R. Rodriguez-Kabana

Biological control of nematodes: Soil amendments and microbial antagonists

Organic matter amendments to soil can be used to manage phytoparasitic nematodes. The most effective amendments are those with narrow C:N ratios and high protein or amine-type N content. For soil with 1.0% (w/w) organic matter amendment there is a direct relation between extent of nematode control and the N content of amendments. A special group of amendments are those containing chitinous materials. Chitin addition to soil results in stimulation of a select microflora capable of degrading the polymer. Several microbial species are known to destroy the eggs of phytonematodes (Meloidogyne spp.). Organic matter can be modified by addition of specific compounds or by inoculation with particular microbial species to produce an amendment that will induce suppressiveness.

The determination of soil chitinase activity: conditions for assay and ecological studies

SummaryChitinase activity was determined by incubating a mixture of toluene-treated soil with 1% (w/w) colloidal chitin suspension for 18 h at 37°C and then, after dilution, assaying the amount of N-acetyl-glucosamine released. Maximal chitinase activity was observed at 45°C and optimal pH for enzymatic reaction was 5.0–5.5. Soil chitinase activity decreased with increasing soil depth and was significantly affected by crop cover and fertilization regime. Chitin added to soil stimulated chitinase activity. Enzyme activity was correlated with the soil fungal population but not with numbers of actinomycetes or bacteria. A specialized mycoflora was associated with chitin decomposition.

Rhizosphere bacteria antagonistic to soybean cyst (Heterodera glycines) and root-knot (Meloidogyne incognita) nematodes : identification by fatty acid analysis and frequency of biological control activity

Rhizosphere bacteria were isolated from roots of young and mature plants with known antagonism to phytopathogenic nematodes, including velvet bean (Mucuna deeringiana), castor bean (Ricinus communis), sword bean (Cannavalia ensiformis), and Abruzzi rye (Secale cereale). Isolates from antagonistic plants were compared to soybean isolates for the frequency of antagonism to the root-knot (Meloidogyne incognita) and soybean cyst (Heterodera schachtii) nematodes in a disease assay with soybean. Bacterial isolates were identified using fatty acid analysis, and isolates which exhibited a significant reduction in incidence of soybean damage from both nematodes were characterized physiologically. The bacterial taxa associated with antagonistic plants were markedly different from soybean bacteria. Isolates from soybean were predominantly Bacillus spp., while those from antagonistic plants included more coryneform and Gram-negative genera. Pseudomonas cepacia and Pseudomonas gladioli were predominant among Gram-negative bacteria on antagonistic plants but were not isolated from soybean. Four to six times the number of bacteria from antagonistic plants, compared to soybean, significantly reduced disease incidence of both nematodes. No single pattern of physiological reactions was common among all these bacteria, suggesting that multiple mechanisms accounted for the observed biological control. The results suggest that rhizospheres of antagonistic plants may be useful sources of potential biological control agents for phytopathogenic nematodes.

Effects of free-air CO2 enrichment on microbial populations in the rhizosphere and phyllosphere of cotton

Abstract Cotton ( Gossypium hirsutum L.) plants were exposed to free-air CO 2 enriched (FACE = 550 μmol mol −1 ) or ambient (CONTROL = 370 μmol mol −1 ) levels of atmospheric CO 2 and to wet (100% of evapotranspiration replaced) or dry (67% of ET replaced) soil water content treatments. Foliar, soil and root samples were collected in June and August 1991 to determine the effects of elevated CO 2 on selected groups of phyllosphere and rhizosphere microorganisms. Foliage and rhizosphere soil were analyzed for bacteria and/or fungi using dilution plating. Mycorrhizal colonization of cotton roots was assessed. Root-zone soil was analyzed for populations of nematodes, microarthropods and Rhizoctonia using various extraction methods. A dehydrogenase assay for total microbial respiration and a bioassay for cotton root infecting organisms were also conducted using root-zone soil. Populations of fungi on cotton leaves varied, by genera, in response to CO 2 enrichment, but none was affected by soil water content treatments; populations of foliar bacteria were not affected by either CO 2 or soil water content treatments. In August, higher total numbers of rhizosphere fungi were found under the wet compared with the dry soil water treatment, but differences related to CO 2 were not detected. There was a trend for infestation by Rhizoctonia solani to be higher under FACE in the August sample, but the soil bioassay demonstrated no increase in damping-off potential. There was a significant interaction between CO 2 concentration and soil water content for populations of saprophagous nematodes; populations were different between the CO 2 levels in the dry soil treatment only, with higher numbers under FACE. Microarthropod numbers were low; however, there was a trend for Collembola populations to be higher under FACE in the August sample and more fungi were isolated from Collembola in June. Total microbial activity was higher under FACE at both sample dates. Effects of elevated atmospheric CO 2 on plant microbe interactions could have profound influence on the productivity of agro-ecosystems, and deserve further research.

Analysis of populations and physiological characterization of microorganisms in rhizospheres of plants with antagonistic properties to phytopathogenic nematodes

Populations of rhizosphere microflora of plants which have demonstrated an antagonism toward phytopathogenic nematodes, including velvet bean (Mucuna deeringiana), castor bean (Ricinus communis), sword bean (Cannavalia ensiformis), and Abruzzi rye (Secale cereale)., were compared to the rhizosphere microflora of soybean. Population densities of total bacteria were significantly lower for young Abruzzi rye, mature velvet bean, and mature castor bean, and fungi from mature velvet bean than for soybean. Population densities of spore-forming bacilli were significantly higher for Abruzzi rye than for soybean. Population densities of coryneform bacteria for mature sword bean and velvet bean were significantly higher than for soybean. All seedling test poants supported significantly higher population densities of chitinolytic fungi than soybean. On mature plants, chitinolytic bacteria were significantly higher on all test plants except velvet bean. Populations of endophytic root bacteria for three of the four test plants were significantly higher than for soybean. Fifty randomly, selected bacterial strains from seedlings and mature plants of soybean and each test plant were characterized for various physiological traits associated with rhizosphere competence, including chitinolytic activity, gelatin hydrolysis, production of hydrogen cyanide, starch hydrolysis, phenol oxidation, siderophore production, and production of antifungal compounds (inhibition ofPythium ultimum and/orRhizoctonia solani). There was a strong trend to increased frequency in each of the physiological tests with bacteria from test plants in comparison to those from soybean. The frequency of starch hydrolysis was up to 24 times greater for strains from test plants than for soybean strains, and siderophore production was up to 22 times more frequent for test plants. These results demonstrate that, compared to soybean, plants with properties antagonistic to phytopathogenic nematodes have a distinct rhizosphere microflora.

Potential for biological control of early leafspot of peanut using Bacillus cereus and chitin as foliar amendments

Abstract Chitin applied as an amendment to peanut leaves increased the populations of indigenous chitinolytic epiphytes from 40% of the total microflora. The total epiphytic bacterial populations increased by up to 0.3 log with the addition of chitin. When the chitinolytic Bacillus cereus strain 304 isolated from chitin-amended leaves was reapplied to peanut foliage with chitin, its populations were sustained as actively growing vegetative cells for a longer period than when applied to nonamended leaves. Eight days after application, B. cereus populations were > 1 log higher on chitin-amended leaves than on nonamended leaves. In two of three field trials, significant reductions in the severity of early leafspot caused by Cercospora arachidicola were seen on chitin-amended leaves, and a trend toward additional control (P

Cropping systems for the management of phytonematodes.

Damage caused by nematodes is one of the limiting factors in crop production. Traditional nematode management is based on the use of crop rotations, resistant cultivars, nematicides, or combinations of these methods. For a crop like peanut (Arachis hypogaea), cultivars resistant to root-knot nematodes are not available. There are soybean (Glycine max) cultivars resistant to some of the species of root-knot nematodes (Meloidogyne spp.); however, most fields have nematode infestations composed of mixtures of species. Research at Auburn has shown that tropical crops can be used effectively in rotation to manage nematode problems. Rotations with American jointvetch (Aeschynomene americana), castor (Ricinus communis), hairy indigo (Indigofera hirsuta), partridge pea (Cassia fasciculata), sesame (Sesamum indicum), and velvetbean (Mucuna deeringiana) have resulted in good nematode control and increased yields of peanut and soybean. Some crops (castor, sesame) are considered ‘active’ in that they produce compounds that are nematicidal, whereas others (e.g. corn, sorghum) are simply non-host, that is, ‘passive’.

Effects of crop rotation and fertilization on catalase activity in a soil of the southeastern United States

SummaryCatalase activity of a loamy sand under a 3-year crop rotation in the southeastern U.S.A. was monitored. Corn (Zea mays L.), cotton (Gossypium hirsutum L.), and soybean [Glycine max (L.) Merr.] were the summer crops in the rotation. Winter wheat (Triticum aestivum L.) was planted after corn, and soybean was followed by a winter fallow period. Cotton was followed by a mixture of common vetch (Vicia sativa L.) and crimson clover (Trifolium incarnatum Gibelli & Belli) which was eventually plow-incorporated as a green manure. Highest mean catalase activities were recorded in soil under the wheat, soybean, and winter legume crops; lowest activities were found in soil bearing corn and cotton, and during the winter fallow period. The fertilization regime influenced soil catalase activity independently of the crop. Soil deficient in any of the major elements showed low enzyme activity. Highest activity was found in soil fertilized with P and K, and with N supplied by a winter legume crop. Addition of supplementary mineral nitrogen to this regime reduced catalase activity. Elimination of the winter legume crop from an otherwise complete fertilization regime resulted in a drastic reduction in enzyme activity. In soil receiving a complete fertilization regime there was a close correlation between soil catalase and xylanase activities. A similar correlation between these two enzymes was not found in soil receiving incomplete fertilization.

Integration of soil solarization with chemical, biological and cultural control for the management of soilborne diseases of vegetables

The long-term effectiveness of soil solarization integrated with (integration of pest management [IPM]) a biological control agent (Trichoderma virens), chemical fungicide (pentachloronitrobenzene [PCNB]), organic amendment (chicken litter) or physical method (black agriplastic mulch) to reduce southern blight (Sclerotium rolfsii) and southern root-knot diseases (Meloidogyne incognita) were evaluated on vegetable production. Results showed that the long-term effectiveness of IPM plus soil solarization reduced soilborne diseases of vegetables more than two years following the termination of solarization. These disease management strategies in 1991 and 1992, following soil solarization in 1990, reduced the numbers of sclerotia in the soil, and the number of plants killed by southern blight and root-knot of tomatoes, compared to nonsolarized bare soil treatment. The integration of a reduced dosage level of PCNB or T. virens in field plots, reduced southern blight of tomatoes by 100% and 71%, respectively, in solarized soil, compared to nonsolarized bare soil two years following soil solarization. PCNB effectively controlled southern blight in nonsolarized bare soil both years. All solarized treatments, except PCNB plus solarized soil increased tomato yields compared to nonsolarized bare soil plots. In the second study (1992) following soil solarization in 1991, the effectiveness of solarized bare soil, and nonsolarized bare soil mulched with black agriplastic film, with or without Reemay spunbounded polyester row cover, were effective in reducing root-knot of tomatoes as indicated by the root-knot gall index. Following a one year fallow period in 1994 three years following soil solarization, the root-knot gall index for severity of tomato roots grown in solarized bare soil, nonsolarized bare soil, black agriplastic mulched bare nonsolarized soil and black agriplastic mulched solarized bare soil, were 1.0, 3.0, 3.0 and 2.0, respectively, on a 0–5 scale, where 0=0% and 5=100% root-knot galled. In the third study 1992 and 1993, different dosage levels of chicken litter were used to amend soil artificially infested with sclerotia of S. rolfsii at different depths following solarization, decreased the number of viable sclerotia by 85–100%. All solarized treatments and nonsolarized bare soil amended with 18.8 MT/ha of chicken litter, were effective in controlling southern root-knot damage, and postharvest storage root rots of sweetpotato storage roots (Fusarium root rot [Fusarium solani] and Java black rot [Diplodia tubericola]). Our study showed that all soil solarization treatments, and soils amended with chicken litter, stimulated a shift in the soil microbial population dynamics. Rhizobacteria of Bacillus spp. and fluorescent pseudomonads increased significantly in the rhizosphere, rhizoplane, and interior root tissues of tomatoes and sweetpoatoes, grown in solarized soil compared to nonsolarized soil. These microorganisms may have contributed to the increased growth response of vegetables and some were probably suppressive to soilborne diseases

Evaluation of Natural Chemical Compounds Against Root-lesion and Root-knot Nematodes and Side-effects on the Infectivity of Arbuscular Mycorrhizal Fungi

The survival of two species of plant parasitic nematodes: the root-lesion nematode Pratylenchus brachyurus, and the root-knot nematode Meloidogyne javanica, was evaluated in saturated atmospheres of 12 natural chemical compounds. The infectivity of two isolates of arbuscular mycorrhizal fungi: Glomus mosseae and Glomus intraradices, under identical experimental conditions, was also determined. All the compounds tested exerted a highly significant control against M. javanica and among them, benzaldehyde, salicilaldehyde, borneol, p-anisaldehyde and cinnamaldehyde caused a mortality rate above 50% over P. brachyurus. The infectivity of G. intraradices was inhibited by cinnamaldehyde, salicilaldehyde, thymol, carvacrol, p-anisaldehyde, and benzaldehyde, while only cinnamaldehyde and thymol significantly inhibited mycorrhizal colonization by G. mosseae.