Myrothins A–F from Endophytic Fungus Myrothecium sp. BS-31 Harbored in Panax notoginseng

Endophytic fungi play important roles for host's stress tolerance including invasion by pathogenic microbes. Small molecules are common weapons in the microbe–microbe interactions. Panax notoginseng is a widely used traditional Chinese medicinal plant and harbors many endophytes, some exert functions against pathogens. Here, we report six new compounds named myrothins A–F ( 1 – 6 ) produced by Myrothecium sp. BS-31, an endophyte isolated from P. notoginseng, and their antifungal activities against pathogenic fungi causing host root-rot disease. Their structures were elucidated with analysis of spectroscopic data including 1D and 2D NMR, HR-ESI-MS. Myrothins B ( 2 ) and E ( 5 ) showed the weak activity against Fusarium oxysporum and Phoma herbarum, and myrothins F ( 6 ) showed weak activity against F. oxysporum.     

Antifungal Effect of Essential Oils from Five Kinds of Rutaceae Plants – Avoiding Pesticide Residue and Resistance

Panax notoginseng root is a traditional Chinese herb, of which the yield and quality have been seriously affected by microorganisms, and is commonly used to treat various kinds of bleeding. In this experiment, the effects of the antifungal properties of essential oils (EOs) from five kinds of Rutaceae plants on the growth of three kinds of pathogens were studied to develop natural, environmentally friendly antifungal agents. Citrus medica EO was found to have stronger inhibitory effects on the growth of pathogenic fungi in vitro than other EOs with the Oxford cup method, of which the chemical composition was further investigated by GC/MS. The major components were d ‐limonene (22.79 %) and γ‐terpinene (9.71 %). The antifungal activities were evaluated by MIC and FIC assays. In these assays, C. medica EO, d ‐limonene and γ‐terpinene were effective against three pathogens of P. notoginseng with MIC values ranging from 0.12 to 12.05 mg/mL. The association between hymexazol and C. medica EO showed a high synergistic effect with lower FIC index values (FICi=0.31–2.00). Furthermore, C. medica EO was further assessed in P. notoginseng planted in a continuous cropping soil (CCS) and was found to reduce the disease incidence and disease severity compared with P. notoginseng planted in CCS only without EO addition. This finding suggested that C. medica EO has potential as a natural environmentally antifungal agent against pathogens of P. notoginseng, ensuring its safety.     

First Report of <i>Fusarium striatum</i> Causing Root Rot Disease of <i>Panax notoginseng</i> in Yunnan,China

Panax notoginseng is a traditional Chinese medicinal plant. Root rot of P. notoginseng is one of the most serious diseases affecting P. notoginseng growth and causes wilted leaves, fewer lateral roots and rotten roots. Root rot is a soil-borne disease, and mainly occurs from June to August in Yunnan Province when the temperatures are high and the air is humid. In this study, the endophytic fungal genus Fusarium isolate E-2018.1.22-#3.2 was obtained from a P. notoginseng embryo. Fusarium isolate E-2018.1.22-#3.2 was identified as Fusarium striatum based on morphological characteristics and molecular analysis. The fungus was found to have conidiophores and macroconidia, and its ITS, LSU and TEF-1α genes shared 100%, 99.2% and 99% identities with the homologous genes of Fusarium striatum, respectively. Isolate F. striatum E-2018.1.22-#3.2 can cause root rot symptoms, including black, soft roots, fewer lateral roots and leaf wilt, in 93% of the experimental P. notoginseng plants, and could be re-isolated, fulfilling Koch’s postulates. When the P. notoginseng plants were treated with the fungicide pyraclostrobin, isolate F. striatum E-2018.1.22-#3.2 was unable to cause root rot. We have therefore demonstrated that F. striatum E-2018.1.22-#3.2 is able to cause root rot disease in P. notoginseng. This is the first report of root rot disease caused by F. striatum on P. notoginseng in China.     

Fungicidal Activity of Essential Oils from Cinnamomum cassia against the Pathogenic Fungi of Panax notoginseng Diseases

The frequent disease of Panax notoginseng caused by the pathogenic fungi in field cultivation has become the major threaten to the sustainable development of it. The present study was conducted to find natural agent with potential inhibition against pathogen. Therefore, the inhibitory effects of Cinnamomum cassia (L.) J.Presl essential oils (EOs) against P. notoginseng associated pathogenic fungi were conducted both in vitro and in vivo experiments. The results of the Oxford cup test revealed that C. cassia dry bark EO (50 mg/mL) had significant inhibitory activity on the growth of all tested fungi, and the growth of various pathogens was completely inhibited, except for that of Fusarium solani. Therefore, the constituents of C. cassia EOs were analyzed by GC/MS, and the research demonstrated that the main constituents of C. cassia dry bark EO were trans‐cinnamaldehyde (75.65 %), (E)‐2‐methoxycinnamaldehyde (6.08 %), cinnamaldehyde (3.47 %) and cinnamyl acetate (1.02 %). The MIC results showed that C. cassia dry bark EO and the main compounds had good antifungal effect on the tested strains, and the inhibitory effect was similar to that of hymexazol (chemical pesticide). By analyzing the value of the fraction inhibitory concentration index (FICI), additive effects, irrelevant effects and synergistic effects were observed after the mixture of hymexazol against various pathogens. Moreover, in vivo model showed that C. cassia dry bark EO could reduce the occurrence of anthrax in P. notoginseng. To widen the resources of C. cassia available, the compositions of both C. cassia fresh bark and leaf EOs were also tested and many common compositions existed among them. Taken together, it was concluded that C. cassia EO had the potential use in the field to reduce the pathogenic disease.     

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.     

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。     

Pathogenic fungi in garlic seed cloves and first report of Fusarium proliferatum causing cloves rot of stored bulbs in upper Egypt

Isolation trails from garlic seed cloves of certain cultivars commercially distributed and rotted cloves of stored Baladi cultivar during 2010 and 2011 in different regions of Sohag, Egypt resulted in detection of four fungal genera Aspergillus, Botrytis, Fusarium and Penicillium. Moreover, Fusarium spp. was the most dominant fungi. Koch’ postulates were performed on cloves, seedlings and potted garlic plants. Results showed that isolates of F. oxysporum, F. proliferatum and F. solani were superior to other tested fungi and induced the highest cloves rot (CR). F. oxysporum highly reduced clove germination (CG), produced extensive seedlings damping-off and induced highly disease severity index of rotted roots/cloves followed by F. solani. Unexpectedly, it was found that F. proliferatum is contaminated and colonised seed cloves, extremely reduced CG and following harvesting caused the highest CR of stored bulbs. To our knowledge, this is first notice for occurrence of F. proliferatum causing CR of stored garlic bulbs in Egypt.     

Molecular characterisation of Fusarium oxysporum causing rot diseases in small cardamom (Elettaria cardamomum Maton)

Incidence of root rot and foliar yellowing, rhizome rot, panicle wilt and stem rot diseases of small cardamom (Elettaria cardamomum Maton) are caused by Fusarium oxysporum Schlecht., and were surveyed in the high ranges of Idukki district, Kerala during 2010–2011. The diseases were noticed in different areas to varying degrees. Root rot was found to be most severe, followed by pseudostem rot, rhizome rot and panicle wilt. The Fusarium infections were prevalent throughout the year (January–December) and varied from 1.5 to 10.6%. Even though the pathogen was isolated from different plant parts, during pathogenicity studies, all the isolates could cross-infect other plant parts too. Twenty different isolates of F. oxysporum were obtained from diseased samples, and five morphologically distinct isolates were analysed with Randomly Amplified Polymorphic DNA (RAPD) markers to study the genetic variability, if any, among them. PCR amplification of total genomic DNA with random oligonucleotide primers generated unique banding patterns, depending upon primers and isolates. Nine oligunucleotide primers were selected for the RAPD assays, which resulted in 221 bands for the five isolates of F. oxysporum. The number of bands obtained was entered into an NTSYS, and the results showed moderate genetic variability among F. oxysporum isolates causing root rot, rhizome rot, panicle wilt and pseudostem rot, collected from different locations. The dendrogram of different isolates into groups resulted in one major cluster at 0.61 similarity index comprising of four isolates (CRT 3, CRR 3, CPW 2 and CSR 1) and one isolate (CRT 5) formed in a separate cluster. Among the five isolates of F. oxysporum, CRT 5 was entirely different from the other four isolates. The isolates also differ according to the geographical area, as revealed from the genetic variability observed in different root rot isolates (CRT 3 and CRT 5). It is inferred that despite moderate variability, F. oxysporum, infecting small cardamom in Idukki district of Kerala, consists of a single clonal lineage.     

Analysis of interspecific relationships between Funneliformis mosseae and Fusarium oxysporum in the continuous cropping of soybean rhizosphere soil during the branching period

This study analysed the interspecific relationships between the dominant arbuscular mycorrhizal (AM) fungus, Funneliformis mosseae, and the major soybean root rot pathogen, Fusarium oxysporum, in the rhizosphere soil of continuous cropped soybean. Our aim was to provide theoretical evidence on the AM fungi to overcome the obstacles of soybean continuous cropping. We selected soybean cultivars, including Kenfeng 16 (an intermediate cultivar), Heinong 44 (a high-fat cultivar) and Heinong 48 (a high-protein cultivar), and sowed in the soybean continuous cropping soil under different treatments. The infection status of the soybean roots during the branching period by Fu. mosseae and F. oxysporum was estimated using the standard polymerase chain reaction method, as well as their colonisation status in rhizosphere soil. The AM fungal colonisation rates and F. oxysporum disease incidence of soybean roots were determined, respectively. Quantitative polymerase chain reaction was applied to analyse the DNA content of Fu. mosseae and F. oxysporum to investigate the relationship between Fu. mosseae and F. oxysporum. The results show that both Fu. mosseae and F. oxysporum can infect the soybean roots during the branching period and colonise the rhizosphere. However, the DNA content of F. oxysporum clearly decreased in soybean root and rhizosphere samples after the inoculation with Fu. mosseae. In addition, the disease incidence of F. oxysporum significantly decreased after inoculation with Fu. mosseae, which might indicate inhibitive effects of Fu. mosseae over F. oxysporum.     

Radiometric assessment of tillage and seed treatment effect on soybean root rot caused by Fusarium spp. in central Minnesota

Soybean root rot, caused primarily by Fusarium solani f. sp. phaseoli in a complex with F. oxysporum and Rhizoctonia solani, has become an increasing problem for soybeans, dry beans, and other rotation crops in central Minnesota due to soil conditions associated with reduced tillage. This study was conducted, in two field sites in central Minnesota located near Staples and Verndale, to develop methods for nondestructive assessment of root rot severity using plant radiometric properties. Soybean canopy reflectance was measured with a hand-held multi-spectral radiometer. Prior to the radiometer measurements, attempts were made to create differing root rot situations with moldboard or chisel tillage, and with or without a biological seed treatment. Root rot severity was estimated using a visual disease severity scale. Colony-forming units (CFU) were determined to estimate soil populations of pathogenic F. solani and F. oxysporum. Results from the Verndale site consistently showed significant treatment effects in the measured canopy radiometric parameters, and in the visual disease rating and yield (significant for seed treatment). Values of a simple ratio vegetation index from this site exhibited negative relationships with disease rating and F. oxysporum CFU, and a positive linear relationship with yield. Treatment effects were generally not significant at the Staples site because of low initial F. oxysporum populations. The results indicate that remote sensing is potentially a rapid, nondestructive means for assessment of root rot diseases in soybean.     

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.     

Microconidia germination of the tomato pathogen Fusarium oxysporum in the presence of root exudates

Abstract

In this study we assessed microconidia germination of the tomato pathogens F. oxysporum f. sp. lycopersici (Fol) and F. oxysporum f. sp. radicis-lycopersici (Forl) in the presence of root exudates. Tomato root exudates stimulated microconidia germination and the level of stimulation was affected by plant age. Treatment of root exudates with insoluble polyvinylpolypyrrolidone, which binds phenolic compounds, indicated that tomato root exudates contain phenolic compounds inhibitory to F. oxysporum microconidia germination. Our study indicates that tomato root exudates similarly stimulate microconidia germination of both Fol and Forl. However, individual F. oxysporum strains differ in the degree of germination response to the root exudates. Furthermore, root exudates from non-host plants also contain compounds that stimulate microconidia germination of Fol. In general, the effects of root exudates from non-host plants did not differ considerably from those of tomato. The ability of phenolic compounds to inhibit germination of Fol seems not to be plant-specific.     

Isolation and characterization of mutants of <Emphasis Type="Italic">Pseudomonas maltophila</Emphasis> PM-4 altered in chitinolytic activity and antagonistic activity against root rot pathogens of clusterbean (<Emphasis Type="Italic">Cyamopsis tetragonoloba</Emphasis>)

Pseudomonas maltophila PM-4, an antagonist of pathogenic fungi including Rhizoctonia bataticola, R. solani, Fusarium oxysporum and Sclerotinia sclerotiorum associated with root rot of clusterbean (Cyamopsis tetragonoloba) was mutagenized with Tn5. Hyperchitinase producing mutants showing large zone of colloidal chitin dissolution were identified on medium containing calcoflor dye as an indicator. A mutant P-48 producing 137% higher chitinase activity than the parent strain PM-4 was identified. Seed bacterization of clusterbean (Cyamopsis tetragonoloba) with P-48 controlled the root rot upto 40.8% in the presence of conglomerate of all the four fungal pathogens Rhizoctonia bataticola, R. solani, F. oxysporum and Sclerotinia sclerotiorum.     

中药提取物对黄芪根腐病菌的抑制效果

打破了传统的化学药剂(特别是农药)防治黄芪根腐病菌的局限,以发展绿色防治中药材病害为前提,采用蛇床子、知母的提取液制备含药培养基,并以灭菌水作为对照,分别接种黄芪根腐病菌,探讨这两种中药对黄芪根腐病菌—尖孢镰刀菌(Fusarium oxysporum)的抑制作用。研究结果表明,2种中药提取物对抑制黄芪根腐病菌均有不同程度的影响,实验结果表明,无论从药剂类型还是药剂浓度来看,蛇床子防治黄芪根腐病菌的效果最佳,具有显著水平,其最高抑制率为96.13%。     

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.     

Effect of mycorrhiza and biofertilisers on reducing the incidence of Fusarium root and pod rot diseases of peanut

Pathogenicity tests of twenty-six fungal isolates were tested on peanut plants (Giza 5 cv.) and the results revealed that, Fusarium oxysporum isolate (No. I) followed by F. solani (No. II) then F. moniliforme (No III) significantly caused highest incidence of root rot disease. Also, F. moniliforme (No III) followed by F. solani (No II) then F. oxysporum (No I) gave the highest incidence of pod rot disease. The effectiveness of vescular arbuscular-mycorrhiza (VAM) at different application rates on the incidence of root rot, pod rot diseases and plant growth parameters of peanut was studied. All soil treatments with each rate of VAM significantly reduced root and pod rot diseases compared with control (rate 0%). The best reduction in the severity of both diseases with VAM was found at the rate of 3%. Application of rhizobacterin, microbin and cerialin biofertilisers at the different concentrations decreased the severity of both root rot and pod rot severity diseases compared with non-treated seeds. The greatest reduction in both diseases was achieved at a concentration of 8/100?g seeds. The highest number of pods and fresh weight (g) was achieved in seed supplemented with each biofertiliser at concentration of 8/100?g seed.     

Utilisation of biocides for controlling pest attacks on potato tubers

Essential oils of fennel, peppermint and caraway were formulated in the form of emulsifiable concentrates. Different concentrations of the formulated oils were prepared. The prepared concentrations were tested in vivo for their inhibitory activity against the growth of Fusarium oxysporum (the casual agent of dry rot on potato). The results showed that different concentrations of the formulated fennel, peppermint and caraway oils exerted highly toxicity against the inoculated fungus. In addition, all formulated oils with different forms showed high activity for controlling the decay when applied as protective and therapeutic agents.     

The synthetic strigolactone GR24 influences the growth pattern of phytopathogenic fungi

Strigolactones that are released by plant roots to the rhizosphere are involved in both plant symbiosis with arbuscular mycorrhizal fungi and in plant infection by root parasitic plants. In this paper, we describe the response of various phytopathogenic fungi to the synthetic strigolactone GR24. When GR24 was embedded in the growth medium, it inhibited the growth of the root pathogens Fusarium oxysporum f. sp. melonis, Fusarium solani f. sp. mango, Sclerotinia sclerotiorum and Macrophomina phaseolina, and of the foliar pathogens Alternaria alternata, Colletotrichum acutatum and Botrytis cinerea. In the presence of this synthetic strigolactone, intense branching activity was exhibited by S. sclerotiorum, C. acutatum and F. oxysporum f. sp. melonis. Slightly increased hyphal branching was observed for A. alternata, F. solani f. sp. mango and B. cinerea, whereas suppression of hyphal branching by GR24 was observed in M. phaseolina. These results suggest that strigolactones not only affect mycorrhizal fungi and parasitic plants, but they also have a more general effect on phytopathogenic fungi.     

Cloning and Sequence Analysis of the Endopolygalacturonase Gene from the Pitch Canker Fungus, Fusarium circinatum

The fungus Fusarium circinatum causes pitch canker disease on mature pine trees and root rot and damping-off of pine seedlings. Endopolygalacturonases (endoPGs) play a major role during penetration of plants by fungi. Digestion of the pectic polysaccharides in the plant primary cell walls is one of the earliest functions of endoPGs during infection. The research objective was to clone and characterize the gene encoding endopolygalacturonase in F. circinatum. A 970-bp DNA fragment was cloned by using degenerate PCR amplification from F. circinatum DNA. Sequence data for this fragment were used to design specific primers for use in genome walking to amplify and sequence the remaining portion of the F. circinatum endoPG gene (Fcpg). The amino acid sequence predicted from this gene showed 90% and 87% similarity to Fusarium oxysporum and Fusarium moniliforme endoPGs, respectively. Received: 10 August 2000 / Accepted: 30 October 2000     

Determination of a Bacillus velezensis strain for controlling soybean root rot

Bacteria were obtained from soybean rhizosphere soil. And control effect of a tested isolate for controlling soybean root rot infected by Fusarium oxysporum was evaluated. The selected bacterial isolate exhibited the greatest inhibition of F. oxysporum in the laboratory and substantially reduced soybean root rot in pot-controlled assays.