Effect of label and sublabel rates of metam sodium in combination with<Emphasis Type="Italic">Trichoderma hamatum,T. harzianum,T. virens,T. viride</Emphasis> on survival and growth of<Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

This work was undertaken to determine the effects ofTrichoderma spp. combined with label and sublabel rates of metam sodium on survival ofRhizoctonia solani in soil. Soils were infested with wheat bran preparations ofTrichoderma hamatum Tri-4,T. harzianum Th-58,T. virens Gl-3, andT. viride Ts-1-R3. Soil was also infested with sterile beet seeds that were colonized withR. solani. Beet seeds were later recovered, plated onto water agar plus antibiotics, and the growth ofR. solani was recorded. Preliminary experiments showed thatT. hamatum andT. virens reduced survival and saprophytic activity ofR. solani when the biocontrol fungi were incorporated into soil at 1.5% (w:w) or greater. Based on these data, biocontrol fungi in subsequent experiments were incorporated into soil at 2%. Metam sodium at label rate killed all biocontrol fungi andR. solani. At 1:2 and 1:5 dilutions, metam sodium reduced survival ofR. solani and allTrichoderma spp. When biocontrol fungi plus the label rate of metam sodium and 1:5, 1:10, 1:50 or 1:100 dilutions of the label rate were tested together, there were no interactions between any biocontrol agent and the fumigant with respect to colony diameter, reflecting that allTrichoderma isolates tested reacted similarly to increasing concentrations of metam sodium. At the label rate of metam sodium, allTrichoderma spp. significantly reduced colony diameter, but not growth rate, ofR. solani from beet seed. For the levels of metam sodium tested in combination withTrichoderma, it does not appear feasible to use a reduced rate of metam sodium to controlR. solani. However, the combination ofTrichoderma with metam sodium does reduce growth ofR. solani in comparison with that provided by metam sodium at the label rate. http://www.phytoparasitica.org posting Feb. 11, 2004.     

Selective accumulation of Trichoderma species in soils suppressive to radish damping-off disease after repeated inoculations with Rhizoctonia solani, binucleate Rhizoctonia and Sclerotium rolfsii

Artificial suppression of radish damping-off disease was induced by repeated soil inoculations with Rhizoctonia solani, binucleate Rhizoctonia (BNR) and Sclerotium rolfsii in pot systems. Soils repeatedly inoculated with R. solani and BNR showed suppressive to disease caused by R. solani and S. rolfsii, while soils repeatedly inoculated with S. rolfsii were suppressive to disease caused by S. rolfsii but not by R. solani. Species of Trichoderma were consistently isolated from soils repeatedly inoculated with R. solani, BNR and S. rolfsii. These Trichoderma spp. accumulated selectively in relation to the fungal species that was repeatedly added to the soils. The ratios of the frequencies of T. viride, T. harzianum and T. hamatum were 5:2:2 and 8:5:2 in soils repeatedly inoculated with R. solani and BNR, respectively. In S. rolfsii- inoculated soils, T. koningii was predominantly isolated. T. viride, T. harzianum and T. hamatum isolates obtained from either R. solani or BNR after repeated additions to the soils suppressed radish damping-off disease caused after challenge inoculations with R. solani or S. rolfsii. Among the Trichoderma species, T. viride consistently yielded high levels of suppression. However, isolates of T. koningii obtained from S. rolfsii-infested soils suppressed disease caused by S. rolfsii but failed to suppress disease caused by R. solani. Generally, the species of Trichoderma accumulated in a selective pattern that was closely related to the species of fungal pathogen used to induce the suppressive soil.     

Effect of GRAS compounds on mycelial growth, pectic enzyme activity and disease severity of postharvest pathogens on rambutan (Nephelium lappaceum)

The GRAS compounds cinnamaldehyde (at 30 ppm), acetaldehyde (at 70 ppm), benzalde-hyde (at 50 ppm) and potassium metabisulphite (at 250 ppm) either completely inhibited or significantly reduced thein vitro mycelial growth ofGliocephalotrichum microchlamydosponim,Colletotrichum gloeosporioides andBotryodiplodia theobromae, the causative fungi of brown spot, anthracnose and stem end rot, respectively, of rambutan fruits,Nephelium lappaceum. The four compounds also significantly reduced the severity of all three diseases and the activity of pectic lyase and polygalacturonase enzymes secreted by the three fungi.     

Control ofRhizoctonia solani andSclerotium rolfsii in the greenhouse using endophyticBacillus spp.

Isolates of different endophytic bacteria were recovered from surface-disinfected seeds obtained from commercial companies, plants in the field and tissue culture. The bacteria were isolated from seeds after stringent surfacedisinfection.Pseudomonas fluorescens (isolate no. 14) from bean inhibited growth of all fungi tested and was fluorescent on King B medium.Bacillus cereus fromSinapis (isolate no. 65) inhibited growth ofRhizoctonia solani, Pythium ultimum andSclerotium rolfsii and also exhibited chitinase activity.Bacillus subtilis from onion tissue culture (isolate no. 72) inhibitedR. solani andP. ultimum growth.B. cereus from cauliflower (isolate no. 78) inhibited growth ofR. solani. B. pumilus from sunflower (isolate no. 85) inhibited growth ofR. solani andS. rolfsii. B. cereus (isolate no. 65) was introduced into cotton, and by using radioactive labelling we found that it was present for 16 days in the root-stem junction. It is most likely that these bacteria were still found 72 days after their introduction in the root and stem, at levels of 2.8·10⁵ and 5·10⁴ cfu g^–1 fresh weight, respectively, when selective medium was used. There was no difference between control and treated plants in their height or in the fresh weight of roots, stems and leaves.When cotton seedlings were inoculated withB. cereus (isolate no. 65),B. subtilis (isolate no. 72) orB. pumilus (isolate no. 85), disease incidence caused byRhizoctonia solani was reduced in the greenhouse by 51%, 46% and 56%, respectively. In bean seedlings inoculated withB. subtilis (isolate no. 72),B. cereus (isolate no. 78) orB. pumilus (isolate no. 65), disease incidence caused bySclerotium rolfsii was reduced by 72%, 79% and 26%, respectively, as compared to control. In both cotton and bean seedlings, these endophytes reduced the disease index more than 50%. These results indicate that endophytic bacteria can survive inside cotton plants and are efficient agents for biological control against plant pathogens under greenhouse conditions.     

Biological control of Rhizoctonia solani on potatoes by antagonists. 3. Inoculation of seed potatoes with different fungi

Inoculation of seed potatoes withVerticillium biguttatum and three other hyperparasitic fungi, alone or in combination, resulted in statistically significant reduction of infestation of potato plants byRhizoctonia solani. Gliocladium roseum, Trichoderma hamatum andHormiactis fimicola did not show prolonged protection againstR. solani under farming conditions.H. fimicola, however, inhibited mycelial growth ofR. solani in vitro, particularly in the lower temperature range whereV. biguttatum did not show any growth. Combining these two antagonistic fungi may be advantageous as they cover the entire temperature range in whichR. solani is active. At the end of the vegetation period,V. biguttatum was superseded byG. roseum.Production of sclerotia on newly formed tubers from seed potatoes inoculated withV. biguttatum (alone or in combination with the other three antagonists) was significantly reduced.Samenvatting Inoculative van pootaardappelen metVerticillium biguttatum, apart of sament met drie andere hyperparasieten, had een gunstig effect op het onderdrukken vanRhizoctonia solani op de plant;Trichoderma hamatum, Gliocaldium roseum enHormiactis fimicola, ieder apart toegediend, boden de plant op lange termijn geen bescherming tegenR. solani onder praktijkomstandigheden.H. fimicola bleek bij lage temperatuur, waarbijV. biguttatum geen groei-activiteit meer vertoonde, op hyfen en sclerotiën vanR. solani te kunnen groeien. Toepassing van deze schimmel enV. biguttatum in een gemengde inoculatie zou over een breder temperatuurtraject effectief kunnen zijn.Tegen het einde van het groeiseizoen vond er op de ondergrondse plantedelen een verschuiving plaats, waarbijG. roseum meer op de voorgrond trad. Verondersteld wordt dat de afname vanV. biguttatum op stolonen hiervan een gevolg was.De produktie van sclerotia op nieuwe aardappelen afkomstig van metV. biguttatum behandeld pootgoed (alleen of met andere hyperparasitaire schimmels) bleek sterk verminderd te zijn. Vooral dit gegeven maakt de biologische bestrijding vanR. solani interessant:V. biguttatum blijkt ook op lange termijn effectief te zijn.     

Insecticidal activity of crude seed extracts of<Emphasis Type="Italic">Annona</Emphasis> spp.,<Emphasis Type="Italic">Lansium domesticum</Emphasis> and<Emphasis Type="Italic">Sandoricum koetjape</Emphasis> against lepidopteran larvae

Crude ethanolic seed extracts ofAnnona muricata, A. squamosa (Annonaceae),Lansium domesticum andSandoricum koetjape (Meliaceae) collected from different locations and years in Maluku, Indonesia, were screened for inhibition of larval growth against the polyphagous lepidopteranSpodoptera litura (Noctuidae). Extracts ofA. squamosa were significantly more active (20-fold) than those ofA. muricata. A. squamosa collected from Namlea yielded the extracts with the greatest inhibitory activity. There were significant differences among locations for bothA. squamosa andA. muricata but not forL. domesticum andS. koetjape. Extracts ofA. squamosa, collected from Namlea, inhibited larval growth in a dose-dependent manner, with a dietary EC₅₀ (effective concentration to inhibit growth by 50% relative to controls) of 191.7 ppm fresh weight. Extracts ofA. squamosa collected from individual trees in Namlea also varied in growth inhibitory effect againstS. litura andTrichoplusia ni larvae. This species is a candidate for development of a botanical insecticide for local use in Indonesia. http://www.phytoparasitica.org posting Dec. 1, 2003.     

Potential inhibitors against <Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis>, produced by the fungus <Emphasis Type="Italic">Myrothecium</Emphasis> sp. associated with the marine sponge <Emphasis Type="Italic">Axinella</Emphasis> sp.

Sclerotinia sclerotiorum is a worldwide ascomycete fungal plant pathogen, which causes enormous yield losses on major economic crops such as crucifers, grain legumes and several other plant families. The objective of this research was to isolate and characterise some bioactive products from cultures of fungi associated with the marine sponge Axinella sp. In total, nine fungal isolates were obtained from the marine sponge Axinella sp. collected from the South China Sea. A group of test strains, including two G⁺ strains (Bacillus subtilis and Staphylococcus aureus), two G⁻ strains (Escherichia coli and Pseudomonas aeruginosa) and three fungi including two plant pathogenic fungi Sclerotinia sclerotiorum and Magnaporthe grisea and Saccharomyces cerevisiae, were employed as the indicator organisms for bioactivity screening. Using antagonistic tests and bioactive screening of the ethyl acetate (EtOAc) extracts of the corresponding cultures, fungal isolate JS9 showed the stronger efficacy against the test indicator strains, especially the indicator fungal pathogens. Isolate JS9 was further identified as Myrothecium sp. by a combination of morphological features and 18S rDNA BLAST on GenBank. Two macrocyclic trichothecenes, roridin A (compound 1) and roridin D (compound 2) were purified by tracking the activity of the EtOAc extract fractions and characterised with spectral analyses including MS, ¹H-NMR, ¹³C-NMR and disortionless enhancement by polarization transfer (DEPT). In vitro antifungal tests showed that the two macrocyclic trichothecenes were bioactive against S. cerevisiae, M. grisea and S. sclerotiorum with minimal inhibitory concentrations of 31.25, 125 and 31.25 μg ml⁻¹ for roridin A, and 62.5, 250 and 31.25 μg ml⁻¹ for roridin D, respectively. The present investigation demonstrated that two antifungal trichothecenes including roridin A and roridin D produced by the fungus Myrothecium sp. isolated from the marine sponge Axinella sp. could be potential inhibitors against the plant pathogen S. sclerotiorum. Lian Wu Xie and Shu Mei Jiang contributed equally to this work.     

Note:In vitro antagonism of actinomycetes isolated from fungus-growing ants against plant pathogenic fungi

Fungus-growing ants have been found recently to be symbiotic with actinomycetes living on the ant’s cuticle; these bacteria are inhibitory to soil fungi that are detrimental to the ants’ fungus gardens. In order to investigate whether actinomycetes found on the cuticle of attine ants also had inhibitory properties against plant pathogenic fungi, we isolated 32 strains of actinomycetes from fungus-growing ants (Atta, Trachymyrmex, andCyphomyrmex), from the Mexican states of Coahuila, Nuevo León and Tamaulipas. Of the actinomycetes tested against selected plant pathogenic fungi (Alternaria solani, Aspergillus flavus, Colletotrichum lindemuthianum, Rhizoctonia solani, Sclerotium sp.) on Czapek-Dox agar medium, 13 isolates inhibited at least one of the fungi.C. lindemuthianum was inhibited by 11 actinomycetes, andRhizoctonia by three. An actinomycete strain isolated fromCyphomyrmex rimosus inhibited all five fungi tested. http://www.phytoparasitica.org posting July 30, 2008.     

Identification and pathogenicity of<Emphasis Type="Italic">Rhizoctonia solani</Emphasis> and binucleate<Emphasis Type="Italic">Rhizoctonia</Emphasis> anastomosis groups isolated from forage legumes in Erzurum,Turkey

Abstrast  Three-hundred-twenty-five isolates ofRhizoctonia (215R. solani and 110 binucleateRhizoctonia) were obtained from roots and crowns of alfalfa, sainfoin and common vetch grown in Erzurum, Turkey. The isolates were assigned to five anastomosis groups (AG) ofR. solani (AG-2-1, AG-3, AG-4, AG-5, and AG-10) and two anastomosis groups of binucleateRhizoctonia (AG-I and AG-K). In pathogenicity tests on alfalfa, sainfoin and common vetch, the highest disease severities were caused by isolates of AG-4 and AG-5. Isolates of AG-10 and AG-I were not pathogenic on the three tested forage legumes, whereas isolates of AG-K on alfalfa and sainfoin, and of AG-2-1 on sainfoin, were moderately virulent. Alfalfa isolate AG-3 was moderately virulent on sainfoin. This is the first report ofR. solani AG-3, AG-5, AG-10 and binucleateRhizoctonia AG-I on alfalfa. In addition, all theR. solani and binucleateRhizoctonia groups isolated from sainfoin and common vetch were recovered from these crops for the first time in Turkey. http://www.phytoparasitica.org posting Dec. 16, 2002.     

The use of Solacol® (validamycin) as a growth retardant in the isolation of soil fungi

Solacol^®, a formulation of the antibiotic validamycin, at 0.33% in 2% malt extract agar, reduced the spread of fungi on dilution plates drastically and allowed twice as much incubation time before subculturing; this resulted in an elevated number of species isolated. Using pure cultures of 62 common soil fungi, it was shown that all fast-growing species (exceptPythium ultimum) were efficiently inhibited but not completely suppressed. Inhibition was comparable to that by 0.5% oxgall, though, while this substance completely suppressed several species, Solacol very strongly inhibited onlyGaeumannomyces graminis, Gerlachia nivalis, Harzia acremonioides, Verticillium biguttatum andRhizoctonia solani. In a further experiment each separate constituent of Solacol was tested against 22 fungi at equivalent concentrations. Validamycin strongly inhibitedChaetomium globosum and two Basidiomycetes, though hardly more than the non-ionic detergent which mainly inhibited the other fungi. A few species were, however, more inhibited by Solacol than by the detergent alone. Solacol at 0.33% is a suitable aid in dilution plating of soil fungi, by increasing the number of colonies and species observed.Samenvatting Solacol^®, een formulering van het antibioticum validamycine, remde de groei van schimmels in verdunningsplaten met een concentratie van 0.33% in 2% moutagar en maakte het mogelijk de periode tot afenten met een factor 2 te verlengen; daardoor was het aantal geïsoleerde soorten duidelijk toegenomen. Met reincultures van 62 algemene grondschimmelsoorten werd aangetoond, dat alle snelgroeiende soorten (met uitzondering vanPythium ultimum) voldoende geremd, maar niet volkomen onderdrukt werden. Het remmingspercentage was vergelijkbaar met dat van 0.5% ossegal, hoewel dit laatste sommige soorten volkomen onderdrukte; Solacol remde alleenGaeumannomyces graminis, Gerlachia nivalis, Harzia acremonioides, Verticillium biguttatum enRhizoctonia solani zeer sterk. In een volgend experiment werden de componenten van Solacol t.o.v. 22 fungi apart getoetst in concentraties equivalent aan 0.33% Solacol. Validamycine remde alleenChaetomium globosum en twee basidiomyceten behoorlijk, maar nauwelijks meer dan de niet-ionische uitvloeier, die in hoofdzaak de overige remeffecten veroorzaakte. Enkele soorten werden echter door het complete Solacol veel sterker geremd dan door de uitvloeier alleen. Solacol in een verdunning van 0,33% wordt aanbevolen bij verdunningsplaten voor het isoleren van grondschimmels ten einde het aantal kolonies en soorten te verhogen.     

Identification and pathogenicity of<Emphasis Type="Italic">Fusarium</Emphasis> spp. from stem bases of winter wheat in Erzurum,Turkey

Four-hundred-sixty-eightFusarium andFusarium-like isolates were obtained from crowns and subcrown internodes of winter wheat grown in Erzurum, Turkey. Of these isolates, 34.8% wereFusarium acuminatum, 32.3% wereF. equiseti, 16.9% wereF. oxysporum, 15.0% wereMicrodochium nivale (formerlyFusarium nivale), 0.6% wereF. tabacinum and 0.4% wereF. solani. In pathogenicity tests on wheat, the highest disease severity was caused by isolates ofM. nivale, whereas isolates ofF. acuminatum, F. equiseti, F. oxysporum andF. solani were slightly virulent; isolates ofF. tabacinum were nonpathogenic. This is the first report ofM. nivale andF. tabacinum from wheat in Turkey. http://www.phytoparasitica.org posting Jan. 29, 2003.     

First report of southern blight of common barley in Puerto Rico

In 1990–91, 96% of the test plants in a barley (Hordeum vulgare L.) winter nursery in Puerto Rico was lost toSclerotium rolfsii. Small necrotic lesions (1–3 mm long) developed at the base of the stem, followed by wilting and general chlorosis as the lesions progressed and eventually girdled the base of the stem. Sclerotia then developed on the plants at the stem base. Two morphologically different strains ofS. rolfsii that were mycelial incompatible were isolated. This is the first report of a destructive blight due toS. rolfsii on field barley in the Caribbean. The apparent rate of disease increase (r) was estimated to be 12.2% per day using the logistic model, with an initial disease incidence of 0.042% (R²=0.89, p<-0.001).     

The potential for using cyanobacteria (blue-green algae) and algae in the biological control of plant pathogenic bacteria and fungi

Cyanobacteria (blue-green algae) and eukaryote algae occur in freshwater, marine, and terrestrial (soil) habitats. In fact, these microorganisms comprise most of the world's biomass. Although the cyanobacteria are mostly photoautotrophic, some are facultative heterotrophs, capable of growing on certain substrates in darkness. Also, some are non-phototrophic and hence, are obligate heterotrophs. A number of cyanobacteria and eukaryote algae, particularly macroalgae, produce various, biologically active compounds. These include antibiotics which in laboratory tests inhibited bacteria and fungi that incite diseases of humans. In addition, the following fungi which are of interest to plant pathologists, were inhibitedin vitro by substances produced by various cyanobacteria: The saprophytesChaetomium globosum, Cunninghamella blakesleeana, andAspergillus oryzae and the plant pathogensRhizoctonia solani andSclerotinia sclerotiorum. Extracts from seaweeds (macroalgae) sprayed on plants have been reported to reduce the incidence ofBotrytis cinerea (gray mold) on strawberries,Erysiphe polygoni (powdery mildew) on turnips, and damping-off of tomato seedlings. Because many cyanobacteria and algae produce a large number of antibacterial and antifungal materials, are almost never a threat to the environment, and many can be grown in quantity in mass culture, they are suitable candidates for exploitation as biocontrol agents of plant pathogenic bacteria and fungi. Much additional work remains to be done however, to thoroughly evaluate cyanobacteria and algae and their products for this role.     

Note: Evaluation of antibacterial activity of essential oil of<Emphasis Type="Italic">Rosa damascena</Emphasis> on<Emphasis Type="Italic">Erwinia amylovora</Emphasis>

Significant antibacterial activity was observed in the essential oil (E.O.) ofRosa damascena Mill. and the Minimum Bactericidal Concentration (MBC) of the E.O. was determined as 1386.5 μg ml⁻¹ forErwinia amylovora, the causal agent of fire blight disease. Ooze formation on immature pears and lesion formation in artificially inoculated shoots were completely (100%) prevented by the essential oil ofR. damascena (1500 μg ml⁻¹), essential oil ofThymbra spicata var.spicata (500 μg ml⁻¹) and streptomycin (100 μg ml⁻¹). Copper oxychloride plus maneb significantly reduced ooze formation and lesion formation, but the control was less than that obtained with the essential oils or streptomycin. The essential oil ofR. damascena may be a useful natural bactericide for the control of the fire blight pathogen,E. amylovora. http://www.phytoparasitica.org posting July 14, 2004.     

Inhibition of in vitro growth of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> by liquid residue derived from steam-treated grass clippings

The effects of liquid residue derived from grass clipping after steam-treatment on the in vitro growth of Rhizoctonia solani SN-1, AG-4 were evaluated to verify the antifungal activities of the residue. The liquid residue inhibited R. solani growth, with increasing inhibition with higher concentrations and lower pH. Test of individual organic acids (acetic, formic and propionic acids) present in the residue indicate that the inhibitory effect may be attributed to the organic acids, and the effect of the organic acids seems to depend on pH, especially at lower concentrations.     

Control of postharvest diseases of rambutan using controlled atmosphere storage and potassium metabisulphite or<Emphasis Type="Italic">Trichoderma harzianum</Emphasis>

Botryodiplodia theobromae, Colletotrichum gloeosporioides andGliocephalotrichum microchlamydosporum are the causal fungi of three rambutan postharvest diseases, stemend rot, anthracnose and brown spot, respectively. Two different treatments of rambutan fruits to control the diseases were investigated: application of potassium metabisulphite (250 ppm) or culture filtrate ofTrichoderma harzianum (TrH 40) followed by controlled atmosphere storage (CA) at 13.5°C and 95% r.h. Potassium metabisulphite at 250 ppm under CA effectively controlled the incidence and severity of the three postharvest diseases and maintained the eating quality and color of the fruit for 21 days. The greatest effect of this treatment was on brown spot disease, caused byG. microchlamydosporum. Application of TrH 40 was less effective than potassium metabisuphite. http://www.phytoparasitica.org posting July 8, 2002.     

Biotrophic mycoparasitism byVerticillium biguttatum onRhizoctonia solani

Verticillium biguttatum cannot utilise cellulose or nitrate-nitrogen and it requires biotin for growth, yet it grew and sporulated abundantly onRhizoctonia solani on cellulose, obtaining at least organic carbon, nitrogen and biotin fromR. solani. Videomicroscopy of inter-hyphal interactions on films of water agar showed thatV. biguttatum behaved as a biotrophic mycoparasite. From germinating spores, it penetrated the hyphae ofR. solani and formed haustorium-like branches without killing the host cells, and the haustoria supported an external mycelial network of the mycoparasite. Later the mycoparasite sporulated, and the infected host cells died. On cellulosic substrataV. biguttatum did not reduce the growth ofR. solani, and often enhanced the rate of cellulose degradation. However,V. biguttatum drastically reduced the production of sclerotia byR. solani, often completely suppressing sclerotium production when the mycoparasite infected only a localized region of the host colony. This is ascribed to the creation of a nutrient sink by the parasite, consistent with biotrophy. On plates of cellulose agar the suppression of sclerotia was not confined to parasitized colonies but extended to adjacent colonies ofR. solani that had successfully anastomosed with the parasitized colony. There was no effect on adjacent vegetatively incompatible colonies, where attempted anastomoses caused cytoplasmic death. In comparable experiments the necrotrophic mycoparasiteGliocladium roseum had no long-distance effect on sclerotium production byR. solani.Suppression of sclerotium production may explain the reported success ofV. biguttatum in biocontrol of black scurf of potato in experimental field conditions.     

Pathogenic fungi involved in root rot of peas in the Netherlands and their physiological specialization

Research on root rot pathogens of peas in the Netherlands has confirmed the prevalence ofFusarium solani, F. oxysporum, Pythium spp.,Mycosphaerella pinodes andPhoma medicaginis var.pinodella. Aphanomyces euteiches andThielaviopsis basicola were identified for the first time as pea pathogens in the Netherlands. Other pathogens such asRhizoctonia solani andCylindrocarpon destructans were also found on diseased parts of roots. F. solani existed in different degrees of pathogenicity, and was sometimes highly specific to pea, dwarf bean of field bean, depending on the cropping history of the field.A. euteiches was specific to peas, whereasT. basicola showed some degree of physiological specialization.     

In vitro characterization andin vivo detection ofRigidoporus lignosus,the causal agent of white root disease inHevea brasiliensis,by ELISA techniques

The aim of these studies is to develop a method for early detection ofRigidopoms lignosus (Basidiomycete, Polyporaceae), the causative agent of white root disease of rubber tree. Two polyclonal sera were produced against soluble mycelial proteins of twoR. lignosus isolates, one from Africa (FCI2), the other from Asia (FID2). The specificity of the antisera was tested using isoelectric focusing (IEF)/Western-blot and DAS-ELISA. The two sera recognized all 20R. lignosus isolates from various geographical origins. The banding patterns obtained by Western-blot enabled a distinction to be made between isolates from Africa and those from Asia. In DAS-ELISA and Western-blot analyses, strong cross reactions were observed withR. ulmarius. Only slight reactions were observed in Western-blot analysis toR. lineatus andP. noxius, both causative agents of root rot inHevea. These cross reactions were not observed under our DAS-ELISA analysis conditions. Finally, no cross reactions were obtained with 9 otherPolyporaceae orHevea root pathogen species. The sensitivity threshold of the DAS-ELISA method was 5 ng ml^–1 ofR. lignosus protein. An initial approach to using the DAS-ELISA test for the detection ofR. lignosus in infected plants was carried out on artificially inoculated root samples. The DAS-ELISA protocol enabled detection ofR. lignosus in the root systems of diseased plants. Moreover, no cross reaction was observed with healthy plant extracts.     

Toxicity of fungal endophyte secondary metabolites to plant parasitic nematodes and soil-borne plant pathogenic fungi

Fungi isolated from the cortical tissue of surface sterilized tomato roots collected from field plots produced secondary metabolites in nutrition broth that were highly toxic toMeloidogyne incognita. Especially strains ofFusarium oxysporum were highly active with 13 of 15 strains producing culture filtrates toxic to nematodes. The mechanism of action of the toxic metabolites produced by the non-pathogenicF. oxysporum strain 162 with proven biological control ofM. incognita in pot experiments was investigated. These metabolites reducedM. incognita mobility within 10 min of exposure. After 60 min, 98% of juveniles were inactivated. Juveniles were initially inactivated within a few minutes of exposure, but with exposure of 5 h 50% of the juveniles were dead and 24 h exposure resulted in 100% mortality. In a bioassay with lettuce seedlings metabolite concentrations > 100 mg/l reduced the number ofM. incognita juveniles on the roots comparing to the water control. TheF. oxysporum toxins were highly effective towards sedentary parasites and less effective towards migratory endoparasites. Nonparasitic nematodes were not influenced at all. Metabolites of strain 162 also reduced significantly the growth ofPhytophthora cactorum, Pythium ultimum andRhizoctonia solani in vitro. Secondary metabolites of endophytic fungi on plant-parasitic nematodes and soil-borne fungi should be considered for control of plant parasitic nematodes and plant pathogenic fungi. The results also show the need for proper selection of target nematodes inin vitro bioassays.