Combination of Pseudomonas aeruginosa and Pochonia chlamydosporia for Control of Root-Infecting Fungi in Tomato

A plant growth‐promoting rhizobacterium, Pseudomonas aeruginosa strain IE‐6, and a fungal antagonist, Pochonia chlamydosporia, were tested for their ability to inhibit mycelial growth of root‐infecting fungi under laboratory conditions including Macrophomina phaseolina, Fusarium oxysporum, F. solani and Rhizoctonia solani. Biocontrol effectiveness of the bacterium and the fungus alone or in combination was also determined for the control of root‐infecting fungi under field conditions. In a dual‐culture plate assay, the colonies of P. chlamydosporia and P. aeruginosa met each other and no further growth of either organism occurred. Against M. phaseolina, F. solani and R. solani, an ethyl acetate extract of the culture filtrates of P. aeruginosa inhibited fungal growth greater than the hexane extract, but against F. oxysporum the hexane extract caused greater inhibition of fungal growth. By contrast, against M. phaseolina, F. oxysporum and F. solani, the hexane extract of P. chlamydosporia was more effective in the inhibition of fungal growth than the ethyl acetate fraction. Ethyl acetate extracts of P. aeruginosa at 1.0 mg/ml not only inhibited the radial colony growth of R. solani but also lysed the fungal mycelium. P. aeruginosa produced siderophores and hydrogen cyanide under laboratory conditions. Field experiments conducted in 1997 and repeated in 1998 revealed that Pochonia chlamydosporia and P. aeruginosa significantly suppressed the root‐infecting fungi M. phaseolina, F. oxysporum, F. solani and R. solani and that the combination of the two caused greater inhibition of the fungal pathogens than either alone. Application of P. chlamydosporia and P. aeruginosa as a soil drench also resulted in enhanced growth of tomato plants.     

Fungal Parasitism of Heterodera glycines Eggs as Influenced by Egg Age and Pre-colonization of Cysts by Other Fungi

The objective of this study was to determine the effect of egg age and pre-colonization of cysts by a saprophytic or parasitic fungus on parasitism of Heterodera glycines eggs by other parasitic fungi. In agar and in soil tests, fungi generally parasitized more eggs in early developmental stages than eggs containing a juvenile. The effect of pre-colonization of cysts by a fungus on parasitism of eggs by other fungi depended on the fungi involved. In most cases, pre-colonization of cysts by an unidentified, saprophytic fungal isolate (A-1-24) did not affect parasitism of eggs in the cysts subsequently treated with other fungi. However, pre-colonization of cysts by A-1-24 reduced fungal parasitism of eggs in cysts subsequently treated with Cylindrocarpon destructans isolate 3. In agar tests, pre-colonization of cysts by Chaetomium cochliodes, a saprophytic or weakly parasitic fungus, reduced parasitism of eggs in cysts subsequently treated with Verticillium chlamydosporium Florida isolate, Fusarium oxysporum, Fusarium solani, ARF18, and another sterile fungus. However, in soil tests, pre-colonization of cysts by C. cochliodes had no effect on parasitism of eggs by subsequent fungal parasites. In another test, parasitism of eggs by V. chlamydosporium in cysts was not affected by pre-colonizing fungi C. destructans, F. oxysporum, and F. solani but was reduced by Mortierella sp., Pyrenochaeta terrestris, and C. cochliodes. Parasitism of eggs in cysts by ARF18 was reduced by pre-colonizing fungi C. destructans, F. oxysporum, F. solani, P. terrestris, and C. cochliodes but not Mortierella sp.     

Mycoparasitism ofFusarium SPP. onRhizoctonia solani Kühn

Summary Experiments on nutrient and staled agar were carried out to investigate the mycoparasitic activity of some fusaria againstRhizoctonia solani Kühn. Penetration and coiling byFusarium oxysporum Sch.,F. semitectum Berk & Rav. andF. udum Butler in and around theR. solani hyphae was observed. Lysis ofF. udum hyphae was observed inside theR. solani hyphae showing the reverse of the normal direction of necrotrophic mycoparasitic relationships. The mycoparasitic activity ofFusarium spp. was much affected in staled agar plates.     

Phytopathogenic Fusarium oxysporum Schlecht, as a Necrotrophic Mycoparasite

Fusarium oxysporum f. sp. dianthi, f. sp. lycopersici, f. sp. cepae, f. sp. niveum and one unidentified F. oxysporum isolate proved to be active necrotrophic mycoparasites. In dual cultures hyphae of Trichoderma hamatum, T. longibrachiatum, T. pseudokoningii, T. harzianum, Botrytis cinerea and Rhizoctonia solani were parasitized and destroyed by F. oxysporum. One isolate of Phytophthora sp. was not affected. Mutual parasitism between F. oxysporum and T. pseudokoningii and T. longibrachiatum has been observed, too. Details of parasitic hyphal interactions: hyphal coiling, penetration sites, resistance sheat formation, hyphal invasion and internal growing are described. The mycoparasitic feature as well as antimicrobial metabolic production of F. oxysporum is probably a common phenomenon to ensure this important plant pathogenic species to compete successfully against other soil-borne fungal pathogens and saprophytes.     

Microsatellite marker-based detection of Fusarium wilt pathogens of Psidium guajava L.

Wilt of Psidium guajava L., incited by Fusarium oxysporum f. sp. psidii and Fusarium solani is a serious soil-borne disease of guava in India. Forty-two isolates each of F. oxysporum f. sp. psidii (Fop) and F. solani (Fs) collected from different agro climatic zones of India showing pathogenicity were subjected to estimate the genetic and molecular characterisation in terms of analysis of microsatellite marker studies. Out of eight microsatellite markers, only four microsatellite markers, viz. MB 13, MB 17, RE 102 and AY212027 were amplified with single band pattern showing the character of identical marker for molecular characterisation and genetic identification. Microsatellite marker MB 13 was amplified in F. oxysporum f. sp. psidii and F. solani isolates. Product size of 296 bps and 1018 bps were exactly amplified with a single banding pattern in all the isolates of F. oxysporum f. sp. psidii and F. solani, respectively. Microsatellite markers, viz. MB 17, RE 102 and AY212027 were also exactly amplified with a single banding pattern. MB 17 was amplified in F. oxysporum f. sp. psidii isolates with a product size of 300 bp. RE 102 and AY212027 were amplified in F. solani isolates with the product size of 153 bp and 300 bp, respectively. Therefore, amplified microsatellite marker may be used as identifying DNA marker.     

Decomposition of cellulose strips in relation to climate,litterfall nitrogen,phosphorus and C/N ratio in natural boreal forests

Isolates of Alternaria alternata, Botrytis cinerea, Fusarium oxysporum, Penicillium sp., Rhizoctonia solani, Stemphylium sp., Thielaviopsis basicola, and Verticillium dahliae were cultured on potato–dextrose agar (PDA), barley-sand and alfalfa-sand substrates in petri-dish or in column microcosms. N-mineralization by fungi and fungal-feeding nematodes in combination or fungi alone was assessed. Numbers of Aphelenchus avenae or Aphelenchoides composticola supported by the fungi were measured every 7 days. Times for full colonization of the substrates by fungi ranged from 5 to 15 days. Rhizoctonia solani and B. cinerea on PDA supported the largest A. avenae and A. composticola populations, respectively. Penicillium sp. was a nonhost for A. composticola and A. avenae. Rhizoctonia solani, B. cinerea, V. dahliae, and F. oxysporum supported significantly more nematodes than the other four fungal species. The ranked order of fungi based on the amount of N mineralized in columns free of nematodes was A. alternata (with a rate of 0.052 μg N/g-sand per day), Stemphylium sp., V. dahliae, T. basicola, B. cinerea, F. oxysporum, R. solani, and Penicillium sp. (with a rate of 0.0045 μg N/g-sand perday). The presence of A. avenae resulted in significant increases in mineral N, compared to nematode-free columns colonized by F. oxysporum, R. solani, and T. basicola alone. The presence of A. composticola resulted in significant increases in mineral N, compared to nematode-free columns colonized by A. alternata, B. cinerea, F. oxysporum, and R. solani alone. There was more mineral N incolumns in the presence of A. composticola than A. avenae in most cases. This revised version was published online in August 2006 with corrections to the Cover Date.     

Peroxidase, catalase, amine oxidase and acid phosphatase activities in Pisum sativum during infection with Fusarium oxysporum and F. solani

Two genotypes (cv. Smaragd and line DP1059) of Pisum sativum with different susceptibility to Fusarium oxysporum and F. solani and influence of pathogenesis on enzyme activities were studied. The increase of activity of studied enzymes was mostly observed in both roots and shoots during pathogenesis. Only activity of acid phosphatase decreased in the root and increased in shoots. The correlation between enzyme activity change and susceptibility of pea cultivars to F. oxysporum or F. solani was observed.     

<Emphasis Type="Italic">Exiguobacterium acetylicum</Emphasis> strain 1P (MTCC 8707) a novel bacterial antagonist from the North Western Indian Himalayas

Exiguobacterium acetylicum strain 1P (MTCC 8707) is a rhizospheric, Gram positive, rod shaped, yellow pigmented bacterium isolated from an apple orchard rhizospheric soil, on nutrient agar plates incubated at 4°C. The species level identification was arrived on the basis of 16S rRNA gene sequencing. The sequence showed 98% similarity with sequences of E. acetylicum available in the public domain. The strain was positive for siderophore and HCN production. In separate invitro assays it was found to inhibit the growth and development of Rhizoctonia solani, Sclerotium rolfsii, Pythium and Fusarium oxysporum. The volatile compound produced by the bacterium was found to be the most potent in inhibiting the hyphal development of R. solani, S. rolfsii, Pythium and F. oxysporum by 45.55, 41.38, 28.92 and 39.74% respectively. Commonly observed deformities caused by the diffusible and volatile compounds produced by the bacterium included hyphal inhibition, constriction and deformation. Under pot culture conditions the bacterium improved the germination and early growth parameters of pea (Pisum sativum) in the presence of R. solani and S. rolfsii.     

Frequency and diversity of Fusarium spp. colonizing roots of Egyptian cottons

Abstract

Fusarium species are known to play a role in several diseases of cotton including the seedling disease complex, wilt, and boll rot. Therefore, a mycoflora study was conducted in 1998 in order to identify Fusarium species found in association with cotton roots. A total of 109 samples of cotton seedlings infected with post-emergence damping-off or rotted roots of adult plants were obtained from different cotton-growing areas in Egypt. Forty-six isolates were recovered and were identified as follows: F. oxysporum (28 isolates), F. moniliforme (9), F. solani (6), F. avenaceum (2), F. chlamydosporum (1). F. oxysporum, F. moniliforme and F. solani, the dominant species, accounted for 60.9%, 19.6% and 13% of the total isolates, respectively in 1998. F. oxysporum showed the highest isolation frequency in Beharia and Minufiya while F. moniliforme showed the most isolation frequency in Minufiya and Gharbiya. F. oxysporum was one of the major taxa of the Fusarium assemblage from Giza 70. F. oxysporum showed the most frequently isolated fungus in May while F. moniliforme and F. solani were the most frequently isolated fungi in August. Isolation frequency of Fusarium spp. during July and August was significantly greater than that of April or June. This implies that cotton roots are subjected more to colonization by Fusarium spp. as plants mature. Regarding pathogenicity, of the 46 isolates of Fusarium spp. tested under greenhouse conditions, 38 isolates (82.4%) were pathogenic to seedlings of Giza 89. This study indicates that F. oxysporum and F. moniliforme are important pathogens in the etiology of cotton damping-off in Egypt.     

Root Rot and Wilt of Kangaroo Paw (Anigozanthos manglesii) Caused by Pythium myriotylum (Drechs.) in Israel

A root rot and wilt disease of Anigozanthos manglesii (Kangaroo Paw) grown in greenhouses in Israel, for exporting as cut flowers to Europe, was characterized. Pythium myriotylum (Drechs.) and Rhizoctonia solani (Kühn) were the prevalent pathogens in diseased plants collected from commercial greenhouses. Fusarium oxysporum, Fusarium spp. and Myrothecium sp. were also isolated, but P. myriotylum or R. solani were not detected in samples from symptomless plants in tissue cultures (Australian origin) or plants at different stages in the nursery; non‐pathogenic F. oxysporum and Fusarium spp. were detected in several samples. In pathogenicity tests carried out in pots, plant mortality occurred 7 days after inoculation with P. myriotylum. In a field experiment carried out in methyl bromide‐fumigated soil, the incidence of dead plants following inoculation with P. myriotylum alone was 22% 10 days after inoculation, increasing to 78% after an additional 25 days. The incidence of dead plants following inoculation with R. solani alone was only 5% and in plants inoculated simultaneously with both pathogens, disease incidence was 88% 35 days after inoculation. Mortality reached 90–100% in plants inoculated with P. myriotylum, either singly or combined with R. solani 60 days after inoculation, whereas in plants inoculated with R. solani it was 5%. The maximum mortality in plants inoculated with R. solani was 25%, 76 days after inoculation. These results clearly demonstrate that P. myriotylum was the dominant pathogen in the root rot and wilt of A. manglesii.     

Prevalence of wilt-root rot and their associated pathogens at reproductive phase in lentil

A survey of 116 districts of nine lentil growing states covering 603 farmers' fields revealed a range of 0.7–9.3% mean plant mortality at reproductive stages in different lentil growing states of the country. The overall mean mortality was 6.3%. The main pathogens found associated with plant mortality at this stage were Fusarium oxysporum f. sp. lentis (62.0%), Rhizoctonia bataticola (25.2%) and Sclerotium rolfsii (9.8%). The minor involvement of 1.8% was that of F. solani, F. chlamydosporum. F. equisetii, and R. solani. For the first time a national scenario of lentil wilt-root rot incidence at the crucial reproductive stage and their associated pathogens is reported here.     

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.     

Enzymes Associated with Defence against Toxic Oxygen Species in PCNB-Tolerant and Sensitive Soil Fungi

The specific activities of enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPOX) and glutathione reductase (GR), which are involved in protection against toxic species of oxygen, were determined in mycelia extracts of pentachloronitrobenzene (PCNB)-tolerant and susceptible soil fungi. The organisms assayed were the highly PCNB-sensitive Rhizoctonia solani and Rhizopus arrhizus; Sclerotium rolfsii and Trichoderma harzianum, which are moderately susceptible to PCNB, and the fungicide-tolerant Fusarium oxysporum f. sp. melonis and Pythium aphanidermatum. No GPOX activity was detected in the six examined fungi. Significant differences in the specific activities of the other enzyme systems among the fungi were evident. Remarkably low levels of CAT activities were measured in R. solani. Except for T. harzianum, no meaningful differences regarding SOD, CAT and GR activities with age of the fungi cultures were observed. The electrophoretic patterns of SOD and CAT displayed dissimilarities among the fungi under study. P. aphanidermatum is more polymorphic with respect to both SOD and CAT enzyme systems as compared to the other fungi. The SOD of F. oxysporum f. sp. melonis, R. arrhizus and T. harzianum is a cuprozinc enzyme, while the mangano-SOD species was detected in S. rolfsii, R. solani and T. harzianum.     

Evaluation of antifungal potential of Marchantia polymorpha L., Dryopteris filix-mas (L.) Schott and Ephedra foliata Boiss. against phyto fungal pathogens

Methanol and flavonoid extracts (free and bound) of Marchantia polymorpha L., Dryopteris filix-mas (L.) Schott and Ephedra foliata Boiss. were screened against three fungal plant pathogens: Alternaria solani, Fusarium oxysporum and Rhizoctonia solani. The extracts from D. filix-mas and E. foliata showed >80% of mycelial inhibition of A. solani whereas M. polymorpha and D. filix-mas (rhizome) completely inhibited the mycelial growth of R. solani when tested at highest concentration (5 mg/ml). Inhibition of spore germination of fungi (A. solani and F. oxysporum) was observed to be 100% by most of the extracts at 10 mg/ml. Moreover, plant extracts were found effective in increasing seed germination and seed vigour simultaneously thereby decreasing the percentage of pathogen infection. The results of the present study reveal that the plants screened possess the potential to inhibit the crop fungal pathogens and further investigation is required to explore the biologically active constituents of these plants and to use them as natural plant protectants for agriculture.     

Studies on the genus Fusarium in Egypt

Five Fusarium species were recovered from the rhizoplane of healthy and damped-off cotton, pea, tomato, maize and wheat seedlings raised in the field during a 12-month experiment: F. solani, F. oxysporum, F. moniliforme, F. acuminatum and F. equiseti. F. solani and F. oxysporum were the most common species in the rhizoplane of healthy and damped-off seedlings of cotton, pea, wheat and tomato. In the case of maize, they were surpassed by F. moniliforme which was very scarce in the roots of the other test plants. There was some regular periodicity in the occurrence of Fusarium species in the rhizoplane of test plants. F. oxysporum showed its highest records usually in winter months, F. solani usually in moderate and high temperature months, and F. moniliforme, in maize rhizoplane, in winter months.     

Etiology of a Root Rot Disease Complex of Alstroemeria in Alberta, Canada

Fusarium oxysporum, Pythiu-m ultimum, and Rhizoctonia solani were isolated from the basal stems of diseased alstroemeria showing symptoms of dark brown stripes along leaf margins, leaf chlorosis, plant wilting, browning or rotting of basal stem, rhizome, and storage and fibrous roots. The pathogen isolated most frequently was Fusarium spp. (40.5 % of plants examined). Pythium spp. and R. solani were isolated less frequently (5.5 % and 6.8 % of plants examined, respectively). F. oxysporum caused the highest mortality in alstroemeria when rhizomes were grown in unsterilized soil-less mix medium. This is the first report in North America of a root-rot disease complex affecting alstroemeria.     

Interaction of Rotylenchulus reniformis with Seedling Disease Pathogens of Cotton

The impact of 10 Fusarium species in concomitant association with Rotylenchulus reniformis on cotton seedling disease was examined under greenhouse conditions. In experiment 1, fungal treatments consisted of Fusarium chlamydosporum, F. equiseti, F. lateritium, F. moniliforme, F. oxysporum, F. oxysporum f.sp. vasinfectum, F. proliferatum, F. semitectum, F. solani, and F. sporotrichioides; Rhizoctonia solani; and Thielaviopsis basicola. The experimental design was a 2 × 14 factorial consisting of the presence or absence of R. reniformis and the 12 fungal treatments plus two controls in autoclaved field soil. In experiment 2, the same fungal and nematode treatments were examined in autoclaved or non-autoclaved soil. This experimental design was a 2 × 2 × 14 factorial consisting of field or autoclaved soil, presence or absence of R. reniformis, and the 12 fungal treatments plus two controls. In both tests, Fusarium oxysporum f. sp. vasinfectum, F. solani, R. solani, and T. basicola consistently displayed extensive root and hypocotyl necrosis that was more severe (P ≤ 0.05) in the presence of R. reniformis. Soil treatment (autoclaved vs. non-autoclaved) influenced the impact of the Fusarium species on cotton seedling disease, with disease being more severe in the autoclaved soil. Rotylenchulus reniformis reproduction on cotton seedlings was greater in field soil compared to autoclaved soil (P ≤ 0.05). This study suggests the importance of Fusarium species and R. reniformis in cotton seedling disease.     

POPULATION DIFFERENCES IN INDIGENOUS FUSARIUM SPECIES BY CORN CULTURE OF PRAIRIE SOIL

Six species of Fusarium were identified among approximately 7,000 cultures isolated from roots and soils collected from prairies and cornfields in southern Minnesota. In both soil sources, F. oxysporum and F. solani predominated, followed in order by F. roseum, F. episphaeria, F. tricinctum, and F. moniliforme. The same order was obtained for the 28 species of prairie grasses and forbs as for corn. From prairie plant and corn roots, respectively, F. oxysporum was isolated from 64 and 90%; F. solani, 26 and 80%; F. roseum, 8 and 35%; and F. tricinctum, 6 % each. In 10 of 17 companion soil pairs, populations of Fusarium species were higher in cornfield than in prairie soils. Populations of F. roseum and F. moniliforme especially, were higher in cornfield soils of sample pairs, but populations of F. solani were higher in prairie soils. Fusarium roseum ‘Equiseti’ was the predominant cultivar of this species in both prairie and cornfield soils. Thus all six species of Fusarium appeared to be indigenous to the prairie, and some, especially F. roseum, apparently had increased in prevalence in soil and roots by corn culture.     

Identification of Fusarium species causing basal rot of onion in East Azarbaijan province,Iran and evaluation of their virulence on onion bulbs and seedlings

One of the economically important diseases of onion is the basal rot caused by various Fusarium species. Identification of the pathogenic species prevalent in a region is indispensable for designing management strategies, especially to develop resistant cultivars. Eighty Fusarium isolates are obtained from red onion bulbs on infected fields of East Azarbaijan province. Inoculating the onion bulbs with 38 selective isolates indicated that 17 isolates were pathogenic on onion. According to the morphological and molecular characteristics, these isolates were identified as F. oxysporum, F. solani, F. proliferatum and F. redolens. This is the first report of F. redolens on onion in Iran. On the other hand, the virulence of each pathogenic isolate was evaluated on onion bulbs and seedlings. F. oxysporum which causes severe rot and damping-off was considered as a highly virulent species in both conditions. While, F. proliferatum was considered as the most destructive on onion bulbs. Rot ability of F. solani was not considerable, and only the 4S isolate caused pre- and post-emergence damping-off more than 50%. Finally, F. redolens with less pathogenicity on onion bulbs was identified as the most virulent isolate on onion seedlings, which was explanatory of its importance on farm.     

Effect of the high polycyclic aromatic hydrocarbon, benzo[a]pyrene, on the lipid content of Fusarium solani

The purpose of the present paper was to study the effect of the high polycyclic aromatic hydrocarbon (PAH), benzo[a]pyrene, on the lipid [fatty acid (FA) and sterol] composition and content of the fungi Fusarium solani and F. oxysporum, respectively recognized as good and poor PAH degraders. The major FAs and the major sterol that characterized the tested Fusarium strains were C16:0, C18:1, C18:2, and ergosterol. Lipid profiles of F. solani remained unchanged with the addition of benzo[a]pyrene in the culture media at all concentrations and duration of treatment. However, in the presence of benzo[a]pyrene, significant decreases in FA content, which reached 18 % in young cultures and 28 % in mature colonies, were registered. Similarly, the sterol content of F. solani was reduced by 27 % in the presence of benzo[a]pyrene. In contrast, no modification in lipid profile and lipid content were observed with F. oxysporum, a strain recognized as a low benzo[a]pyrene degrader.