Search results for: filamentous fungi
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 471

Search results for: filamentous fungi

471 Enhanced Enzymes Production through Immobilization of Filamentous Fungi

Authors: Zhanara B. Suleimenova, Zhazira K. Saduyeva

Abstract:

Filamentous fungi are major producers of enzymes that have important applications in the food and beverage industries. The overall objective of this research is a strain improvement technology for efficient industrial enzymes production. The new way of filamentous fungi cultivation method has been developed. Such technology prolong producers’ cultivation period up to 60 days and create the opportunity to obtain enzymes repeatedly in every 2-3 days of fungal cultivation. This method is based on immobilizing enzymes producers with solid support in submerged conditions of growth. Immobilizing has a range of advantages: Decreasing the price of the final product, absence of foreign substances, controlled process of enzyme-genesis, ability of various enzymes simultaneous production, etc. Design of proposed technology gives the opportunity to increase the activity of immobilized cells culture filtrate comparing to free cells, growing in periodic culture conditions. Thus, proposed research focuses on new, more versatile, microorganisms capable of squeezing more end-products as well as proposed cultivation technology led to increased enzymatic productivity by several times.

Keywords: filamentous fungi, immobilization, industrial enzymes production, strain improvement

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470 Ethanol and Biomass Production from Spent Sulfite Liquor by Filamentous Fungi

Authors: M. T. Asadollahzadeh, A. Ghasemian, A. R. Saraeian, H. Resalati, P. R. Lennartsson, M. J. Taherzadeh

Abstract:

Since filamentous fungi are capable of assimilating several types of sugars (hexoses and pentoses), they are potential candidates for bioconversion of spent sulfite liquor (SSL). Three filamentous fungi such as Aspergillus oryzae, Mucor indicus, and Rhizopus oryzae were investigated in this work. The SSL was diluted in order to obtain concentrations of 50, 60, 70, 80, and 90% and supplemented with two types of nutrients. The results from cultivations in shake flask showed that A. oryzae and M. indicus were not able to grow in pure SSL and SSL90% while R. oryzae could grow only in SSL50% and SSL60%. Cultivation with A. oryzae resulted in the highest yield of produced fungal biomass, while R. oryzae cultivation resulted in the lowest fungal biomass yield. Although, the mediums containing yeast extract, (NH4)2SO4, KH2PO4, CaCl2∙2H2O, and MgSO4∙7H2O as nutrients supplementations produced higher fungal biomass compared to the mediums containing NH4H2PO4 and ammonia, but there was no significant difference between two types of nutrients in terms of sugars and acetic acid consumption rate. The sugars consumption in M. indicus cultivation was faster than A. oryzae and R. oryzae cultivation. Acetic acid present in SSL was completely consumed during cultivation of all fungi. M. indicus was the best and fastest ethanol producer from SSL among the fungi examined, when yeast extract and salts were used as nutrients supplementations. Furthermore, no further improvement in ethanol concentration and rate of sugars consumption was obtained in medium supplemented with NH4H2PO4 and ammonia compared to medium containing yeast extract, (NH4)2SO4, KH2PO4, CaCl2∙2H2O, and MgSO4∙7H2O. On the other hand, the higher dilution of SSL resulted in a better fermentability, and better consumption of sugars and acetic acid.

Keywords: ethanol, filamentous fungi, fungal biomass, spent sulfite liquor

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469 Investigating the Biosorption Potential of Indigenous Filamentous Fungi from Copperbelt Tailing Dams in Zambia with Copper and Cobalt Tolerance

Authors: Leonce Dusengemungu

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Filamentous fungi indigenous to heavy metals (HMs) contaminated environments have a considerable biosorption potential yet are currently under-investigated in developing countries. In the work presented herein, the biosorption potential of three indigenous filamentous fungi (Aspergillus transmontanensis, Cladosporium cladosporioides, and Geotrichum candidum) isolated from copper and cobalt mining wasteland sites in Zambia's Copperbelt province was investigated. In Cu and Co tolerance tests, all the fungal isolates were shown to be tolerant, with mycelial growth at HMs concentrations of up to 7000 ppm. However, exposure to high Cu and Co concentrations hindered the growth of the three strains to varying degrees, resulting in reduced mycelial biomass (evidenced by loss of the infrared bands at 887 and 930 cm-1 of the 1,3-glucans backbone) as well as morphological alterations, sporulation, and pigment synthesis. In addition, gas chromatography-mass spectrometry characterization of the fungal biomass extracts allowed to detect changes in the chemical constituents upon exposure to HMs, with profiles poorer in maltol, 1,2-cyclopentadione, and n-hexadecanoic acid, and richer in furaldehydes. Biosorption tests showed that A. transmontanensis and G. candidum showed better performance as bioremediators than C. cladosporioides, with biosorption efficiencies of 1645, 1853 and 1253 ppm at pH 3, respectively, and may deserve further research in field conditions.

Keywords: bioremediation, fungi, biosorption, heavy metal

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468 Influence of Culturing Conditions on Biomass Yield, Total Lipid and Fatty Acid Composition of Some Filamentous Fungi

Authors: Alla V. Goncharova, Tatyana A. Karpenyuk, Yana S. Tsurkan, Rosa U. Beisembaeva, Togzhan D. Mukasheva, Ludmila V. Ignatova, Ramza Z. Berzhanova

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In this work the effect of culturing conditions of filamentous fungi Penicillium raistrickii, Penicillium anatolicum, Fusarium sp. on biomass yield, the content of total lipids and fatty acids was studied. It has been established that in time the process of lipids accumulation correlated with biomass growth of cultures, reaching maximum values in stationary growth phase. Biomass yield and accumulation of general lipids was increased by adding zinc to the culture medium. The more intensive accumulation of biomass and general lipids was observed at temperature 18°C. Lowering the temperature of culturing has changed the ratio of saturated: Unsaturated fatty acids in the direction of increasing the latter.

Keywords: biomass, culturing conditions, fungi, fatty acids (FA), growth dynamics, lipids

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467 Microbial Diversity of El-Baida Marsh: Setif, Algeria

Authors: H. Necef, A. Benayad

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Fungi are becoming more and more important in our life. Therefore, as a start for the symposium on filamentous fungi in biotechnology a short survey of the role of fungi in biotechnology. Salin soils occupy about 7% of land area; they are characterized by unsuitable physical conditions for the growth of living organisms. However, researches showed that some microorganisms especially fungi are able to grow and adapt to such extreme conditions; it is due to their ability to develop different physiological mechanisms in their adaptation. This is the first study on the physiological and biological characteristics of El-Beida marsh. Nine soil samples were taken at different points in two steps, the first was in winter (low temperature), and the second was in summer (high temperature). The physicochemical analyses of the soil were conducted, then the isolation process was applied using two methods, direct method and dilution method (10-1, 10-2, 10-3, 10-4). Different species of fungi were identified belong to 21 genera in addition to 3 yeast species, Aspergillus showed the highest proportion by 43%, then Penicillium by 20% then Alternaria by 7%, in addition to various genera in different proportions. As for the sampling periods, it was observed that the spread of fungi in winter was higher than in summer with the proportion 75.47% and 24.53% respectively. Some halotolerant fungi have a biotechnological importance especially if the salinity of the medium is necessary for the fermentation, and if the halotolerance genes of the fungus will define, this will open the research to study and improve this property for the industrial important micro-organisms.

Keywords: salinity, identification, aspergillus oryzae, halotolerance, fungi

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466 Distribution and Taxonomy of Marine Fungi in Nha Trang Bay and Van Phong Bay, Vietnam

Authors: Thu Thuy Pham, Thi Chau Loan Tran, Van Duy Nguyen

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Marine fungi play an important role in the marine ecosystems. Marine fungi also supply biomass and metabolic products of industrial value. Currently, the biodiversity of marine fungi along the coastal areas of Vietnam has not yet been studied fully. The objective of this study is to assess the spatial and temporal diversity of planktonic fungi from the coastal waters of Nha Trang Bay and Van Phong Bay in Central Vietnam using culture-dependent and independent approach. Using culture-dependent approach, filamentous fungi and yeasts were isolated on selective media and then classified by phenotype and genotype based on the sequencing of ITS (internal transcribed spacers) regions of rDNA with two primer pairs (ITS1F_KYO2 and ITS4; NS1 and NS8). Using culture-independent approach, environmental DNA samples were isolated and amplified using fungal-specific ITS primer pairs. A total of over 160 strains were isolated from 10 seawater sampling stations at 50 cm depth. They were classified into diverse genera and species of both yeast and mold. At least 5 strains could be potentially novel species. Our results also revealed that planktonic fungi were molecularly diverse with hundreds of phylotypes recovered across these two bays. The results of the study provide data about the distribution and taxonomy of mycoplankton in this area, thereby allowing assessment of their positive role in the biogeochemical cycle of coastal ecosystems and the development of new bioactive compounds for industrial applications.

Keywords: biodiversity, ITS, marine fungi, Nha Trang Bay, Van Phong Bay

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465 Extracellular Laccase Production by Co-culture between Galactomyces reesii IFO 10823 and Filamentous Fungal Strains Isolated from Fungus Comb Using Natural Inducer

Authors: P. Chaijak, M. Lertworapreecha, C. Sukkasem

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Extracellular laccases are copper-containing microbial enzymes with many industrial biotechnological applications. This study evaluated the ability of nutrients in coconut coir to enhance the yield of extracellular laccase of Galactomyces reesii IFO 10823 and develop a co-culture between this yeast and other filamentous fungi isolated from the fungus comb of Macrotermes sp. The co-culture between G. reesii IFO 10823 and M. indicus FJ-M-5 (G3) gave the highest activity at 580.20 U/mL. When grown in fermentation media prepared from coconut coir and distilled water at 70% of initial moisture without supplement addition, G3 produced extracellular laccase of 113.99 U/mL.

Keywords: extracellular laccase, production, yeast, natural inducer

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464 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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463 Biodiesel Production and Heavy Metal Removal by Aspergillus fumigatus sp.

Authors: Ahmed M. Haddad, Hadeel S. El-Shaal, Gadallah M. Abu-Elreesh

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Some of filamentous fungi can be used for biodiesel production as they are able to accumulate high amounts of intracellular lipids when grown at stress conditions. Aspergillus fumigatus sp. was isolated from Nile delta soil in Egypt. The fungus was primarily screened for its capacity to accumulate lipids using Nile red staining assay. The fungus could accumulate more than 20% of its biomass as lipids when grown at optimized minimal medium. After lipid extraction, we could use fungal cell debris to remove some heavy metals from contaminated waste water. The fungal cell debris could remove Cd, Cr, and Zn with absorption efficiency of 73%, 83.43%, and 69.39% respectively. In conclusion, the Aspergillus fumigatus isolate may be considered as a promising biodiesel producer, and its biomass waste can be further used for bioremediation of wastewater contaminated with heavy metals.

Keywords: biodiesel, bioremediation, fungi, heavy metals, lipids, oleaginous

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462 Cross-Linked Amyloglucosidase Aggregates: A New Carrier Free Immobilization Strategy for Continuous Saccharification of Starch

Authors: Sidra Pervez, Afsheen Aman, Shah Ali Ul Qader

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The importance of attaining an optimum performance of an enzyme is often a question of devising an effective method for its immobilization. Cross-linked enzyme aggregate (CLEAs) is a new approach for immobilization of enzymes using carrier free strategy. This method is exquisitely simple (involving precipitation of the enzyme from aqueous buffer followed by cross-linking of the resulting physical aggregates of enzyme molecules) and amenable to rapid optimization. Among many industrial enzymes, amyloglucosidase is an important amylolytic enzyme that hydrolyzes alpha (1→4) and alpha (1→6) glycosidic bonds in starch molecule and produce glucose as a sole end product. Glucose liberated by amyloglucosidase can be used for the production of ethanol and glucose syrups. Besides this amyloglucosidase can be widely used in various food and pharmaceuticals industries. For production of amyloglucosidase on commercial scale, filamentous fungi of genera Aspergillus are mostly used because they secrete large amount of enzymes extracellularly. The current investigation was based on isolation and identification of filamentous fungi from genus Aspergillus for the production of amyloglucosidase in submerged fermentation and optimization of cultivation parameters for starch saccharification. Natural isolates were identified as Aspergillus niger KIBGE-IB36, Aspergillus fumigatus KIBGE-IB33, Aspergillus flavus KIBGE-IB34 and Aspergillus terreus KIBGE-IB35 on taxonomical basis and 18S rDNA analysis and their sequence were submitted to GenBank. Among them, Aspergillus fumigatus KIBGE-IB33 was selected on the basis of maximum enzyme production. After optimization of fermentation conditions enzyme was immobilized on CLEA. Different parameters were optimized for maximum immobilization of amyloglucosidase. Data of enzyme stability (thermal and Storage) and reusability suggested the applicability of immobilized amyloglucosidase for continuous saccharification of starch in industrial processes.

Keywords: aspergillus, immobilization, industrial processes, starch saccharification

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461 Isolation and Identification of Fungi from Different Types of Medicinal Plants Cultivated in Ecuador

Authors: Ana Paola Echavarria, Mariuxi Medina, Haydelba D'Armas, Carmita Jaramillo, Diana San Martin

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The use of medicinal plants is one of the oldest and most extended medical therapies that goes back to prehistoric times, and nowadays, they are also used in the preparation of phytopharmaceuticals with options to cure diseases. The test for the determination of fungi was carried out in the Pharmacy Pilot Plant (treatment of the leaves of the plant species) and the Microbiology Laboratory (determination of fungi of the plant species, using growth medium called Sabouraud agar plus the vegetal sample), of the Academic Unit of Chemical Sciences and Health, of the Universidad Tecnica de Machala. Subsequently, colony counting was performed, both macroscopic, which is determined in the growth medium of the seeding, and microscopic, to identify the germinative forms using blue lactophenol. The procedure was repeated in duplicate to replicate the results data. The determination of the total fungal content of the following plant species was evaluated: Cymbopogon citratus (lemon verbena), Melissa officinalis (lemon balm), Taraxacum officinale (dandelion), Artemisia absinthium (absinthe), Piper carpunya (guaviduca), Moringa oleifera (moringa), Coriandrum sativum (coriander), Momordica charantia (achochilla), Borago officinalis (borage), Aloysia citriodora (cedron), Ambrosia artemisifolia (altamisa) and Ageratum conyzoides (mastrante). The results obtained showed that all the samples of the twelve plant species studied developed filamentous fungi, with great variability of them, within the permissible limits and contemplated by the Ecuadorian Institute of Normalization (INEN), being suitable as raw material for its use in the preparation of nutraceuticals and medicinal products or phytodrugs; with the exception of A. conyzoides (mastranto) which is the only species that exceeds the regulation in the average of dilutions.

Keywords: colonies, fungi, medicinal plants, microbiological quality, Sabouraud agar

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460 Potential Role of Arbuscular Mycorrhizal (AM) Fungi in CO₂-Sequestration During Bipartite Interaction with Host Plant Oryza Sativa

Authors: Sadhana Shukla, Pushplata Singh, Nidhi Didwania

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Arbuscular mycorrhizal (AM) fungi are a highly advantageous and versatile group of fungi that significantly contribute to the formation of soil organic matter by creating a demand for plant carbon (C) and distributing it through below-ground hyphal biomass, regardless of their substantial contribution in enhancing net primary productivity and accumulating additional photosynthetic fixed C in the soil. The genetic role of AM fungi in carbon cycling is largely unexplored. In our study, we propose that AM fungi significantly interact with the soil, particularly: the provision of photosynthates by plants. We have studied the expression of AM fungi genes involved in CO₂ sequestration during host-plant interaction was investigated by qPCR studies. We selected Rhizophagus proliferus (AM fungi) and Oryza sativa (Rice) (inoculated with or without 200ppg AMF inoculums per plant) and investigated the effect of AM fungi on soil organic carbon (SOC) and rice growth under field conditions. Results thus provided faster SOC turnover, 35% increased nutrient uptake in plants and pronounced hyphal biomass of AM fungi which enhanced soil carbon storage by 15% in comparison to uninoculated plants. This study will offer a foundation for delving into various carbon-soil studies while also advancing our comprehension of the relationship between AM fungi and the sustainability of agricultural ecosystems.

Keywords: arbuscular mycorrhizal (AM) fungi, carbon sequestration, gene expression, soil health, plant development.

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459 The Importance of Fungi and Plants for a More Sustainable on Our Planet Earth

Authors: Njabe Christelle

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Fungal products are essential building blocks for change towards a more sustainable future for our planet. In nature, fungi are special in breaking down plant material by means of a rich spectrum of plant cell wall degrading enzymes. Enzymes serve as catalysts in organic synthesis. Imagine the immense benefits that the known 250000 plant genes might provide in the future through scientific investigation. Plants are the primary basis for human sustenance, used directly for food, clothing, and shelter or indirectly in processed form and through animal feeding. Fungi are the only organisms known to extensively degrade lignin, a major component of wood. Although humans cannot digest cellulose and lignin, many fungi, through their assimilation of these substances, produce food in the form of edible mushrooms.

Keywords: plants, fungi, sustainable use, planet earth

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458 Biological Control of Tuta absoluta (Meyrick) (Lep: Gelechiidae) with Enthomopathogenic Fungi

Authors: Dahliz Abderrahmène, Lakhdari Wassim, Bouchikh Yamina, Hammi Hamida, Soud Adila, M’lik Randa, Benglia Sara

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Devastating insects constitute one of strains for cultivate tomato. Among this vandal insects, the tomato leafminer (T. absoluta), which has been introduced in Algeria constitute a challenge for both agricultures and scientists. Firstly, this insect is introduced without their natural enemies which may reduce their damage. Secondly, this species has developed insecticide resistance to many active matters. To contribute to establish a control strategy for T. absoluta we have mad an inventory for their enthomopathogenic fungi. Two fungi were identified among others taken from adults and pupae. These fungi are Aspergillus flavus and Metarhizium sp. A study was conducted in laboratory to recognize the efficiency of these antagonists. These species had unregistered a mortality mounts of 42% and 56% respectively.

Keywords: Tuta absoluta, enthomopathogenic fungi, Aspergillus flavus, Metarhizium sp, control strategy

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457 Antifungal Nature of Bacillus Subtilis in Controlling Post Harvest Fungal Rot of Yam

Authors: Ifueko Oghogho Ukponmwan, Mike O. Orji

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This study investigated the antifungal activity of Bacilluss subtilis in the control of postharvest fungal rot of white yam (Dioscorea spp). Bacillus subtilis was isolated from the soil and fungi (Aspergillus spp, Mucor and yeasts) were isolated from rotten yam. The organisms were paired in yam nutrient agar (YNA) and yam Sabourraud dextrose agar media. In the yam dextrose agar media (YSDA) plates, the Bacillus grew rapidly and established itself and restricted the growth of the fungi organisms, but there was no zone of inhibition. This behaviour of Bacillus on the plates of YSDA was also observed in the yams where the fungi caused rot but the rot was suppressed by the presence of the Bacillus as compared to the degree of rot observed in the control that had only spoilage fungi. The control yam showed greater rot than other yams that contained a combination of Bacillus and fungi. The t-Test analysis showed that the difference in the rot between the treated samples and the control sample is significant and this implies that the presence of Bacillus significantly reduced the growth of fungi in the samples (yams). It was revealed from this study that Bacillus subtilis treatment can be successfully used to preserve white yams in storage. Its fast growth and early establishment in the sample accounts for its antifungal strength.

Keywords: Bacillus subtilis, rot, fungi, yam

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456 Epiphytic Growth on Filamentous Bacteria Found in Activated Sludge: A Morphological Approach

Authors: Thobela Conco, Sheena Kumari, Thor Stenstrom, Simona Rosetti, Valter Tandoi, Faizal Bux

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Filamentous bacteria are well documented as causative agents of bulking and foaming in the biological wastewater treatment process. These filamentous bacteria are however closely associated with other non-filamentous organism forming a micro-niche. Among these specific epiphytic bacteria attach to filaments in the consortium of organisms that make up the floc. Neither the eco-physiological role of the epiphytes nor the nature of the interaction between the epiphytic bacteria and the filament hosts they colonize is well understood and in need of in-depth investigations. The focus of this presentation is on the interaction between the epiphytic bacteria and the filament host. Samples from the activated sludge treatment have been repeatedly collected from several wastewater treatment plants in KwaZulu Natal. Extensive investigations have been performed with SEM and TEM electron microscopy, Polarized Light Microscopy with Congo red staining, and Thioflavin T staining to document the interaction. SEM was used to document the morphology of both the filament host and their epiphytes counterparts with the focus on the interface/point of contact between the two, while the main focus of the TEM investigations with the higher magnification aimed to document the ultra-structure features of two organisms relating to the interaction. The interaction of the perpendicular attachment partly seems to be governed by the physiological status of the filaments. The attachment further seems to trigger a response in the filaments with distinct internal visible structures at the attachment sites. It is postulated that these structures most likely are amyloid fibrils. Amyloid fibrils may play an overarching role in different types of attachments and has earlier been noted to play a significant role in biofilm formation in activated sludge. They also play a medical role in degenerative diseases such as Alzheimer’s and Diabetes. Further studies aims to define the eco-physiological role of amyloid fibrils in filamentous bacteria, based on their observed presence at interaction sites in this study. This will also relate to additional findings where selectivity within the species of epiphytes attaching to the selected filaments has been noted. The practical implications of the research findings is still to be determined, but the ecophysiological interaction between two closely associated species or groups may have significant impact in the future understanding of wastewater treatment processes and broaden existing knowledge on population dynamics.

Keywords: activated sludge, amyloid proteins, epiphytic bacteria, filamentous bacteria

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455 Secondary Metabolites from Turkish Marine-Derived Fungi Hypocrea nigricans

Authors: H. Heydari, B. Konuklugil, P. Proksch

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Marine-derived fungi can produce interesting bioactive secondary metabolites that can be considered the potential for drug development. Turkey is a country of a peninsula surrounded by the Black Sea at the north, the Aegean Sea at the west, and the Mediterranean Sea at the south. Despite the approximately 8400 km of coastline, studies on marine secondary metabolites and their biological activity are limited. In our ongoing search for new natural products with different bioactivities produced by the marine-derived fungi, we have investigated secondary metabolites of Turkish collection of the marine sea slug (Peltodoris atromaculata) associated fungi Hypocrea nigricans collected from Seferihisar in the Egean sea. According to the author’s best knowledge, no study was found on this fungal species in terms of secondary metabolites. Isolated from ethyl acetate extract of the culture of Hypocrea nigricans were (isodihydroauroglaucin,tetrahydroauroglaucin and dihydroauroglaucin. The structures of the compounds were established based on an NMR and MS analysis. Structural elucidation of another isolated secondary metabolite/s continues.

Keywords: Hypocrea nigricans, isolation, marine fungi, secondary metabolites

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454 Test of Biological Control against Brachytrupes Megacephalus Lefèbre, 1827 (Orthoptera, Gryllinae) by Using Entomopathogenic Fungi

Authors: W. Lakhdari, B. Doumendji-Mitich, A. Dahliz, S. Doumendji, Y. Bouchikh, R. M'lik, H. Hammi, A. Soud

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This work was done in order to fight against Brachytrupes megacephalus, a major pest in the Algerian oasis and promote one aspect of biological control against it. He wears a hand on the isolation and identification of indigenous fungi on imagos of this insect harvested in the station of INRAA Touggourt and secondly, the study of the pathogenicity of these strains fungal on this orthoptère adults. The results obtained showed the presence of six different species of entomopathogenic fungi, it is: Aspergillus flavus, Fusarium sp, Beauveria bassiana, Penicillium sp, Metharizium anisopliae and Aspergillus Niger. The pathogenicity test using fungi Beauveria bassiana strains and Metharizium anisopliae. On adult of B. megacephalus highlights the effectiveness of these strains of predatory adults, with a mortality rate approaching 100% after 11 days.

Keywords: biological control, brachytrupes megacephalus, entomopathogenic fungi, Southeastern Algeria

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453 Media Manipulations and the Culture of Beneficial Endophytic Fungi in the Leaves and Stem Bark of Grewia lasiocarpa E. Mey. Ex Harv

Authors: Akwu A. Nneka, Naidoo, Yougasphree

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A significantly high number of microbes exist in higher plants; these microbes include bacteria, fungi, and actinomycetes. There are reports on the benefits of endophytic fungi and their products of metabolism to the host plant and man, consequently, it is expedient to explore the changes that could arise as a result of manipulating their growth media. Grewia lasiocarpa E. Mey. ex Harv. (Malvaceae) is an indigenous Southern African plant, that belongs to a genus with known medicinal properties. Three media were used to culture the endophytic fungi viz., Potato Dextrose Agar (PDA), Malt Extract Agar (MEA), and Bacteriological Agar (BA) were used singly, and supplemented with three dilutions of the leaves and stem bark extracts. The manipulated growth media composition had a significant effect on the diversity of the isolated fungal populations. Several endophytic fungi were isolated; their distribution and diversity revealed a significant relatedness with the manipulated media. The media supplemented with the plant extracts was observed to give a significant increase in the growth rate and yield of the endophytes. To the best of our knowledge, this is the first study describing the endophytic fungi present in the leaves and stem bark of G. lasiocarpa E. Mey. ex Harv.

Keywords: Grewia lasiocarpa, plant-based extracts, endophytic fungi, Malvaceae

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452 Treatment of Grey Water from Different Restaurants in FUTA Using Fungi

Authors: F. A. Ogundolie, F. Okogue, D. V. Adegunloye

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Greywater samples were obtained from three restaurants in the Federal University of Technology; Akure coded SSR, MGR and GGR. Fungi isolates obtained include Rhizopus stolonifer, Aspergillus niger, Mucor mucedo, Aspergillus flavus, Saccharomyces cerevisiae. Of these fungi isolates obtained, R. stolonifer, A. niger and A. flavus showed significant degradation ability on grey water and was used for this research. A simple bioreactor was constructed using biodegradation process in purification of waste water samples. Waste water undergoes primary treatment; secondary treatment involves the introduction of the isolated organisms into the waste water sample and the tertiary treatment which involved the use of filter candle and the sand bed filtration process to achieve the end product without the use of chemicals. A. niger brought about significant reduction in both the bacterial load and the fungi load of the greywater samples of the three respective restaurants with a reduction of (1.29 × 108 to 1.57 × 102 cfu/ml; 1.04 × 108 to 1.12 × 102 cfu/ml and 1.72 × 108 to 1.60 × 102 cfu/ml) for bacterial load in SSR, MGR and GGR respectively. Reduction of 2.01 × 104 to 1.2 × 101; 1.72 × 104 to 1.1 × 101, and 2.50 × 104 to 1.5 × 101 in fungi load from SSR, MGR and GGR respectively. Result of degradation of these selected waste water by the fungi showed that A. niger was probably more potent in the degradation of organic matter and hence, A. niger could be used in the treatment of wastewater.

Keywords: Aspergillus niger, greywater, bacterial, fungi, microbial load, bioreactor, biodegradation, purification, organic matter and filtration

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451 Degradation of Poly -β- Hydroxybutyrate by Trichoderma asperellum

Authors: Nuha Mansour Alhazmi

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Replacement of petro-based plastics by a biodegradable plastic are vastly growing process. Poly-β-hydroxybutyrate (PHB) is a biodegradable biopolymer, synthesized by some bacterial genera. The objective of the current study is to explore the ability of some fungi to biodegrade PHB. The degradation of (PHB) was detected in Petri dish by the formation of a clear zone around the fungal colonies due to the production of depolymerase enzyme which has an interesting role in the PHB degradation process. Among 10 tested fungi, the most active PHB biodegraded fungi were identified as Trichoderma asperellum using morphological and molecular characters. The highest PHB degradation was at 25°C, pH 7.5 after 7 days of incubation for the tested fungi. Finally, the depolymerase enzyme was isolated, purified using column chromatography and characterized. In conclusion, PHB can be biodegraded in solid and liquid medium using depolymerase enzyme from T. asperellum.

Keywords: degradation, depolymerase enzyme, PHB, Trichoderma asperellum

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450 Screening for Antibacterial Activity of Fungi from Indian Marine Environments: A Possible Alternative for New Antibiotics for the Treatment of Skin Microbial Infections

Authors: Shivankar Agrawal, Sunil Kumar Deshmukh, Colin Barrow, Alok Adholeya

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Millions of people worldwide are affected by infectious diseases caused by bacteria and fungi. Skin and skin structure infections (SSSI) represent a significant category of infectious disease. Unexpectedly, many pathogens have developed resistance towards current antibiotics and over the time this problem has become more and more serious. All these new problems necessitate the continuous search for novel and alternative antibiotics and antifungals. The aim of our research is the screening of extracts of marine fungi for their antibacterial activity against bacteria causing skin and wound infection in humans. A total of 40 marine samples were collected from west coast and Andaman Island of India and 35 morphologically different marine fungi were isolated using natural sea water medium. Among 35 marine fungi, eight isolates exhibited significant antimicrobial activity against human pathogens. In the course of systematic screening program for bioactive marine fungi, strain 'D5' was found to be most potent strain with MIC value of 1 mg/mL, which was morphologically identified as Simplicillium lamellicola. The effects of the most active crude extracts against their susceptible test microorganisms were also investigated by SEM analysis. Purification and characterization of crude extracts for identification of active lead molecule is under process. The results of diversity and antimicrobial activity have increased the scope of finding industrially important marine fungi from Indian marine environments and these organisms could be vital sources for the discovery of pharmaceutically useful molecules.

Keywords: antimicrobial activity, antibiotic, marine fungi, skin infections

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449 Combined Aplication of Indigenous Pseudomonas fluorescens and the AM Fungi as the Potential Biocontrol Agents of Banana Fusarium wilt

Authors: Eri Sulyanti, Trimurti Habazar, Eti Farda Husen, Abdi Dharma, Nasril Nasir

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In this study, combination of some biocontrol agents with different mechanisms was an alternative to improve the effectiveness of the biological control agents. Single and combined applications of indigenous Pseudomonas fluorescens and Arbuscular Mychorrhizae Fungi (AM Fungi) isolates were tested to induce the resistance on susceptible Cavendish banana against F.oxysporum f. sp. cubense race 4 under greenhouse conditions. These isolates originally isolated from healthy banana rhizosphere at endemic Fusarium wilt areas in the centre of production banana in West Sumatra. These researches were conducted with Randomized Block Design with 16 treatments and 10 replications. The treatments were three indigenous isolates of Pseudomonas fluorescens (Par1-Cv, Par4-Rj1, Par2-Jt1) and 3 isolates of AM Fungi (Gl1BuA4, Gl2BuA6, and Gl1KeP3. The biocontrol agents were applied as single agents and combination two of them. This study demonstrated that the application of combination biocontrol organisms Pseudomonas fluorescens and AM Fungi provided were more effective than single application. The combination of Par1-Cv and Gl1BuA4 isolates was the most effective to control Fusarium wilt and followed by the combination of Par1-Cv and Gl2BuA6 and Par2-Jt1 and Gl1P3.

Keywords: pseudomonad fluorescens (Pf), arbuscular mychorrhizae fungi (AM Fungi) indigenous isolates, fusarium oxysporum f. sp. cubense, soil rhizosphere

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448 Lifestyle Switching Phenomenon of Plant Associated Fungi

Authors: Gauravi Agarkar, Mahendra Rai

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Fungi are closely associated with the plants in various types of interactions such as mycorrhizal, parasitic or endophytic. Some of these interactions are beneficial and a few are harmful to the host plants. It has been suggested that these plant-associated fungi are able to change their lifestyle abd this means endophyte may become parasite or vice versa. This phenomenon may have profound effect on plant-fungal interactions and various ecological niches. Therefore, it is necessary to identify the factors that trigger the change in fungal lifestyle and understand whether these different lifestyles are interconnected at some points either by physiological, biochemical or molecular routes. This review summarizes the factors affecting plant fungal interactions and discusses the possible mechanisms for lifestyles switching of fungi based on available experimental evidences. Research should be boosted in this direction to fetch more advantages in future and to avoid the severe consequences in agriculture and other related fields.

Keywords: endophytic, lifestyle switching, mycorrhizal, parasitic, plant-fungal interactions

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447 Screening of Four Malaysian Isolated Endophytes with Candesartan in a Microtiter Plate

Authors: Rasha Saad, Jean Frederic Weber, Fatimah Bebe, Sadia Sultan

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The goal of study was to screen the effects of candesartan and four endophytic fungi for their potential in microbial biotransformation. In this experiment, four types of unidentified fungi with the codes of TH2L1, TH2R10, TH1P35 and TH1S46 were used in screening process by MECFUS (Microtiter plate, Elicitors, Combination, Freeze-drying, UHPLC, Statistical analysis) protocol. The experiment was carried out by using 96-well microtiter plate (MTP) with different media and elicitors. Various media with two concentrations of Potato Dextrose Broth (PDB) and elicitors used were to induce the production of secondary metabolites from the fungi as well as the biotransformation of the drug compound. After incubation, cultures were extracted by freeze drying method and finally analyzed by ultra-High performance Liquid Chromatography (uHPLC). The extracts analyzed by uHPLC followed by LC/Ms, demonstrated the presence of biotransformation products from the drug compound and elicitation of the secondary metabolism from the fungi by the occurrence of the additional peaks. From the four fungi, TH1S46 showed highly potential produced secondary metabolites as well as the biotransformation of candesartan. For other fungi, they responded when candesartan was introduced. Moreover, the additional peaks produced in uHPLC need to be further investigation by using LC-MS or NMR.

Keywords: biotransformation, candesartan, endophytes, secondary metabolites

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446 Biotransformation of Monoterpenes by Whole Cells of Eleven Praxelis clematidea-Derived Endophytic Fungi

Authors: Daomao Yang, Qizhi Wang

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Monoterpenoids are mainly found in plant essential oils and they are ideal substrates for biotransformation into oxygen-containing derivatives with important commercial value due to their low price and simple structure. In this paper, eleven strains of endophytic fungi from Praxelis clematidea were used as test strains to conduct the whole cell biotransformation of the monoterpenoids: (+)-limonene, (-)-limonene and myrcene. The fungi were inoculated in 50 ml Sabouraud medium and incubated at 30 ℃ with the agitation of 150 r/min for 6 d, and then 0.5% (v/v) substrates were added into the medium and biotransformed for further 3 d. Afterwards the cultures were filtered, and extracted using equal volume of ethyl acetate. The metabolites were analyzed by GC-MS technique with NIST database. The Total Ion Chromatogram of the extractions from the eleven strains showed that the main product of (+)- and (-)-limonene biotransformation was limonene-1,2-diol, while it is limonene and linalool oxide for biotransformation of myrcene. This work will help screen the microorganisms to biotransform the monoterpenes.

Keywords: endophytic fungi, (+)–limonene, (-)–limonene, myrcene

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445 Comparision of Neutrophil Response to Curvularia, Bipolaris and Aspergillus Species

Authors: Eszter J. Tóth, Alexandra Hoffmann, Csaba Vágvölgyi, Tamás Papp

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Members of the genera Curvularia and Bipolaris are closely related melanin producing filamentous fungi; both of them have the teleomorph states in genus Cochliobolus. While Bipolaris species infect only plants and may cause serious agriculture damages, some Curvularia species was recovered from opportunistic human infections. The human pathogenic species typically cause phaeohyphomycoses, i.e. mould infections caused by melanised fungi, which can manifest as invasive mycoses with frequent involvement of the central nervous system in immunocompromised patients or as local infections (e.g. keratitis, sinusitis, and cutaneous lesions) in immunocompetent people. Although their plant-fungal interactions have been intensively studied, there is only little information available about the human pathogenic feature of these fungi. The aim of this study was to investigate the neutrophil granulocytes’ response to hyphal forms of Curvularia and Bipolaris in comparison with the response to Aspergillus. In the present study Curvularia lunata SZMC 23759 and Aspergillus fumigatus SZMC 23245 both isolated from human eye infection, and Bipolaris zeicola BRIP 19582b isolated from plant leaf were examined. Neutrophils were isolated from heparinised venous blood of healthy donors with dextran sedimentation followed by centrifugation over Ficoll and hypotonic lysis of erythrocytes. Viability and purity of the cells were checked with trypan blue and Wright staining, respectively. Infection of neutrophils was carried out with germinated conidia in a ratio of 5:1. Production of hydrogen peroxide, superoxide anion, and nitrogen monoxide was measured both intracellularly and extracellularly in response to the germinated spores with or without the supernatant and after serum treatment. ROS and NOS production of neutrophils in interaction with the three fungi were compared. It is already known that Aspergillus species induce ROS production of neutrophils only after serum treatment. Although, in case of C. lunata, serum opsonisation also induced an intensive production of reactive species, lower level of production was measured in the lack of serum as well. After interaction with the plant pathogenic B. zeicola, amount of reactive species found to be similar with and without serum treatment. The presence of germination supernatant decreased the reactive species production in case of each fungus. Interaction with Curvularia, Bipolaris and Aspergillus species induced different response of neutrophils. It seems that recognition of C. lunata and B. zeicola is independent of serum opsonisation, albeit it increases the level of the produced reactive species in response for C. lunata. The study was supported by the grant LP2016-8/2016.

Keywords: Curvularia, neutrophils, NOS, ROS, serum opsonisation

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444 Efficiency for Enzyme Production of Fungi Isolated from the Stomach of Buffalo

Authors: Suphalucksana, Wichai, Sangsoponjit Settasit, Soytong Kasem

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A study on the efficiency for enzyme production of fungi isolated from stomach of buffalo was conducted. The fungi were collected from 4 parts of stomach as rumen, reticulum, omasum and abomasums. The objective to study the efficiency of fungi from stomach of buffalo had effected to produced enzyme and to selected fungi for their ability to produced enzyme cellulase, hemicellulase and ligninase. Results shown that the fungi isolated from rumen were: Eupenicillium sp. (B-RU-01-1), Eupenicillium sp. (B-RU-02-3G), Rhyzopus stolonifer (B-RU-01-4) and Trichoderma sp. (B-RU-01-2). From the reticulum, Aspergillus glaucus (B-RET-02-3), Aspergillus orchraceus (B-RET-02-2) and Penicillium sp. (B-RET-02-4) were found. In the omasum Aspergillus fumigatus (B-OMA-01-1G), Eurotium sp. (B-OMA-01-4) and Rhizopus stolonifer (B-OMA-02-3) were isolated and in the abomasums Aspergillus flavas (B-ABO-02-3), Aspergillus fumigatus (B-ABO-02-1), Aspergillus niger (B-ABO-01-3G), Aspergillius terreus (B-ABO-02-4) and Mucor sp. (B-ABO-02-4G). Results of enzyme analysis revealed that cellulase was produced by isolated: Eupenicillium sp. (B-RU-02-3G), Eupenicillium sp. (B-RU-01-1), Penicillium sp. (B-RET-02-4), Aspergillius glaucus (B-RET-02-3), Aspergillus ochraceus (B-RET-02-2), Aspergillius fumigatus (B-OMA-01-1G), Eurotium sp. (B-OMA-01-4), Aspergillius flavus (B-ABO-02-3), Aspergillius fumigatus (B-ABO-02-1), Aspergillius niger (B-ABO-01-3G), Aspergillius terreus (B-ABO-02-4). Hemicellulase was produced Eupenicillium sp. (B-RU-02-3G), Eupenicillium sp. (B-RU-01-1), Rhizopus stolonifer (B-RU-01-4), Trichoderma sp. (B-RU-01-2), Aspergillius glaucus (B-RET-02-3), Aspergillus ochraceus (B-RET-02-2), Penicillium sp. (B-RET-02-4), Aspergillius fumigatus (B-OMA-01-1G), Eurotium sp. (B-OMA -01-4), Aspergillius flavus (B-ABO-02-3), Aspergillius fumigatus (B-ABO-02-1) Aspergillius niger (B-ABO-01-3G), Aspergillius terreus (B-ABO-02-4), Mucor sp. (B-ABO-02-4G). For the enzyme ligninase, two isolates were found to produced this enzyme namely : Trichoderma sp. (B-RU-01-2) and Mucor sp. (B-ABO-02-4G).

Keywords: enzyme production from fungi, enzyme production, fungi, agricultural technology

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443 Molecular Profiling and Potential Bioactive Characteristics of Endophytic Fungi Isolated from Leptadenia Pyrotechnica

Authors: Walaa Al-Maghraby

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Endophytes are organisms that colonize internal plant tissues without causing apparent harm to their host. Almost all groups of microorganisms have been found in endophytic association with plants may be fungi. They stimulate the production of secondary metabolites with a diverse range of biological activities. Leptadenia pyrotechnica is a more or less leafless, erect shrub with straight stems which is highly distributed in Saudi Arabia. Four endophytes fungi were isolated from Leptadenia pyrotechnica and identified using 18S ribosomal RNA sequences, which revealed four fungi genuses, namely Aspergillus terreus; Aspergillus welwitschiae; Aspergillus fumigatus and Aspergillus flavus. In this present study, four endophytic fungi from Leptadenia pyrotechnica were used for obtaining crude aqueous and ethyl acetate extracts for antimicrobial screening against 6 human pathogens, the antibacterial tests presented satisfactory results, where the pathogenic bacteria were inhibited by the four extracts tested, except for Escherichia coli that was inhibited by all extracts except ethyl acetate extract of Aspergillus terreus. Analysis of variance showed that the extract produced by endophyte Leptadenia pyrotechnica was the most effective against all bacteria, either gram-negative or positive. However, the extract was not efficient against pathogenic fungi. Therefore, this study indicates that endophytes from medicinal plant Leptadenia pyrotechnica could be potential sources of antibacterial substances.

Keywords: antimicrobial activity, Aspergillus sp, endophytes, Leptadenia pyrotechnica

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442 Growth of Albizia in vitro: Endophytic Fungi as Plant Growth Promote of Albizia

Authors: Reine Suci Wulandari, Rosa Suryantini

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Albizia (Paraserianthes falcataria) is a woody plant species that has a high economic value and multifunctional. Albizia is important timber, medicinal plants and can also be used as a plant to rehabilitate critical lands. The demand value of Albizia is increased so that the large quantities and high quality of seeds are required. In vitro propagation techniques are seed propagation that can produce more seeds and quality in a short time. In vitro cultures require growth regulators that can be obtained from biological agents such as endophytic fungi. Endophytic fungi are micro fungi that colonize live plant tissue without producing symptoms or other negative effects on host plants and increase plant growth. The purposes of this research were to isolate and identify endophytic fungi isolated from the root of Albizia and to study the effect of endophytic fungus on the growth of Albizia in vitro. The methods were root isolation, endophytic fungal identification, and inoculation of endophytic fungi to Albizia plants in vitro. Endophytic fungus isolates were grown on PDA media before being inoculated with Albizia sprouts. Incubation is done for 4 (four) weeks. The observed growth parameters were live explant percentage, percentage of explant shoot, and percentage of explant rooted. The results of the research showed that 6 (six) endophytic fungal isolates obtained from the root of Albizia, namely Aspergillus sp., Verticillium sp, Penicillium sp., Trichoderma sp., Fusarium sp., and Acremonium sp. Statistical analysis found that Trichoderma sp. and Fusarium sp. affect in vitro growth of Albizia. Endophytic fungi from the results of this research were potential as plant growth promoting. It can be applied to increase productivity either through increased plant growth and increased endurance of Albizia seedlings to pests and diseases.

Keywords: Albizia, endophytic fungi, propagation, in vitro

Procedia PDF Downloads 266