Efficacy of Talaromyces flavus Alone or in Combination with other Antagonists in Controlling Verticillium dahliae in Growth Chamber Experiments

Talaromyces flat-US reduced viability of microsclerotia of Verticitlium dahliae on senescent potato stems collected from the field when applied as ascospores in carboxy-methylcellulose or in talcum powder. Incorporating an alginate wheat-bran preparation of T. fiavus in soil at a rate of 0.5% (w/w) was followed by a decrease of > 90% of the population of V. dahliae in soil at both 15 and 25 C. Population densities of V. dahliae were negatively correlated (r = -0.50; P = 0.001) with those of T. flavus. However, the population of V. dahliae was also reduced in soil with alginate wheat-bran alone. When incorporated in soil in alginate wheat-bran and simultaneously coated on seeds in ta.lcum powder. T. flavus reduced colonization of roots and infection of eggplants by K. dahliae. Although to a lesser extent than with the antagonist, alginate wheat-bran without T. flavus also reduced infection by the pathogen. Treatment with combinations of T. flavus with other biocontrol agents, viz. Bacillus subtilis, Fusarium oxysporum or Gliocladium roseum , containing half of the inoculum of the single application of each antagonist, gave similar control of root colonization and stem infection by V. dahliae as application of the single antagonists. Population densities on the root of each antagonist were not or only slightly affected by the presence of the co-inoculated antagonist suggesting that the combinations were compatible.     

Colonization of cotton roots by soil micro-organisms

An experiment was carried out to study the colonization of soil microorganisms on cotton roots buried in the soil.Fusarium culmorum, F. solani, F. oxysporum, Macrophomina phaseoli andBacillus sp. were found to be the predominant colonizers followed byBotryodiplodia theobromae, Fusarium acuminatum, Aspergillus niger, A. flavus, Stachybotrys atra, Tilachlidium humicola andRhizopus sp. The practice of leaving cotton roots in the soil would therefore add to the building up of the inoculum potential of the soil inhabiting fungi several of which are serious root pathogens.     

The modes of action of two benzimidazoles and thiophanate-methyl used for the control of Verticillium wilt of strawberry

Benomyl, thiabendazole and thiophanate-methyl were found to be fungistatic against Verticillium dahliae in vitro. All materials suppressed wilt symptoms on artificially inoculated strawberry plants when applied as a root dip, soil drench or a foliar application. Quantitative bioassays of the amount of toxic residues and fungal colonization in tissues of treated plants after root treatment revealed that these materials or their toxic breakdown products were readily translocated to crown and leaves: they suppressed the internal spread of the pathogen throughout the plant. After foliar application there was no marked basipetal movement of material within the plant: fungal colonization was greatly reduced in treated leaves, but the roots and crowns were colonized to a similar extent to those of untreated plants.     

Nematode Response to Carbofuran

Higher populations of Meloidogyne incognita larvae and Pratylenchus penetrans were recovered from soil treated with carbofuran 10 and 15 days after treatment, respectively, than were recovered from untreated control soil. The number of P. penetrans, however, was lower 50 days after treatment, and symptoms developed only occasionally on the root systems of host plants. Populations of Tylenchorhynchus claytoni inoculated at different distances from the base of corn seedlings growing in carbofuran-treated soil did not move toward the plant, whereas they were attracted in untreated soil from a distance of 12 cm. P. penetrans moved at random in treated agar medium when inoculations occurred 4 cm away from the root tips of tomato seedlings under aseptic conditions. Those nematodes that reached the roots were never observed feeding during a 20-day observation period. Specimens of P. penetrans placed on the developing roots moved at random and never penetrated. In contrast, numerous P. penetrans penetrated roots of seedlings growing in untreated medium.     

Co-inoculation with antibiotic-producing bacteria to increase colonization and nodulation by rhizobia

A study was conducted to determine whether colonization of legume roots and nodulation byRhizobium meliloti andBradyrhizobium japonicum could be enhanced by using inocula containing microorganisms that produce antibiotics suppressing soil or rhizosphere inhabitants but not the root-nodule bacteria. An antibiotic-producing strain of Pseudomonas and one of Bacillus were isolated, and mutants ofR. meliloti andB. japonicum sp. resistant to the antibiotics were used. The colonization of the alfalfa rhizosphere and nodulation byR. meliloti were enhanced by inoculation of soil withPseudomonas sp. in soil initially containing 2.7×10⁵ R. meliloti per g. The colonization of soybean roots byB. japonicum was enhanced by inoculating soil with three cell densities ofBacillus sp., and nodulation was stimulated byBacillus sp. added at two cell densities. In some tests, the dry weights of soybeans and seed yield increased as a result of these treatments, and co-inoculation with Bacillus also increased pod formation. Inoculation of seeds withBacillus sp. and the root-nodule bacterium enhanced nodulation of soybeans and alfalfa, but colonization byB. japonicum andR. meliloti was stimulated only during the early period of plant growth. Studies were also conducted withStreptomyces griseus and isolates ofR. meliloti andB. japonicum resistant to products of the actinomycete. Nodulation of alfalfa byR. meliloti was little or not affected by the actinomycete alone; however, both nodulation and colonization were enhanced if the soil was initially amended with chitin andS. griseus was also added. Chitin itself did not affectR. meliloti. Treatments of seeds with chitin orS. griseus alone did not enhance colonization of alfalfa roots byR. meliloti or soybean roots byB. japonicum, but the early colonization of the roots by both bacterial species was promoted if the seeds received both chitin andS. griseus; this treatment also increased nodulation and dry weights of alfalfa and soybeans and the N content of alfalfa. It is suggested that co-inoculation of legumes with antibiotic-producing microorganisms and root-nodule bacteria resistant to those antibiotics is a promising means of promoting nodulation and possibly nitrogen fixation.     

Effects of inorganic amendments (N,P and K) to soil on the rhizosphere microflora of antirrhinum plants infected withVerticillium dahliae Kleb

Summary A study of the inorganic amendments (N, P and K) to soil, and their effect on the rhizosphere microflora, as well as their relation to the control of wilt of antirrhinum plants caused byVerticillium dahliae Kleb. was done. Ammonium sulphate was the only chemical found to be significantly inhibitory toV. dahliae in vitro. Soil amendments (NPK) affected the rhizosphere microorganisms of the antirrhinum plants. Higher concentration of the chemicals were phytotoxic. It was further observed that ammonium sulphate, and the combined chemicals (NPK 25%) in soil delayed the senescence in healthy plants, suggests that chemical fertilisers affected the host plants directly. Addition of ammonium sulphate (0.25%), calcium nitrate (0.25%, 0.5%) combined NPK (0.25%) to soil caused considerable reduction in disease severity. It is assumed that this reduction may be caused by the (1) fungitoxic nature of the chemicali.e. ammonium sulphate, (2) antagonistic environment for the pathogen in the rhizosphere was boostedi.e. where calcium nitrate was added as soil amendments and (3) reduction in disease severity in soil-amended with combined NPK, may be due to the fact that antagonistic actinomycete population was boosted in the rhizosphere.     

Long-Term Effects of Reclamation Treatments on Plant Succession in Iceland

The long‐term effects (20–45 years) of reclamation treatments on plant succession are examined at two localities in Iceland that were fertilized and seeded from 1954 to 1979 with perennial grasses or annual grasses, or left untreated. The areas that underwent reclamation treatments had significantly higher total plant cover (7–100%) than the untreated control plots (<5%), and floristic composition was usually significantly different between treated and untreated plots. Dwarf‐shrubs (Calluna vulgaris and Empetrum nigrum), bryophytes, biological soil crust, grasses, and shrubs characterized the vegetation in the treated plots, but low‐growing herbs that have negligible effects on the environment, such as Cardaminopsis petraea and Minuartia rubella, and grasses characterized the control plots. The seeded grass species had declined (<10%, the perennials) or disappeared (the annuals) but acted as nurse species that facilitated the colonization of native plants. It seems that by seeding, some factors that limit plant colonization were overcome. Soil nutrients, vegetation cover, litter, and biological soil crust were greater in the treated areas than the control plots. This may have enhanced colonization through an increase in soil stability and fertility, increased availability of safe microsites, increased moisture, and the capture of wind‐blown seeds. This study demonstrates the importance of looking at the long‐term effects of reclamation treatments to understand their impact on vegetation succession.     

Studies on antagonism betweenFusarium udum Butler and root region microflora of pigeon-pea

Antagonism betweenFusarium udum Butler causing wilt of pigeon-pea (Cajanus cajan (L.) Millsp.) and the saprophytic microflora of the root region of the host was studied with reference to colony interaction, hyphal interference, volatile and non-volatile metabolites and staling growth products. Studies were extended to screen potential antagonists against the wilt pathogen in soil. Aspergillus flavus, A. niger, A. terreus, Penicillium citrinum andMicromonospora globosa (an actinomycete) were antagonistic againstF. udum, whereas the pathogen parasitized and killedAspergillus luchuensis, Cunninghamella echinulata, Curvularia lunata, Mortierella subtilissima andSyncephalastrum racemosum. The pattern of growth of microorganisms on nutrient agar staled by rhizosphere soil inocula of healthy or wilted pigeon-pea plants was found to be different.F. udum colonized and grew on nutrient agar staled by the rhizosphere inoculum of the wilted plants upto 120h of incubation. However, it could not colonise and grow on the nutrient agar staled by rhizosphere microflora of healthy plants after 48h of incubation because of the presence of antagonists likeA. niger, A. flavus, A. terreus and a few species ofPenicillium in the soil inoculum. When pure cultures in soil ofF. udum was mixed with those of antagonists in different ratios,A. niger, A. flavus andM. globosa significantly suppressed the population ofF. udum, whereasA. terreus markedly reduced the population. When inoculated in soil, the antagonists exhibited a high fungistatic activity againstF. udum.     

Cellulose decomposing ability of Trichoderma in relation to their saprophytic survival

Abstract

Forty one strains of Trichoderma were isolated from four provinces in the Philippines and tested for their cellulose adequacy indices (CAI). T. harzianum strain no. 94-016 with the highest CAI was further tested for its saprophytic colonization in pots relative to the amount of inoculum. The higher the amount of inoculum of T. harzianum strain no. 94-016, the higher percentage colonization of rice straw in the soil. Further tests were done to compare the decomposition of rice straw on the surface and buried in the soil and the effect of moisture on soil decomposition. Buried rice straw inoculated with T. harzianum strain no. 94-016 watered on a daily basis provided better decomposition.     

Survival of Phytophthora cinnamomi in Buried Eucalyptus Roots

Colonization and survival of Phytophthora cinnamomi in roots was tested in 3 months old, axenically grown seedlings of Eucalyptus maculata (field resistant) and E. sieberi (susceptible). The roots were inoculated, then one week later were excised and buried in three non-sterile, conducive soils; a lateritic gravel, an infertile duplex soil, a loamy sand as well as in a fertile, suppressive krasnozem. Pathogen viability, percentage root colonization and chlamydospore numbers were examined at matric potentials of ?1/3, ?5 and ?10 bar after periods of 10, 100 and 200 days at 21°C. At 10 days, survival was 100% in the form of mycelium and the only significant difference was between the two Eucalyptus species. At 100 days survival was solely due to chlamydospores, but the pathogen was viable in all inoculated roots and at each matric potential. At 200 days soils had dried to less than ?10 bars and the pathogen failed to survive. No significant differences were found between the two pathogen isolates but significant differences were obtained between the susceptible and field resistant Eucalyptus species. Pathogen viability, percentage root colonization and chlamydospore number were highly correlated with soil types and matric potential. These components declined with decreasing soil matric potential. The Krasnozem was only suppressive at relatively high soil matric potentials (?1/3 bar). At lower values (?5, ?10 bar) survival of the pathogen, chlamydospore numbers and percentage colonization of the roots in the Krasnozem were comparable with that of the 3 conducive soils tested. Chlamydospores were present, but in low numbers in roots buried in the suppressive soil at ?1/3 bar.     

Phytophthora cinnamomi: the Dynamics of Chlamydospore Formation and Survival

Chlamydospore activity was investigated by inoculating 10 day old seedlings of Lupinus angustifolius with Phytophthora cinnamomi. Infected roots were excised and buried in 4 non-sterile media: – glass beads, gravel, sand or soil at different water potentials and incubated at 22°C. Roots were examined and chlamydospores counted initially and after 10, 20 and 30 days. Large numbers of spores formed in the roots buried in the various media. Spore formation was rapid in glass beads and gravel, and slower in soil, but more chlamydospores survived the 30 day period when roots were buried in soil. Nylon mesh squares 1 cm²were inoculated with 38 chlamydospores and buried in the same 4 media. 70–100% of chlamydospores germinated and colonised 90–100% of the mesh, producing mycelia and an increased number of chlamydospores. In some instances there was a ten-fold increase in spore numbers and numbers were still increasing at 28 days. Maximum numbers and maximum survival occurred on mesh buried in soil. The role of the chlamydospores is considered, as that of a dynamic unit with a saprobic phase in the life cycle of the fungus which is independent of host, and has the capacity to increase both population density and distribution of the pathogen.     

Fungal pathogen and ethanol affect host selection and colonization success in ambrosia beetles

1. Ambrosia beetles exhibit broad host ranges but a narrow preference based on the condition of the host. Tissues infected by pathogens or containing ethanol can facilitate attacks by ambrosia beetles, although it still remains unclear how these factors interact.
2. The present study aimed to examine how (i) chestnut logs infected with the fungal pathogen Cryphonectria parasitica and treated with ethanol (i.e. baited with ethanol lure, soaked in ethanol or untreated) and (ii) hornbeam logs soaked in different ethanol concentrations (3–12.5%) affect host selection and colonization success of ambrosia beetles.
3. Ethanol‐soaked logs were more attractive to Anisandrus dispar than ethanol‐baited logs or untreated logs, although this difference was more evident in uninfected than infected logs. Increasing ethanol concentration in host tissues was differentially attractive to Xyleborinus saxesenii and Xylosandrus germanus. A nonlinear relationship was also documented between ethanol concentration and emergence of X. germanus adults.
4. Overall, the results obtained suggest that the presence of C. parasitica in chestnut logs can affect host selection in ambrosia beetles. In addition, the ethanol concentration in tree tissues affects host selection and colonization success, although the effect varies depending on the beetle species. This contrasting response could be a niche‐partitioning mechanism based on ethanol within host tissues.

     

Effect of liming on spore germination,germ tube growth and root colonization by vesicular-arbuscular mycorrhizal fungi

Summary The effect of soil acidity on spore germination, germ tube growth and root colonization of vesicular-arbuscular mycorrhizal (VAM) fungi was examined using a Florida Ultisol. Soil samples were treated with 0, 4, 8 and 12 meq Ca/MgCO₃/100 g soil and each lime level received 0, 240, and 720 ppm P as superphosphate. Corn (Zea mays L.) was planted in the soil treatments, inoculated with eitherGlomus mosseae orGigaspora margarita spores and grown for 31 days. Acid soil inhibits mycorrhizal formation byG. mosseae through its strong fungistatic effect against the spores. The dolomitic lime increased mycorrhizal formation by both fungal species.G. margarita is much less sensitive to acidic conditions thanG. mosseae. Al ions are a very important component of the fungistatic property against the VAM symbiosis. VAM fungus adaptation may be important for plants growing on infertile acid soils if soil inoculation with these fungi is to contribute significantly to low-input technology for tropical agricultural systems.     

The effect of fumigants on soil recolonization in the glasshouse by organisms colonizing the spermosphere

Germinating turnip seeds were used as baits to measure changes in colonizing floras of the spermosphere in glasshouse soil boxes following fumigation. The spermosphere mycoflora of the untreated soil was always dominated by Fusarium, Pythium, and Gliocladium spp. Methyl bromide, chloropicrin, and MBC₃₃ (a 67:33 mixture of methyl bromide and chloropicrin), applied in polyethylene bags at a rate of 2·5 ml/cubic foot of soil, greatly reduced colonization by Fusarium and Pythium spp. for 120 days. Fungi and actinomycetes recolonizing treated soils were more active than in untreated soils, and each fumigant induced a characteristic recolonization pattern. The spermosphere of methyl bromide-treated soil was initially dominated by actinomycetes and later by Penicillium spp. In chloropicrin-treated soil, Trichoderma and later Penicillium spp. dominated, and in soil treated with MBC ₃₃, Penicillium and Trichoderma spp. were co-dominant. Dilution plates, made at the same time from the same soil, indicated a close correlation between inoculum density of different fungi in the soil and their inoculum potentials towards the spermosphere.     

Field experiments in the control of Verticillium dahliae and Heterodera rostochiensis on potatoes

Potatoes were grown in soil infested with Verticillium dahliae and Heterodera rostochiensis and treated with methyl bromide, aldicarb or benomyl. Successive crops were grown in subsequent years on the same plots but without further treatment. Largest yields in the year of treatment were from plots fumigated with methyl bromide but in the second crop benomyl-treated plots yielded most. Neither methyl bromide nor aldicarb affected yields from trie third crop. After lifting the first crop, soil from plots treated with methyl bromide or benomyl contained less V. dahliae than that from plots receiving aldicarb or nothing. Soil was also less infective after the second crop following methyl bromide fumigation, but not the third. Plots treated with methyl bromide or aldicarb contained many fewer H. rostochiensis than untreated plots after the first crop, slightly less after the second and equal numbers after the third crop. H. rostochiensis were also fewer in benomyl-treated plots than in untreated after the first crop, but, surprisingly, were fewest after the second crop. The increased yield after applying benomyl to soil seems to depend more on its effect on H. rostochiensis than on V. dahliae. Fumigating with methyl bromide decreased common scab and black scurf on progeny tubers but increased infections by Oospora pustulans.     

Inoculum dynamics ofGliocladium virens associated with roots of cotton seedlings

A system was developed to evaluate the effects of root growth of cotton seedlings on the inoculum dynamics ofGliocladium virens in nonsterile soil. In soil infested withG. virens, inoculum densities of the fungus increased when plants remained alive. After 30 days, shoots were excised and the roots allowed to deteriorate. During this portion of the experiment (30–60 days) soil inoculum densities ofG. virens declined. In infested soil without a seedling, inoculum densities remained constant throughout the duration of the experiments. Colonization of roots byG. virens was found to increase throughout the duration of the experiments. At 60 daysG. virens was recovered from approximately 60% of the root pieces (1-cm) sampled. The percentage of primary, secondary, or tertiary roots colonized was different (P = 0.01), but the total colonization of roots at three depths (0–10, 10–20, and 20–30 cm) was not different (P = 0.64). In noninfested soil, colonization of roots by indigenous propagules ofG. virens was never greater than 3%. Offprint requests to: C. M. Kenerley.     

Rapid in vitro ectomycorrhizal infection onPinus densiflora roots byTricholoma matsutake

The root systems of 11-wk-oldPinus densiflora seedlings were inoculated with a hyphal suspension ofTricholoma matsutake and aseptically incubated for 4 wk in a forest soil without supplying exogenous carbohydrates. One week following inoculation, fungal hyphae had colonized the root surface and bound soil particles together establishing a root-substrate continuum. Fungal hyphae were visible within the main root cortex following clearing bleaching and staining. In the ensuing days, fungal colonization was observed within elongating lateral roots in which Hartig net formation was confirmed 4 wk after inoculation. This is the first report of rapid ectomycorrhizal infection ofP. densiflora seedings byT. matsutake.     

Interrelationship betweenCoprinus lagopus andFusarium udum in soil in relation to competitive saprophytic ability and microbial antagonism

Summary The interrelationship between the growth ofCoprinus lagopus andFusarium udum on pigeon pea substrates in soil was studied at 22-2°C and 30±2°C in relation to their competitive saprophytic ability and antagonism.C. lagopus was observed to be a potent and frequent colonizer of pigeon pea substrates in soil along withF. udum. Saprophytic colonization of pigeon pea substrate byF. udum precolonized byC. lagopus was inhibited in all inoculum soil mixtures. Saprophytic colonization of the substrate byF. udum was better at 22°C than at 30°C and that byC. lagopus better at 30°C than at 22°C. The colonization of substrate by each fungus was suppressed by the other fungus depending upon the temperature used. Hyphal parasitism and colony interactions between these fungi and also with a number of other saprophytic fungi were investigated. No antagonistic reaction was observed betweenF. udum andC. lagopus. However, these fungi were observed to be potent antagonists against other microfungi tested. The formation of fruiting bodies ofC. lagopus was also observed in the laboratory on nutrient media but more frequently on pigeon pea substrates.     

Saprophytic colonization ofFusarium oxysporum f. sp.ciceri in soil

Competitive saprophytic colonization (CSC) of soybean and chickpea stem pieces byFusarium oxysporum f.sp.ciceri increased with the increase in inoculum density in inoculum soil mixtures. The colonization was higher even at loeer concentration of inoculum. Progressive dilutions of autoclaved soils with unsterilized soil decreased the CSC. Lower temperatures favoured the colonization in both red sandy loam and black soils. Maximum colonization occurred at 40°C indicating an inverse relation between colonization and temperature.     

Colonization capacity of the soil micro-organisms in relation to fungistasis

Summary Evans' tube and cellophane agar-disc techniques were used to evaluate the colonization capacity of the soil microbes and fungistasis, respectively. Spore germination ofAlternaria tenuis, Curvularia geniculata, Helminthosporium rostratum andPestalotia sp. was studied on a natural soil. Aspergillus fumigatus was the pioneer colonizer, followed in the order of decreasing colonizing capacity by,A. flavus, A. niger, A. terreus andPenicillium nigricans. Sensitivity of test spores to soil fungistasis declined in the order,C. geniculata → Pestalotia sp. →A. tenuis → H. rostratum. The fungistatic level of the soil samples decreased gradually from side arms 1 to 7 of the Evans tube, inoculated with unsterilized soil from one end. Studies conducted with an inoculum of four soil fungi (A. fumigatus, A. flavus, A. niger, andP. nigricans) as well as those involving fumigation of Evans tube by CS₂ also proved the greater colonizing capacity ofA. fumigatus. The level of fungistasis followed closely the colonization pattern of this fungus. When added individually to the soil, all fungi were able to inhibit spore germination of test fungi to some degree or other.