Colony Color Research Articles

Sarocladium kiliense: A first report of phytopathogenic fungus causing maize stalk rot in India

Post-flowering stalk rot (PFSR) disease drastically affects maize yields, leading to substantial economic losses. Fusarium verticillioides was widely recognized as a major pathogen responsible for maize PFSR disease. Astonishingly, apart from F. verticillioides, another pathogen was also observed infecting the stalk rot and thus, further pathogen purification was initiated. In this study, we revealed the first report of Sarocladium kiliense, as a maize stalk rot pathogen. This research study was validated by isolation, purification, and morphological examination, which revealed the variability among S. kiliense isolates in traits such as pigmentation, colony color, mycelial characteristics, and conidial morphology. Notably, isolates exhibited distinct pigmentation patterns: 3 were orange and 3 were yellow, with consistent white to dirty white colony colors. Pathogenicity assays using toothpick inoculation demonstrated that all the S. kiliense isolates caused significant damage to the maize stalks, with brown discoloration, conversion of pith into a powdery texture, and subsequent stalk lodging. Isolate PKSK55 exhibited the highest pathogenicity, producing the longest lesions among tested isolates. Molecular characterization using ITS rDNA and RPB-2 genes confirmed the identity of S. kiliense across all isolates. Conclusively, this research uncovered and documents the first report of S. kiliense causing stalk rot in maize in India.

Morphological and Molecular Diversity of Colletotrichum gloeosporioides Causing Yam (Dioscorea alata L.) Anthracnose Disease in Côte d’Ivoire

Yam is a plant of great importance for the Ivorian people in terms of food, nutrition, economy, and socio-cultural value. However, its production is threatened by several biotic factors, including anthracnose, one of the most destructive diseases affecting yams. The objective of this study was to assess the morphological and molecular diversity of Colletotrichum gloeosporioides isolates responsible for anthracnose in yam (Dioscorea alata) in Côte d'Ivoire. Twenty-seven previously isolated strains were collected from Plant Health Laboratory of Nangui ABROGOUA University and analyzed. The study examined the cultural (coloration, growth mode, texture, and colony diameter), microscopic (conidial shape, size, and spore concentration), and molecular characteristics of these isolates. Significant morphological diversity was observed among the isolates. Macroscopically, three major groups were identified based on colony coloration (whitish, orange, and grayish). Colony textures varied, presenting cottony, flaky, downy, or filamentous appearances, with growth patterns ranging from concentric and radiating to regular rings. Colony diameters ranged from 37 mm to 82 mm. Microscopically, sporulation varied significantly among isolates, ranging from 1.1 × 10⁵ to 85.4 × 10⁵ spores, as did conidial size, which ranged from 10.54 to 19.29 µm in length and 3.93 to 6.43 µm in diameter. Molecular analysis through comparison with GenBank nucleotide sequences revealed homology rates of 99-100% and varying proximities between the rDNA sequences of the isolates. In conclusion, the characterization of these isolates provides valuable insights for improving disease control strategies for yam anthracnose.

Vancomycin Resistant Enterococci and Trend of Antimicrobial Susceptibility in Urine Cultures

Background: Enterococci are emerging nosocomial pathogen showing resistance to broad spectrum antibiotics. As reported by the center for disease control and prevention, enterococci (13.9%) are the second leading cause of urinary tract infection, after Escherichia coli. After the first report of vancomycin-resistant enterococci in England (1988), India reported its first vancomycin resistant enterococci isolate from New Delhi (1999). The prevalence of vancomycin resistant enterococci ranges from 1-9%, in India. To evaluate the prevalence of vancomycin resistant enterococci among urinary isolates and to check their antimicrobial resistance pattern. Methods: A retrospective observational study conducted at a tertiary care hospital. Data were analysed from November, 2017 to October, 2018. A total of 12,129 urine samples were received and subjected to culture on HiCrome UTI agar (Hi-Media laboratories, Mumbai) and incubated at 37 °C for a period of 18-24 hours. Antimicrobial susceptibility was performed using Kirby–Bauer disc diffusion method. Identification was done on the basis of colony colour and species identification was done by using biochemical test as per standard laboratory protocol. In one case automated siemens microscan walkaway identification system was used for confirmation. Results: Enterococci were isolated in 4.06% from urine cultures out of which 7.52% were vancomycin resistant. Most of the vancomycin resistant enterococci isolate (86.49%) were multi drug resistant. Conclusion: In vancomycin resistant enterococci related urinary tract infection, nitrofurantoin can be used as an appropriate first choice in uncomplicated cases and linezolid should be reserved for serious and complicated urinary tract infections.

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.

Study on the Cordyceps militaris Cultivation in Differential Media and Substrates Formulation

The study was study on the Cordyceps militaris Cultivation in Differential Media in plant diseases and weeds laboratory, Faculty of Agriculture, National University of Laos. The experiment was conducted for six months. This is of study have two parts, part one to testing fibers of growing Cordyceps militaris in Differential Media and Substrates Formulation. The experiment was set up as Completely Randomized Design (CRD) layout was used with four replications and three treatments as they are: T1 PDA (control), T2 supplements one + Silk worm pupae, T3 supplements two + egg. To testing fibers of growing Cordyceps militaris in three kind differentials of twenty days had different significant (P<0.05), T2 have medium testing fibers of growing Cordyceps militaris with amount of 8.71 cm, by T3 is 8.32 cm, the lowest by T1 8.06 cm, and to notice T2 have dark orange in color colony, by T3 and T1 light color yellow. Part two to testing substrates and supplements Cordyceps militaris in three kind differentials to yield. The experiment was arranged in a Completely Randomized Design (CRD) layout was used with four replications and five treatments as they are: T1 Rice bean + potato dextrose broth PDB (control), T2 Rice rations + potato dextrose broth (PDB), T3 Rice bean + liquid broth Silk worm pupae 30 gram, T4 Rice bean + liquid broth Silk worm pupae 60 gram, T5 Rice rations + liquid broth Silk worm pupae 60 gram. The study was use twenty-four. The results found that there are different significant (P<0.05), T4 fibers of growing Cordyceps militaris by 5.93 days,by T5, T3and T2 is 6.25, 8.00 and 10.18 days respectively. By T1 is 10.00 days. The developmenting of fibers to substitute flower mushroom in 5 treatments. The results found that there are different significant (P<0.05), T5. The developmenting of fibers to substitute flower mushroom 22.18 days, by T2, T3 and T4 is 28.56, 25.06 and 21.81 days respectively. T1 is 29.31 days. However, weight flower mushroom T5 weight 27.25 gram/glass, by T4, T3 and T1 weight 26.88, 23.44 and 18.13 gram/glass respectively. By T2 weight 17.81 gram/glass.

Morphology of Bacteria and Fungi From The Fermentation of Asam Drien as a Local Food Ingredient of West Aceh

A typical food ingredient of the Aceh region, especially the western and southern, is Asam drien (language from the West Aceh region). Asam drien is made from durian fruit meat, usually due to the abundance of durian fruit, or because the durian fruit is of poor quality such as the taste of the bland fruit meat, then most of the people utilize the durian fruit into Asam drien which can be used as a seasoning for cooking. The purpose of this study was to identify the morphological characteristics of bacterial and fungal colonies in Asam drien processing, this study used a qualitative descriptive method for colony characteristics, and biochemical tests. The results of research on bacteria and fungi in Asam drien found 14 colonies from the entire fermentation both at home and in the laboratory, namely irregular and diffuse, elongated, round, and threaded shapes; colony color characteristics obtained only one color, namely cream color; characteristics of colony edges there are 5, namely: wavy, squiggly, smooth, serrated, and threadbare. Colony elevation characteristics obtained 2 forms, namely flat and raised. There are 2 types of colony surface characteristics, namely rough and smooth shiny. Then reisolated on selective media DeMan Rogosa Sharpe Agar (MRS Agar) for lactic acid bacteria. The results of observations found 11 colonies from the entire fermentation each consisting of 2 types of isolates Ka and Kb which are Gram-positive bacteria bacillus and coccus cell forms. As well as fungi that grow on Asam drien, namely the genus Rhizopus and the type of yeast found from the genus Saccharomyces. The catalase test on Asam drien bacteria is positive, has proteolytic, amylolytic, and cellulolytic potential, but is not able to hydrolyze fat and alcohol.

Morphological and Molecular Characterization of Trichoderma Isolates from Vegetable Crop Rhizospheres in Nepal

Background Trichoderma spp. hold significant potential as biocontrol agents in agriculture due to their antagonistic properties against plant pathogens. The study aimed to characterize and identify Trichoderma isolates from rhizospheric regions of vegetable crops. Methods In this study, Trichoderma isolates were collected from rhizospheric soil samples of vegetable crops from different ecological zones and were selected for comprehensive morphological and molecular characterization. The isolates were visually assessed for colony color, growth pattern, aerial mycelium presence, phialide and conidial morphology, and chlamydospore presence. Molecular analysis was employed based on ITS and tef-1α sequences. Diversity indices were also computed for different ecological zones. Results The morphological characteristics and phylogenetic trees for both regions provided a clear species resolution, with four main clades: Harzianum, Viride, Brevicompactum and Longibrachiatum with 12 species T. harzinaum, T. afroharzianum, T. lentiforme, T. inhamatum, T. camerunense, T. azevedoi, T. atroviride, T. asperellum, T. asperelloides, T. koningii, T. longibrachiatum and T. brevicompactum and nine species as a new country record. Diversity indices indicated that high mountain regions displayed the highest species diversity and evenness (H = 1.724 [0.28], J = 0.84, D = 0.28), followed by hilly regions (H = 1.563 [0.28], J = 0.72, D = 0.28). Plains, on the other hand, exhibited lower species diversity (H = 1.515, J = 0.66, D = 0.33). The calculated species abundance values showed that plains (E = 2.11), mid-hills (E = 1.95), and high mountains (E = 1.99) each had their unique diversity profiles. Notably, T. afroharzianum and T. asperellum were predominant. Conclusions Overall, the study unveiled a rich diversity of Trichoderma species in different agricultural zones of Nepal. These findings shed light on the ecological distribution and diversity of Trichoderma spp., which could have significant implications for sustainable agriculture and biological control strategies.

Acptp2,3 participates in the regulation of spore production, stress response, and pigments synthesis in Aspergillus cirstatus.

Aspergillus cristatus was a filamentous fungus that produced sexual spores under hypotonic stress and asexual spores under hypertonic stress. It could be useful for understanding filamentous fungi's sporulation mechanism. Previously, we conducted functional studies on Achog1, which regulated the hyperosmotic glycerol signaling (HOG) pathway and found that SI65_02513 was significantly downregulated in the transcriptomics data of ΔAchog1 knockout strain. This gene was located at multiple locations in the HOG pathway, indicating that it might play an important role in the HOG pathway of A. cristatus. Furthermore, the function of this gene had not been identified in Aspergillus fungi, necessitating further investigation. This gene's conserved domain study revealed that it has the same protein tyrosine phosphatases (PTPs) functional domain as Saccharomyces cerevisiae, hence SI65_02513 was named Acptp2,3. The function of this gene was mostly validated using gene knockout and gene complementation approaches. Knockout strains exhibited sexual and asexual development, as well as pigments synthesis. Morphological observations of the knockout strain were carried out under several stress conditions (osmotic stress, oxidative stress, Congo Red, and sodium dodecyl sulfate (SDS). Real-time fluorescence polymerase chain reaction (PCR) identified the expression of genes involved in sporulation, stress response, and pigments synthesis. The deletion of Acptp2,3 reduced sexual and asexual spore production by 4.4 and 4.6 times, demonstrating that Acptp2,3 positively regulated the sporulation of A. cristatus. The sensitivity tests to osmotic stress revealed that ΔAcptp2,3 strains did not respond to sorbitol-induced osmotic stress. However, ΔAcptp2.3 strains grew considerably slower than the wild type in high concentration sucrose medium. The ΔAcptp2,3 strains grew slower than the wild type on media containing hydrogen peroxide, Congo red, and SDS. These findings showed that Acptp2,3 favorably controlled osmotic stress, oxidative stress, and cell wall-damaging chemical stress in A. cristatus. Deleting Acptp2,3 resulted in a deeper colony color, demonstrating that Apctp2,3 regulated pigment synthesis in A. cistatus. The expression levels of numerous stress-and pigments-related genes matched the phenotypic data. According to our findings, Acptp2,3 played an important role in the regulation of sporulation, stress response, and pigments synthesis in A. cristatus. This was the first study on the function of PTPs in Aspergillus fungi.

Aurantiochytrium mutant strains exhibiting different colony colors altered the contents of squalene.

Aurantiochytrium sp. 18W-13a, a marine heterotrophic protist belonging to the genus thraustochytrid, is known to accumulate high levels of squalene and carotenoids. Nowadays, the mutagenesis breeding of microorganisms is still widely practiced because the induced mutations of DNA do not involve the permanent integration of heterologous DNA sequences. Therefore, in this study, we focused on the improvement of squalene yield by mutagenesis breeding using Aurantiochytrium sp. 18W-13a. To bypass the massively laborious screening, we propose to use colony colors as the first criterion to screen mutants with high squalene accumulation, since the carotenoid and squalene synthetic pathways share an intermediate. We selected pale (white)-colored mutants after carbon ion irradiation. The white mutants exhibited larger squalene yields than twice as much of the original strain. The results clearly indicate that the present screening method with colony colors promises to obtain productive strains of squalene.

Yunchengibacter salinarum gen. nov., sp. nov., a novel bacterium of the family Kordiimonadaceae isolated from sediment in Yuncheng salt lake.

A Gram-stain-negative, aerobic, motile and rod-shaped bacterium, the color of the bacterial colony ranges from light yellow to yellow, designated YC-2023-2T, was isolated from sediment sample of Yuncheng salt lake. Growth occurred at 15-45℃ (optimum 37℃), pH 6.0-9.0 (optimum pH 7.0-8.0) and with 0-8.0% NaCl (w/v, optimum 2.0%). The phylogenetic analysis based on 16S rRNA gene sequences showed that strain YC-2023-2T belonged to the family Kordiimonadaceae. The closely related members were Gimibacter soli 6D33T (92.38%), Kordiimonas lipolytica M41T (91.88%), Eilatimonas milleporae DSM 25217T (91.88%) and Kordiimonas gwangyangensis JCM 12864T (91.84%). The genome of strain YC-2023-2T was 2957513 bp, and the genomic DNA G+C content was 63.91%. The main respiratory quinone was Q-10 and the major fatty acids (>10%) were iso-C15:0, C16:0, C19:0 cyclo ω8c, Summed Feature 8 (C18:1 ω6c or C18:1 ω7c) and Summed Feature 9 (iso-C17:1 ω9c or C16:0 10-methyl). The major polar lipids consisted of phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, unidentified glycolipid, unidentified lipid, and two unidentified aminolipids. Based on the phylogenetic, phenotypic and chemotaxonomic characteristics, strain YC-2023-2T is proposed to represent a novel species of a novel genus named Yunchengibacter salinarum gen. nov., sp. nov., within the family Kordiimonadaceae. The type strain is YC-2023-2T (= GDMCC 1.4502T = KCTC 8546T).

Isolation, morphological characterization, and screening virulence of Beauveria bassiana and Metarhizium robertsii fungal isolates on Galleria mellonella

Background Entomopathogenic fungi exists naturally in plants as an asymptote and have the potential to reduce the population of insect pests through indirect interactions. This study was conducted to detect and characterize the endophytic fungi Beauveria bassiana and Metarhizium robertsii from the rhizosphere soil of tomato plants and their virulence effect on Galleria melonella. Methods From the rhizosphere soil of 40 tomato fields, three Beauveria bassiana and seven Metarhizium robertsii isolates were isolated using the galleria bait method. All fungi isolate were morphologically characterized by their colony color, shape, and surface texture. Isolates with the highest percentages of germination, conidial yield, and radial growth were selected, and their virulence was evaluated on second instar larvae of Galleria melonella under laboratory conditions. Results In this study, Beauveria bassiana showed white colony color and aseptate hyphae, whereas Metarhizium robertsii showed dark green to light green colony color and septate hyphal structures. Maximum spore production and conidial length were obtained by Beauveria bassiana isolate APPRC-27 with 2.67x107 spores ml-1 and 3.24 µm, respectively. Colony radial growth rates ranged from 1.73 to 3.24 mm day-1. The results revealed that the highest mortality rate of Galleria melonella (100%) was obtained by Metarhizium robertsii isolates K-61 and K-102 at a concentration of 1x108 conidial ml-1 at 7 days post-inoculation. The lowest mortality rate was registered by Metarhizium robertsii isolate RST-11. Conclusions In the present study, isolates that produced the most spores and had the highest germination rates were the most virulent to Galleria mellonella second instar larvae. Therefore, virulent entomopathogenic fungi, Beauveria bassiana and Metarhizium robertsii, are promising bioagents for the control of insect pests.

First Report of Burkholderia glumae Causing Bacterial Panicle Blight in Rice in Bangladesh.

Bacterial panicle blight (BPB) is one of the emerging diseases occurring in different Agro-Ecological Zones (AEZ) of Bangladesh and can cause up to 75% yield loss. In Bangladesh, the typical symptoms of BPB include sheath rot, panicle blight, grain spotting, and grain rot in both inbred and hybrid rice varieties, which resemble those reported by Zhou (2019). To confirm, 300 field samples of 20 panicles each with typical BPB symptoms from 20 districts (3 locations each district and 5 fields per location) were collected during mid-November 2022 for the causal pathogen(s) isolation. Nearly 70% of the panicles showed a dark brown chaffy appearance in the fields. For identification of the causal pathogen(s), 1 g of rice grains with typical BPB symptoms was surface sterilized by immersing for 15 seconds in 70% ethanol, 1 min in 3% sodium hypochlorite solution followed by rinsing the grains three times, and soaked in 1 mL sterile distilled water for 10 min (Mirghasempour et al. 2018). During grinding using mortar and pestle, 5 mL water was added (Islam et al. 2023) after which the suspension (20 μL) was then streaked onto the selective medium (S-PG) (Tsushima et al. 1986). Purple color colonies on the S-PG medium were selected and purified as candidate pathogens. For further confirmation, the genomic DNA of the bacterial isolates was extracted and amplified by PCR using 16SF (5'-AGAGTTTGATCCTGGCTCAG-3') and 16SR (5'-GGCTACCTTGTTACGACTT-3') (Nandakumar et al. 2009), and glu-FW (5'-GAAGTGTCGCCGATGGAG-3') and glu-RV (5'-CCTTCACCGACAGCACGCAT-3') primers (Maeda et al. 2006). The PCR products were visualized on 1% agarose gel resulting amplicons of 1494bp for 16S-rDNA and 529bp for gyrB. The PCR results revealed 529bp amplification for gyrB gene in one sample that was collected from a field in Natore (24°21'0.00" N 89°04'59.88" E) district cultivating Swarna (a local rice variety), primarily indicating the causal pathogen is Burkholderia glumae. The PCR products were sequenced using both primers and sequence data was analyzed by the BLAST nucleotide program. The obtained partial sequences of 16S rDNA and gyrB were deposited in Genbank (OR573691 and PP332812 respectively). The homology of 16S rDNA resulted over 98% with B. glumae (OK559611 and ON870618.1) and 100% with B. glumae (PP332812 and KX213523) for gyrB gene. To confirm B. glumae by pathogenicity test, 10 mL (108 UFC/ml) suspension of the representative strains, 0.5 mL was then injected into the panicles and sheaths of Horidhan (a susceptible local variety) in greenhouse condition and a control was inoculated with distilled water (Nandakumar et al. 2009). Typical BPB like symptoms were observed after 3 weeks post inoculation. The pathogen was again confirmed by reisolating from the infected spots as B. glumae to fulfill Koch's postulates. This report confirms the presence of B. glumae causing BPB of rice in Bangladesh. Future research for the investigation of BPB and the evolutionary origins of its causal bacteria is necessary to reduce the emergence of the disease and its management in Bangladesh.

Morphological, genetic and pathogenic variability among Botrytis cinerea species complex causing gray mold of strawberry

The necrotrophic fungus Botrytis cinerea is capable of infecting more than 1400 plant species, encompassing both dicotyledons and monocotyledons. Recognized as the world's second most destructive fungal pathogen, B. cinerea is responsible for significant pre- and postharvest diseases, especially gray mold on a variety of economically important crops, vegetables, and fruits. This study focused on characterizing the variability of ten B. cinerea isolates from strawberry fruits harvested across various regions in Egypt. The differentiation among these isolates was evaluated through morphological, genetic, pathogenic, and biochemical methods. The isolates exhibited variation in cultural development, colony color, shape, margin, and texture. The study highlighted morphological differences in conidiophores, variations in conidia size, and sclerotia production on three different media. The ten Botrytis isolates have differed in mycelial growth rate and pathogenicity on strawberry fruits and positively correlated with oxalic acid production. Isolate B3 of B. cinerea was the most aggressive, while isolate B5 demonstrated the lowest. Two isolates of Botrytis represented the first occurrence of Botrytis species in Egypt, B. californica and B. eucalypti, which were identified using ITS sequencing and combined with morphological characteristics. They were also responsible for causing gray mold on strawberry fruits. These results are expected to greatly contribute to the knowledge of morphological and genetic variability in B. cinerea isolates, as well as offer valuable insights for the development of disease management strategies.

Isolation and Characterization of Azotobacter and Phosphate Solubilizing Bacterial Isolates

Isolation from 20 soil samples collected from rhizosphere of watermelon was done on Ashby's medium and Pikovskaya's agar media.8 Azotobacter and 6 Phosphate solubilizing bacteria PSB isolates were obtained from all samples. All the obtained isolates of Azotobacter sp and PSB were gram negative. On morphological study it was observed that, all Azotobacter isolates were motile and positive for KOH test. In case of PSB, the colony shapes of 4 obtained isolates were circular and irregular in 2. All the isolates showed smooth appearance on the media with white colony colour. The biochemical results for both Azotobacter spp and PSB revealed that, most of them were positive for methyl red test, catalase test, starch hydrolysis, gelatine hydrolase test, oxidase test and indol test. Out of eight Azotobacter isolates, Isolate 4 (Azob-1) showed highest (14.25 mg/ml) N fixing ability whereas, from of six PSB isolates, Isolate1 (PSB-1) showed highest solubilising index (4.13) therefore these two isolates were found most efficient in their roles and therefore recommended for use for field experiments.

Termite gut microbiome being a rich source of potential lignocellulolytic bacterial strains

A study was carried out to identify prospective cellulose degrading bacteria isolated from termite gut collected from various agriculture sites. Eight of the bacterial isolates displayed a very distinct zone of clearing on CMC (Carboxylmethyl cellulose) agar media after staining with 1% Congo red, indicating that they may have cellulose degrading ability. On a CMC agar plate, CRDB6 had the largest colony diameter (3.08 cm) as well as clear zone diameter (0.96 cm), however CRDB 1 has maximum cellulolytic activity (1.45 cm) followed by CRDB5 and CRDB6 (1.33 cm and 1.28 cm) respectively. CRDB3, CRDB4 and CRDB9 showed Gram negative reaction among crop residue degrading bacterial isolates, while the remainder showed Gram positive reaction. The morphological characteristics of the bacteria isolates, such as colony shape, elevation, margin, colour and opacity, differed. Isolates CRDB6 produced highest β-glucosidase activity (12.66 m L-1) followed by CRDB1 (12.20 mg m L-1) and CRDB5 (10.21 m L-1) in Luria broth culture. Indole acetic acid production (with tryptophan) was greater in CRDB1 (389.33 mg/ml), CRDB6 (356.9 m L-1) and CRDB3 (315.96 m L-1) while as P-solubilization potential was peaked in CRDB6 (2.41 m L-1) followed by CRDB4 (1.76 m L-1) and CRDB3. Overall, isolate CRDB6 exhibited the highest cellulolytic activity as compared to rest of the bacterial isolates. These results will help in preparing bacterial consortia for the management of crop residues.

Fungal diversity in the temple with an Internal Transcribed Spacer (ITS) molecular and morphological approach

Abstract. Wijayanti C, Kuswytasari ND, Shovitri M, Danilyan E, Zulaika E. 2024. Fungal diversity in the temple with an Internal Transcribed Spacer (ITS) molecular and morphological approach. Biodiversitas 25: 2896-2906. Red bricks and andesite stones comprise most of Indonesia's temple architecture. Red brick is a highly porous building material that weathers rapidly in temples. Physical and biological weathering are the main causes of temple weathering. Rain and other physical elements can raise the temple's humidity, encouraging the growth of lichen and fungus. Many studies have been conducted to investigate the fungal variety in temples made of andesite stones, but the diversity found in these studies has only been identified at the genus level. This research aimed to determine the variety of fungi in the temples by employing morphological and molecular approaches like ITS biomarkers. Macroscopic characterization involves visual inspection of the colony's color on its surface and back and its edges, texture, diameter, shape, and surface to perform morphological identification. Meanwhile, the slide culture method was used for microscopic characterization, utilizing character observations such as reproductive and hyphal structures. Several steps were involved in the molecular identification process: DNA extraction using the Genomic ex Promega Corp Wizard kit, ITS marker-based DNA amplification visualized with Gel Documenter, DNA sequencing, BLAST searches to determine the isolate species, and MEGA 11 phylogenetic tree reconstruction. Class Eurotiomycetes, which includes Talaromyces funiculosus (CB L1B isolate), Talaromyces purpureogenus (CB L3A isolate), Penicillium oxalicum (CB L4A isolate), Penicillium cataractarum (CT TL3 isolate), and Aspergillus aflatoxiformans (CT TL4 isolate), was the most prevalent among the identified species from the classes Sordariomycetes and Eurotiomycetes. Purpureocillium lilacinum (CBL1C isolate), Trichoderma anaharzianum (CB L3B isolate), and Fusarium equiseti (CT L41 isolate) were the discovered species from the Sordariomycetes class.

Bridging Diagnostic Gaps: Utilising HiCrome Agar and Tetrazolium Reduction Medium for the Rapid and Presumptive Identification and Speciation of Candida Species in Vulvovaginal Candidiasis in Low-Resource Environments.

Background Vulvovaginal candidiasis (VVC) is a common fungal infection caused by an overgrowth of Candida species, primarily Candida albicans (C. albicans). Using HiCrome agar and tetrazolium reduction medium offers cost-effectiveness in Candida detection by eliminating the need for additional tests, reducing equipment costs compared to automated systems, and simplifying workflow with direct species identification while maintaining high specificity. They expedite detection by directly identifying Candida species based on colony colour, bypassing the multiple steps of phenotypic methods. This efficiency saves time in the laboratory, providing rapid results without the extended processing times associated with automated systems and facilitating prompt diagnosis and treatment decisions. These diagnostic tools are especially valuable in low-resource environments where a quick and accurate diagnosis of VVC is crucial for effective treatment and management of antifungal resistance. Aims and objectives This study aims to evaluate the efficacy of HiCrome agar and tetrazolium reduction medium's efficacy in speciating Candida species in VVC cases. Materials and methods A cross-sectional observational study was conducted at Saveetha Medical College and Hospitals, Chennai, India, over six months. High vaginal swabs from 126 patients suspected of VVC were collected and plated on Sabouraud dextrose agar (SDA), HiCrome Candida differential agar (Himedia, Mumbai, India), and tetrazolium reduction medium. The results were compared with those obtained from the VITEK2 compact system (bioMérieux, Marcy-l'Étoile, France). Results Of the 126 samples, 74.6% showed single yeast infections, 7.9% displayed mixed yeast infections, and 17.5% showed no growth. A total of 114 Candida isolates were identified. Both HiCrome agar and tetrazolium reduction medium accurately identified all isolates, with complete concordance with the VITEK2 compact system. The most commonly isolated species were C. albicans (55.2%), Candida tropicalis (32.4%), Candida glabrata (8.8%), and Candida parapsilosis (3.6%). Both media provided rapid and accurate presumptive identification in low-resource settings. Conclusions HiCrome agar and tetrazolium reduction medium demonstrated high sensitivity and specificity in identifying Candida species. These methods are reliable for rapid and accurate diagnosis, particularly in resource-limited settings. However, they may require supplementary tests for definitive species identification. The adoption of these diagnostic tools represents a significant advancement in clinical microbiology, improving VVC management and addressing antifungal resistance.

Molecular Characterisation and Toxicity Analysis of Indigenous Bacillus thuringiensis Berliner Isolates against Cucurbit Fruit Fly Maggots, Zeugodacus cucurbitae (Coquilett) (Diptera: Tephritidae)

Background: Indiscriminate use of insecticides against fruit flies (Tephritidae: Diptera) has led to the development of residue and resistance and there is a need to try alternate eco-friendly management practices. Bacillus thuringiensis (Bt) is the most widely used bacterium forming proteins active against insects of different orders. Thus, the present study was framed to characterize and evaluate 50 indigenous Bt isolates against cucurbit fruit fly maggots, Zeugodacus cucurbitae. Methods: Colony morphology was observed visually, while crystal morphology was observed through a microscope. The cry gene content of the Bt isolates was screened by Polymerase Chain Reaction and the proteins harbored were assessed by SDS- PAGE analysis. Full diet contamination method was used to evaluate the toxicity of Bt isolates against cucurbit fruit fly maggots. Result: Uniform full white, off-white and creamy white colony colours were found except one isolate each, exhibiting full white at centre surrounded by off white and off white at centre surrounded by full white. Spherical shape crystal (45.10%) was predominant followed by cuboidal (29.41%), bipyramidal (17.65%), rectangular (3.92%) and minute crystal attached to spore (3.92%). Insecticidal proteins varied from ~15 to greater than 200kDa in size with one to four and more distinct bands per isolate. PCR screening revealed cry4Aa in 2 isolates, cry4Ba and cry11Aa in one isolate each, cyt1 in 1 isolate. The standard strain Bti 4Q2 (100% mortality of maggots) and the indigenous Bt isolates, T166 (92.59%), T184 (91.11%) and T60 (90.48%) were found to be significantly toxic.

Isolation and Characterization of Native Isolates of Metarhizium sp. as a Biocontrol Agent of Hypothenemus hampei in Rodríguez de Mendoza Province—Peru

Entomopathogenic fungi represent the pinnacle of efficacy among biological control agents when combating insect pests within natural ecosystems, combating them without altering the environment. This study aimed to isolate and characterize the morphology, physiology, and pathogenicity of native isolates of Metarhizium sp. isolated from soil samples from the province of Rodriguez de Mendoza. Eighteen native isolates of Metarhizium sp. were isolated and characterized. Colony coloration varied between yellow-gray, white, brown, and olive, with feathery or wavy edges. As for radial growth, the highest averages were obtained by isolates LLM-M2 and TOR-M16, with 43.15 mm and 42.85 mm, respectively. Conidia production at 15 days was higher for isolate LLM-M2 with 9.8 × 107 conidia/mL; in the percentage of germination at 14 h, the treatment that reached 100% germination was TOR-M16. Isolate CMR-M7 reached 97.49% mycelial growth percentage at 288 h, being the best result; in the percentage of mortality in CBB adults, the treatments that reached 100% were TOR-M9, TOR-M16, and MNR-M1. In general, the results demonstrate the entomopathogenic potential of native isolates of Metarhizium sp. acting as biocontrol agents of Hypothenemus hampei, being a low-cost, easily accessible, and environmentally friendly alternative.

In planta ANALYSIS OF SALICYLIC ACID MEDIATED MANAGEMENT OF

Bacterial leaf blight caused by Xanthomonas oryzae pv. oryzae (Xoo) is a catastrophic bacterial disease of rice, and researchers are facing difficulties in developing the management practices, whereas the continued use of bactericides viz, streptomycin, copper oxychloride leads to the development of resistant Xoo strains. As a consequence, the development of an environmentally safe bactericide for the management of this devastating disease is needed. The present study focused on identification of effective synthetic activators against bacterial leaf blight in rice during their interaction. Survey was conducted to collect different isolates of Xoo and their morphological characters and biochemical properties were determined. All the 10 isolates exhibited circular to flattened slimy yellow colour colonies of the pathogen and showed positive reaction for citrate utilization, lysine utilization and ornithine utilization test. All the ten isolates showed negative reaction for sorbitol utilization. The collected isolates were subjected to PCR for the amplification of 16S-23S intergenic region with the species specific primer and obtained amplicon at ⁓470 bp for the confirmation at molecular level. The virulent isolate was identified by measuring biofilm formation and xanthomonadin pigment which revealed that the isolate ADT 53 strain T3 with the accession number OR121377 recorded maximum OD value of 0.356 for biofilm formation at 570 nm and 0.686 for xanthomonadin pigment at 445 nm. In planta studies revealed that salicylic acid at 100 ppm has the maximum inhibitory effect against bacterial leaf blight disease with the minimal lesion size of 0.7 cm compared to negative control (water). Keywords: Activators, BLB, Biofilm, Characterization, in planta, Xanthomonadin.