Search results for: fusarium solani
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 110

Search results for: fusarium solani

80 Bacillus thuringiensis CHGP12 Uses a Multifaceted Strategy to Suppress Fusarium Wilt of Chickpea and to Enhance the Total Biomass of Chickpea Plants

Authors: Muhammad Naveed Aslam, Rida Fatima, Anam Moosa, Muhammad Taimoor Shakeel

Abstract:

Bacillus strains produce antifungal secondary metabolites making them potential candidates for suppressing Fusarium wilt of chickpea disease. In this study, eighteen Bacillus strains were evaluated for their antagonistic effect against Fusarium oxysporum f. sp. ciceris causing Fusarium wilt of chickpea disease. In a direct antifungal assay, thirteen strains showed significant inhibition zones while the remaining five strains did not produce inhibition zones of FOC. Bacillus thuringiensis CHGP12 was the most promising strain exhibiting the highest inhibition of FOC. Antifungal lipopeptides were extracted from CHGP12 strain which showed significant inhibition of the pathogen. Liquid chromatography mass spectrometry (LCMS) analysis revealed that CHGP12 was positive for the presence of iturin, fengycin, surfactin, bacillaene, bacillibactin, plantazolicin, and bacilysin. CHGP12 was tested for biochemical determinants in an in vitro qualitative test where it showed the ability to produce lipase, amylase, cellulase, protease, siderophores, and indole 3-acetic acid (IAA). Furthermore, in a greenhouse experiment CHGP12 also showed a significant decrease in the disease severity in treated plants compared to control. Moreover, CHGP12 also exhibited a significant increase in plant growth parameters viz, root and shoot growth parameters, stomatal conductance, and photosynthesis rate. Conclusively, our findings present the promising potential of Bacillus strain CHGP12 to suppress Fusarium wilt of chickpea and to promote plant growth.

Keywords: liquid chromatography mass spectrometry, growth promotion, antagonism, hydrolytic enzymes, inhibition, lipopeptides.

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79 Isolation and Identification Fibrinolytic Protease Endophytic Fungi from Hibiscus Leaves in Shah Alam

Authors: Mohd Sidek Ahmad, Zainon Mohd Noor, Zaidah Zainal Ariffin

Abstract:

Fibrin degradation is an important part in prevention or treatment of intravascular thrombosis and cardiovascular diseases. Plasmin like fibrinolytic enzymes has given new hope to patient with cardiovascular diseases by treating fibrin aggregation related diseases with traditional plasminogen activator which have many side effects. Various researches involving wide range of sources for production of fibrinolytic proteases, from bacteria, fungi, insects and fermented foods. But few have looked into endophytic fungi as a potential source. Sixteen (16) endophytic fungi were isolated from Hibiscus sp. leaves from six different locations in Shah Alam, Selangor. Only two endophytic fungi, FH3 and S13 showed positive fibrinolytic protease activities. FH3 produced 5.78cm and S13 produced 4.48cm on Skim Milk Agar after 4 days of incubation at 27°C. Fibrinolytic activity was observed; 3.87cm and 1.82cm diameter clear zone on fibrin plate of FH3 and S13 respectively. 18srRNA was done for identification of the isolated fungi with positive fibrinolytic protease. S13 had the highest similarity (100%) to that of Penicillium citrinum strain TG2 and FH3 had the highest similarity (99%) to that of Fusarium sp. FW2PhC1, Fusarium sp. 13002, Fusarium sp. 08006, Fusarium equiseti strain Salicorn 8 and Fungal sp. FCASAn-2. Media composition variation showed the effects of carbon nitrogen on protein concentration, where the decrement of 50% of media composition caused drastic decrease in protease of FH3 from 1.081 to 0.056 and also S13 from 2.946 to 0.198.

Keywords: isolation, identification, fibrinolytic protease, endophytic fungi, Hibiscus leaves

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78 Differentially Response of Superoxide Dismutase in Wheat Susceptible and Resistant Cultivars against FHB

Authors: M. Sorahi Nobar, V. Niknam, H. Ebrahimzadeh, H. Soltanloo

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Fusarium graminearum is one of the most destructive crop diseases in the world. Infection occurs during the flowering period in warm and humid conditions. It causes reduction in yield. Moreover, harvested grain is often contaminated with mycotoxins and its acetylated derivatives. Fusarium mycotoxines are potent inhibitor of protein synthesis, and thereby presents hazards for both human and animal health. A rapid production of reactive oxygen intermediates, primarily superoxide and hydrogen peroxide at the site of attempted infection considered as key feature underlying successful pathogen recognition. Here, we compared the time course activity of superoxide dismutase (SOD) as a first line of defenses against ROS- induced oxidative burst between FHB- resistant Sumai3 and susceptible Falat at 48, 96 and 144 hours after infection. Our results showed that Sumai3 SOD activity increased with time and reached the highest-level 4 days after infection while in susceptible cultivar Falat, SOD activity decreased during the first 96 h. after infection. Decreased was followed by an increased at 6 days after infection. According to our results rapid induction of SOD activity in resistant cultivar may play an important role in resistance against FHB in wheat.

Keywords: Fusarium graminearum, mycotoxins, resistant cultivar, superoxide dismutase

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77 Antifungal Potential of the Plant Growth-Promoting Rhizobacteria Infecting Kidney Beans

Authors: Zhazira Shemsheyeva, Zhanara Suleimenova, Olga Shemshura, Gulnaz Mombekova, Zhanar Rakhmetova

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Bacteria that colonize plant roots and promote plant growth are referred to as plant growth-promoting rhizobacteria (PGPR). They not only provide nutrients to the plants (direct plant growth promotion) and protect plants against the phytopathogens (indirect plant growth promotion) but also increase the soil fertility. Indirectly PGPRs improve the plant growth by becoming a biocontrol agent for a fungal pathogen. The antifungal activities of the PGPrhizobacteria were assayed against different species of phytopathogenic fungi such as Fusarium tricinctum, Fusarium oxysporum, Sclerotiniasclerotiorum, and Botrytis cinerea. Pseudomonas putidaSM-1, Azotobacter sp., and Bacillus thuringiensis AKS/16 strains have been used in experimental tests on growth inhibition of phytopathogenic fungi infecting Kidney beans. Agar well diffusion method was used in this study. Diameters of the zones of inhibition were measured in millimeters. It was found that Bacillus thuringiensis AKS/16 strain showed the lowest antifungal activity against all fungal pathogens tested. Zones of inhibition were 15-18 mm. In contrast, Pseudomonas putida SM-1 exhibited good antifungal activity against Fusarium oxysporum and Fusarium tricinctum by producing 29-30 mm clear zones of inhibition. The moderate inhibitory effect was shown by Azotobacter sp. against all fungal pathogens tested with zones of inhibition from24 to 26 mm. In summary, Pseudomonas putida SM-1 strain demonstrated the potential of controlling root rot diseases in kidney beans.

Keywords: PGPR, pseudomonas putida, kindey beans, antifungal activity

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76 Tomato Endophytes Trichoderma asperellum AAUTLF and Stenotrophomonas maltophilia D1B Exhibits Plant Growth-Promotion and Fusarium Wilt Suppression

Authors: Bandana Saikia, Ashok Bhattacharyya

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Endophytic microbes and their metabolites positively impact overall plant health, which may have a potential implication in agriculture. In the present study, 177 bacterial endophytes and 57 fungal endophytes were isolated, with the highest recovery rate from tomato roots. A maximum of 112 endophytes were isolated during monsoon, followed by 64 isolates and 58 isolates isolated during pre-monsoon and post-monsoon periods, respectively, indicating the rich diversity in bacterial and fungal endophytes of tomato crops from different locations of Assam, India. Further, the endophytes were evaluated for their antagonistic potential against Fusarium oxysporum f. sp. lycopersici. Fungal endophytic isolate AAUTLF (Endophytic Fungi of Tomato Leaf from Assam Agricultural University, Assam, India area) and bacterial endophyte D1B (Endophytic bacteria of tomato from Dhemiji, India district) showed the highest antifungal activity against the pathogen both in vitro and in vivo. Based on 5.8 rDNA sequence analysis of fungal and 16S rDNA sequence of bacteria endophytes, the most effective fungal and bacterial isolates against FOL were identified as Trichoderma asperellum AAUTLF and Stenotrophomonas maltophilia D1B, respectively. The isolates showed an antagonistic effect against Fusarium oxysporum f.sp. lycopersici in-vitro and reduced the disease index of Fusarium wilt in tomatoes by 64.4% under pot conditions. Trichoderma asperellum AAUTLF produced an antifungal compound viz., 6-pentyl-2H-pyran-2-one, which also possesses growth-promoting characteristics. The bacteria Stenotrophomonas maltophilia D1B produced antifungal compounds, including benzothiazole, oleic acid, phenylacetic acid, and 3-(Hydroxy-phenyl-methyl)-2,3-dimethyl-octan-4-one. This would be of high importance for the source of antagonistic strains and biocontrol of tomato Fusarium wilt, as well as other plant fungal diseases.

Keywords: root endophytes, Stemotrophomonas, Trichoderma, benzothiazole, 6-pentyl-2H-pyran-2-one

Procedia PDF Downloads 44
75 Chemical Control Management Strategies for Corm Rot in Gladiolus communis L. under Field Conditions

Authors: Shahbaz Ahmad, Muhammad Ali, Sahar Naz

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Corm rot is caused by the fungus Fusarium oxysporum f.sp. gladioli and it causes remarkable losses to the growers. Experiment was conducted in order to find some viable recommendations for this agronomically as well as economically important problem. Four fungicides, namely Carbendazim, Mancozeb, Thiophanate methyl and Chlorothalonil were used to control corm rot in gladiolus field. Fungicides were applied singly as foliar, in irrigation as well as with sulphuric acid in variable doses. The results revealed that application of all fungicides was variably effective to control corm rot in acid mixed irrigation followed by fungicide in irrigation. The application of all fungicides in various combinations was observed to be ineffective at all three doses.

Keywords: gladiolus, corm rot, Fusarium oxysporum, fungicides

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74 Biocontrol of Fusarium Crown and Root Rot and Enhancement of Tomato Solanum lycopersicum L. Growth Using Solanum linnaeanum L. Extracts

Authors: Ahlem Nefzi, Rania Aydi Ben Abdallah, Hayfa Jabnoun-Khiareddine, Nawaim Ammar, Sined Medimagh-Saidana, Mejda Daami-Remadi

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In the present study, leaf, stem, and fruit aqueous extracts of native wild Solanum linnaeanum L. were screened for their ability to suppress Fusarium Crown and Root Rot disease and to enhance tomato (Solanum lycopersicum L.) growth under greenhouse conditions. Leaf extract used at 30% w/v was the most effective in reducing leaf and root damage index by 92.3% and the extent of vascular discoloration by 97.56% compared to Fusarium oxyxporum f. sp radicis lycopersici -inoculated and untreated control. A significant promotion of growth parameters (root length, shoot height, root and shoot biomass and stem diameter) was recorded on tomato cv. Rio Grande seedlings by 40.3-94.1% as compared to FORL inoculated control and by 9.6-88.8% over pathogen-free control. All S. linnaeanum aqueous extracts tested significantly stimulated the germination by 10.2 to 80.1% relative to the untreated control. FORL mycelial growth, assessed using the poisoned food technique, varied depending on plant organs, extracts, and concentrations used. Butanolic extracts were the most active, leading to 60.81% decrease in FORL mycelial growth. HPLC analysis of butanolic extract revealed the presence of thirteen phenolic compounds. Thus, S. linnaeanum can be explored as a potential natural source of antifungal and biofertilizing compounds.

Keywords: antifungal activity, HPLC-MS analysis, Fusarium oxysporum f. sp. radicis-lycopersici, tomato growth

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73 Multilocus Phylogenetic Approach Reveals Informative DNA Barcodes for Studying Evolution and Taxonomy of Fusarium Fungi

Authors: Alexander A. Stakheev, Larisa V. Samokhvalova, Sergey K. Zavriev

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Fusarium fungi are among the most devastating plant pathogens distributed all over the world. Significant reduction of grain yield and quality caused by Fusarium leads to multi-billion dollar annual losses to the world agricultural production. These organisms can also cause infections in immunocompromised persons and produce the wide range of mycotoxins, such as trichothecenes, fumonisins, and zearalenone, which are hazardous to human and animal health. Identification of Fusarium fungi based on the morphology of spores and spore-forming structures, colony color and appearance on specific culture media is often very complicated due to the high similarity of these features for closely related species. Modern Fusarium taxonomy increasingly uses data of crossing experiments (biological species concept) and genetic polymorphism analysis (phylogenetic species concept). A number of novel Fusarium sibling species has been established using DNA barcoding techniques. Species recognition is best made with the combined phylogeny of intron-rich protein coding genes and ribosomal DNA sequences. However, the internal transcribed spacer of (ITS), which is considered to be universal DNA barcode for Fungi, is not suitable for genus Fusarium, because of its insufficient variability between closely related species and the presence of non-orthologous copies in the genome. Nowadays, the translation elongation factor 1 alpha (TEF1α) gene is the “gold standard” of Fusarium taxonomy, but the search for novel informative markers is still needed. In this study, we used two novel DNA markers, frataxin (FXN) and heat shock protein 90 (HSP90) to discover phylogenetic relationships between Fusarium species. Multilocus phylogenetic analysis based on partial sequences of TEF1α, FXN, HSP90, as well as intergenic spacer of ribosomal DNA (IGS), beta-tubulin (β-TUB) and phosphate permease (PHO) genes has been conducted for 120 isolates of 19 Fusarium species from different climatic zones of Russia and neighboring countries using maximum likelihood (ML) and maximum parsimony (MP) algorithms. Our analyses revealed that FXN and HSP90 genes could be considered as informative phylogenetic markers, suitable for evolutionary and taxonomic studies of Fusarium genus. It has been shown that PHO gene possesses more variable (22 %) and parsimony informative (19 %) characters than other markers, including TEF1α (12 % and 9 %, correspondingly) when used for elucidating phylogenetic relationships between F. avenaceum and its closest relatives – F. tricinctum, F. acuminatum, F. torulosum. Application of novel DNA barcodes confirmed the fact that F. arthrosporioides do not represent a separate species but only a subspecies of F. avenaceum. Phylogeny based on partial PHO and FXN sequences revealed the presence of separate cluster of four F. avenaceum strains which were closer to F. torulosum than to major F. avenaceum clade. The strain F-846 from Moldova, morphologically identified as F. poae, formed a separate lineage in all the constructed dendrograms, and could potentially be considered as a separate species, but more information is needed to confirm this conclusion. Variable sites in PHO sequences were used for the first-time development of specific qPCR-based diagnostic assays for F. acuminatum and F. torulosum. This work was supported by Russian Foundation for Basic Research (grant № 15-29-02527).

Keywords: DNA barcode, fusarium, identification, phylogenetics, taxonomy

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72 Bioprotective Role of Soil Borne Bacillus Strains against Selected Fungal Pathogens of Agriculture Relevance

Authors: Asad Ali, Asif Jamal

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The agriculture productivity losses due to microbial pathogens have been a serious issue in Pakistan and rest of the world. Present work was designed to isolate soil borne microorganisms having the antagonistic ability against notorious phytopathogens. From the initial collection of 23 bacterial isolates, two potent strains of Bacillus were screened on the basis of their comparative efficacy against devastating fungal pathogens. The strains AK-1 and AK-5 showed excellent inhibitory indexes against the majority of tested fungal strains. It was noted that both strains of Bacillus showed significant biocontrolling activity against Aspergillus flavus, Fusarium moniliforme, Colletotricum falcatum, Botrytis cinerea, Aspergillus niger, Fusarium oxysporum, Phythopthora capsici and Rhizopus oryzae. The strain AK-1 was efficient to suppress Aspergillus species and Rhizopus oryzae while AK-5 expressed significant antagonistic activity against Fusarium, Botrytis and Colletotricum species. On the basis of in vitro assay, it can be postulated that the Bacillus strains AK-1 and AK-5 can be used as bio-protective agent against various plant diseases. In addition, their applications as natural pesticides could be very helpful to prevent the adverse effects of chemical pesticides.

Keywords: bacillus species, biocontrol agent, biopesticides, phytopathogens

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71 Bioprotective Role of Soil Borne Bacillus Strain against Selected Fungal Pathogens of Agriculture Relevance

Authors: Asif Jamal, Asad Ali, Muhammad Ishtiaq Ali

Abstract:

The agriculture productivity losses due to microbial pathogens have been a serious issue in Pakistan and rest of the world. Present work was designed to isolate soil borne microorganisms having the antagonistic ability against notorious phytopathogens. From the initial collection of 23 bacterial isolates, two potent strains of Bacillus were screened on the basis of their comparative efficacy against devastating fungal pathogens. The strains AK-1 and AK-5 showed excellent inhibitory indexes against the majority of tested fungal strains. It was noted that both strains of Bacillus showed significant biocontrolling activity against Aspergillus flavus, Fusarium moniliforme, Colletotricum falcatum, Botrytis cinerea, Aspergillus niger, Fusarium oxysporum, Phythopthora capsici and Rhizopus oryzae. The strain AK-1 was efficient to suppress Aspergillus species and Rhizopus oryzae while AK-5 expressed significant antagonistic activity against Fusarium, Botrytis, and Colletotricum species. On the basis of in vitro assay, it can be postulated that the Bacillus strains AK-1 and AK-5 can be used as a bio-protective agent against various plant diseases. In addition, their applications as natural pesticides could be very helpful to prevent the adverse effects of chemical pesticides.

Keywords: biological control, Bacillus spp, fungal pathogens, agriculture

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70 Antifungal Potential of Higher Basidiomycetes Mushrooms

Authors: Tamar Khardziani, Violeta Berikashvili, Mariam Rusitashvili, Eva Kachlishvili, Vladimir Elisashvili, Mikheil Asatiani

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Last years, the search for natural sources of novel and effective antifungal substances became a scientific and technological challenge. In the present research, thirty basidiomycetes isolated from various ecological niches of Georgia and belonging to different taxonomic groups were screened for their antifungal activities against pathogenic fungi such as Aspergillus, Fusarium, and Guignardia bidwellii. Among mushroom tested, several potential producers of antifungal substances have been revealed, such as Schizophyllum commune, Lentinula edodes, Ganoderma abietinum, Fomes fomentarius, Hericium erinaceus, and Trametes versicolor. For mushroom cultivation and expression of antifungal potential, submerged and solid-state fermentations of different plant raw materials were performed and various approaches and strategies have been exploited. Sch. commune appeared as a most promising producer of antifungal compounds. It was established that among different agro-industrial wastes, the presence of mandarin juice production waste in a nutrient medium, causing the significant increase of antifungal activity Sch. commune (growth inhibition: Aspergillus – 59 %, Fusarium – 55 %, G. bidwellii – 78 %, after 3, 2 and 4 days of cultivation, respectively). Besides this, Sch. commune demonstrate similar antifungal activities in the presence of glucose, glycerol, maltose, mannitol, and xylose, and growth inhibition of Fusarium ranged in 41 % - 49 % during 6 days of cultivation. Inhibition of Aspergillus growth inhibition varied in 27 % - 36 %, and inhibition of G. bidwellii was in the range 49 % - 61 %, respectively. Sch. commune under solid-state fermentation of mandarin peels at 13 days of cultivation demonstrates powerful growth inhibition of pathogenic fungi (growth inhibition: Aspergillus – 50 %, Fusarium – 61 %, G. bidwellii – 68 %, after 3, 4, and 4 days of cultivation, respectively) as well as at 20 days old mushroom (growth inhibition: Aspergillus – 41 %, Fusarium – 54 %, G. bidwellii – 66 %, after 3 days of cultivation). It was established that Sch. commune was effective as a producer of antifungal compounds in submerged as well as in solid-state fermentation. Finally, performed study confirms that the higher basidiomycetes possess antifungal potential, which strongly depends on the physiological factors of growth. Acknowledgments: The work was implemented with the financial support of fundamental science project FR-19-3719 by the Shota Rustaveli National Science Foundation of Georgia.

Keywords: antifungal potential, higher basidiomycetes, pathogenic fungi, submerged and solid-state fermentation

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69 Induced Systemic Resistance in Tomato Plants against Fusarium Wilt Disease Using Biotic Inducers

Authors: Mostafa A. Amer, I. A. El-Samra, I. I. Abou-ElSeoud, S. M. El-Abd, N. K. Shawertamimi

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Tomato Fusarium wilt disease caused by Fusarium oxysporum f. sp. Lycopercisi (FOL) is considered one of the most destructive diseases in Egypt. Effect of some biotic inducers such as Bacillus megaterium var. phosphaticum, Glomus intraradices and Glomus macrocarpum at seven different mixed treatments, was tested for their ability to induce resistance in tomato plants against the disease. According to pathogenicity tests, all the tested isolates of FOL showed wilt symptoms on both of the tested cultivars; however, they considerably varied in percentages of disease incidence (DI) and disease severity (DS). Castle Rock was more susceptible than Peto 86, which was relatively resistant. Pretreatment of both cultivars, under greenhouse conditions, with the tested biotic inducers alone or in combination with each other's, significantly increased the induction of chitinase, β-1,3-glucanase, peroxidase, and polyphenoloxidase and reduced disease incidence and severity, compared with untreated noninoculated (C1) and untreated inoculated (C2) controls. Application of a combination of BMP, with GI and GM was the most effective in increasing the induction rated of the tested enzymes, compared with the other treatments. Induction of enzymes in most of the tested bioinducers treatments gradually increased, attaining maximum values after 48 or/and 72 hrs after challenging with FOL, then gradually declined. GI was the least effective bioinducer.

Keywords: F. oxysporum f. sp. lycopersici, defense enzymes, induced systemic resistance, ISR, B. megaterium var. phosphaticum, G. macrocarpum, G. intraradices

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68 Investigation of the Effect of Anaerobic Digestate on Antifungal Activity and in Different Parameters of Maize

Authors: Nazia Zaffar, Alam Khan, Abdul Haq, Malik Badshah

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Pakistan is an agricultural country. The increasing population leads to an increase in demand for food. A large number of crops are infected by different microbes, and nutrient deficiency of soil adversely affects the yield of crops. Furthermore, the use of chemical fertilizers like Nitrogen, Phosphorus, Potassium (NPK) Urea, and Diammonium phosphate (DAP) and pesticides have environmental consequences. Therefore, there is an urgent need to explore alternative renewable and sustainable biofertilizers. Maize is one of the top growing crops in Pakistan, but it has low yield compared to other countries due to deficiency of organic matter, widespread nutrients deficiency (phosphorus and nitrogen), unbalanced use of fertilizers and various fungal diseases. In order to get rid of all these disadvantages, Digestate emerged as a win-win opportunity for the control of a few plant diseases and a replacement for the chemical fertilizers. The present study was designed to investigate the effect of Anerobic digestate on Antifungal Activity and in different parameters of Maize. The antifungal activity, minimum inhibitory concentration (MIC), and minimum fungicidal concentration (MFC) against selected phytopathogens (Colletotrichum coccodis, Pythium ultimum, Phytophthora capsci, Rhizoctonia solani, Bipolaris oryzae and Fusarium Fujikuroi) were determined by microtiter plate method. The effect of various fertilizers in different growth parameters height, diameter, chlorophyll, leaf area, biomass, and yield were studied in field experiments. The extracts from anaerobic digestate have shown antifungal activity against selected phytopathogens, the highest activity was noted against P. ultimum, the MIC activity was high in case of P. ultimum and B. oryzae. The present study concludes that anaerobic digestate have a positive effect on maize growth and yield as well as an antifungal activity which can be potentially a good biofertilizer.

Keywords: anaerobic digestate, antifungal activity, MIC, phytopathogens

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67 Chemical-Induced Mutation for Development of Resistance in Banana cv. Nanjangud rasabale

Authors: H. Kishor, G. Prabhuling, D. S. Ambika, D. P. Prakash

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The chemical mutagens have become important tool to enhance agronomic traits of banana crop. It is being used to develop fusarium resistance lines in various susceptible banana cultivars. There are several mutagens like EMS and NaN3 available for banana crop improvement and each mutagen has its own important role as positive or negative effects on growth and development of banana plants. Explants from shoot tip culture were treated with various EMS (0.30, 0.60, 0.90 and 0.12%) and NaN3 (0.01, 0.02 and 0.03%) concentrations. The putative mutants obtained after in vitro rooting were subjected for artificial inoculation of Fusarium oxysporum f.sp. cubense. Screening putative mutants resistance to Panama disease was carried out by using syringe method of inoculation. It was observed that, EMS treated mutants were more susceptible compared to NaN3 treatment. Among the NaN3 doses 0.01% found to produce 3 resistant lines during preliminary screening under greenhouse conditions.

Keywords: Nanjangud rasabale, EMS, NaN3, putative mutants

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66 The Biofumigation Activity of Volatile Compounds Produced from Trichoderma afroharzianum MFLUCC19-0090 and Trichoderma afroharzianum MFLUCC19-0091 against Fusarium Infections in Fresh Chilies

Authors: Sarunpron Khruengsai, Patcharee Pripdeevech

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This study aimed to investigate the fumigation activities of the volatile compounds produced by Trichoderma spp. against Fusarium oxysporum and F. proliferatum fungi that cause significant rot in fresh chilies. Two Trichoderma spp. were isolated from the leaves of Schefflera leucantha grown in Thailand and later identified as T. afroharzianum MFLUCC19-0090 and T. afroharzianum MFLUCC19-0091. Both in vitro and in vivo dual culture volatile assays were used to study the effects of the produced volatile compounds on mycelial growth. In vitro results showed that the volatile compounds produced by T. afroharzianum MFLUCC19-0090 significantly inhibited the growth of F. oxysporum, while the volatile compounds produced by T. afroharzianum MFLUCC19-0091 significantly inhibited the growth of F. proliferatum. The effectiveness of Trichoderma-derived volatile compounds in inhibiting the mycelial growth of the selected pathogens in the inoculated, fresh chili samples was further demonstrated in vivo. The volatile profiles of both Trichoderma spp. were characterized using gas chromatography-mass spectrometry. Seventy-three volatile compounds were detected from both strains. Among the major volatile compounds detected, phenyl ethyl alcohol was found to possess the strongest antifungal activity against both pathogens. The results support the possibility of using volatile compounds produced by T. afroharzianum MFLUCC19-0090 and T. afroharzianum MFLUCC19-0091 as alternative fumigants for preventing Fusarium rot of fresh chilies during the post-harvest period.

Keywords: antifungal activity, biocontrol, endophytic fungi, post-harvest

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65 Anticandidal and Antibacterial Silver and Silver(Core)-Gold(Shell) Bimetallic Nanoparticles by Fusarium graminearum

Authors: Dipali Nagaonkar, Mahendra Rai

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Nanotechnology has experienced significant developments in engineered nanomaterials in the core-shell arrangement. Nanomaterials having nanolayers of silver and gold are of primary interest due to their wide applications in catalytical and biomedical fields. Further, mycosynthesis of nanoparticles has been proved as a sustainable synthetic approach of nanobiotechnology. In this context, we have synthesized silver and silver (core)-gold (shell) bimetallic nanoparticles using a fungal extract of Fusarium graminearum by sequential reduction. The core-shell deposition of nanoparticles was confirmed by the red shift in the surface plasmon resonance from 434 nm to 530 nm with the aid of the UV-Visible spectrophotometer. The mean particle size of Ag and Ag-Au nanoparticles was confirmed by nanoparticle tracking analysis as 37 nm and 50 nm respectively. Quite polydispersed and spherical nanoparticles are evident by TEM analysis. These mycosynthesized bimetallic nanoparticles were tested against some pathogenic bacteria and Candida sp. The antimicrobial analysis confirmed enhanced anticandidal and antibacterial potential of bimetallic nanoparticles over their monometallic counterparts.

Keywords: bimetallic nanoparticles, core-shell arrangement, mycosynthesis, sequential reduction

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64 Physiological Insight into an Age Old Biocontrol Practice in Banana Cultivation

Authors: Susmita Goswami, Joyeeta Mitra, Indu Gaur, Neha Bhadauria, Shilpi Shilpi, Prabir K. Paul

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'Malbhog’, an indigenous banana variety, much prized for its flavour and delicacy suffers production losses due to Fusarium oxysporum f.sp. cubense. The pathogen enters young plants through feeder roots causing wilting of plants ultimately leading to death of plants. The pathogen spreads rapidly to other plants in the field. In eastern part of India, this variety escapes the onslaught of the pathogen when either co-cultivated or rotated with Amorphophallus campanulatus (yam). The present study provides an insight into the physiological aspect of the biocontrol by yam. In vitro application of sterile aqueous extract of yam tuber (100gm/100ml distilled water and its 1:10 and 1:100 dilutions) were mixed with PDA media which was substantially inoculated with spores of Fusarium oxysporum f.sp. cubense. The extract could significantly reduce germination of pathogen spores. Banana variety susceptible to Fusarium sp was raised in soil rite under aseptic conditions. Spores of the pathogen (106 spores/ml) were inoculated into the soil rite. The plants were spread with aqueous extract of yam. The control plants were treated with sterilized distilled water. The activity of phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO) and peroxidase (POX) were estimated in leaves and roots at interval of 24 hours for 5 days after treatment. The incidence of wilt disease was recorded after two weeks. The results demonstrated that yam extract could induce significant activity of PAL, PPO and POX along with accumulation of phenols in both roots and leaves of banana plants. However, significantly high activity of enzymes and phenol accumulation was observed in roots. The disease incidence was significantly low in yam treated plants. The results clearly demonstrated the control of the pathogen due to induction of defense mechanism in the host by the extract. The observed control of the pathogen in the field could possibly be due to induction of such defense responses in host by exudates leached into the soil from yam tubers. Yam extract could be a potential source of environment-friendly biocide against Panama wilt of banana.

Keywords: Amorphophallus campanulatus, banana, Fusarium oxysporum f.sp. cubense, phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), peroxidase (POX)

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63 Antibacterial and Antifungal Activity of Essential Oil of Eucalyptus camendulensis on a Few Bacteria and Fungi

Authors: M. Mehani, N. Salhi, T. Valeria, S. Ladjel

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Red River Gum (Eucalyptus camaldulensis) is a tree of the genus Eucalyptus widely distributed in Algeria and in the world. The value of its aromatic secondary metabolites offers new perspectives in the pharmaceutical industry. This strategy can contribute to the sustainable development of our country. Preliminary tests performed on the essential oil of Eucalyptus camendulensis showed that this oil has antibacterial activity vis-à-vis the bacterial strains (Enterococcus feacalis, Enterobacter cloaceai, Proteus microsilis, Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa) and antifungic (Fusarium sporotrichioide and Fusarium graminearum). The culture medium used was nutrient broth Muller Hinton. The interaction between the bacteria and the essential oil is expressed by a zone of inhibition with diameters of MIC indirectly expression of. And we used the PDA medium to determine the fungal activity. The extraction of the aromatic fraction (essentially oil- hydrolat) of the fresh aerian part of the Eucalyptus camendulensis was performed by hydrodistillation. The average essential oil yield is 0.99%. The antimicrobial and fungal study of the essential oil and hydrosol showed a high inhibitory effect on the growth of pathogens.

Keywords: essential oil, Eucalyptus camendulensis, bacteria and fungi, red river gum

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62 Characterization of a Broad Range Antimicrobial Substance from Pseudozyma aphidis

Authors: Raviv Harris, Maggie Levy

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Natural product-based pesticides may serve as an alternative to the traditional synthetic pesticides, which have a potentially damaging effect, both to human health and for the environment. Along with plants, microorganisms are a prospective source of such biological pesticides. A unique and active strain of P. aphidis (designated isolate L12, Israel 2004), an epiphytic and non-pathogenic basidiomycete yeast, was isolated in our lab from strawberry leaves. P. aphidis L12 secretions were found to inhibit broad range of plant pathogens. This work demonstrates that metabolites isolated from the biocontrol agent P. aphidis (isolate L12) can inhibit varied fungal and bacterial phytopathogens. Biologically active metabolites were extracted from P. aphidis biomass, using the organic solvent ethyl acetate. The antimicrobial activity of the extract was demonstrated, both in vitro and in planta. Using disk diffusion assays, the following inhibition zones were obtained: 43cm² for Pseudomonas syringae pv. tomato, 28.5cm² for Xanthomonas campestris pv. vesicatoria, 59cm² for Clavibacter michiganensis subsp. michiganensis, 34cm² for Erwinia amylovora and 34cm² for Agrobacterium tumefaciens. Additionally, strong inhibitory activity of the extract against fungi mycelial growth was established, with IC₅₀ values of 606µg ml⁻¹ for Botrytis cinerea, 221µg ml⁻¹ for Pythium spp., 519µg ml⁻¹ for Rhizoctonia solani, 455µg ml⁻¹ for Sclerotinia sclerotiorum, 2270µg ml⁻¹ for Fusarium oxysporum f. sp. lycopersici, and 2038µg ml⁻¹ for Alternaria alternata. The results of the in planta experiments demonstrated a dose-dependent reduction in disease infection. Significant inhibition of B. cinerea lesions on tomato plants was obtained when a spore suspension of this pathogen was treated with extract concentrations higher than 4.2mg ml⁻¹. Concentration of 7mg ml⁻¹ caused a reduction of over 95% in the lesion size of B. cinerea on tomato plants. The strong antimicrobial activity demonstrated both in vitro and in planta against varied phytopathogens, may indicate that the extracted antimicrobial metabolites have potential to serve as natural pesticides in the field.

Keywords: antimicrobial, B. cinerea, metabolites, natural pesticides, P. aphidis

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61 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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60 The Molecular Analysis of Effect of Phytohormones and Spermidine on Tomato Growth under Biotic Stress

Authors: Rumana Keyani, Haleema Sadia, Asia Nosheen, Rabia Naz, Humaira Yasmin, Sidra Zahoor

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Tomato is a significant crop of the world and is one of the staple foods of Pakistan. A vast number of plant pathogens from simple viruses to complex parasites cause diseases in tomatoes but fungal infection in our country is quite high. Sometimes the symptoms are too harsh destroying the crop altogether. Countries like our own with continuously increasing massive population and limited resources cannot afford such an economic loss. There is an array of morphological, genetic, biochemical and molecular processes involved in plant resistance mechanisms to biotic stress. The study of different metabolic pathways like Jasmonic acid (JA) pathways and most importantly signaling molecules like ROS/RNS and their redoxin enzymes i.e. TRX and NRX is crucial to disease management, contributing to healthy plant growth. So, improving tolerance in crop plants against biotic stresses is a dire need of our country and world as whole. In the current study, fungal pathogenic strains Alternaria solani and Rhizoctonia solani were used to inoculate tomatoes to check the defense responses of tomato plant against these pathogens at molecular as well as phenotypic level with jasmonic acid and spermidine pretreatment. All the growth parameters (root and shoot length, dry and weight root, shoot weight measured 7 days post-inoculation, exhibited that infection drastically declined the growth of the plant whereas jasmonic acid and spermidine assisted the plants to cope up with the infection. Thus, JA and Spermidine treatments maintained comparatively better growth factors. Antioxidant assays and expression analysis through real time quantitative PCR following time course experiment at 24, 48 and 72 hours intervals also exhibited that activation of JA defense genes and a polyamine Spermidine helps in mediating tomato responses against fungal infection when used alone but the two treatments combined mask the effect of each other.

Keywords: fungal infection, jasmonic acid defence, tomato, spermidine

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59 Design, Development and Application of a Green Manure Fertilizer Based on Mucuna Pruriens (L.) in Pelletized Presentation

Authors: Andres Zuñiga Orozco

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Green manure fertilizers have special importance in the development of organic and sustainable agriculture as a substitute or complement to chemical fertilization. They have many advantages, but they have application limitations in greenhouse crops and in open field crops that have low growing size. On the other hand, the logistics of sowing, harvesting and applying have been difficult for producers to adopt. For this reason, a pelletized presentation was designed in conjunction with Trichoderma harzianum. The biopellet was applied in pineapple as the first experience, managing to improve carbon levels in the soil and some nutrients. Then it was applied to tomatoes where it was proven that, nutritionally, it is possible to nourish the crop up to day 60 only with the biopellet, improve carbon levels in soil and control the fungus Fusarium oxysporum. Subsequently, it was applied to coffee seedlings with an organo-mineral formulation. Here, the improvement in the growth and nutrition of the plants was notable, as well as the increase in the microbial activity of the soil. M. pruriens biopellets allow crops to be nourished, allow biocontrolers to be added, improve soil conditions to promote greater microbial activity, reincorporate carbon and CO2 into the soil, are easily applicable, allow dosing and have a favorable shelf-life. They can be applied to all types of crops, both in the greenhouse and in the field.

Keywords: Mucuna pruriens, pellets, carbon, Trichoderma, Fusarium

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58 Fusarium Wilt of Tomato: Plant Growth, Physiology and Biological Disease Management

Authors: Amna Shoaib, Sidrah Hanif, Rashid Mehmood

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Current research work was carried out to check influence of farmyard manure (FYM) in Lycopersicon esculentum L. against Fusarium oxysporum f. sp. lycopersici (FO) in copper polluted soil. Silt-loam soil naturally enriched with 70 ppm of Cu was inoculated with 1 x 106 spore suspensions of FO and incorporated with 0%, 1%, 1.5% or 2% FYM. The multilateral interaction of host-pathogen-metal-organic amendment was assessed in terms of morphology, growth, yield, physiology, biochemistry and metal uptake in tomato plant after 30 and 60 days of sowing. When soil was inoculated with FO, plant growth and biomass were significantly increased during vegetative stage, while declining during flowering stage with substantial increase in productivity over control. Infected plants exhibited late wilting and disease severity was found on 26-50% of plant during reproductive stage. Incorporation of up to 1% FYM suppressed disease severity, improved plant growth and biomass, while it decreased yield. Rest of manure doses was found ineffective in suppressing disease. Content of total chlorophyll, sugar and protein were significantly declined in FO inoculated plants and incorporation of FYM caused significant reduction or no influence on sugar and chlorophyll content, and no pronounced difference among different FYM doses were observed. On the other hand, proline, peroxidase, catalase and nitrate reductase activity were found to be increased in infected plants and incorporation of 1-2% FYM further enhanced the activity of these enzymes. Tomato plant uptake of 30-40% of copper naturally present in the soil and incorporation of 1-2% FYM markedly decreased plant uptake of metal by 15-30%, while increased Cu retention in soil. Present study concludes that lower dose (1%) of FYM could be used to manage disease, increase growth and biomass, while being ineffective for yield and productivity in Cu-polluted soil. Altered physiology/biochemistry of plant in response to any treatment could be served as basis for resistant against pathogen and metal homeostasis in plants.

Keywords: Lycopersicon esculentum, copper, Fusarium wilt, farm yard manure

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57 Isolation and Identification of Novel Escherichia Marmotae Spp.: Their Enzymatic Biodegradation of Zearalenone and Deep-oxidation of Deoxynivalenol

Authors: Bilal Murtaza, Xiaoyu Li, Liming Dong, Muhammad Kashif Saleemi, Gen Li, Bowen Jin, Lili Wang, Yongping Xu

Abstract:

Fusarium spp. produce numerous mycotoxins, such as zearalenone (ZEN), deoxynivalenol (DON), and its acetylated compounds, 3-acetyl-deoxynivalenol (3-ADON) and 15-acetyl-deoxynivalenol (15-ADON) (15-ADON). In a co-culture system, the soil-derived Escherichia marmotae strain degrades ZEN and DON into 3-keto-DON and DOM-1 via enzymatic deep-oxidation. When pure mycotoxins were subjected to Escherichia marmotae in culture flasks, degradation, and detoxification were also attained. DON and ZEN concentrations, ambient pH, incubation temperatures, bacterium concentrations, and the impact of acid treatment on degradation were all evaluated. The results of the ELISA and high-performance liquid chromatography-electrospray ionization-high resolution mass spectrometry (HPLC-ESI-HRMS) tests demonstrated that the concentration of mycotoxins exposed to Escherichia marmotae was significantly lower than the control. ZEN levels were reduced by 43.9%, while zearalenone sulfate ([M/z 397.1052 C18H21O8S1) was discovered as a derivative of ZEN converted by microbes to a less toxic molecule. Furthermore, Escherichia marmotae appeared to metabolize DON 35.10% into less toxic derivatives (DOM-1 at m/z 281 of [DON - O]+ and 3-keto-DON at m/z 295 of [DON - 2H]+). These results show that Escherichia marmotae can reduce Fusarium mycotoxins production, degrade pure mycotoxins, and convert them to less harmful compounds, opening up new possibilities for study and innovation in mycotoxin detoxification.

Keywords: mycotoxins, zearalenone, deoxynivalenol, bacterial degradation

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56 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

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55 Combined Use of Microbial Consortia for the Enhanced Degradation of Type-IIx Pyrethroids

Authors: Parminder Kaur, Chandrajit B. Majumder

Abstract:

The unrestrained usage of pesticides to meet the burgeoning demand of enhanced crop productivity has led to the serious contamination of both terrestrial and aquatic ecosystem. The remediation of mixture of pesticides is a challenging affair regarding inadvertent mixture of pesticides from agricultural lands treated with various compounds. Global concerns about the excessive use of pesticides have driven the need to develop more effective and safer alternatives for their remediation. We focused our work on the microbial degradation of a mixture of three Type II-pyrethroids, namely Cypermethrin, Cyhalothrin and Deltamethrin commonly applied for both agricultural and domestic purposes. The fungal strains (Fusarium strain 8-11P and Fusarium sp. zzz1124) had previously been isolated from agricultural soils and their ability to biotransform this amalgam was studied. In brief, the experiment was conducted in two growth systems (added carbon and carbon-free) enriched with variable concentrations of pyrethroids between 100 to 300 mgL⁻¹. Parameter optimization (pH, temperature, concentration and time) was done using a central composite design matrix of Response Surface Methodology (RSM). At concentrations below 200 mgL⁻¹, complete removal was observed; however, degradation of 95.6%/97.4 and 92.27%/95.65% (in carbon-free/added carbon) was observed for 250 and 300 mgL⁻¹ respectively. The consortium has been shown to degrade the pyrethroid mixture (300 mg L⁻¹) within 120 h. After 5 day incubation, the residual pyrethroids concentration in unsterilized soil were much lower than in sterilized soil, indicating that microbial degradation predominates in pyrethroids elimination with the half-life (t₁/₂) of 1.6 d and R² ranging from 0.992-0.999. Overall, these results showed that microbial consortia might be more efficient than single degrader strains. The findings will complement our current understanding of the bioremediation of mixture of Type II pyrethroids with microbial consortia and potentially heighten the importance for considering bioremediation as an effective alternative for the remediation of such pollutants.

Keywords: bioremediation, fungi, pyrethroids, soil

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54 Post Harvest Fungi Diversity and Level of Aflatoxin Contamination in Stored Maize: Cases of Kitui, Nakuru and Trans-Nzoia Counties in Kenya

Authors: Gachara Grace, Kebira Anthony, Harvey Jagger, Wainaina James

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Aflatoxin contamination of maize in Africa poses a major threat to food security and the health of many African people. In Kenya, aflatoxin contamination of maize is high due to the environmental, agricultural and socio-economic factors. Many studies have been conducted to understand the scope of the problem, especially at pre-harvest level. This research was carried out to gather scientific information on the fungi population, diversity and aflatoxin level during the post-harvest period. The study was conducted in three geographical locations of; Kitui, Kitale and Nakuru. Samples were collected from storage structures of farmers and transported to the Biosciences eastern and central Africa (BecA), International Livestock and Research Institute (ILRI) hub laboratories. Mycoflora was recovered using the direct plating method. A total of five fungal genera (Aspergillus, Penicillium, Fusarium, Rhizopus and Bssyochlamys spp.) were isolated from the stored maize samples. The most common fungal species that were isolated from the three study sites included A. flavus at 82.03% followed by A.niger and F.solani at 49% and 26% respectively. The aflatoxin producing fungi A. flavus was recovered in 82.03% of the samples. Aflatoxin levels were analysed on both the maize samples and in vitro. Most of the A. flavus isolates recorded a high level of aflatoxin when they were analysed for presence of aflatoxin B1 using ELISA. In Kitui, all the samples (100%) had aflatoxin levels above 10ppb with a total aflatoxin mean of 219.2ppb. In Kitale, only 3 samples (n=39) had their aflatoxin levels less than 10ppb while in Nakuru, the total aflatoxin mean level of this region was 239.7ppb. When individual samples were analysed using Vicam fluorometer method, aflatoxin analysis revealed that most of the samples (58.4%) had been contaminated. The means were significantly different (p=0.00<0.05) in all the three locations. Genetic relationships of A. flavus isolates were determined using 13 Simple Sequence Repeats (SSRs) markers. The results were used to generate a phylogenetic tree using DARwin5 software program. A total of 5 distinct clusters were revealed among the genotypes. The isolates appeared to cluster separately according to the geographical locations. Principal Coordinates Analysis (PCoA) of the genetic distances among the 91 A. flavus isolates explained over 50.3% of the total variation when two coordinates were used to cluster the isolates. Analysis of Molecular Variance (AMOVA) showed a high variation of 87% within populations and 13% among populations. This research has shown that A. flavus is the main fungal species infecting maize grains in Kenya. The influence of aflatoxins on human populations in Kenya demonstrates a clear need for tools to manage contamination of locally produced maize. Food basket surveys for aflatoxin contamination should be conducted on a regular basis. This would assist in obtaining reliable data on aflatoxin incidence in different food crops. This would go a long way in defining control strategies for this menace.

Keywords: aflatoxin, Aspergillus flavus, genotyping, Kenya

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53 The Genus Bacillus, Effect on Commercial Crops of Colombia

Authors: L. C. Sánchez, L. C. Corrales, A. G. Lancheros, E. Castañeda, Y. Ariza, L. S. Fuentes, L. Sierra, J. L. Cuervo

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The importance of environment friendly alternatives in agricultural processes is the reason why the research group Ceparium, the Colegio Mayor de Cundinamarca University, Colombia, investigated the genus Bacillus and its applicability for improving crops of economic importance in Colombia. In this investigation, we presented a study in which the genus Bacillus plays a leading role as beneficial microorganism. The objective was to identify the biochemical potential of three indigenous species of Bacillus, which were able to carry out actions for biological control against pathogens and pests or promoted growth to improve productivity of crops in Colombia. The procedures were performed in three phases: first, the production of biomass of an indigenous strain and a reference strain starting from culture media for production of spores and toxins were made. Spore count was done in a Neubauer chamber, concentrations of spores of Bacillus sphaericus were prepared and a bioassay was done at the Laboratory of Entomology at the University Jorge Tadeo Lozano of Plutella xylostella larvae, insect pest of crucifers in several Colombian regions. The second phase included the extraction in the liquid state fermentation, a secondary metabolite that has antibiosis action against fungi, call iturin B, and was obtained from strains of Bacillus subtilis. The molecule was identified using High Resolution Chromatography (HPLC) and its biocontrol effect on Fusarium sp fungus causes vascular wilt in economically important plant varieties, was confirmed using testing of antagonism in Petri dish. In the third phase, an initial procedure in that let recover and identify microorganisms of the genus Bacillus from the rhizosphere in two aromatic herbs, Rosmarinus officinalis and Thymus vulgaris L. was used. Subsequently, testing of antagonism against Fusarium sp were made and an assay was done under greenhouse conditions to observe biocontrol and growth promoting action by comparing growth in length and dry weight. In the first experiment, native Bacillus sphaericus was lethal to 92% Plutella xylostella larvae in 10 DDA. In the second experiment, iturin B was identified and biological control of Fusarium sp was demonstrated. In the third study, all strains demonstrated biological control and the B14 strain identified as Bacillus megaterium increased root length and productivity of the two plants in terms of weight. It was concluded that the native microorganisms of the genus Bacillus has a great biochemical potential that provides a beneficial interactions with plants, improve their growth and development and therefore a greater impact on production.

Keywords: genus bacillus, biological control, PGPRs, biochemical potential

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52 Biological Methods to Control Parasitic Weed Phelipanche ramosa L. Pomel in the Field Tomato Crop

Authors: F. Lops, G. Disciglio, A. Carlucci, G. Gatta, L. Frabboni, A. Tarantino, E. Tarantino

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Phelipanche ramosa L. Pomel is a root holoparasitic weed plant of many cultivations, particularly of tomato (Lycopersicum esculentum L.) crop. In Italy, Phelipanche problem is increasing, both in density and in acreage. The biological control of this parasitic weed involves the use of living organisms as numerous fungi and bacteria that can infect the parasitic weed, while it may improve the crop growth. This paper deals with the biocontrol with microorganism, including Arbuscular mycorrhizal (AM) fungi and fungal pathogens as Fusarium oxisporum spp. Colonization of crop roots by AM fungi can provide protection of crops against parasitic weeds because of a reduction in their seed germination and attachment, while F. oxisporum, isolated from diseased broomrape tubercles, proved to be highly virulent on P. ramosa. The experimental trial was carried out in open field at Foggia province (Apulia Region, Southern Italy), during the spring-summer season 2016, in order to evaluate the effect of four biological treatments: AM fungi and Fusarium oxisporum applied in the soil alone or combined together, and Rizosum Max® product, compared with the untreated control, to reduce the P. ramosa infestation in processing tomato crop. The principal results to be drawn from this study under field condition, in contrast of those reported previously under laboratory and greenhouse conditions, show that both AM fungi and F. oxisporum do not provide the reduction of the number of emerged shoots of P. ramosa. This can arise probably from the low efficacy seedling of the agent pathogens for the control of this parasite in the field. On the contrary, the Rizosum Max® product, containing AM fungi and some rizophere bacteria combined with several minerals and organic substances, appears to be most effective for the reduction of P. ramosa infestation.

Keywords: Arbuscular mycorrhized fungi, biocontrol methods, Phelipanche ramosa, tomato crop

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51 Chemical Composition and Antifungal Activity of Selected Essential Oils against Toxigenic Fungi Associated with Maize (Zea mays L.)

Authors: Birhane Atnafu, Chemeda Abedeta Garbaba, Fikre Lemessa, Abdi Mohammed, Alemayehu Chala

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Essential oil is a bio-pesticide plant product used as an alternative to pesticides in managing plant pests, including fungal pathogens. Thus, the current study aims to investigate the chemical composition and antifungal activities of essential oils (EO) extracted from three aromatic plants i.e., Thymus vulgaris, Coriandrum sativum, and Cymbopogon martini. The leaf parts of those selected plants were collected from the Jimma area and their essential oil was extracted by hydro-distillation method in a Clevenger apparatus. The chemical composition of selected plant essential oil was analyzed by using Gas chromatography-mass spectrometry (GC/MS) and their inhibitory effects were tested in vitro on toxigenic fungi isolated from maize kernel. Chemical analysis results revealed the presence of 32 compounds in C. sativum with Hexanedioic acid, bis (2-ethylhexyl) ester (46. 9%), 2-Decenal, (E)- (12.6), and linalool (8.3%) being the dominant ones. T. vulgaris essential oils constituted 25 compounds, of which thymol (34.4%), o-cymene (17.5%), and Gamma-Terpinene (16.8%) were the major components. Twenty-five compounds were detected in C. martinii of which geraniol (51.4%), Geranyl acetate (14.5%), and Trans – ß-Ocimene (11.7%) were dominant. The EOs of the tested plants had very high antifungal activity (up to 100% efficacy) against Aspergillus flavus, Aspergillus niger, Fusarium graminearum and Fusarium verticillioides in vitro and on maize grains. The antifungal activities of these essential oils were dependent on the major components such as thymol, hexanedioic acid, bis (2-ethylhexyl) ester, and geraniol. The study affirmed the potential of these essential oils controlling as bio-fungicides to manage the effects of potentially toxigenic fungi associated with maize under post-harvest stages. This can reduce the consequences of the health impacts of the mold and toxigenic compounds produced in maize.

Keywords: bio-activity, bio-pesticides, maize, mycotoxin

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