Potato Sucrose Research Articles

Efficiency of Penicillium sp. and Aspergillus sp. for bioleaching lithium cobalt oxide from battery wastes in potato dextrose broth and sucrose medium

Lithium-ion batteries, a primary power source, generate electronic hazardous waste at the end of their lifespan, which is a richer source of highly concentrated metals such as cobalt and lithium than those in natural resources. The waste is a potential renewable resource for the extraction of metals. Bioleaching, a process utilizing microbial activity, was used in this study to investigate the cobalt and lithium leaching efficiency from battery wastes. Two fungal strains, Aspergillus sp. strain JMET 15 and Penicillium sp. strain JMET 24 were used with different organic carbon sources (sucrose medium and potato dextrose broth) at 1 % pulp density. The sucrose medium derived a higher acid quantity and leached higher concentration of cobalt and lithium than in potato dextrose broth. Leaching efficiency by JMET 24 was higher than JMET 15. Cobalt (32.57 % and 27.19 %) and lithium (65.07 % and 40.58 %) were bioleached by JMET 24 and JMET 15, respectively, after 30 d cultivation in the sucrose medium. Lower pulp density was conducted to achieved higher leaching efficiency by JMET24, cobalt (77.87 % and 74.5 %) and lithium (99.88 % and 78.4 %) were bioleached at 0.1 % and 0.5 % pulp density, respectively. Cobalt oxide (Co3O4) was the product of the one-step recovery (leaching and precipitation) process. Cobalt was transformed from lithium cobalt oxide to cobalt phosphate-oxalate by JMET 24 in the sucrose medium. Thus, the potential of bioleaching as an effective method for recovering valuable metals from lithium-ion battery waste is demonstrated, presenting a promising approach for both waste management and resource recovery.

Study on the Identification, Biological Characteristics, and Fungicide Sensitivity of the Causal Agent of Strawberry Red Core Root Rot

Strawberry red core root rot disease affects the growth and yield of strawberry (Fragaria ananassa Duch), which leads to economic losses in China. The study employed a tissue separation method to isolate and identify the causal agent responsible for strawberry red core root rot. This was achieved by the observation of its morphological characteristics, sequencing analyses, and pathogenicity tests. The sensitivity of five chemical fungicides against the two species of Fusarium was determined using the mycelial growth rate method, and the biological characteristics of the two species were examined. The pathogens were identified as Fusarium solani and Fusarium oxysporum. The optimal conditions for the mycelial growth of F. solani and F. oxysporum were determined to be potato sucrose agar at 25 °C and pH 6, and potato dextrose agar at 30 °C and pH 8, respectively, with a 24:24 light cycle. The most suitable carbon and nitrogen sources for the mycelial growth of F. solani were sucrose and sodium nitrate (NaNO3), while for F. oxysporum, they were glucose and peptone. A fungicide sensitivity test indicated that Prochloraz had a good inhibitory effect on the growth of F. solani and F. oxysporum with EC50 values of 0.088 mg L−1 and 0.162 mg. The growth inhibition effect of Azoxystrobin to F. solani and Carbendazim to F. oxysporum was not obvious. This study provides a theoretical basis for further research on strawberry red core root disease and its prevention.

Biological and molecular characterization of pydiflumetofen and phenamacril dual-resistant Fusarium graminearum strains.

Fusarium head blight (FHB) caused by Fusarium graminearum species complex (FGSG) remains a major challenge to cereal crops and resistance to key fungicides by the pathogen threatens control efficacy. Pydiflumetofen, a succinate dehydrogenase inhibitor, and phenamacril, a cyanoacrylate fungicide targeting myosin I, have been applied to combat this disease. Nonetheless, emergence of pydiflumetofen resistance in a subset of field isolates alongside laboratory-induced facile generation of phenamacril-resistant isolates signals a critical danger of resistance proliferation. Our study investigates the development of dual resistance to these fungicides in F. graminearum. Utilizing pydiflumetofen-resistant (PyR) and -sensitive (PyS) isolates, we obtained dual-resistant (PyRPhR) and phenamacril-resistant (PySPhR) mutants on potato sucrose agar containing phenamacril. Mutation rates for phenamacril resistance were comparable between pydiflumetofen-resistant and -sensitive isolates, implying independent pathways for resistance development. The mutants compromised in fungal growth, competitive viability and deoxynivalenol production, suggesting fitness penalties for the dual-resistant mutants. However, no cross-resistance was found with tebuconazole or fludioxonil. In addition, we characterized four critical amino acid changes (S217L, C423R, K537T, E420G) in the Myo1 that were verified to confer phenamacril resistance in F. graminearum. This research indicates the possibility of resistance development for both pydiflumetofen and phenamacril in F. graminearum and emphasizes the need for fungicide resistance management for FHB. © 2024 Society of Chemical Industry.

Disease prevalence, incidence, morphological and molecular characterisation of Lasiodiplodia pseudotheobromae causing collar rot disease on peanut plants in Turkey

AbstractPeanut (Arachis hypogaea L.) holds significant commercial and dietary importance as a major source of edible oil and protein in Turkey. Stem, collar or root rot, caused by several fungal disease agent, are serious soil-borne diseases of peanut. Accurate and precise identification of the disease agent provides fundamental and precise information for integrated plant management. During the period from June to September 2021, symptoms consistent with collar rot disease, including dark-brown stem rot, chlorotic leaves, wilting, and eventual whole plant death, were observed on peanut plants cultivated in the different districts of Osmaniye Province of Turkey. The disease incidence ranged from 8.0 to 45.0% in the inspected fields with an average of 3.4% overall. Twenty-four single-spore representative isolates were obtained from surface-disinfected symptomatic tissues. Morphological characteristics of fungal mycelium, conidial and pycnidial structures on potato sucrose agar (PSA) and water agar (WA) closely resembled those described for Lasiodiplodia spp. All isolates caused typical collar rot symptoms upon artificial inoculation of peanut seedlings. Morphological identification of Lasiodiplodia spp. isolates was corroborated by MALDI-TOF and molecular analyses utilizing sequences from the internal transcribed spacer (ITS), β-tubulin 2 (tub2) and translation elongation factor-1 alpha (TEF1-α) loci. Phylogenetic analysis confirmed that the representative fungal isolates (MKUBK-B1 and MKUBK-K22) belong to Lasiodiplodia pseudotheobromae. To the best of our knowledge, this is the first report of L. pseudotheobromae infecting peanut plants in Turkey. This work is expected to contribute to previously limited knowledge regarding the host range, incidence and prevalence of L. pseudotheobromae as a soilborne pathogen of peanuts. Due to the potential destructiveness and broad host range of this pathogen, it is essential to develop new strategies to establish more reliable, environmentally sustainable, and cost-effective management approaches for this disease.

First Report of Colletotrichum gloeosporioides Causing Anthracnose on Euphorbia lathyris in China.

Euphorbia lathyris L. is a biennial herb in the Euphorbiaceae that has been used as a medicinal plant. It is distributed or cultivated worldwide, and the seeds of E. lathyris are the main source of ingenol, which is the precursor of Picato, the first medicine approved by USFDA for the treatment of solar keratosis (Abramovits et al. 2013). However, the production of E. lathyris can be severely hampered by the occurrence of plant diseases. Between 2020-2022 (specifically in October-November of each year), anthracnose-like symptoms were observed on E. lathyris in fields (E 118°49'50″, N 32°3'33″) in Nanjing, Jiangsu Province, China. The incidence of E. lathyris with disease symptoms was between 25%-30% (n = 100). The lesions on the leaves were evident initially as dark brown spots, which expanded into larger necrotic spots, finally resulting in leaves withering and dropping off. In severe cases, stem wilting was also observed. To determine the causal agent, we collected diseased leaf samples (n = 20) from different E. lathyris plants in the field (~ 1800 m2). After cleaning, the junctions of the diseased and healthy parts were excised and sterilized in 75% ethanol for 20-25 seconds, and rinsed with sterile water. After that, they were transferred onto potato sucrose agar (PSA) plates and placed at 25℃ for 3-4 days, until fungal growth was evident. The fungus was purified by recovering single conidia and growing them on PSA (Hu et al. 2015). A consistent fungal colony, based on morphological characteristics, was recovered from 17 samples. The colony color was initially white, green in the middle, and gradually changed into gray green as the colony matured. Conidia were transparent and cylindrical (22-28 µm × 7-9 µm, n = 50). Five loci informative (ITS, TUB, ACT, GAPDH, and CHS-1) (Weir et al. 2012) for Colletotrichum spp. identification were sequenced from two isolates ELC-1 and ELC-2 obtained from different plant individuals. Compared with a reference isolate (Colletotrichum gloeosporioides ZH3), the GAPDH, CHS-1, and TUB2 sequences of ELC-1 and ELC-2 showed 95% (263 bp out of 275 bp), 98% (295 bp out of 299 bp), and 99% (711 bp out of 712 bp and 717 bp out of 719 bp) similarity, respectively. The ITS sequence identities were 100% (577 bp out of 577 bp) and 99% (594 bp out of 597 bp), while the ACT sequence identities were 100% (281 bp out of 281 bp) and 98% (279 bp out of 284 bp). All sequences have been deposited in Genbank database (OR865865-OR865866 and OR873625-OR873632). After performing phylogenetic analysis with Mega 11, the pathogen was confirmed as C. gloeosporioides. To fulfil Koch's postulates, we sprayed six-week-old healthy plants with a conidia suspension of C. gloeosporioides (106 spores/mL) or sterile water (serve as control). The inoculated plants were placed at 25℃, 100% relative humidity, and 12-h photoperiod (Zhang et al. 2021). Six plants were inoculated for each treatment, and the experiment was repeated three times. After 6-8 days, the plants inoculated with C. gloeosporioides showed similar symptoms to those observed on diseased plants in the field, while the control plants remained healthy and free of disease. The pathogens were then re-isolated and identified as C. gloeosporioides. To our knowledge, this is the first report of C. gloeosporioides causing anthracnose on E. lathyris. Anthracnose may cause significant yield losses in E. lathyris production, and our results will provide experimental and theoretical basis for the management of the disease.

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.

Evaluation of Tolerance and Uptake of Cd and Mn for Microfungi Aspergillus flavus, Aspergillus oryzae, and Aspergillus terreus Isolated from Landfill Soil Collected from Bangar, La Union Philippines

Excessive deposition of heavy metals into the environment due to anthropogenic activities necessitates an eco-friendly clean-up strategy. Among microorganisms, limited studies have been made on the mycoremediation potential of microfungi. This paper evaluated three landfill microfungal isolates of Aspergillus species for tolerance and uptake to Cd and Mn. Culture media optimization was also performed for the evaluation of the tolerance index and heavy metal analysis of soil samples from the landfill site. Among the nine heavy metals analyzed, Mn and Fe were detected in relatively high amounts, while Cd, Ni, and Cu were detected in a moderate range. Luxuriant mycelial growth of A. oryzae (MK120548.1) and A. flavus (MH864264.1) was observed in potato dextrose agar while A. terreus (MH047280.1) grew best in potato sucrose agar. In terms of tolerance index, A. oryzae (MK120548.1) and A. flavus (MH864264.1) demonstrated high tolerance to Cd up to 10 mg/kg. A. oryzae (MK120548.1) showed high tolerance to Mn up to 1,000 mg/kg while A. flavus (MH864264.1) exhibited a very high 10,000 mg/kg tolerance. In terms of metal uptake, A. oryzae (MK120548.1) showed the highest metal uptake of up to 654 mg/kg of Cd, while A. terreus (MH047280.1) exhibited the highest metal uptake of 997 mg/kg ofMn. With these findings, A. oryzae (MK120548.1), A. flavus (MH864264.1), and A. terreus (MH047280.1) have considerable mycoremediation potential. Bioremediation studies in conjunction with plants can be explored to further assess the potential of these Aspergillus species.

Antifungal evaluation of turmeric rhizome extract against Colletotrichum capsici, the causal agent of anthracnose on red-chili peppers (Capsicum annum L.)

This study was conducted to evaluate the antifungal activity of the ethanolic extract of turmeric rhizome against Colletotrichum capsici Syd. and its efficacy in reducing anthracnose disease development on red chili peppers. The in vitro experiment aimed to determine the effect of the extract on inhibiting C. capsici growth in potato sucrose agar (PSA) medium incorporating the turmeric rhizome extract. The variables examined were mycelial growth, fungal sporulation, and spore germination. The in vivo experiment aimed to evaluate the effectiveness of the extract in reducing disease development on red chili peppers. The treatments were conducted using a completely randomized design with five concentration treatments and three replicates. The variables examined were radial mycelial growth zone diameter, fungal sporulation, spore germination, and disease intensity. Data were analyzed by analysis of variance, and the least significant difference was used for comparing means between treatments. The results of these experiments showed that ethanolic turmeric rhizome extract exhibited antifungal activity against C. capsici both in vitro and in vivo conditions. Turmeric rhizome extracts with concentrations of 1%, 2%, 3%, and 4% significantly inhibited mycelial growth, and even at a 4% concentration, C. capsici fungal colonies did not grow. Evaluation of the effect of the extract on fungal sporulation showed that inhibition occurred at a concentration of 3%, while inhibition of spore germination occurred at concentrations of 1%, 2%, and 3%. Application of turmeric extract at concentrations of 1%, 2%, 3%, and 4% effectively reduced the intensity of anthracnose on red chili.

The Potential of Endophytic Bacteria in Controlling Cylindrocladium sp., The Cause of Leaf Blight on Eucalyptus pellita Seedlings by In-Vitro

Cylindrocladium sp is a fungus that causes leaf blight on Eucalyptus pellita seedlings. Endophytic bacteria are an alternative biological control that can reduce the risk of environmental pollution and reduce resistance by pathogens. This research aims to obtain endophytic bacteria that have potential as biocontrol agents and to characterize isolates that inhibit more than 30%. Testing was carried out using the dual culture method on Potato Sucrose Agar (PSA) media. The results of the antagonism test showed that the endophytic bacterial isolates E320 and E323 were able to form an inhibition zone of more than 30%, namely 38.79% and 41.25% and had the potential to act as biocontrol agents for Cylindrocladium sp., which causes leaf blight on eucalyptus. The characteristics of isolate E320 were that it was able to produce protease, cellulase and siderophore activities, while isolate E323 was only able to produce protease

MEDIA DASAR LIMBAH DAUN KAKAO UNTUK PERTUMBUHAN MISELIUM JAMUR TIRAM PUTIH (Pleurotus ostreatus)

Cocoa leaf wastes as a basic medium are the nutrition that needed for the growth of white oyster mushroom mycelium (Pleurotus ostreatus). The objectives of this study were determine the cocoa leaf wastes as a basic medium for the growth of white oyster mushroom mycelium and the dosage of basic medium to encourage the best fungal growth. This research was designed in a Completely Randomized Design (CRD), consisting of seven treatments and three replications, namely P1 (Basic Media of 100% cocoa leaf waste), P2 (Basic Media 90% + Suplement 10%), P3 (Basic Media 80% + Suplement 20 %), P4 (Basic Media 70% + Suplement 30%), P5 (Basic Media 60% + Suplement 40%), P6 (Basic Media 50% + Suplement 50%), P7 (Sawdust 70%, 20% rice bran + flour corn 10%). The method that used in this study were media preparation, drying, grinding and formulation. The parameters that observed were the basic media, microscopic characteristics of hyphae, mycelium growth curve and macroscopic characteristics of colonies. The results of P1 and P7 showed the best growth for mycelium, observation of mycelium growth were done every 3 days. The isolation stage was carried out on Potato Sucrose Agar (PSA) media, the results obtained that P1 was 0,1 x 1012 CFU/g, P2 was 0,4 x 1012 CFU/g, P3 was 0,3 x 1012 CFU/g, P4 as much as 0,7 x 1012 CFU/g, P5 as much as 0,6 x 1012 CFU/g, P6 as much as 0,4 x 1012 CFU/g, P7 as much as 2,4 x 1012 CFU/g, respectively. Cocoa leaf waste are potentially as a growth medium for mycelium of white oyster mushroom.

Exploring the influence of maitake (Grifola frondosa) on Mus musculus learning-memory and anxiety behavior

Progressive neurodegeneration affect memory and behavior. One solution to increase neuroprotection is the consumption of medicinal mushrooms such as Grifola frondosa. This study aims to analyze the effect of G. frondosa extract on learning-memory and anxiety levels in mice. Mycelium was cultured in Potato Sucrose Broth medium at room temperature for 14 days. Dried mycelial was extracted using a solvent mixture (22% ethyl acetate and 11% methanol). Learning-memory and anxiety levels were observed using Y-Maze Alternation (Y-Maze) test, Morris Water Maze (MWM) test, Novel Object Recognition test, and Light and Dark Transition test. A total of 18 male mice were divided into three groups and given oral treatment for 14 days: control (distilled water), GF100 (extract of 100 mg/kg BW), and GF200 (crude extract of 200 mg/kg BW). The GF100 and GF200 groups increased spatial learning-memory in Y-Maze and novel object recognition memory, but both treatment groups did not significantly affect anxiety and learning-memory in MWM.

Morphological and molecular characterization of Fusarium incarnatum as a causal disease agent of pepper (Capsicum annuum) fruit rot

AbstractChilli pepper (Capsicum annuum L.) is one of the most important commercially cultivated and consumed vegetables in Turkey. During a disease survey, typical symptoms of fruit rot were observed on mature chilli pepper fruits in several surveyed fields and on dried pepper fruits obtained from local retailers/bazaars in Hatay Province, Turkey. Disease incidence varied from 15 to 45% of the plants in the inspected fields. Following standard isolation procedures, 40 fungal isolates were isolated, purified and single‐spore cultures were obtained from surface‐disinfected, rotted dried pepper tissue. Of these isolates, six fungal isolates with dense, cottony white aerial mycelia that became beige with age, were isolated on a potato sucrose agar. All isolates were found to be pathogenic on artificially inoculated chilli pepper fruit. Based on morphological characteristics, the isolates were initially identified as Fusarium incarnatum (Desm.) Sacc. 1886. Morphological identification of F. incarnatum isolates was further confirmed by MALDI‐TOF and molecular analyses using the sequences of the internal transcribed spacer (ITS), partial translation elongation factor‐1α (TEF‐1α) and second largest subunit of RNA polymerase II (RPB2) loci. Phylogenetic analysis based on ITS, TEF‐1α, RPB2 and concatenation of TEF‐1α, RPB2 loci sequences performed with several isolates of Fusarium spp. confirmed that representative fungal isolates (MKUZF1 and MKUZF4) belong to F. incarnatum. To our knowledge, this is the first report of F. incarnatum causing fruit rot in chilli peppers grown in Turkey.

Survey and Isolation of Rice False Smut Pathogen Ustilaginoidea virens (Cooke) Takahashi

Roving survey has been conducted during first quarter of 2023 at selected paddy fields of Cauvery Delta zone (Tanjore, Perambalur) and Western zone (Coimbatore, Erode) of Tamil Nadu to assess the Percent Disease Incidence for rice false smut disease in which the maximum disease incidence was recorded in Erode district (40%). The infected spikelet were observed with the characteristic symptoms where the individual rice grains turned into large, fluffy and velvety green balls, later these balls turns hard during maturity. The pathogen Ustilaginoidea virens was isolated from the infected smut balls in potato sucrose agar medium in which the fungus appears as small white colonies after seven days of isolation.

Cultural, Morphological and Molecular Characterization of Ustilaginoidea virens Causing Rice False Smut in Tamil Nadu, India

Rice false smut, caused by the fungus Ustilaginoidea virens is one of the most destructive and re-emerging disease in many rice growing areas of Tamil Nadu, India. The current study was carried out to characterize different isolates of Ustilaginoidea virens collected from major rice growing areas in Tamil Nadu, India. The pathogen was isolated by different isolation methods, out of which inner spore dust was effective. The colony was milky white to yellowish white, fluffy, leathery and compact. The colony diameter ranged between 35.33 mm and 65.33mm. The mycelium was hyaline, septate and width of the hyphae ranged between 1.84 and 3.10 µm. The size of the conidia was also varied among the isolates and the maximum size (6.35 µm) was observed in FS 6 isolate and minimum in FS 2 (4.10 µm). The growth of fungus in different media was examined. The maximum colony diameter was observed in Potato Sucrose Agar (PSA) medium and minimum growth in Czapek Agar (CA) medium. The molecular confirmation was done through U. virens-specific internal transcribed spacer (ITS) region amplification using the primers, US 1-5/US3-3. The primers amplified a product of 380 bp which are specific for the fungus U. virens. The identity of the fungus was further confirmed through ITS sequencing which showed 98-99% sequence homology. The dendrogram showed two main clusters; cluster I comprised of five isolates (FS1, FS2, FS9, FS10 and FS3), while Cluster II consisted of five isolates (FS4, FS8, FS6, FS5 and FS7).

Study of Optimal Conditions to Grow Thai Ganoderma, Fruiting Test, Proximate and Their Alpha Glucosidase Inhibitory Activity.

Ganoderma (Ganodermataceae) has a worldwide distribution and has been widely used in traditional medicines. In this study, we report wild strains of Ganoderma that include two G. sichuanense and one G. orbiforme from northern Thailand. Optimal conditions for mycelium growth were ensured. The most favourable medium was potato sucrose agar for G. sichuanense and oatmeal agar for G. orbiforme and at 25 °C and 30 °C and pH 4-8. All types of cereal grains can be used to promote the growth of the mycelia of Ganoderma species. Fruiting tests were performed. All strains of Ganoderma produce fruiting bodies successfully in bag culture at 28 ± 1 °C with 75-85% relative humidity. Only G. orbiforme produced fruiting bodies in field cultivation at the laboratory scale. In the first flush yields, the G. sichuanense strain MFLUCC 22-0064 gave better production (the B.E was 152.35 ± 6.98 g). This study is the first to document the bag and field cultivation of wild Thai G. orbiforme. Ganoderma species are revealed to contain high amounts of fiber (47.90-52.45% d.b.), protein (12.80-14.67% d.b.), fat (4.90-5.70% d.b.), and carbohydrates (3.16-4.02% d.b.). Additionally, G. sichuanense and G. orbiforme were preliminarily screened for biological activity for inhibition of alpha-glucosidase enzyme activity. The IC50 values of G. orbiforme (MFLUCC 22-0066) was 105.97 ± 1.36 µg/mL and G. sichuanense (MFLUCC 22-0064) was 126.94 ± 0.87 µg/mL. Both strains had better inhibition than acarbose (168.18 ± 0.89 µM). These results on wild strains of Ganoderma will be useful for further studies on the applications of Ganoderma. Later the species can be introduced to domestic markets for cultivation and medicinal use.

Metode Isolasi Jamur Patogen Serangga (Aschersonia Placenta) Dengan Menggunakan Media Potato Sucrose Agar (PSA)

Cultivating oranges is one of the main fruit commodities in Bali. Citrus cultivation often experiences problems, one of which is insect pests and plant diseases. Pests that often attack are whitefly. Whiteflies cause damage directly or indirectly. It directly results in stunted plant leaves because the whitefly sucks on the young shoots and leaves. Indirectly, the whitefly has been reported as a vector of various diseases caused by viruses. Based on this, it is necessary to control efforts, one of which is by using biological control of insect pathogenic fungi. The pathogenic fungus found in association with the citrus whitefly is Aschersonia placenta. The insect pathogen A. placenta naturally exists in nature and has potential as a biological control agent, however special handling is required in its utilization. One such treatment is the isolation of the fungus from the field for mass cultivation in the laboratory. Isolation poetry using Potato Sucrose Agar (PSA) was the best, of the four isolation methods performed. The method is detailed as follows: the fungal stroma is rinsed with sterile distilled water, with 70% alcohol, then rinsed again with sterile distilled water before being planted in PSA media.

Metabolite Analysis of Alternaria Mycotoxins by LC-MS/MS and Multiple Tools

Alternaria fungi are widely distributed plant pathogens that invade crop products, causing significant economic damage. In addition, toxic secondary metabolites produced by the fungi can also endanger consumers. Many of these secondary metabolites are chemically characterized as mycotoxins. In this study, Q Exactive Orbitrap mass spectrometry was used for the non-targeted analysis of the metabolome of seven Alternaria isolates cultured on Potato Carrot Agar (PCA), Potato Dextrose Agar (PDA) and Potato Sucrose Agar (PSA) medium. Due to the difficulty of detecting modified toxins, an analytical strategy with multiple visual analysis tools was also used to determine the presence of sulfate conjugated toxins, as well as to visualize the molecular network of Alternaria toxins. The results show that PSA medium exhibits more advantageous properties for the culture of Alternaria, with more toxigenic species and quantities and more obvious metabolic pathways. Based on high-resolution tandem mass spectrometry (MS/MS) data, the mycotoxins and their metabolites were mainly clustered into four groups: alternariol (AOH)/alternariol monomethyl ether (AME)/altenusin (ALU)/altenuene (ALT)/dehydroaltenusin (DHA)/Desmethyldehydroaltenusin (DMDA) families, Altertoxin-I (ATX-I) family, tentoxin (TEN) family and tenuazonic acid (TeA) family. Moreover, the PSA medium is more suitable for the accumulation of AOH, AME, ALU, ALT, DHA and DMDA, while the PDA medium is more suitable for the accumulation of ATX-I, TEN and TeA. This research may provide theoretical support for the metabolomics study of Alternaria.

Morpho-molecular variability of different isolates of Ustilagonoidea virens causing false smut of rice

Spore balls of false smut of rice caused by Ustilagonoidea virens were collected from different major rice belts of eastern part of India. Ten isolates from ten different locations were collected and isolated in the laboratory for their, cultural, morphological and molecular characterization. Potato sucrose agar media was used for their cultural studies and observation on colony diameter, colour and sporulation were made. Spores of all the isolates were observed under scanning electron microscope (SEM) and light microscope for spore size, spine length and colour. Differences in colony characters were observed where the highest mycelial growth was observed in isolate Uv2 with 5.33 cm colony diameter and lowest mycelial growth was observed in Uv9 (3.70 cm). Most of the isolates produced white and circular colony. Spores of all the isolates observed under a light microscope appeared to be olive green in colour, however, a significant variation in spore size and spike length under SEM was observed. Spores size and spike length of different isolates ranged between 4.533 μm to 6.169 μm and 362.567 nm to 799.533 nm, respectively. For genetic identification and variability, DNAs of isolates were amplified using ITS1 and ITS 4 primers. The length of nucleotide sequences of different isolates varied from 583 to 606 bp. Phylogenetic analysis revealed genetic variation among the isolates, dividing them into two major groups.

Molecular characterization of endophytic fungi-Daldinia eschscholtzii from Aloe vera plants in Bangladesh

Aloe vera (L.) Burm. f. is a plant of health, beauty and medicine that is widely used in Bangladesh with high demand. An experiment was performed to find out the association of endophytic fungus with Aloe vera leaves and to assess their growth behavior on different culture media, temperatures and pH. The endophytic fungus-Daldinia eschscholtzii (Ehrenb.) Rehm. was identified based on the morphological and molecular characterization using the ITS-rDNA sequence. Various solid culture media were used to assess the fungal growth, and temperature regimes i.e., 15℃, 20℃, 25℃, 30℃, 35℃ were investigated. The fungal endophyte showed maximum mycelial growth on Potato Sucrose Agar followed by Potato Dextrose Agar and Carrot Agar media. Optimum growth and development of the endophyte was recorded at temperature 25 to 35℃. We observed the effect of five different pH conditions, viz., 5, 6, 7, 8, 9 on the mycelial growth of the fungus. However, the fungal endophyte was very insensitive to the pH of the substrate. To the best of our knowledge, this is the first known occurrence of D. eschscholtzii as an endophyte in Aloe vera in Bangladesh.
 Jahangirnagar University J. Biol. Sci. 11(1 & 2): 1-11, 2022 (June & December)

First Report of Leaf Spot and Blight on Crown Daisy Caused by Cercospora apii in China.

Crown daisy (Glebionis coronaria L.), also known as chrysanthemum greens, is a popular vegetable in Asia, especially in China. The leaves have been used in folk medicine as a tonic for the liver, blood, intestines and to control anemia and high blood pressure. In November 2020, severe leaf spot and blight was observed with 80% to 95% incidence on crown daisy growing in greenhouses in Fengxian, Shanghai, China(121°22'E, 30°53'N). Irregular rounded spots appeared with a light gray center and water-soaked margins. Round lesions enlarged and merged with age, followed by the development of a necrotic area resulting in the typical "frog-eye" and causing a continuous deterioration of crown daisy. Diseased leaves were washed in running water for 30 min. Small fragments (5 × 5 mm) taken from the margin of lesions were disinfected with 1% NaClO for 3 min, rinsed three times with sterile water, cultured on potato sucrose agar (PSA) augmented with 50 mg streptomycin/liter at 26 oC,and incubated in the dark. Colonies had identical morphology, and TH11290202 was selected and deposited in the plant pathology lab of Shanghai Academy of Agricultural Sciences. Mycelium was initially cottony and white and became appressed to the medium and dark brown with time. Conidia did not form on any media, including PSA, PDA, V8 agar (V8A), maize leaf carbonate agar (MLPCA), pepper leaf carbonate agar (PLPCA), etc. To confirm the identity of the pathogen, genomic DNA was extracted from TH11290202 with the cetyltrimethylammonium ammonium bromide (CTAB) method from the mycelia. Five loci were PCR amplified, namely, the internal transcribed spacer (ITS), translation elongation factor (TEF), calmodulin (cmdA), histone (H3) and actin (ACT), using primers ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Jaklitsch et al. 2005), CAL-F/CAL-R (O'Donnell et al. 2000), cylh3f/cylh3r (Glass and Donaldson 1995), and ACT-512F/ACT-783R (Carbone and Kohn 1999), respectively. The resulting sequences were deposited in GenBank (MW819910, MW981277, MW981278, ON798723, and MW981279). Analysis of the ITS, TEF, cmdA, H3 and ACT gene sequences of isolate TH11290202 revealed that it was a member of the genus Cercospora, sharing 99.79%, 99.66%, 98.10% 99.74% and 100% sequence similarity with type strain of Cercospora apii CBS 116455. A multilocus phylogenetic analysis was performed using sequences from other closely related taxa obtained from GenBank. Based on morphological and molecular characteristics, TH11290202 was identified as C. apii (Crous and Braun 2003; Groenewald et al. 2006; Milosavljević et al. 2014). To confirm pathogenicity, Koch's postulates were fulfilled on 30 mature plants, which were maintained in a growth chamber (at 26 °C, relative humidity 90%, 12/12 h light/dark). Surface-sterilized leaves were sprayed with a mycelial suspension. Brown lesions were formed 7 days after inoculation on 15 plants, whereas the noninoculated controls remained asymptomatic on the other 15 plants. To our knowledge, this is the first report of C. apii causing leaf spot and blight on G. coronaria in China and will provide useful information for developing effective control strategies.