In vitro biopesticide effect of alkaloids and flavonoids of some plants against Fusarium oxysporum

Alkaloids and flavonoids of three plants, namely Withania somnifera, Euphorbia hirta and Terminalia chebula, were tested for their antifungal potential against plant pathogenic fungi Fusarium oxysporum. The spore counting technique was followed for the antibiotic properties of the extracts tested at three different concentrations (2.5, 5 and 10?mg/ml). Results revealed that the bound flavonoid of root of W. somnifera and free flavonoids of stem bark and fruits of T. chebula totally inhibited spore germination of the fungi (100%). The range of minimum inhibitory concentration and minimum fungicidal concentration of extracts recorded was 0.039–0.625 and 0.039–1.25?mg/ml, respectively. Bound flavonoids of selected plants were more effective than free flavonoids and alkaloids. The study suggests that both alkaloids and flavonoids extracts of selected plants have a potential antifungal properties for the control of diseases of various crops.     

Molecular characterization of fungal endophytes from Calotropis procera plants in Taif region (Saudi Arabia) and their antifungal activities

Fungal endophytes were isolated from the leaves of Calotropis procera (Apocynaceae) collected from Taif region (Saudi Arabia). Thirty-three different taxa were recovered. The overall foliar colonization rate was 35.1%. A total of 161 isolates were obtained and identified into 33 distinct operational taxonomic units based on the sequencing of the internal transcribed spacer regions of the rRNA gene. The most prevalent fungi were Aspergillus flavus, Chaetomium globosum, Cochliobolus lunatus, Fusarium dimerum, F. oxysporum, and Penicillium chrysogenum. A total of 161 isolates were tested for antifungal activities against four plant pathogenic fungi (Alternaria alternata, Fusarium oxysporum, Botrytis cinerea, and Pythium ultimum), of which 33 isolates showed antifungal activity against at least one plant pathogenic fungi. Four isolates of Chaetomium globosum and three isolates of Myrothecium verrucaria showed the strongest antifungal activity. This study reported the occurrence of a much wider spectrum of fungi, when compared with previous work. Also, it confirmed the variation of different isolates from the same species in terms of antifungal activity.     

Antifungal activity of herbal extracts against plant pathogenic fungi

The present study was designed to evaluate in vitro antifungal activity of herbal extracts against three plant pathogenic fungi (viz. Rhizopus stolonifer, Botrytis cinerea and Colletotrichum coccodes). Extracts of leaves and rinds of Garcinia indica, rhizomes of Curcuma aromatica, roots of Glycyrrhiza gahliae, leaves of Nyctanthes arbour-tristis and seeds of Vernonia anthelmintica were used for screening. Screening was done using poisoned food technique. Relatively potent extracts were shortlisted from this study and were further assayed to find out their minimum fungicidal concentration (MFC). From the above studies, it was observed that ethyl acetate extract of rhizomes of C. aromatica and unripe fruit rinds of G. indica have shown the lowest MFC values amongst the other tested plant extracts. This study indicates that the potential of these plant extracts in the management of diseases caused by plant pathogenic fungi.     

Antifungal activity of six Saudi medicinal plant extracts against five phyopathogenic fungi

Six medicinal plants such as Amaranthus spinosus, Barbeya oleoides, Clutia lanceolata, Lavandula pubescens, Maerua oblongifolia and Withania somnifera collected from different locations in the southwestern part of Saudi Arabia were tested for antifungal activities against five plant pathogenic fungi causing serious diseases of vegetable crops. These fungi were Alternaria brassicae, Alternaria solani, Botrytis fabae, Fusarium solani and Phytophthora infestans. Aqueous plant extracts reduced mycelial growth and inhibited spore germination of all fungi tested. It is clear that the aqueous extract of Lavandula pubescens leaves was the best for controlling all phytopathogenic fungi under study. These results suggested that medicinal plant extracts play an important role in controlling the phytopathogenic fungi.     

Microbial conversion and in vitro and in vivo antifungal assessment of bioconverted docosahexaenoic acid (bDHA) used against agricultural plant pathogenic fungi

Microbial modification of polyunsaturated fatty acids can often lead to special changes in their structure and in biological potential. Therefore, the aim of this study was to develop potential antifungal agents through the microbial conversion of docosahexaenoic acid (DHA). Bioconverted oil extract of docosahexaenoic acid (bDHA), obtained from the microbial conversion of docosahexaenoic acid (DHA) by Pseudomonas aeruginosa PR3, was assessed for its in vitro and in vivo antifungal potential. Mycelial growth inhibition of test plant pathogens, such as Botrytis cinerea, Colletotrichum capsici, Fusarium oxysporum, Fusarium solani, Phytophthora capsici, Rhizoctonia solani and Sclerotinia sclerotiorum, was measured in vitro. bDHA (5 μl disc⁻¹) inhibited 55.30–65.90% fungal mycelium radial growth of all the tested plant pathogens. Minimum inhibitory concentrations (MICs) of bDHA against the tested plant pathogens were found in the range of 125–500 μg ml⁻¹. Also, bDHA had a strong detrimental effect on spore germination for all the tested plant pathogens. Further, three plant pathogenic fungi, namely C. capsici, F. oxysporum and P. capsici, were subjected to an in vivo antifungal screening. bDHA at higher concentrations revealed a promising antifungal effect in vivo as compared to the positive control oligochitosan. Furthermore, elaborative study of GC-MS analysis was conducted on bioconverted oil extract of DHA to identify the transformation products present in bDHA. The results of this study indicate that the oil extract of bDHA has potential value of industrial significance to control plant pathogenic fungi.     

小桐子枝叶提取物对植物病原真菌的生物活性

采用菌丝生长抑制法,测定了小桐子枝叶6种不同溶剂提取物对小麦赤霉病菌、稻瘟病菌、烟草疫霉菌和辣椒疫霉菌的抑制作用,从中筛选出抑制作用最强的粗提物进行进一步的活性组分分离和抑菌活性测定。结果表明小桐子枝叶的乙醇提取物对4种植物病原菌抑制活性最高,在浓度为0.8 g·L^-1时,小桐子枝叶的乙醇提取物对小麦赤霉病菌、稻瘟病菌、烟草疫霉菌、辣椒疫霉菌菌丝生长抑制率分别为：87.1%、90.3%、86.4%、77.9%,其抗菌活性与农药世高均无显著差异;在小桐子枝叶乙醇提取物的不同溶剂萃取物对稻瘟病菌和烟草疫霉病菌进行生物活性追踪测试中发现,石油醚和水萃取物都具有较好的活性,当浓度为0.8 g·L^-1时,石油醚和水萃取物对两种病菌抑制率都达50%以上。表明小桐子枝叶含有丰富的抗植物病原真菌活性物质,且主要存在于小桐子枝叶乙醇提取物的石油醚相和水相中。     

Effect of extracts from in vitro-grown shoots of <Emphasis Type="Italic">Quillaja saponaria</Emphasis> Mol. on <Emphasis Type="Italic">Botrytis cinerea</Emphasis> Pers.

The ethanolic and aqueous extracts from in vitro shoots of Quillaja saponaria Mol. (Quillay) were studied for their antifungal activity against the phytopathogenic fungus Botrytis cinerea Pers. These extracts reduced conidial germination and mycelial growth of B. cinerea, ethanolic extracts being more active than aqueous extracts. In addition, the damage areas produced by this fungus on tomato leaves and strawberry fruits pre-treated with quillay extracts were diminished. The fungitoxic effect of in vitro-grown quillay extract was similar to those obtained with commercial fungicides of both natural (BC-1000) and synthetic (iprodione–dicarboximide) origin. On the other hand, the antifungal action of quillay extracts obtained from adult trees naturally grown was only slightly superior to the fungitoxic activity of the extract from in vitro plants. HPLC analysis of the extract showed that it contained saponins and some phenolic compounds such as chlorogenic, caffeic, vanillic, and salicylic acids, and scopoletin, which have been identified as antifungal agents on phytopathogenic fungi. The results obtained in this work, suggests that extracts of in vitro-grown quillay have an important protective effect against B. cinerea and support the use of an in vitro culture system as a biotechnological alternative to obtain environmental safe antifungal quillay extracts to control B. cinerea, contributing to the preservation of this indigenous Chilean species.     

Purification and characterization of antifungal compounds from Bacillus coagulans TQ33 isolated from skimmed milk powder

Bacillus coagulans TQ33, isolated from skimmed milk powder, displays strong antifungal activity against plant pathogenic fungi. The antifungal compound of the B. coagulans TQ33 culture was extracted by thin-layer chromatography and column chromatography, and its structure was elucidated based on HPLC, LC-MS, and NMR analysie. The antifungal compound was identified as phenyllactic acid (PLA), and it was found to have a minimum inhibitory concentration on Phytophthora drechsleri Tucker of 18 mg/mL. Bio-control activity tests indicated that PLA has a wide spectrum of antagonistic effects against Fusarium oxysporum, Botrytis cinerea, Glomerella cingulata, Penicillium citrinum, Penicillium digitatum, particularly against F. oxysporum. PLA is the most notable antimicrobial compound with broad and effective antimicrobial activity against both bacteria and fungi that has been isolated and identified to date. These results indicate that B. coagulans TQ33 has the potential for application in biological pesticides.     

Molecular Phylogeny,Diversity, and Bioprospecting of Endophytic Fungi Associated with wild Ethnomedicinal North American Plant Echinacea purpurea (Asteraceae)

The endophytic fungal community associated with the ethnomedicinal plant Echinacea purpurea was investigated as well as its potential for providing antifungal compounds against plant pathogenic fungi. A total of 233 endophytic fungal isolates were obtained and classified into 42 different taxa of 16 genera, of which Alternaria alternata, Colletotrichum dematium, and Stagonosporopsis sp. 2 are the most frequent colonizers. The extracts of 29 endophytic fungi displayed activities against important phytopathogenic fungi. Eight antifungal extracts were selected for chemical analysis. Forty fatty acids were identified by gas chromatography‐flame‐ionization detection (GC‐FID) analysis. The compounds (–)‐5‐methylmellein and (–)‐(3R)‐8‐hydroxy‐6‐methoxy‐3,5‐dimethyl‐3,4‐dihydroisocoumarin were isolated from Biscogniauxia mediterraneaEPU38CA crude extract. (–)‐5‐Methylmellein showed weak activity against Phomopsis obscurans, P. viticola, and Fusarium oxysporum, and caused growth stimulation of C. fragariae, C. acutatum, C. gloeosporioides, and Botrytis cinerea. (–)‐(3R)‐8‐Hydroxy‐6‐methoxy‐3,5‐dimethyl‐3,4‐dihydroisocoumarin appeared slightly more active in the microtiter environment than 5‐methylmellein. Our results indicate that E. purpurea lives symbiotically with different endophytic fungi, which are able to produce bioactive fatty acids and aromatic compounds active against important phytopathogenic fungi. The detection of the different fatty acids and aromatic compounds produced by the endophytic community associated with wild E. purpurea suggests that it may have intrinsic mutualistic resistance against phytopathogen attacks in its natural environment.     

Essential oil composition of Angelica archangelica L. (Apiaceae) roots and its antifungal activity against plant pathogenic fungi

This study reports the results of gas chromatography–mass spectrometry (GC–MS) analyses of the essential oil of Angelica archangelica L. (Apiaceae) roots, as well as its in vitro antifungal activity against 10 plant pathogenic fungi. Moreover, the essential oil was evaluated for its antifungal activity using the agar dilution method, and also minimum inhibitory concentrations and minimum fungicidal concentrations were determined. The major compounds identified by GC–MS were α-pinene (21.3%), δ-3-carene (16.5%), limonene (16.4%), and α-phellandrene (8.7%). The oil showed in vitro antifungal activity against some species of the Fusarium genus, Botrytis cinerea, and Alternaria solani. Our study indicates that the oil of A. archangelica could be used as a control agent for plant pathogenic fungi in natural formulations.     

Antimicrobial activities of extracts of Cyperus rotundus L. rhizomes against some bacterial and fungal pathogens of strawberry and tomato

Antimicrobial activities of rhizome extracts of Cyperus rotundus were investigated on selected plant pathogenic bacteria and fungi. Ethyl acetate and hexane extracts showed antibacterial activity against three isolates of Clavibacter michiganensis subsp. michiganensis at concentrations of 900 and 1000 μg/ml. However, Gram-negative bacterial pathogens of tomato; Pseudomonas syringae pv. tomato and Ralstonia solanacearum were not inhibited from the extracts. Ethyl acetate extracts at 100 μg/ml inhibited mycelial growth and spore germination of the two strawberry isolates of Botrytis cinerea; however, no significant inhibition was found in tomato fungal pathogen, Fusarium oxysporum f. sp. radicis lycopersici. Minimum inhibitory concentrations were determined as 0.0625 and 0.125 mg/ml against C. michiganensis subsp. michiganensis for ethyl acetate and hexane extracts of rhizomes, respectively. This study shows the potentials of extracts of C. rotundus rhizomes as antimicrobial agents that are effective against the tested plant pathogenic bacteria and fungi.     

Antimycotic activities of Cinnamon-derived compounds against <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> in vitro

In this study, the effects of medicinal plant extracts on the development of mycelium in the following phytopathogenic fungi were evaluated: Phytophthora capsici, Rhizoctonia solani, Fusarium solani, Colletotrichum gloeosprorioides, and Botrytis cinera. Of the 26 medicinal plants tested, six plant extracts showed antifungal activity against phytopathogenic fungi. The highest antifungal activity was exerted against R. solani by the n-hexane fraction of a Cinnamon (Cinnamomum cassia Blume) solvent extract. Therefore, the antifungal compound fractions I and II were purified from the n-hexane fraction by TLC on silica gel plates. When treated with solutions containing compound fractions I or II at a concentration of 2%, the mycelia growth rate of R. solani was reduced to 0.19 and 0.18, respectively. In addition, microscopic observation of the hyphal morphology of R. solani following treatment with compound fraction I revealed the presence of severely damaged hyphae. Specifically, the hyphal tips became swollen, collapsed or were completely destroyed in response to treatment with solution containing compound fraction I at concentration of 1%.     

Assessment of the bioregulatory activity of the leaf juices of higher plants in Al-Taif,Saudi Arabia against Fusarium solani,Phytophthora spp. and Rhizoctonia solani

Resistance to conventional fungicides causes poor disease control in agriculture. Natural products from plants have great potential as novel fungicide sources for controlling pathogenic fungi. In this study the antipathogenic activity of the leaf juices of 11 plant species (Chenopodium ambrosioides, Pulicaria vulgaris, Lavandula pubescens, Lavandula dentata, Ageratum conyzoides, Ficus retusa, Zizyphus nummularia, Acacia tortilis, Phragmanthera sp. Aff. Rufescens, Lawsonia alba and Olea europaea) were evaluated in vitro against three plant pathogenic fungi (Fusarium solani, Phytophthora spp. and Rhizoctonia solani). Plate assays showed that the leaf aqueous juices have antifungal activity against these fungi. The aqueous extracts of Pulicaria vulgaris, Lavandula dentata, Ageratum conyzoides, Ficus retusa, Zizyphus nummularia, Acacia tortilis, Phragmanthera sp. Aff. Rufescens (when associated with Acacia tortilis), Lawsonia alba and Olea europaea exhibited antifungal properties against Fusarium solani, Phytophthora spp. and Rhizoctonia solani with variable degrees. On the other hand, Chenopodium ambrosioides, Lavandula pubescens and Phragmanthera sp. Aff. Rufescens (when associated with Zizyphus nummularia) did not exhibit any fungitoxicity. All these observations suggest the possible exploitation of Chenopodium oil as a potential botanical fungitoxicant in ecofriendly control of post-harvest biodeterioration of food commodities from storage fungi.     

黄花倒水莲内生真菌生物活性评价及HNLF-44菌株鉴定

为充分开发黄花倒水莲(Polygala fallax)的内生真菌资源，获得具有抗植物病原真菌、抗氧化活性的内生真菌，该文以黄花倒水莲内生真菌为研究对象，使用平板对峙法检测内生真菌对6种植物病原真菌的抑菌活性，测定内生真菌发酵液的DPPH清除自由基能力和总还原能力，评价内生真菌的抗氧化活性，并对具有强抑菌活性和抗氧化活性的菌株进行形态和ITS鉴定。结果表明：(1)黄花倒水莲内生真菌中有2株内生真菌对香蕉专化尖孢镰刀菌、柑橘树脂病菌、叶点霉菌、香蕉具条叶斑病菌、茄病镰刀菌、三七根腐病菌具有明显的抑菌活性，抑菌率在50.3%～91.4%之间，其中HNLF-5对柑橘树脂病菌的抑菌率为73.2%,HNLF-44对香蕉专化尖孢镰刀菌抑菌率为91.4%。(2)内生真菌发酵液具有良好的抗氧化活性，DPPH清除率均在80%以上，总还原能力吸光值范围为0.279 2～0.748 8。(3)HNLF-44菌株为链格孢属真菌。该研究表明，药用植物黄花倒水莲内生真菌具有较好的生物活性，为后续从黄花倒水莲内生真菌中挖掘潜在新型抑菌活性和抗氧化活性物质奠定了基础。     

Synthesis,Antifungal Activity,DFT Study and Molecular Dynamics Simulation of Novel 4-(1,2,4-Oxadiazol-3-yl)-N-(4-phenoxyphenyl)benzamide Derivatives

In order to find novel potential antifungal agrochemicals, a series of new 4-(1,2,4-oxadiazol-3-yl)-N-(4-phenoxyphenyl)benzamide derivatives 3a – j were designed, synthesized and characterized by their ¹H - , ¹³C-NMR and HRMS spectra. The preliminary antifungal assay in vitro revealed that compounds 3a – j exhibited moderate to good antifungal activity against five plant pathogenic fungi. Especially, compound 3e presented significant antifungal activity against Alternaria solani, Botrytis cinerea and Sclerotinia sclerotiorum, superior to positive control boscalid. In the in vivo antifungal assay on tomato plants and cucumber leaves, compound 3e presented good inhibition rate against B. cinerea at 200 mg/L. Molecular dynamics simulation revealed that compound 3e could bind with the active site of class II histone deacetylase (HDAC).     

In vitro assessment of fungicidal activity of Ageratum conyzoides and Cybopogon flexuosus weed extracts against some phytopathogenic fungi associated with fruit rot of water melon (Citrullus lunatus (Thunb.))

The ethanolic extracts of Cybopogon flexuosus and Ageratum conyzoides were tested at concentrations of 20, 40, 60, 80, 100 and 120?mg/ml for their in vitro fungicidal activities against five phytopathogenic fungi isolated from diseased watermelon fruits. The pathogens were Fusarium verticillioides, Aspergillus flavus, Botryodiplodia theobromae, Curvularia lunata and Alternaria cucumeria – Amans as confirmed by pathogenicity tests. All isolated pathogenic fungi were significantly (p?≥?0.05) highly pathogenic with the exception of A. cucumeria which had the least significant (p?≥?0.05) pathogenicity. The inhibitory effects of the extracts increased significantly (p?≥?0.05) with increase in concentrations. Some of the concentrations reduced the mycelial growth of the pathogens to a significant (p?≥?0.05) level. Very strong fungicidal activity was produced by extracts of A. conyzoides at 100?mg/ml against all the fungi. The inhibitory effects of C. flexuosus extracts at 20, 40 and 60?mg/ml were greater than those of C. flexuosus on A. flavus, F. verticillioides and A. cucumeria. The results of the investigation indicated that plant extracts possess antifungal activity that can be exploited as an ideal treatment for future plant disease management in the control of rot of water melon.     

1‐Phenyl‐3‐pentanone,a Volatile Compound from the Edible Mushroom Mycoleptodonoides aitchisonii Active Against Some Phytopathogenic Fungi

Volatiles produced by mycelia of mushrooms with aromatic odour were investigated for their antifungal activity against plant‐pathogenic fungi. The results of the screening of 23 species of basidiomycetes revealed that volatile substances from mycelia of Mycoleptodonoides aitchisonii (TUFC10099), an edible mushroom, strongly inhibited the mycelial growth, spore germination and lesion formation on host leaves of some plant‐pathogenic fungi including Alternaria alternata, A. brassicicola, A. brassicae, Colletotrichum orbiculare and Corynespora cassiicola. The volatile compounds were isolated from the culture filtrate of M. aitchisonii, and 1‐phenyl‐3‐pentanone was identified as a major antifungal volatile. The compound had significantly inhibitory activity against plant‐pathogenic fungi at 35 ppm. This is the first report that the volatile compound produced by mycelia of M. aitchisonii has antifungal activity against plant‐pathogenic fungi.     

Effect of seven Indian plant extracts on Fenton reaction-mediated damage to DNA constituents

The influences of substoichiometric amounts of seven plant extracts in the Fenton reaction-mediated damage to deoxynucleosides, deoxynucleoside monophosphates, deoxynucleoside triphosphates, and supercoiled plasmid DNA were studied to rationalize anticancer properties reported in some of these extracts. Extracts from Acacia catechu, Emblica officinalis, Spondias dulcis, Terminalia belerica, Terminalia chebula, as well as gallic acid, epicatechin, chebulagic acid and chebulinic acid enhance the extent of damage in Fenton reactions with all monomeric substrates but protect supercoiled plasmid DNA, compared to standard Fenton reactions. The damage to pyrimidine nucleosides/nucleotides is enhanced by these extracts and compounds to a greater extent than for purine ones in a concentration dependent manner. Dolichos biflorus and Hemidesmus indicus extracts generally do not show this enhancement for the monomeric substrates though they protect plasmid DNA. Compared to standard Fenton reactions for deoxynucleosides with ethanol, the presence of these five plant extracts render ethanol scavenging less effective as the radical is generated in the vicinity of the target. Since substoichiometric amounts of these extracts and the four compounds produce this effect, a catalytic mechanism involving the presence of a ternary complex of the nucleoside/nucleotide substrate, a plant compound and the hydroxyl radical is proposed. Such a mechanism cannot operate for plasmid DNA as the planar rings in the extract compounds cannot stack with the duplex DNA bases. These plant extracts, by enhancing Fenton reaction-mediated damage to deoxynucleoside triphosphates, slow down DNA replication in rapidly dividing cancer cells, thus contributing to their anticancer properties.     

Anti-fungal properties of 43 plant species against Alternaria solani and Botrytis cinerea

Investigation of plants containing natural anti-microbial metabolites for plant protection has been identified as a desirable method of disease control. Crude methanolic extracts of 43 plant species belonging to 27 families, which most of them are medicinal plants, mostly collected from the west of Iran were screened for anti-fungal activity against two economically important phytopathogenic fungi, Alternaria solani and Botrytis cinerea during 2010–2012. Bioassay of the extracts was conducted by agar diffusion method on agar plate cultures with five replications. Among all the 43 plant methanolic extracts, mycelia growth of A. solani and B. cinerea was reduced by 28 (65%) and 30 (70%) plant extracts when compared to the control, respectively. The strongest extracts with more than 50% inhibition against A. solani were Elaeagnus angustifolia, Dodonaea viscosa, Haplophyllum perforatum and inflorescence of Allium hirtifolium, respectively. Leaves of A. hirtifolium, H. perforatum, inflorescence of A. hirtifolium and D. viscosa showed highest inhibitory effect (≥50%) against B. cinerea. Moreover, complete inhibition of leaves of A. hirtifolium against B. cinerea was due to their fungistatic activity. The results of this experiment and high number of plants with anti-fungal activity showed that the flora in the west of Iran could be regarded as a rich source of plants with anti-fungal activity. Therefore, further screening of other plant species, identifying active fractions or metabolites and in vivo application of active extracts are warranted.