable agriculture by fortifying cool-season grasses such as Elymus spp. against various stresses. Elymus spp. are widely distributed in Northwest China with a high incidence of endophyte infections. In this study, we identified 20 Epichloë endophytic fungal strains carried by five Elymus spp. from five areas of Northwest China and systematically characterized their morphology, molecular phylogeny, mating type, and alkaloid diversity for the first time. The morphological characterization underscores strain diversity, with variable colony textures and growth rates. A phylogenetic analysis confirms all strains are E. bromicola, emphasizing their taxonomic status. Alkaloid-encoding gene profiling delineates distinct alkaloid synthesis capabilities among the strains, which are crucial for host adaptability and resistance. A mating-type analysis reveals uniformity (mtAC) across the Epichloë strains, simplifying breeding strategies. Notably, the Epichloë strains exhibit diverse alkaloid synthesis gene profiles, impacting host interactions. This research emphasizes the ecological significance of Epichloë endophytes in Elymus spp. ecosystems, offering insights into their genetic diversity and potential applications in sustainable agriculture.

Study on the Biological Characteristics of Dark Septate Endophytes under Drought and Cadmium Stress and Their Effects on Regulating the Stress Resistance of Astragalus membranaceus.

Astragalus membranaceus is a famous traditional medicinal plant. However, drought and cadmium (Cd) pollution are the main abiotic stress factors that affect plant growth and yield and the ability to improve the host's stress resistance through the use of beneficial endophytic fungi. To evaluate the tolerance of dark septate endophytes (DSE) to various abiotic stresses, 10 DSE strains [Microsphaeropsis cytisi (Mc), Alternaria alstroemeriae (Aa), Stagonosporopsis lupini (Sl), Neocamarosporium phragmitis (Np), Paraphoma chlamydocopiosa (Pc), Macrophomina phaseolina (Mp'), Papulaspora equi (Pe), Alternaria tellustris (At), Macrophomina pseudophaseolina (Mp), and Paraphoma radicina (Pr)] were investigated under different drought and Cd stressors in vitro by using solid-plate cultures and liquid-shaker cultures in the current study. The experiments involved using varying concentrations of PEG (0, 9, 18, and 27%) and Cd2+ (0, 25, 50, and 100 mg/L) to simulate different stress conditions on DSE. Additionally, the effect of DSE (Np and At) on the growth of A. membranaceus at different field water capacities (70% and 40%) and at different CdCl2 concentrations (0, 5, 10, and 15 mg Cd/kg) in soil was studied. The results demonstrated that the colony growth rates of Aa, Np, Pc, Mp', and Mp were the first to reach the maximum diameter at a PEG concentration of 18%. Aa, Np, and At remained growth-active at 100 mg Cd/L. In addition, Aa, Np, and At were selected for drought and Cd stress tests. The results of the drought-combined-with-Cd-stress solid culture indicated that the growth rate of Np was significantly superior to that of the other strains. In the liquid culture condition, the biomasses of Np and Aa were the highest, with biomasses of 1.39 g and 1.23 g under the concentration of 18% + 25 mg Cd/L, and At had the highest biomass of 1.71 g at 18% + 50 mg Cd/L concentration, respectively. The CAT and POD activities of Np reached their peak levels at concentrations of 27% + 50 mg Cd/L and 27% + 25 mg Cd/L, respectively. Compared to the control, these levels indicated increases of 416.97% and 573.12%, respectively. Aa, Np, and At positively influenced SOD activity. The glutathione (GSH) contents of Aa, Np, and At were increased under different combined stressors of drought and Cd. The structural-equation-modeling (SEM) analysis revealed that Aa positively influenced biomass and negatively affected Cd content, while Np and At positively influenced Cd content. Under the stress of 40% field-water capacity and the synergistic stress of 40% field-water capacity and 5 mg Cd/kg soil, Np and At significantly increased root weight of A. membranaceus. This study provides guidance for the establishment of agricultural planting systems and has good development and utilization value.

Evaluating Linear Coral Growth Estimation Using Photogrammetry and Alternative Point Cloud Comparison Methods

Abstract. Corals are critical reef-building organisms, providing essential habitat and ecosystem services. Tracking coral growth over time indicates coral reef health, which can be measured using various established techniques. Several coral growth-related studies have successfully applied photogrammetry to a particular coral of various types. While the focus of previous work was on standardised data processing and, to a certain degree, on the assessment of different point cloud comparison methods (Lange et al. 2022), little attention has been given to the impact of camera calibration. This study measured the annual linear extension of five Acropora spp. colonies using photogrammetry and evaluated all stages of imagery processing. A high focus was given to the analysis of the camera calibration method and the validation of camera parameters derived using an in-situ calibration of coral images with scale bars placed in the camera's field of view. We demonstrate that this method is as reliable as the calibration using a calibration frame. This study also examined the impact of the different point cloud comparison methods for Acropora spp. More specifically, the derived point clouds are compared by applying the point-to-point and point-to-model methods and manually selecting 12 coral branch tips. Histograms derived from the comparison methods were analysed and deemed a suitable and efficient alternative approach for measuring the maximum growth rate of mature colonies over shorter time periods (1 year or less).

A Novel Strain of Fusarium oxysporum Alternavirus 1 Isolated from Fusarium oxysporum f. sp. melonis Strain T-BJ17 Confers Hypovirulence and Increases the Sensitivity of Its Host Fungus to Difenoconazole and Pydiflumetofen.

In the current study, a novel strain of Fusarium oxysporum alternavirus 1 (FoAV1) was identified from the Fusarium oxysporum f. sp. melonis (FOM) strain T-BJ17 and was designated as Fusarium oxysporum alternavirus 1-FOM (FoAV1-FOM). Its genome consists of four dsRNA segments of 3515 bp (dsRNA1), 2663 bp (dsRNA2), 2368 bp (dsRNA3), and 1776 bp (dsRNA4) in length. Open reading frame 1 (ORF1) in dsRNA1 was found to encode a putative RNA-dependent RNA polymerase (RdRp), whose amino acid sequence was 99.02% identical to that of its counterpart in FoAV1; while ORF2 in dsRNA2, ORF3 in dsRNA3, and ORF4 in dsRNA4 were all found to encode hypothetical proteins. Strain T-BJ17-VF, which was verified to FoAV1-FOM-free, was obtained using single-hyphal-tip culture combined with high-temperature treatment to eliminate FoAV1-FOM from strain T-BJ17. The colony growth rate, ability to produce spores, and virulence of strain T-BJ17 were significantly lower than those of T-BJ17-VF, while the dry weight of the mycelial biomass and the sensitivity to difenoconazole and pydiflumetofen of strain T-BJ17 were greater than those of T-BJ17-VF. FoAV1-FOM was capable of 100% vertical transmission via spores. To our knowledge, this is the first time that an alternavirus has infected FOM, and this is the first report of hypovirulence and increased sensitivity to difenoconazole and pydiflumetofen induced by FoAV1-FOM infection in FOM.

Cultural, Morphological and Pathogenic Variability in the Colletotrichum capsici Isolates, Inciting Anthracnose and Fruit rot of Chilli (Capsicum annuum L.) in different Agro-Climatic Zones of Kerala, India

Aims: To study the cultural, morphological and pathogenic variability existing among the pathogen isolates of Colletotrichum capsici, inciting anthracnose and fruit rot of chilli in Kerala. Study Design: Completely randomized design (CRD). Place and Duration of Study: Department of Plant Pathology, College of Agriculture (Kerala Agricultural University), Vellayani, Thiruvananthapuram during 2018-2019. Methodology: A survey was conducted in the five agro-climatic zones (ACZs) of Kerala viz., Northern zone, Central zone, Special problem zone, High range zone and Southern zone to assess the incidence and severity of anthracnose disease. The various symptoms in the surveyed locations were also noted. Pathogen isolates were grown in PDA medium in the laboratory and colony characters (colour and mycelial growth rate of pathogen colonies) were observed. Also, the morphological characters viz., mycelial width, shape and size of conidia, acervular diameter, number and size of setae, appressorial size were recorded. Results: Maximum anthracnose incidence of 90 per cent and severity of 52.60 per cent were noticed at Regional Agricultural Research Station (RARS), Pilicode (Northern zone). Characteristic leaf spot, fruit rot and die-back symptoms were observed in the survey locations. Pathogen colonies showed different shades of white, off-white to grey turning brown or black with regular or irregular margins and concentric rings of black acervuli in the potato dextrose agar (PDA) medium. The diameter of growth of C. capsici isolates ranged from 7.20 to 8.60 cm at 7 days after incubation. All the C. capsici isolates initiated hyaline mycelium having a width of 1.73 - 2.36 µm and produced numerous sickle shaped conidia with a size range of 19.42 - 20.46 µm × 2.16 - 3.09 µm. Black coloured, circular or ellipsoidal acervuli (122.14 - 189.08 µm dia.) had 20 – 46 setae of length 74.13 to 107.30 µm. Also, brown or black appressoria of size 8.64 - 12.64 µm × 5.54 - 7.84 µm were formed in slide culture. Cc3 from College of Agriculture, Vellanikkara was obtained as the most virulent isolate producing a lesion size of 1.13 cm and PDI of 45.33 in the artificially inoculated chilli fruits (var. Vellayani Athulya) at 5 days after inoculation. Conclusion: All the nine C. capsici isolates exhibited variability in their cultural, morphological and pathogenic characteristics. Variability in phytopathogenic fungi helps in identifying novel and sustainable control methods to tackle the diseases incited by them. Understanding pathogen variability provides new insights for developing effective disease management methods. This will help in promoting sustainable agriculture and preventing severe crop losses.

Enhancement of soil aggregation and physical properties through fungal amendments under varying moisture conditions.

Soil structure and aggregation are crucial for soil functionality, particularly under drought conditions. Saprobic soil fungi, known for their resilience in low moisture conditions, are recognized for their influence on soil aggregate dynamics. In this study, we explored the potential of fungal amendments to enhance soil aggregation and hydrological properties across different moisture regimes. We used a selection of 29 fungal isolates, recovered from soils treated under drought conditions and varying in colony density and growth rate, for single-strain inoculation into sterilized soil microcosms under either low or high moisture (≤-0.96 and -0.03 MPa, respectively). After 8 weeks, we assessed soil aggregate formation and stability, along with soil properties such as soil water content, water hydrophobicity, sorptivity, total fungal biomass and water potential. Our findings indicate that fungal inoculation altered soil hydrological properties and improved soil aggregation, with effects varying based on the fungal strains and soil moisture levels. We found a positive correlation between fungal biomass and enhanced soil aggregate formation and stabilization, achieved by connecting soil particles via hyphae and modifying soil aggregate sorptivity. The improvement in soil water potential was observed only when the initial moisture level was not critical for fungal activity. Overall, our results highlight the potential of using fungal inoculation to improve the structure of agricultural soil under drought conditions, thereby introducing new possibilities for soil management in the context of climate change.

Optimal fissioning strategies of social insects with respect to colony dynamics and nest founding probability

Social insects demonstrate two fundamentally different modes of reproduction, independent colony foundation (ICF) by single fertilized queens or dependent colony foundation (DCF) by fissioning of existing colonies into two or several new colonies (swarms). In some species, both reproductive modes occur in parallel. The benefits and disadvantages of DCF vs. ICF have been widely discussed and been subject to empirical studies, but a formal theoretical treatment of the topic is still incomplete. Taking honey bees as example, we provide a resource allocation model of colony dynamics to analyze the ecological conditions under which DCF may be favored over ICF. Using mathematical and numerical methods, we show that it critically depends on the survivorship function linking swarm size to the probability of swarm establishment whether ICF or DCF results in a higher output of surviving new colonies. Because building larger swarms requires larger inter-swarm time intervals, DCF can only be a better strategy if this disadvantage is over-compensated for by a strong size-dependent swarm survivorship and survival of single queens is very low. Colony growth rate has no effect on this decision and the impact of maximum possible colony size is negligible. Further, there is a discontinuity in the optimal swarm size, so that either a swarm size of 1 (ICF) is the best strategy, or emitting swarms of considerable size (DCF). Consequently, a direct evolutionary transition from ICF to DCF appears unlikely and may have been triggered by selective pressures promoting movement of complete nests or distributing single colonies over several nests (polydomy).

Resin acid derivatives: fungicidal properties and prediction of the spectrum of biological activity

Biologically active substances that confer antiviral, anti-tumour and antimicrobial effects, which are found among the components of plant raw materials, as well as the products of their chemical modification, are the subject of considerable research interest. These objects include tricyclic diterpenoids – extractives of wood of abietic and pimaric types (resin acids). The presence of two reaction centres (double bonds and a carboxyl group) in the structure of resin acids opens up a wide range of possibilities for synthesising useful compounds on their basis. One of the most promising areas for the study of resin acids and their derivatives consists assessing their fungicidal properties to inform their introduction into compositions as additives, as well as in the application of protective films to increase the resistance of composite building materials against the aggressive effects of microorganisms. In the present work, the fungicidal activity of the N-phenylimide of maleopimaric acid and its polyfluoroalkyl ethers synthesised by us was evaluated on the cells of the filamentous fungi Aspergillus niger, Alternaria alternata and Penicillium sp. by the rate of colony formation and growth. A comparative analysis of the biological activity of resin acids and their in silico derivatives was performed. According to the AntiBag Pred forecast, the test compounds having the maximum values of the probabilities of the presence and absence of each type of activity are characterised by the manifestation of antibacterial activity in relation to strains of gram-positive bacteria. The results of the predictive model are consistent with previous experimental data. However, AntiFun Pred data related to the calculation of fungicidal activity were not confirmed in vitro.

Conformation Techniques for False Smut Pathogen (U. virens) on Rice through Cultural and Molecular Tools in Telangana

The study was conducted during period of January to April, 2022 at the Department of Plant Pathology and Institute of Biotechnology, Professor Jayashankar Telangana State Agricultural University, Rajendranagar, Hyderabad, Telangana state, India. The main objective of this study was to investigate cultural and morphological characteristics of U. virens isolates on potato sucrose medium (PSA) and their molecular confirmation collected from rice growing regions of Telangana during the kharif (rainy season), 2021. The isolates showed varying colony diameters and growth rates, with Uv3 reaching a maximum diameter of 77 mm and a growth rate of 2.85 mm, while Uv11 exhibited a minimum diameter of 40 mm with a growth rate of 1.48 mm. Notably, twenty two isolates showed diverse characteristics, including colony color, elevation, zonation, sectoring, and chlamydospore formation. Cluster analysis revealed the formation of five major groups, each group containing isolates with distinct features and PCR amplification with universal and species-specific primers yielded products of 700 bp and 380 bp, respectively. Phylogenetic tree analysis, indicated, Uv3 and Uv15 belonged to clade X and clade VII, respectively, both comprising Indian isolates. In contrast, Uv4 was grouped under clade II, which included both Chinese and Indian isolates. The proposed technique coupled with information of U. virens will immensely contribute to further studies on this fungus and false smut disease in India.

Human infrastructures correspond to higher Adélie penguin breeding success and growth rate.

Anthropogenic activities generate increasing disturbance in wildlife especially in extreme environments where species have to cope with rapid environmental changes. In Antarctica, while studies on human disturbance have mostly focused on stress response through physiological and behavioral changes, local variability in population dynamics has been addressed more scarcely. In addition, the mechanisms by which breeding communities are affected around research stations remain unclear. Our study aims at pointing out the fine-scale impact of human infrastructures on the spatial variability in Adélie penguin (Pygoscelis adeliae) colonies dynamics. Taking 24 years of population monitoring, we modeled colony breeding success and growth rate in response to both anthropic and land-based environmental variables. Building density around colonies was the second most important variable explaining spatial variability in breeding success after distance from skua nests, the main predators of penguins on land. Building density was positively associated with penguins breeding success. We discuss how buildings may protect penguins from avian predation and environmental conditions. The drivers of colony growth rate included topographical variables and the distance to human infrastructures. A strong correlation between 1-year lagged growth rate and colony breeding success was coherent with the use of public information by penguins to select their initial breeding site. Overall, our study brings new insights about the relative contribution and ecological implications of human presence on the local population dynamics of a sentinel species in Antarctica.

Genome analysis of Pseudomonas strain 4B with broad antagonistic activity against toxigenic fungi.

Pseudomonas sp. 4B isolated from the effluent pond of a bovine abattoir was investigated as antifungal against toxigenic fungi. The complete genome of Pseudomonas 4B was sequenced using the Illumina MiSeq platform. Phylogenetic analysis and genome comparisons indicated that the strain belongs to the Pseudomonas aeruginosa group. In silico investigation revealed gene clusters associated with the biosynthesis of several antifungals, including pyocyanin, rhizomide, thanamycin, and pyochelin. This bacterium was investigated through antifungal assays, showing an inhibitory effect against all toxigenic fungi tested. Bacterial cells reduced the diameter of fungal colonies, colony growth rate, and sporulation of each indicator fungi in 10-day simultaneous growing tests. The co-incubation of bacterial suspension and fungal spores in yeast extract-sucrose broth for 48h resulted in reduced spore germination. During simultaneous growth, decreased production of aflatoxin B1 and ochratoxin A by Aspergillus flavus and Aspergillus carbonarius, respectively, was observed. Genome analysis and in vitro studies showed the ability of P. aeruginosa 4B to reduce fungal growth parameters and mycotoxin levels, indicating the potential of this bacterium to control toxigenic fungi. The broad antifungal activity of this strain may represent a sustainable alternative for the exploration and subsequent use of its possible metabolites in order to control mycotoxin-producing fungi.

Polyethylene glycol–mediated genetic transformation and fluorescent labelling of Colletotrichum sublineola for studying sorghum anthracnose pathogenicity

AbstractAnthracnose, a disease primarily caused by the fungus Colletotrichum sublineola, affects sorghum production in China. However, the infection mechanisms and complex host interactions of C. sublineola remain to be elucidated. Thus, this study aimed to establish and optimize a protoplast transformation system of C. sublineola so as to lay a foundation for further research on the infection mechanisms and complex host interactions of C. sublineola. A genetic transformation system was established and optimized for C. sublineola strain HX‐5‐1 and used to introduce fluorescent reporter genes, namely, green fluorescent protein (GFP) and mCherry. Results showed that the growth of HX‐5‐1 was completely inhibited on a regenerating medium containing 300 μg mL−1 hygromycin B, indicating that hygromycin B acted as a transformant‐selective marker. HX‐5‐1 produced the highest yield of protoplasts when the mycelia were cultured for 60 h and dissolved in a 0.1% lysozyme solution (0.7 M NaCl) for 3 h. Subsequently, a 106 mL−1 protoplast solution transformed using 5 μg of plasmid DNA exhibited the highest transformation efficiency. Laser confocal microscopy showed that GFP and mCherry fluorescent signals were successfully expressed in the mycelia and spores of HX‐5‐1, indicating that they were stably inherited in the transformants and did not affect colony morphology, growth rate, or pathogenicity. In conclusion, we successfully established a genetic transformation system for C. sublineola and employed it to obtain stable fluorescent protein expression. This study may serve as a foundation for understanding the infection process of C. sublineola.

Taxonomy and systematics of the fungus-growing ant associate Escovopsis (Hypocreaceae).

Escovopsis is a symbiont of fungus-growing ant colonies. Unstandardised taxonomy prevented the evaluation of the morphological diversity of Escovopsis for more than a century. The aim of this study is to create a standardised taxonomic framework to assess the morphological and phylogenetic diversity of Escovopsis. Therefore, to set the foundation for Escovopsis taxonomy and allow interspecific comparisons within the genus, we redescribe the ex-type cultures of Escovopsis aspergilloides, E. clavata, E. lentecrescens, E. microspora, E. moelleri, E. multiformis, and E. weberi. Thus, based on the parameters adopted in this study combined with phylogenetic analyses using five molecular markers, we synonymize E. microspora with E. weberi, and introduce 13 new species isolated from attine nests collected in Argentina, Brazil, Costa Rica, Mexico, and Panama: E. breviramosa, E. chlamydosporosa, E. diminuta, E. elongatistipitata, E. gracilis, E. maculosa, E. papillata, E. peniculiformis, E. phialicopiosa, E. pseudocylindrica, E. rectangula, E. rosisimilis, and E. spicaticlavata. Our results revealed a great interspecific morphological diversity throughout Escovopsis. Notwithstanding, colony growth rates at different temperatures, as well as vesicle shape, appear to be the most outstanding features distinguishing species in the genus. This study fills an important gap in the systematics of Escovopsis that will allow future researchers to unravel the genetic and morphological diversity and species diversification of these attine ant symbionts. Taxonomic novelties: New species: Escovopsis breviramosa Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues, E. chlamydosporosa Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues, E. diminuta Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues, E. elongatistipitata Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues, E. gracilis Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues, E. maculosa Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues, E. papillata Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues, E. peniculiformis Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues, E. phialicopiosa Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues, E. pseudocylindrica Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues, E. rectangula Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues, E. rosisimilis Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues, E. spicaticlavata Q.V. Montoya, M.J.S. Martiarena & A. Rodrigues. Citation: Montoya QV, Martiarena MJS, Rodrigues A (2023). Taxonomy and systematics of the fungus-growing ant associate Escovopsis (Hypocreaceae). Studies in Mycology 106: 349-397. doi: 10.3114/sim.2023.106.06.

Biological Characteristics of Beauveria majiangensis Strain MJ1015 and Optimization of Solid Medium Technology for Sporulation.

The entomopathogenic fungus Beauveria majiangensis strain MJ1015, recently isolated from white grubs on a blueberry farm in Guizhou, China, could be used as a biocontrol agent. As a first step toward determining the effect of different solid culture media, temperature, and pH on colony growth rate and sporulation, we evaluated the optimum solid medium for mycelial growth and conidia production on a commercial scale. Subsequently, we also used single-factor analysis and response surface optimization to optimize the composition of the solid culture medium. On potato dextrose agar (PDA) medium, MJ1015 grew fastest and produced the highest spore yield at 29°C and pH 5. The best solid medium for the growth and sporulation of strain MJ1015 comprised 64.70 g/l of rice, 13.00 g/l of wheat, 0.30 g/l of NaNO3, 0.36 g/l of K2HPO4 · 3H2O, and 1.00 g/l of CaCO3. Rice, NaNO3, and K2HPO4 · 3H2O were the main influencing factors. The predicted value of cultured spores using the optimal medium was 4.56 x 1010 conidia/l. The validation test results showed that the average growth rate of strain MJ1015 on the optimal medium was 85% and 96% faster than that on Sabouraud dextrose agar with yeast extracts medium (SDAY) and PDA, respectively. Sporulation was 43.90 times and 9.65 times of that produced on SDAY and PDA, respectively. Our findings provide a theoretical basis for the commercial production of B. majiangensis to control white grubs.

Studi Pertumbuhan Miselia Jamur Merang (Volvariella volvaceae) G2 Isolat FP005 Faperta UNSIKA Pada Beberapa Bahan Alternatif Media In Vitro

Straw mushroom is one of the horticultural commodities that is in great demand by the community. The successful cultivation of straw mushrooms is determined by the quality of the seeds of the molten mushroom. Tissue culture is one of the efforts to get quality seeds. Quality seedlings are seen from the brood of the merang fungus and the quality of pure culture. Making pure culture is still constrained in making in vitro media, especially alternative media materials. This study aims to obtain the best results from the growth of G2 FP005 mycelia on alternative media materials in vitro. This research was conducted at the Biotechnology and Plant Breeding Laboratory, Faculty of Agriculture, Singaperbangsa University, Karawang from April to May 2023. The research method used was a single factor Complete Randomized Design (CRD) with 7 treatments and 5 replications: A (100% PDA); B (100% Charcoal Husk Solution); C (100% Husk Solution); D (80% PDA + 20% Husk Charcoal Solution), E (75% PDA + 25% Husk Solution), F (50% Husk Charcoal Solution + 50% Husk Solution); G (50% PDA + 25% Husk Charcoal Solution + 25% Husk Solution). The effect of the treatment was analyzed using variance and if the F test was significant at 1% level, then to find out which treatment was the best it was continued with the DMRT (Duncan Multiple Range Test) advanced test at the 1% level of significance. The results achieved from this study were that there was a significant effect of using alternative media materials on the growth of the straw mushroom mycelia. Alternative media 80% PDA + 20% rice husk charcoal gave the highest mycelia colony diameter of 7.88 cm and the highest mycelial colony growth rate of 2,878 cm/day. Keywords: Unsika Faperta Isolate, Straw Mushroom, Tissue Culture.

Bacterial colony size growth estimation by deep learning

The bacterial growth rate is important for pathogenicity and food safety. Therefore, the study of bacterial growth rate over time can provide important data from a medical and veterinary point of view. We trained convolutional neural networks (CNNs) on manually annotated solid medium cultures to detect bacterial colonies as accurately as possible. Predictions of bacterial colony size and growth rate were estimated from image sequences of independent Staphylococcus aureus cultures using trained CNNs. A simple linear model for control cultures with less than 150 colonies estimated that the mean growth rate was 60.3 mu m/h for the first 24 h. Analyzing with a mixed effect model that also takes into account the effect of culture, smaller values of change in colony size were obtained (control: 51.0 mu m/h, rifampicin pretreated: 36.5mu m/h). An increase in the number of neighboring colonies clearly reduces the colony growth rate in the control group but less typically in the rifampicin-pretreated group. Based on our results, CNN-based bacterial colony detection and the subsequent analysis of bacterial colony growth dynamics might become an accurate and efficient tool for bacteriological work and research.

A Normalization Protocol Reduces Edge Effect in High-Throughput Analyses of Hydroxyurea Hypersensitivity in Fission Yeast.

Edge effect denotes better growth of microbial organisms situated at the edge of the solid agar media. Although the precise reason underlying edge effect is unresolved, it is generally attributed to greater nutrient availability with less competing neighbors at the edge. Nonetheless, edge effect constitutes an unavoidable confounding factor that results in misinterpretation of cell fitness, especially in high-throughput screening experiments widely employed for genome-wide investigation using microbial gene knockout or mutant libraries. Here, we visualize edge effect in high-throughput high-density pinning arrays and report a normalization approach based on colony growth rate to quantify drug (hydroxyurea)-hypersensitivity in fission yeast strains. This normalization procedure improved the accuracy of fitness measurement by compensating cell growth rate discrepancy at different locations on the plate and reducing false-positive and -negative frequencies. Our work thus provides a simple and coding-free solution for a struggling problem in robotics-based high-throughput screening experiments.

Fluid-based microbial processes modeling in Trichodesmium colony formation

In tropical and subtropical ocean gyres, Trichodesmium colonies account for up to half of the total N2 fixation in the ocean, making it one of the most ecologically significant N2-fixing cyanobacteria. The processes whereby Trichodesmium colonies and large-scale surface blooms form have not been investigated thoroughly. In particular, the effects of fluid motion have not been included in previous studies. As the first step toward understanding the mechanical processes associated with Trichodesmium colony synthesis, we propose a shear-related flow-based growth model to enlighten how fluid dynamics affect bacteria colony formation and growth. To investigate the possibility that early growth characteristics are strongly dependent on the shear rate, a two-way coupled fluid-colony interaction is developed using the lattice Boltzmann method for a porous colony. This model captures the exponential growth trend during the colony formation phase found in experiments. Our results show that the flow field significantly impacts both the colony growth rate and shape. In pure shear conditions, colonies grow in circular shapes, whereas in uniform flow, they take filament-like forms. Additionally, the Reynolds number (Re) plays a crucial role in shaping the colonies, especially in uniform flow conditions where its effect is more profound.