Search results for: mycelia
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 21

Search results for: mycelia

21 The Increase in Functionalities of King Oyster Mushroom (Pleurotus eryngii) Mycelia Depending on the Increase in Nutritional Components

Authors: Hye-Sung Park, Eun-Ji Lee, Chan-Jung Lee, Won-Sik Kong

Abstract:

This study was conducted to research king oyster mushroom (Pleurotus eryngii) mycelia with reinforced functionalities. 0 to 4% of saccharide components, such as glucose (glu), lactose (lac), mannitol (man), xylose (xyl), and fructose (fru) and 0 to 0.04% of amino acid components, such as aspartic acid (asp). Cysteine (cys), threonine (thr), glutamine (gln), and serine (ser) were added to liquid media, and antioxidant activities, nitrite scavenging activities, and total polyphenol contents of the cultured mycelia were measured. In the saccharide-added group, 4 strains except ASI 2887 had high antioxidant activities when 1% of xyl was added and especially, the antioxidant activity of ASI 2839 was 73.9%, which was the highest value. In the amino acid-added group, the antioxidant activity of ASI 2839 was 66.3% that was the highest value when 0.2% of ser was added. But all the 5 strains had lower antioxidant activities than the saccharide-added group overall. In the saccharide-added group, 4 strains except ASI 2887 had higher nitrite scavenging activities than other group when 1% of xyl was added and especially, the nitrite scavenging activity of ASI 2824 was 57.8% that was the highest value. It was revealed that the saccharide-added group and the amino acid-added group had a similar efficiency of nitrite scavenging activity. Although the same component-added group did not show a certain increase or decrease in total polyphenol contents, ASI 2839 with the highest antioxidant activity had 6.8mg/g, which was the highest content when 1% of xyl was added. In conclusion, this study demonstrated that when 1% of xyl was added, functionalities of Pleurotus eryngii mycelia, including antioxidant activities, nitrite scavenging activities, and total polyphenol contents improved.

Keywords: king oyster mushroom, saccharide, amino acid, mycelia

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20 Medicinal Plants and Arbuscular mycorrhizal Colonization

Authors: Ammani K., Glory M.

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Demands of traditional herbal medicines are increasing day by day over the world. Considering the growing demand of medicinal plants in curative treatments and the role of VAM fungi in augmentation of the production of active secondary metabolites by the medicinal plants, the present work has been undertaken to survey the mycorrhizal status in 30 different medicinal plants belonging to various families from Krishna district, Andhra Pradesh. The roots were collected carefully and stained by the Phillips & Hayman technique. Basing on the occurrence of vesicles and arbuscules, categorized into four grades; Excellent: mycelia, vesicles or arbuscules present more than 75% of root bits, Good: mycelia, vesicles or arbuscules present 50-75% in surface of root bits, moderate: mycelia, vesicles or arbuscules present 25-50% in surface of root bits, and poor: mycelia, vesicles or arbuscules present 1-25% in surface of root bits. The study reveals that the roots of all plants were colonized by AM fungi. Percentage of root colonization by AM fungi was more in Aloe vera, Phylanthus emblica, Azadiracta indica and least in plants such as Aerva lanata, Vinca rosea, Crotalaria verrucosa among the 30 medicinal plants in present study. The enhancement of growth and vigour and increased production of bioactive compounds of the medicinal plants is desirable which may be achieved by inoculation of the roots with Arbuscular mycorrhizal fungi. There is a steady increase in the cultivation of medicinal plants to maintain a steady supply to support the increasing demand but corresponding researches of VAM fungi and their association in medicinal plants have received very little attention as compared to the studies on forest species and field crops. So a vast research on this field is necessary for a better tomorrow.

Keywords: Arbuscular mycorrhizae, colonization, categories, medicinal plants

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19 Genetic Divergence and Morphogenic Analysis of Sugarcane Red Rot Pathogen Colletotrichum falcatum under South Gujarat Condition

Authors: Prittesh Patel, Ramar Krishnamurthy

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In the present study, nine strains of C. falcatum obtained from different places and cultivars were characterized for sporulation, growth rate, and 18S rRNA gene sequence. All isolates had characteristic fast-growing sparse and fleecy aerial mycelia on potato dextrose agar with sickle shape conidia (length x width: varied from 20.0 X 3.89 to 25.52 X 5.34 μm) and blackish to orange acervuli with setae (length x width: varied from 112.37X 2.78 to 167.66 X 6.73 μm). They could be divided into two groups on the base of morphology; P1, dense mycelia with concentric growth and P2, sparse mycelia with uneven growth. Genomic DNA isolation followed by PCR amplification with ITS1 and ITS4 primer produced ~550bp amplicons for all isolates. Phylogeny generated by 18S rRNA gene sequence confirmed the variation in isolates and mainly grouped into two clusters; cluster 1 contained CoC671 isolates (cfNAV and cfPAR) and Co86002 isolate (cfTIM). Other isolates cfMAD, cfKAM, and cfMAR were grouped into cluster 2. Remaining isolates did not fall into any cluster. Isolate cfGAN, collected from Co86032 was found highly diverse of all the nine isolates. In a nutshell, we found considerable genetic divergence and morphological variation within C. falcatum accessions collected from different areas of south Gujarat, India and these can be used for the breeding program.

Keywords: Colletotrichum falcatum, ITS, morphology, red rot, sugarcane

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18 Evaluation of Mixing and Oxygen Transfer Performances for a Stirred Bioreactor Containing P. chrysogenum Broths

Authors: A. C. Blaga, A. Cârlescu, M. Turnea, A. I. Galaction, D. Caşcaval

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The performance of an aerobic stirred bioreactor for fungal fermentation was analyzed on the basis of mixing time and oxygen mass transfer coefficient, by quantifying the influence of some specific geometrical and operational parameters of the bioreactor, as well as the rheological behavior of Penicillium chrysogenum broth (free mycelia and mycelia aggregates). The rheological properties of the fungus broth, controlled by the biomass concentration, its growth rate, and morphology strongly affect the performance of the bioreactor. Experimental data showed that for both morphological structures the accumulation of fungus biomass induces a significant increase of broths viscosity and modifies the rheological behavior. For lower P. chrysogenum concentrations (both morphological conformations), the mixing time initially increases with aeration rate, reaches a maximum value and decreases. This variation can be explained by the formation of small bubbles, due to the presence of solid phase which hinders the bubbles coalescence, the rising velocity of bubbles being reduced by the high apparent viscosity of fungus broths. By biomass accumulation, the variation of mixing time with aeration rate is gradually changed, the continuous reduction of mixing time with air input flow increase being obtained for 33.5 g/l d.w. P. chrysogenum. Owing to the superior apparent viscosity, which reduces considerably the relative contribution of mechanical agitation to the broths mixing, these phenomena are more pronounced for P. chrysogenum free mycelia. Due to the increase of broth apparent viscosity, the biomass accumulation induces two significant effects on oxygen transfer rate: the diminution of turbulence and perturbation of bubbles dispersion - coalescence equilibrium. The increase of P. chrysogenum free mycelia concentration leads to the decrease of kla values. Thus, for the considered variation domain of the main parameters taken into account, namely air superficial velocity from 8.36 10-4 to 5.02 10-3 m/s and specific power input from 100 to 500 W/m3, kla was reduced for 3.7 times for biomass concentration increase from 4 to 36.5 g/l d.w. The broth containing P. crysogenum mycelia aggregates exhibits a particular behavior from the point of view of oxygen transfer. Regardless of bioreactor operating conditions, the increase of biomass concentration leads initially to the increase of oxygen mass transfer rate, the phenomenon that can be explained by the interaction of pellets with bubbles. The results are in relation with the increase of apparent viscosity of broths corresponding to the variation of biomass concentration between the mentioned limits. Thus, the apparent viscosity of the suspension of fungus mycelia aggregates increased for 44.2 times and fungus free mycelia for 63.9 times for CX increase from 4 to 36.5 g/l d.w. By means of the experimental data, some mathematical correlations describing the influences of the considered factors on mixing time and kla have been proposed. The proposed correlations can be used in bioreactor performance evaluation, optimization, and scaling-up.

Keywords: biomass concentration, mixing time, oxygen mass transfer, P. chrysogenum broth, stirred bioreactor

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17 Howard Mold Count of Tomato Pulp Commercialized in the State of São Paulo, Brazil

Authors: M. B. Atui, A. M. Silva, M. A. M. Marciano, M. I. Fioravanti, V. A. Franco, L. B. Chasin, A. R. Ferreira, M. D. Nogueira

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Fungi attack large amount of fruits and those who have suffered an injury on the surface are more susceptible to the growth, as they have pectinolytic enzymes that destroy the edible portion forming an amorphous and soft dough. The spores can reach the plant by the wind, rain and insects and fruit may have on its surface, besides the contaminants from the fruit trees, land and water, forming a flora composed mainly of yeasts and molds. Other contamination can occur for the equipment used to harvest, for the use of boxes and contaminated water to the fruit washing, for storage in dirty places. The hyphae in tomato products indicate the use of raw materials contaminated or unsuitable hygiene conditions during processing. Although fungi are inactivated in heat processing step, its hyphae remain in the final product and search for detection and quantification is an indicator of the quality of raw material. Howard Method count of fungi mycelia in industrialized pulps evaluates the amount of decayed fruits existing in raw material. The Brazilian legislation governing processed and packaged products set the limit of 40% of positive fields in tomato pulps. The aim of this study was to evaluate the quality of the tomato pulp sold in greater São Paulo, through a monitoring during the four seasons of the year. All over 2010, 110 samples have been examined; 21 were taking in spring, 31 in summer, 31 in fall and 27 in winter, all from different lots and trademarks. Samples have been picked up in several stores located in the city of São Paulo. Howard method was used, recommended by the AOAC, 19th ed, 2011 16:19:02 technique - method 965.41. Hundred percent of the samples contained fungi mycelia. The count average of fungi mycelia per season was 23%, 28%, 8,2% and 9,9% in spring, summer, fall and winter, respectively. Regarding the spring samples of the 21 samples analyzed, 14.3% were off-limits proposed by the legislation. As for the samples of the fall and winter, all were in accordance with the legislation and the average of mycelial filament count has not exceeded 20%, which can be explained by the low temperatures during this time of the year. The acquired samples in the summer and spring showed high percentage of fungal mycelium in the final product, related to the high temperatures in these seasons. Considering that the limit of 40% of positive fields is accepted for the Brazilian Legislation (RDC nº 14/2014), 3 spring samples (14%) and 6 summer samples (19%) will be over this limit and subject to law penalties. According to gathered data, 82% of manufacturers of this product manage to keep acceptable levels of fungi mycelia in their product. In conclusion, only 9.2% samples were for the limits established by Resolution RDC. 14/2014, showing that the limit of 40% is feasible and can be used by these segment industries. The result of the filament count mycelial by Howard method is an important tool in the microscopic analysis since it measures the quality of raw material used in the production of tomato products.

Keywords: fungi, howard, method, tomato, pulps

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16 The Production of Reinforced Insulation Bricks out of the Concentration of Ganoderma lucidum Fungal Inoculums and Cement Paste

Authors: Jovie Esquivias Nicolas, Ron Aldrin Lontoc Austria, Crisabelle Belleza Bautista, Mariane Chiho Espinosa Bundalian, Owwen Kervy Del Rosario Castillo, Mary Angelyn Mercado Dela Cruz, Heinrich Theraja Recana De Luna, Chriscell Gipanao Eustaquio, Desiree Laine Lauz Gilbas, Jordan Ignacio Legaspi, Larah Denise David Madrid, Charles Linelle Malapote Mendoza, Hazel Maxine Manalad Reyes, Carl Justine Nabora Saberdo, Claire Mae Rendon Santos

Abstract:

In response to the global race in discovering the next advanced sustainable material that will reduce our ecological footprint, the researchers aimed to create a masonry unit which is competent in physical edifices and other constructional facets. From different proven researches, mycelium has been concluded that when dried can be used as a robust and waterproof building material that can be grown into explicit forms, thus reducing the processing requirements. Hypothesizing inclusive measures to attest fungi’s impressive structural qualities and absorbency, the researchers projected to perform comparative analyses in creating mycelium bricks from mushroom spores of G. lucidum. Three treatments were intended to classify the most ideal concentration of clay and substrate fixings. The substrate bags fixed with 30% clay and 70% mixings indicated highest numerical frequencies in terms of full occupation of fungal mycelia. Subsequently, sorted parts of white portions from the treatment were settled in a thermoplastic mold and burnt. Three proportional concentrations of cultivated substrate and cement were also prioritized to gather results of variation focused on the weights of the bricks in the Water Absorption Test and Durability Test. Fungal inoculums with solutions of cement showed small to moderate amounts of decrease and increase in load. This proves that the treatments did not show any significant difference when it comes to strength, efficiency and absorption capacity. Each of the concentration is equally valid and could be used in supporting the worldwide demands of creating numerous bricks while also taking into consideration the recovery of our nature.

Keywords: mycelium, fungi, fungal mycelia, durability test, water absorption test

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15 Cochliobolus sativus: An Important Pathogen of Cereal Crops

Authors: Awet Araya

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Cochliobolus sativus ((anamorphic stage: Bipolaris sorokiniana (synonyms: Helminthosporium sorokinianum, Drechslera sorokiniana, and Helminthosporium sativum)) is an important pathogen of cereal crops. Many other grass species are also hosts for this fungus. Yield losses have been reported from many regions, especially where barley and wheat are commercially cultivated. The fungus has a worldwide distribution. The pathogen causes root rot, seedling blight, spot blotch, head blight, and black point. Environmental conditions affect disease development. Most of the time, fungus survives as mycelia and conidia. Pseudothecium of the fungus is not commonly encountered and probably not important in the epidemiology of the disease. The fungus can be in seed, soil, or in plant parts. Crop rotation, proper fertilization, reducing other stress factors, fungicide treatments, and resistant cultivars may be used for the control of the disease.

Keywords: Cochliobolus sativus, barley, cultivars, root rot

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14 Antifungal Lactobacilli Affect Mycelium Morphology and Protect Apricot Juice against Mold Spoilage

Authors: Nora Laref, Bettache Guessas

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Preservation of foods mainly depends on delaying or inhibiting the growth of spoilage microorganisms, and antifungal activity of lactic acid bacteria is one of the technological properties researched. The antifungal activity was screened with overlay method of six strains of lactic acid bacteria (Lactobacillus plantarum LB54, LB52, LB51, LB20, LB24 Lactobacillus farciminis LB53) isolated from silage, camel milk and carrot against Aspergillus sp. Lactobacillus plantarum and farciminis inhibit spore germination and mycelia growth of Aspergillus sp., the production of antifungal compounds by these strains was detectable after 4h of incubation at 30°C and show total inhibition after 24h in liquid media, but in solid media showed a good inhibition after 96h of incubation, these compounds cause malformations in the thalle, conidiophore and conidia. These strains could be used as agents of biopreservation since have the ability to retard Aspergillus sp., growth in apricot juice with and without sugar conserved in refrigerator but not in bread.

Keywords: lactobacillus, antifungal substances, aspergillus, biopreservation

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13 Histopathological Features of Infections Caused by Fusarium equiseti (Mart.) Sacc. in Onion Plants from Kebbi State, Northern Nigeria

Authors: Wadzani Dauda Palnam, Alao S. Emmanuel Laykay, Afiniki Bawa Zarafi, Olufunmilola Alabi, Dora N. Iortsuun

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Onion production is affected by several diseases including fusariosis. Study was conducted to investigate the histopathological features of different onion tissues infected with Fusarium equiseti by inoculation with soil drench, root dip and mycelia paste methods. This was carried out by fixation, dehydration, clearing, wax embedding, sectioning, staining and mounting of leaf and root sections for microscopical examination at 400x. Once infection occurred in the roots, the pathogen moved through the vascular system to colonize the whole plant. At first, it grew in the intercellular spaces of the root cortex but soon invaded the cells, followed by colonization of the cells by its hyphae and microconidia. At later stages of infection, the cortex tissue became completely disorganized and decomposed as the pathogen advance to the shoot system via the vessel elements; this may be responsible for the early wilting symptom of infected plants arising from the severe water stress due to blockage of the xylem tissues.

Keywords: onion, histopathology, infection, fusaria, inoculation

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12 Antimicrobial, Antioxidant and Enzyme Activities of Geosmithia pallida (KU693285): A Fungal Endophyte Associated with Brucea mollis Wall Ex. Kurz, an Endangered and Medicinal Plant of N. E. India

Authors: Deepanwita Deka, Dhruva Kumar Jha

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Endophytes are the microbes that colonize living, internal tissues of plants without causing any immediate, overt negative effects. Endophytes are rich source of therapeutic substances like antimicrobial, anticancerous, herbicidal, insecticidal, immunomodulatory compounds. Brucea mollis, commonly known as Quinine in Assam, belonging to the family Simaroubaceae, is a shrub or small tree, recorded as endangered species in North East India by CAMP survey in 2003. It is traditionally being used as antimalarial and antimicrobial agent and has antiplasmodial, cytotoxic, anticancer, diuretic, cardiovascular effect etc. Being endangered and medicinal; this plant may host certain noble endophytes which need to be studied in depth. The aim of the present study was isolation and identification of potent endophytic fungi from Brucea mollis, an endangered medicinal plant, to protect it from extinction due to over use for medicinal purposes. Aseptically collected leaves, barks and roots samples of healthy plants were washed and cut into a total of 648 segments of about 2 cm long and 0.5 cm broad with sterile knife, comprising 216 segments each from leaves, barks and roots. These segments were surface sterilized using ethanol, mercuric chloride (HgCl2) and aqueous solution of sodium hypochlorite (NaClO). Different media viz., Czapeck-Dox-Agar (CDA, Himedia), Potato-Dextrose-Agar (PDA, Himedia), Malt Extract Agar (MEA, Himedia), Sabourad Dextrose Agar (SDA, Himedia), V8 juice agar, nutrient agar and water agar media and media amended with plant extracts were used separately for the isolation of the endophytic fungi. A total of 11 fungal species were recovered from leaf, bark and root tissues of B. mollis. The isolates were screened for antimicrobial, antioxidant and enzymatic activities using certain protocols. Cochliobolus geniculatus was identified as the most dominant species. The mycelia sterilia (creamy white) showing highest inhibitory activity against Candida albicans (MTCC 183) was induced to sporulate using modified PDA media. The isolate was identified as Geosmithia pallida. The internal transcribed spacer of rDNA was sequenced for confirmation of the taxonomic identity of the sterile mycelia (creamy white). The internal transcribed spacer r-DNA sequence was submitted to the NCBI (KU693285) for the first time from India. G. pallida and Penicillium showed highest antioxidant activity among all the isolates. The antioxidant activity of G. pallida and Penicillium didn’t show statistically significant difference (P˃0.05). G. pallida, Cochliobolus geniculatus and P. purpurogenum respectively showed highest cellulase, amylase and protease activities. Thus, endopytic fungal isolates may be used as potential natural resource of pharmaceutical importance. The endophytic fungi, Geosmithia pallida, may be used for synthesis of pharmaceutically important natural products and consequently can replace plants hitherto used for the same purpose. This study suggests that endophytes should be investigated more aggressively to better understand the endophyte biology of B. mollis.

Keywords: Antimicrobial activity, antioxidant activity, Brucea mollis, endophytic fungi, enzyme activity, Geosmithia pallida

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11 Effect of Time of Planting on Powdery Mildew Development on Cucumber

Authors: H. Parameshwar Naik, Shripad Kulkarni

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Powdery mildew is a serious disease among the fungal in high humid areas with varied temperature conditions. In recent days disease becomes very severe due to uncertain weather conditions and unique character of the disease is, it produces white mycelia growth on upper and lower leaf surfaces and in severe conditions it leads to defoliation. Results of the experiment revealed that sowing of crop in the I fortnight (FN) of July recorded the minimum mean disease severity (7.96 %) followed by crop sown in II FN of July (13.19 %) as against the crop sown in II FN of August (41.44 %) and I FN of September (33.78 %) and the I fortnight of October (33.77 %). In the first date of sowing infection started at 45 DAS and progressed till 73 DAS and it was up to 14.66 Percent and in second date of sowing disease progressed up to 22.66 percent and in the third date of sowing, it was up to 59.35 percent. Afterward, the disease started earlier and progressed up to 66.15 percent and in sixth and seventh date of sowing disease progressed up to 43.15 percent and 59.85 percent respectively. Disease progress is very fast after 45 days after sowing and highest disease incidence was noticed at 73 DAS irrespective of dates of sowing. From the results of the present study, it is very clear that disease development will be very high if crop sown in between 1st fortnight of August and the 1st fortnight of September.

Keywords: cucumber, India, Karnataka, powdery mildew

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10 Control of Spoilage Fungi by Lactobacilli

Authors: Laref Nora, Guessas Bettache

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Lactic acid bacteria (LAB) have a major potential to be used in biopreservation methods because they are safe to consume (GRAS: generally regarded as safe) and they naturally occurring microflora of many foods. The preservative action of LAB is due to several antimicrobial metabolites, including lactic acid, acetic acid, hydrogen peroxide, bacteriocins, carbon dioxide, diacetyl, and reuterin. Several studies have focused on the antifungal activity compounds from natural sources for biopreservation in alternatives to chemical use. LAB has an antifungal activity which may inhibit food spoilage fungi. Lactobacillus strains isolated from silage prepared in our laboratory by fermentation of grass in anaerobic condition were screened for antifungal activity with overlay assay against Aspergillus spp. The antifungal compounds were originated from organic acids; inhibitory activity did not change after treatment with proteolytic enzymes. Lactobacillus strains were able also to inhibit Trichoderma spp, Penicillium spp, Fusarium roseum, and Stemphylim spp by confrontation assay. The inhibitory activity could be detected against the mould Aspergillus spp in the apricot juice but not in a bakery product. These antifungal compounds have the potential to be used as food biopreservation to inhibit conidia germination, and mycelia growth of spoilage fungi depending on food type, pH of food especially in heat, and cold processed foods.

Keywords: lactic acid bacteria, Lactobacillus, Aspergillus, antifungal activity

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9 The Biofertilizer Effect of Pseudomonas of Salt Soils of the North-West Algerian, Study of Comportment of Bean (Vicia Faba)

Authors: Djoudi Abdelhak, Djibaoui Rachid, Reguieg Yassaad Houcine

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Our study focuses on the identification of some species of Pseudomonas (P4, P5, P7 and P8) isolated from saline soils in northwestern Algeria and the effect of their metabolites on the growth of Alternaria alternata the causative agent of the blight of the bean disease (Vicia faba). We are also interested in stimulating the growth of this plant species in saline conditions (60 mM/l NaCl) and the absence of salts. The analysis focuses on rates of inhibition of mycelial growth of Alternaria alternata strain and the rate of growth of plants inoculated with strains of Pseudomonas expressed by biometrics. According to the results of the in-vitro test, P5 and P8 species and their metabolites showed a significant effect on mycelia growth and production of spores of Alternaria alternata. The in-vivo test shows that the species P8 and P5 were significantly and positively influencing the growth in biometric parameters of the bean in saline and salt-free condition. Inoculation with strain P5 has promoted the growth of the bean in stem height, stem fresh weight and dry weight of stems of 108.59%, 115.28%, 104.33%, respectively, in the presence of salt Inoculation with strain P5 has fostered the growth of the bean stem fresh weight of 112.47% in the presence of salt The effect of Pseudomonas species on the development of Vicia faba and the growth of Alternaria alternata is considering new techniques and methods of biological production and crop protection.

Keywords: pseudomonas, vicia faba, alternaria alternata, promoting of plant growth

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8 Evaluation of Opposite Type Heterologous MAT Genes Transfer in the Filamentous Fungi Neofusicoccum mediterraneum and Verticillium dahliae

Authors: Stavros Palavouzis, Alexandra Triantafyllopoulou, Aliki Tzima, Epaminondas Paplomatas

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Mating-type genes are present in most filamentous fungi, even though teleomorphs for all species have not been recorded. Our study tries to explore the effect of different growth conditions on the expression of MAT genes in Neofusicoccum mediterraneum. As such, selected isolates were grown in potato dextrose broth or in water agar supplemented with pine needles under a 12 h photoperiod, as well as in constant darkness. Mycelia and spores were collected at different time points, and RNA extraction was performed, with the extracted product being used for cDNA synthesis. New primers for MAT gene expression were designed while qPCR results are underway. The second part of the study involved the isolation and cloning in a selected pGEM-T vector of the Botryosphaeria dothidea MAT1 1 1 and MAT1 2 1 mating genes, including flanking regions. As a next step, the genes were amplified using newly designed primers with engineered restriction sites. Amplicons were excised and subsequently sub-cloned in appropriate binary vectors. The constructs were afterward inserted into Agrobacterium tumefaciens and utilized for Agrobacterium-mediated transformation (ATMT) of Neofusicoccum mediterraneum. At the same time, the transformation of a Verticillium dahliae tomato race 1 strain (70V) was performed as a control. While the procedure was successful in regards to V. dahliae, transformed strains of N. mediterraneum could not be obtained. At present, a new transformation protocol, which utilizes a combination of protoplast and Agro transformation, is being evaluated.

Keywords: anamorph, heterothallism, perithecia, pycnidia, sexual stage

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7 Effect of Biostimulants on Downstream Processing of Endophytic Fungi Hosted in Aromatic Plant, Ocimum basicilium

Authors: Kanika Chowdhary, Satyawati Sharma

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Endophytic microbes are hosted inside plants in a symbiotic and hugely benefitting relationship. Exploring agriculturally beneficial endophytes is quite a prospective field of research. In the present work fungal endophytes associated with aromatic plant Ocimum basicilium L. were investigated for biocontrol potential. The anti-plant pathogenic activity of fungal endophytes was tested against causal agent of stem rot Sclerotinia sclerotiorum. 75 endophytic fungi were recovered through culture-dependent approach. Fungal identification was performed both microscopically and by rDNA ITS sequencing. Curvuaria lunata (Sb-6) and Colletotrichum lindemuthianum (Sb-8) inhibited 86% and 72% mycelia growth of S. sclerotinia on Sabouraud dextrose agar medium at 7.4 pH. Small-scale fermentation was carried out on sterilised oatmeal grain medium. In another set of experiment, fungi were grown in oatmeal grain medium amended with certain biostimulants such as aqueous seaweed extract (10% v/w); methanolic seaweed extract (5% v/w); cow urine (20% v/w); biochar (10% w/w) in triplicate along with control of each to ascertain the degree of metabolic difference and anti-plant pathogenic activity induced. Phytochemically extracts of both the fungal isolates showed the presence of flavanoids, phenols, tannins, alkaloids and terpenoids. Ethylacetate extract of C. lunata and C. lindemuthianum suppressed S. sclerotinia conidial germination at IC50 values of 0.514± 0.02 and 0.913± 0.04 mg/ml. Therefore, fungal endophytes of O. basicilium are highly promising bio-resource agent, which can be developed further for sustainable agriculture.

Keywords: endophytic fungi, ocimum basicilium, sclerotinia sclerotiorum, biostimulants

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6 Assessment of Bio-Control Quality of Ethanolic Extracts of Some Tropical Plants on Fruit Rot Pathogens of Pineapple Fruits in Ado Ekiti

Authors: J. Y. Ijato, A. Adewumi, H. O Yakubu, O. O. Olajide, B. O. Ojo, B. A. Adanikin

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Post-harvest fruit rot pathogens are one of the major factors that are responsible for food security challenges in developing countries like Nigeria. These pathogens also cause fruit food poisoning. Biocidal effects of ethanolic extracts of Khaya grandifoliola, Hyptis suaveolens, Zingiber officinale, Calophyllum inophyllum, Datura stramonium on the mycelia growth of fungal rot pathogens of pineapple fruit was investigated, the ethanolic extracts of these test plants exhibited high significant inhibitory effects on the rot pathogens, the highest ethanolic extract inhibition of Zingiber officinale was on Aspergillus flavus (38.40%) at 1.0g/ml while the least inhibitory effect was on Aspergillus fumigatus (23.10%) at 1.0g/ml, the highest ethanol extract inhibition of Datura stramonium was on Aspergillus tubingensis (24.00%) at 1.0g/ml while the least inhibitory effect was 10.00% on Colletotrichum fruticola at 1.0g/ml, the highest ethanol extract inhibition of Calophyllum inophyllum was on Trichoderma harzianum (18.50%) at 1.0g/ml while the least inhibitory effect was on Aspergillus flavus (15.00%) at 1.0g/ml, the highest ethanol extract inhibition of Hyptis suaveolens was on Aspergillus fumigatus (35.00%) at 1.0g/ml while the least inhibitory effect was on Aspergillus niger (20.00%) at 1.0g/ml, the highest ethanol extract inhibition of Khaya grandifoliola was on Aspergillus flavus (35.00%) at 1.00g/ml while the least inhibitory effect was on Aspergillus fumigates (22.00%) at 1.0g/ml, the antifungal capacity of these test plant extracts on rot causing fungi on pineapple fruit reveals the possibility of their use by farmers and fruit traders as alternative to chemical fungicide that portends great threat to human and environmental health.

Keywords: fruit rot, pathogens, plant extracts, pineapple, food poisoning

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5 Determination of Anti-Fungal Activity of Cedrus deodara Oil against Oligoporus placentus, Trametes versicolor and Xylaria acuminata on Populus deltoids

Authors: Sauradipta Ganguly, Akhato Sumi, Sanjeet Kumar Hom, Ajan T. Lotha

Abstract:

Populus deltoides is a hardwood used predominantly for the manufacturing of plywood, matchsticks, and paper in India and hence has a higher economical significance. Wood-decaying fungi cause serious damage to Populus deltoides products, as the wood itself is perishable and vulnerable to decaying agents, decreasing their aesthetical value which in return results in significant monetary loss for the wood industries concerned. The aim of the study was to determine the antifungal activity of Cedrus deodara oil against three primary wood-decaying fungi namely white-rot fungi (Trametes versicolor), brown-rot fungi (Oligoporus placentus) and soft-rot fungi (Xylaria acuminata) on Populus deltoides samples under optimum laboratory conditions. The susceptibility of Populus deltoides samples on the fungal attack and the ability of deodar oil to control colonization of the wood rotting fungi on the samples were assessed. Three concentrations of deodar oil were considered for the study as treating solutions, i.e., 4%, 5%, and 6%. The Populus deltoides samples were treated with treating solutions, and the ability of the same to prevent a fungal attack on the samples were assessed using accelerated test in the laboratory at Biochemical Oxygen Demand incubator at temperature (25 ± 2°C) and relative humidity 70 ± 4%. Efficacy test and statistical analysis of deodar oil against Trametes versicolor, Oligoporus placentus, and Xylariaacuminataon P. deltoides samples exhibited light, minor and negligible mycelia growth at 4 %, 5% and 6% concentrations of deodar oil, respectively. Whereas, moderate to heavy attack was observed on the surface of the control samples. Statistical analysis further established that the treatments were statistically significant and had significantly inhibited fungal growth of all the three fungus spp by almost 3 to 5 times.

Keywords: populus deltoides, Trametes versicolor, Oligoporus placentus, Xylaria acuminata, Deodar oil, treatment

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4 Biodegradation of Triclosan and Tetracycline in Sewage Sludge by Pleurotus Ostreatus Fungal Pellets

Authors: Ayda Maadani Mallak, Amir lakzian, Elham Khodaverdi, Gholam Hossein Haghnia

Abstract:

The use of pharmaceuticals and personal care products such as antibiotics and antibacterials has been increased in recent years. Since the major part of consumed compounds remains unchanged in the wastewater treatment plant, they will easily find their way into the human food chain following the land use of sewage sludge (SS). Biological treatment of SS is one the most effective methods for expunging contaminants. White rot fungi, due to their ligninolytic enzymes, are extensively used to degrade organic compounds. Among all three different morphological forms and growth patterns of filamentous fungi (mycelia, clumps, and pellets), fungal pellet formation has been the subject of interest in industrial bioprocesses. Therefore this study was aimed to investigate the uptake of tetracycline (TC) and triclosan (TCS) by radish plant (Raphanus sativus) from soil amended with untreated and pretreated SS by P. ostreatus fungal pellets under greenhouse conditions. The experimental soil was amended with 1) Contaminated SS with TC at a concentration of 100 mgkg-1 and pretreated by fungal pellets, 2) Contaminated SS with TC at 100 mgkg-1 and untreated with fungal pellets, 3) Contaminated SS with TCS at a concentration of 50 mgkg-1 and pretreated by fungal pellets, 4) contaminated SS with TCS at 50 mgkg-1 and untreated with fungal pellets. An uncontaminated and untreated SS-amended soil also was considered as control treatment. An AB SCIEX 3200 QTRAP LC-MS/MS system was used in order to analyze the concentration of TC and TCS in plant tissues and soil medium. Results of this study revealed that the presence of TC and TCS in SS-amended soil decreased the radish biomass significantly. The reduction effect of TCS on dry biomass of shoot and root was 39 and 45% compared to controls, whereas for TC, the reduction percentage for shoot and root was 27 and 40.6%, respectively. However, fungal treatment of SS by P. ostreatus pellets reduced the negative effect of both compounds on plant biomass remarkably, as no significant difference was observed compared to control treatments. Pretreatment of SS with P. ostreatus also caused a significant reduction in translocation factor (concentration in shoot/root), especially for TC compound up to 32.3%, whereas this reduction for TCS was less (8%) compared to untreated SS. Generally, the results of this study confirmed the positive effect of using fungal pellets in SS amendment to decrease TC and TCS uptake by radish plants. In conclusion, P. ostreatus fungal pellets might provide future insights into bioaugmentation to remove antibiotics from environmental matrices.

Keywords: antibiotic, fungal pellet, sewage sludge, white-rot fungi

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3 Management Potentialities Of Rice Blast Disease Caused By Magnaporthe Grisae Using New Nanofungicides Derived From Chitosan

Authors: Abdulaziz Bashir Kutawa1, 2, *, Khairulmazmi Ahmad 1, 3, Mohd Zobir Hussein 4, Asgar Ali 5, * Mohd Aswad Abdul Wahab1, Amara Rafi3, Mahesh Tiran Gunasena1, 6, Muhammad Ziaur Rahman1, 7, Md Imam Hossain1, And Syazwan Afif Mohd Zobir1

Abstract:

Various abiotic and biotic stresses have an impact on rice production all around the world. The most serious and prevalent disease in rice plants, known as rice blast, is one of the major obstacles to the production of rice. It is one of the diseases that has the greatest negative effects on rice farming globally, the disease is caused by a fungus called Magnaporthe grisae. Since nanoparticles were shown to have an inhibitory impact on certain types of fungus, nanotechnology is a novel notion to enhance agriculture by battling plant diseases. Utilizing nanocarrier systems enables the active chemicals to be absorbed, attached, and encapsulated to produce efficient nanodelivery formulations. The objectives of this research work were to determine the efficacy and mode of action of the nanofungicides (in-vitro) and in field conditions (in-vivo). Ionic gelation method was used in the development of the nanofungicides. Using the poisoned media method, the synthesized agronanofungicides' in-vitro antifungal activity was assessed against M. grisae. The potato dextrose agar (PDA) was amended in several concentrations; 0.001, 0.005, 0.01, 0.025, 0.05, 0.1, 0.15, 0.20, 0.25, 0.30, and 0.35 ppm for the nanofungicides. Medium with the only solvent served as a control. Every day, mycelial growth was measured, and PIRG (percentage inhibition of radial growth) was also computed. Every day, mycelial growth was measured, and PIRG (percentage inhibition of radial growth) was also computed. Based on the results of the zone of inhibition, the chitosan-hexaconazole agronanofungicide (2g/mL) was the most effective fungicide to inhibit the growth of the fungus with 100% inhibition at 0.2, 0.25, 0.30, and 0.35 ppm, respectively. Then followed by carbendazim analytical fungicide that inhibited the growth of the fungus (100%) at 5, 10, 25, 50, and 100 ppm, respectively. The least were found to be propiconazole and basamid fungicides with 100% inhibition only at 100 ppm. The scanning electron microscope (SEM), confocal laser scanning microscope (CLSM), and transmission electron microscope (TEM) were used to study the mechanisms of action of the M. grisae fungal cells. The results showed that both carbendazim, chitosan-hexaconazole, and HXE were found to be the most effective fungicides in disrupting the mycelia of the fungus, and internal structures of the fungal cells. The results of the field assessment showed that the CHDEN treatment (5g/L, double dosage) was found to be the most effective fungicide to reduce the intensity of the rice blast disease with DSI of 17.56%, lesion length (0.43 cm), DR of 82.44%, AUDPC of 260.54 Unit2, and PI of 65.33%, respectively. The least treatment was found to be chitosan-hexaconazole-dazomet (2.5g/L, MIC). The usage of CHDEN and CHEN nanofungicides will significantly assist in lessening the severity of rice blast in the fields, increasing output and profit for rice farmers.

Keywords: chitosan, hexaconazole, disease incidence, and magnaporthe grisae

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2 Isolation and Identification of Low-Temperature Tolerant-Yeast Strains from Apple with Biocontrol Activity

Authors: Lachin Mikjtarnejad, Mohsen Farzaneh

Abstract:

Various microbes, such as fungi and bacteria species, are naturally found in the fruit microbiota, and some of them act as a pathogen and result in fruit rot. Among non-pathogenic microbes, yeasts (single-celled microorganisms belonging to the fungi kingdom) can colonize fruit tissues and interact with them without causing any damage to them. Although yeasts are part of the plant microbiota, there is little information about their interactions with plants in comparison with bacteria and filamentous fungi. According to several existing studies, some yeasts can colonize different plant species and have the biological control ability to suppress some of the plant pathogens. It means those specific yeast-colonized plants are more resistant to some plant pathogens. The major objective of the present investigation is to isolate yeast strains from apple fruit and screen their ability to control Penicillium expansum, the causal agent of blue mold of fruits. In the present study, psychrotrophic and epiphytic yeasts were isolated from apple fruits that were stored at low temperatures (0–1°C). Totally, 42 yeast isolates were obtained and identified by molecular analysis based on genomic sequences of the D1/D2 and ITS1/ITS4 regions of their rDNA. All isolated yeasts were primarily screened by' in vitro dual culture assay against P. expansum by measuring the fungus' relative growth inhibition after 10 days of incubation. The results showed that the mycelial growth of P. expansum was reduced between 41–53% when challenged by promising yeast strains. The isolates with the strongest antagonistic activity belonged to Metschnikowia pulcherrima A13, Rhodotorula mucilaginosa A41, Leucosporidium Scottii A26, Aureobasidium pullulans A19, Pichia guilliermondii A32, Cryptococcus flavescents A25, and Pichia kluyveri A40. The results of seven superior isolates to inhibit blue mold decay on fruit showed that isolates A. pullulans A19, L. scottii A26, and Pi. guilliermondii A32 could significantly reduce the fruit rot and decay with 26 mm, 22 mm and 20 mm zone diameter, respectively, compared to the control sample with 43 mm. Our results show Pi. guilliermondii strain A13 was the most effective yeast isolates in inhibiting P. expansum on apple fruits. In addition, various biological control mechanisms of promising biological isolates against blue mold have been evaluated to date, including competition for nutrients and space, production of volatile metabolites, reduction of spore germination, production of siderophores and production of extracellular lytic enzymes such as chitinase and β-1,3-glucanase. However, the competition for nutrients and the ability to inhibit P. expansum spore growth have been introduced as the prevailing mechanisms among them. Accordingly, in our study, isolates A13, A41, A40, A25, A32, A19 and A26 inhibited the germination of P. expansum, whereas isolates A13 and A19 were the strongest inhibitors of P. expansum mycelia growth, causing 89.13% and 81.75 % reduction in the mycelial surface, respectively. All the promising isolates produced chitinase and β-1,3-glucanase after 3, 5 and 7 days of cultivation. Finally, based on our findings, we are proposing that, Pi. guilliermondiias as an effective biocontrol agent and alternative to chemical fungicides to control the blue mold of apple fruit.

Keywords: yeast, yeast enzymes, biocontrol, post harvest diseases

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1 Preliminary Characterization of Hericium Species Sampled in Tuscany, Italy

Authors: V. Cesaroni, C. Girometta, A. Bernicchia, M. Brusoni, F. Corana, R. M. Baiguera, C. M. Cusaro, M. L. Guglielminetti, B. Mannucci, H. Kawagishi, C. Perini, A. M. Picco, P. Rossi, E. Salerni, E. Savino

Abstract:

Fungi of the genus Hericium contain various compounds with antibacterial activity, cytotoxic effect on cancer cells and bioactive molecules. Some of the active metabolites stimulate the synthesis of the Nerve Growth Factor (NGF). Recently, the effect of dietary supplement based on Hericium erinaceus on recognition memory and on hippocampal mossy fiber-CA3 neurotransmission was published. The aim of this study was to investigate the presence of Hericium species on Italian territory in order to isolate the strains for further studies and applications. The first step was to collect Hericium sporophores in Tuscany: H. alpestre Pers., H. coralloides (Scop.) Pers. and H. erinaceus (Bull.) Pers. were the species present. The strains of H. alpestre (H.a.1), H. coralloides (H.c.1) and H. erinaceus (H.e.1 & H.e.2) have been isolated in pure culture and preserved in the collection of the University of Pavia (MicUNIPV). The DNA sequences obtained from the strains were compared to other sequences found in international databases. Therefore, it was possible to construct a phylogenetic tree that highlights the clear separation in clades of the sequences and the molecular identification of our strains with the species of Hericium considered. The second step was to cultivate indoor and outdoor H. erinaceus in order to obtain as many sporophores as possible for further chemical analysis. All the procedures for H. erinaceus cultivation have been followed. Among the available recipes for indoor H. erinaceus cultivation, it was used a substrate formulation contained 70% oak sawdust, 20% rice bran, 10% wheat straw, 1% CaCO3 and 1% sucrose. The bioactive compounds present in the mycelia and in the sporophores of H. erinaceus were chemically analyzed in collaboration with the Centro Grandi Strumenti of the University of Pavia using high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS). The materials to be analyzed were previously freeze-dried and then extracted with an alcoholic procedure. Preliminary chromatographic analysis revealed the presence of potentially bioactive and structurally different secondary metabolites such as polysaccharides, erinacins, ericenones, steroids and other terpenoids. Ericenones C and D (in sporophores) and erinacin A (in mycelium) have been identified by comparison with the respective standards. These molecules are known to have effects on the Central Nervous System (CNS) cells, which is the main objective of our studies. Thanks to the high sensitivity in the detection of bioactive compounds of H. erinaceus, it will be possible to use the To obtain lyophilized mycelium and the respective culture broth, 4 small pieces (about 5 mm2) of the respective H.e.1 or H.c.1 strains, taken from the margin of growing cultures (MEA), were inoculated into 1 liter of 2% ME (malt extract, Biokar Diagnostics). The static liquid cultures were kept at 24 °C in the dark chamber and fungi grew for one month. 10 replicates for each strain have been done. The method proposed as an analytical screening protocol to determine the optimal growth conditions of the fungus and to improve the production chain of H. erinaceus. These results encourage to carry out chemical analyzes also on H. alpestre and H. coralloides in order to evaluate the presence of bioactive compounds in these two species.

Keywords: Hericium species, Hercium erinaceus bioactive compounds, medicinal mushrooms, mushroom cultivation

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