Feeding of an "obligate" plant-parasitic nematode (nonfungal feeder), Pratylenchus scribneri, in the absence of plant tissue was demonstrated in an artificial system consisting of liquid media and indicator dyes including amaranth and various nontoxic food colors. Among the compounds tested, sucrose, dextrose, Gamborg's B5 medium, and DL-methionine stimulated a small percentage of feeding (12-36%). A high percentage of feeding (90-100%) occurred in a filtrate from excised corn roots cultured in Gamborg's B5 medium. This feeding system has the potential to develop an artificial medium for plant-parasitic nematodes and to screen novel nematicides that are stomach poisons.
amaranth; dye; feeding stimuli; feeding system; food color; indicator; nematode bait; Pratylenchus scribneri; screening
A drip irrigation delivery system was used to infest field sites with the plant-parasitic root-knot nematodes, Meloidogyne incognita. Juvenile or egg inocula passed through the system without blockage of emitters or harm to the nematodes. Field sites so infested were available for experimentation. Delivery of approximately 5 x 10⁴ to 10⁵ juveniles or 10⁵ to 3 x 10⁵ eggs per emitter through the drip system resulted in heavy root galling of tomatoes planted next to the drip emitters. Nematodes feeding on bacteria (Acrobeloides sp.) and on fungi (Deladenus durus) also were successfully applied through the drip system. This method has potential for uniformly infesting experimental sites with plant-parasitic or entomogenous nematodes and for manipulation of nematode community structure for soil ecological studies.
Acrobeloides sp.; biogation; Deladenus durus; drip irrigation; field infestation; Lycopersion esculentum; Meloidogyne incognita; root-knot nematode
Two grape cultivars, susceptible French Colombard and tolerant Rubired, and four nematodes, Meloidogyne incognita, Pratylenchus vulnus, Tylenchulus semipenetrans, and Xiphinema index, were used to quantify the equilibrium between root (R) and shoot (S) growth. Root and shoot growth of French Colombard was retarded by M. incognita, P. vulnus, and X. index but not by T. semipenetrans. Although the root growth of Rubired was limited by all the nematodes, the shoot growth was limited only by X. index. The R:S ratios of Rubired were higher than those of French Colombard. The reduced R:S ratios of Rubired were primarily an expression of reduction in root systems without an equal reduction in shoot growth, whereas in French Colombard the reduced R:S ratios were due to a reduction in both shoot growth and root growth and to a greater reduction in root growth than shoot growth. All nematodes reproduced equally well on both cultivars. Both foliage and root growth of French Colombard were significantly reduced by M. incognita and P. vulnus. Nematodes reduced the shoot length by reducing the internode length. Accumulative R:S ratios in inoculated plants were significantly smaller than those in controls in all nematode treatments but not at individual harvest dates. Bud break was delayed by X. index and was initiated earlier by P. vulnus and M. incognita. All buds in nematode treatments were less vigorous than in controls.
French Colombard grape; Meloidogyne incognita; Pratylenchus vulnus; Rubired grape; Tylenchulus semipenetrans; Vitis vinifera; Xiphinema index
Pluronic F127 polyol, a block copolymer of propylene oxide and ethylene oxide, was studied as an alternative to agar in culture media for nematodes, bacteria, fungi, actinomycetes, and plant tissues or seedlings, At a polyol concentration of 20% w/v, the culture media, semi-solid at room temperature (22 C) but liquid at lower temperatures, had minimal effects on the test organisms. Most of the fungi and bacteria grew as well in 20% polyol as in 1.5% agar media; however, various species of nematodes and plant seedlings or tissues exhibited differential sensitivities to different concentrations of the polyol. In cases where the organisms were unaffected, the polyol media had certain advantages over agar, including greater transparency and less contamination under nonaseptic conditions. Polyol media have potentially greater ease for recovery of embedded organisms or tissues inside the media by merely shifting to lower temperatures.
plant tissue culture; pluronic polyol; polyglycol; soil micro-organism; solidifying agent
Variability in the reproduction of the four races ofMeloidogyne incognita on the soybean cuhivars Pickett 71 and Centennial was studied in growth chamber experiments. Analysis of variance in the number of eggs produced by the races 6 weeks after the plants had been inoculated with 5,000 eggs of each race revealed that the nematode race by soybean cultivar interaction was highly significant (P = 0.001). Races 1, 3, and 4 produced from about 5,000 to 15,000 eggs per root system on Pickett 71 and only from about 300 to 600 eggs per root system on Centennial. In contrast, race 2 produced about 8,000 eggs per root system on Centennial and about 1,200 eggs per root system on Pickett 71. In a second experiment, in which the plants were inoculated with 2,000 second-stage juveniles, race 1 and race 2 produced about 13,000 and 3,000 eggs per root system, respectively, on Pickett 71 and about 600 and 10,000 eggs per root system, respectively, on Centennial. The results suggest that M. incognita resistance in soybean is race-specific.
Meloidogyne incognita; root-knot nematodes; resistance; Glycine max
The efficacy of avermectins B₁ and B₂ for control of Meloidogyne incognita on tomato was studied in pots and field plots for two seasons. Avermectins were applied as granules and liquid in furrows or by low pressure drip irrigation systems, at rates ranging from 0.093 to 0.34 kg a.i./ha, as single or multiple applications. Levels of control comparable to those obtained by oxamyl and aldicarb at 3.36 kg a.i./ha were achieved by the avermectin with only 1/10 the volume of chemicals applied to the environment. Avermectin protection of the roots remained constant throughout the first 5 weeks giving slightly longer protection than oxamyl or aldicarb.
Lycopersicon esculentum; root-knot nematodes; furrow irrigation; low pressure drip irrigation; PVC injector
The effects of soil temperature and initial inoculum density (Pi) of Meloidogyne incognito and M. javanica on growth of wheat (Triticum aestivum cv. Anza) and nematode reproduction were studied in controlled temperature baths in the glasshouse. Nematode reproduction was directly proportional to temperature between 14 and 30 C for M. incognita and between 18 and 26 C for M. javanica. Reproduction rates (Pf/Pi, where Pf = final number of eggs) for Pi's of 3,000, 9,000, and 30,000 eggs/plant were greatest at each temperature when Pi = 3,000. Maximum M. incognita reproduction rate (Pf/Pi = 51.12) was at 30 C. At 26 C, M. javanica reproduction (Pf/Pi = 14.82, 9.02, and 4.23 for Pi = 3,000, 9,000, and 30,000, respectively) was about half that of M. incognita when Pi = 3,000 or 9,000 but similar when Pi = 30,000. Reproduction of both species was depressed between 14 and 18 C. Shoot and root growth and head numbers were inversely related to soil temperature between 14 and 30 C but were not affected by the Pi of M. incognita when 7 d old seedlings were inoculated. When newly germinated seedlings were inoculated with M. incognita or M. javanica, the Pi did not affect shoot and root fresh weights, shoot/root ratio, and tillering, but it did reduce root dry weight (M. javanica at 26 C) and increase shoot dry weight (M. incognita at 18-22 C). The optimum temperature range is lower for wheat growth than for nematode reproduction. Wheat cv. Anza is a good host for M. incognita and M. javanica, but it is tolerant to both species.
temperature; root-knot nematodes; tolerance; population dynamics
Wheat cultivars Anza and Produra grown in winter in California were planted in Meloidogyne incognita infested and noninfested sandy loam plots in October (soil temperature 21 C) and November (soil temperature 16 C) of 1979. Meloidogyne incognita penetrated roots of mid-October planted Ataza (427 juveniles/g root), developed into adult females by January, and produced 75 eggs/g root by harvest in April. Penetration and development did not occur in late plantings. Anza seedlings grown in infested soil in pots buried in field soil in early spring were not invaded until soil temperature exceeded 18 C. Meloidogyne incognita juveniles can migrate through soil and penetrate roots at temperatures above 18 C (activity threshold), however development can occur at lower temperatures. Grain yields were not significantly different between nematode infested (3,390 kg/ha) and noninfested (2,988 kg/ha) plots. Winter decline of eggs and juveniles in two late plantings anti in fallow soil were 69, 72, and 77%, respectively, but egg and juvenile decline was only 40% in the early Anza plots that supported nematode reproduction in the spring. Delay of planting date until soil temperature is below 18 C is suggested to maximize the use of wheat in rotation as a nematode pest management cultural tactic for suppressing root-knot nematodes.
root-knot nematodes; population dynamics; nematode pest management; Triticum aestivum
The vertical migration of M. incognita juveniles introduced at 20 cm from the roots was studied in five natural soils, 100% silica sand, 95% silica sand with 5% clay, 90% silica sand with 10% clay, and 95% silica sand with 5% clay as a concentrated layer. In natural soils the percentage of juveniles capable of migrating 20 cm and penetrating the roots decreased when the percentage of clay and silt increased. No migration occurred in silica sand without clay particles; when 5 or 10% of clay were mixed to silica sand, 34 and 26%, respectively, of the juveniles were able to migrate 20 cm. Clay separated from silica sand in which tomatoes were grown was attractive for juveniles. It is suggested that clay particles aid in the migration of root-knot juveniles over long distances to plant roots by absorbing and holding root exudates or bacterial by-products which form a concentration gradient enabling nematodes to locate roots.
attraction; nematode movement
Photoperiod influences the migration of M. incognita juveniles toward tomato roots. Approximately 33% migrated vertically 20 cm in 7 days to roots when 12 h dark were alternated with 12 h light. Only 7% migrated when light was constant for 24 h. Vertical migration of M. incognita juveniles was studied at 14, 16, 18, 20, and 22 C. The migration of M. incognita juveniles begins at about 18 C and reaches its maximum at 22 C. The migration of M. hapla and M. incognita juveniles were compared at 14, 18, and 22 C. Juveniles of M. hapla were able to migrate at a lower temperature than those of M. incognita. With M. hapla, there was no significant difference in migration between 18 and 22 C.
Meloidogyne incognita; Meloidogyne hapla; attraction; photoperiod
The effect of methyl bromide (MB) was tested on active and anhydrobiotic Aphelenchus avenae. A. avenae was induced into anhydrobiosis by three different techniques. Both active and anhydrobiotic nematodes were subjected to 3,000 μ1 MB/liter air for 14 periods from 0 to 82 h. Anhydrobiotic nematodes were more resistant to fumigation than active nematodes, regardless of the technique used to induce anhydrobiosis. The percent survival decreased with increasing MB exposures (μ1 MB × h). For an LD₉₅ of 45,000-54,000 μ1/1 × h were required for active nematodes and >279,000 μ1/1 × h for anhydrobiotic nematodes.
anhydrobiosis; Aphelenehus avenae; survival; methyl bromide; soil fumigation
Nematodes of three genera (Acrobeloides sp., Aphelenchus avenae, and Scutellonema brachyurum) were induced to coil and enter anhydrobiosis in drying soil of two types: sandy loam and loamy sand. Coiling was studied in relationship to soil moisture characteristics. Coiling and the physiological state of anhydrobiosis occurred before the water in sandy soils reached a water potential of -15 bars. Coiling was maximum at 3-6 bars, depending on the soil type and nematode species. It appeared that induction of coiling and anhydrohiosis were determined by the physical forces exerted by the water film surrounding the nematode, which, for these three species, was 6-9 monomolecular layers of water, rather than the % moisture and relative humidity of the soil per se.
anhydrobiosis; Aphelenchus avenae; Acrobeloides; Scutellonema brachyururn; soil moisture; survival; monomolecular layers of water
In laboratory testing, egg hatch of Tylenchulus semipenetrans was stimulated at concentrations of 1 and 10 μg/ml aldicarb solution and inhibited at 50 and 100 μg/ml. Aldicarb was more inhibitory to egg hatch than the aldicarb sulfoxide and the aldicarb sulfone. Inhibition of hatch at the high concentration was associated with delays in the molting processes, lack of larval movement within the egg, and delays in embryonic development. Nematode motility was reduced at 10, 50, and 100 μg/ml of aldicarb and aldicarb sulfoxide solution, and at 50 and 100 μg/ml aldicarb sulfone. Male development was retarded at 10 μg/nrl and almost completely inhibited at 50 and 100 μg/ml of the three chemicals. In greenhouse tests, female development antl reproduction on roots of citrus seedlings were suppressed by aldicarb at rates of 2.6 μg/ml and completely inhibited at 10.6 μg/ml of soil solution during a 50-day experimental period. Under field conditions, there was little systemic movement of aldicarb into roots located outside treated areas. Aldicarb reduced the nematode larvae and the female adult population in the second year after the second treatment. There were no differences in egg hatch and sex ratio of citrus nematodes between treated and nontreated roots.
citrus nematode; hatching; development; reproduction; chemical control; nonfumigant nematicide; motility; mortality; mode of action
Anhydrobiotic nematodes were fixed and extracted from dry Mojave desert soils with hot and cold fixatives (5% formalin and 4% gluteraldehyde). Morphologically, extracted nematodes were tightly coiled and shrunken in size. Various concentrations of KCl, ethylene glycol, and sucrose solutions were compared for their effectiveness in extracting viable nematodes still in the anhydrobiotic state. Approximately 80-95% of the anhydrobiotic nematodes extracted with 1.25 M and 1.5 M sucrose were tightly coiled and shrunken in a manner similar to those fixed and extracted in formalin. Anhydrobiotic nematodes can be maintained in this form for up to 24 days in 1.25 M sucrose solution and still be revived. All molarities of ethylene glycol and KC1 tested were ineffective in recovering and maintaining nematodes in the natural anhydrobiotic state. Straight, air-dried, and active nematodes served as controls and did not coil when placed in hot or cold fixatives or in any concentration of KCI, ethylene glycol, or sucrose. Anhydrobiosis, as represented by the coiled form of nematodes from desert soils, was not confined to any particular life stage or trophic group.
cryptobiosis; nematode survival
A severe root rot of tomato caused by Meloidogyne incognita and Rhizoctonia solani was associated with nutrient mobilization into gall tissue and root exudation. Root decay did not develop when root exudates were continuously removed by leaching. When leachates were collected from M. incognita-infected and control roots and applied to roots of tomatoes inoculated with R. solani alone, roots receiving leachates from M. incognita-infected roots developed a severe rot while roots receiving leachates from control roots were free of decay. During the fourth and fifth weeks following nematode infection, an increased mobilization of ¹⁴C labelled compounds to nematode-infected roots occurred. Higher counts of water soluble nonvolatile ¹⁴C labelled exudates leaked from nematode-infected roots than from control roots at each weekly sampling period. Higher concentrations of Ca, Mg, Na, K, Fe, and Cu were found in exudates from nematode-infected roots than from the control roots. During the first 14 days following nematode infection, carbohydrates were the major organic constituents in exudates leaking from nematode-infected roots. Fourteen days after nematode infection, nitrogenous compounds become the major organic constituents leaking from roots. Shifts in C/N ratio of root exudates from nematode-infected roots were associated with parasitic development of R. solani in tomato roots.
fungus-nematode interactions; Meloidogyne incognita; root exudates; C/N ratios
The lesion nematode, Pratylenchus thornei, was clearly demonstrated as a parasite of wheat. It reduced plant stands and stunted plants in the field under the environmental conditions found in Sonora, Mexico. Other soil organisms also may have contributed to the problem. The nematode is widely distributed throughout the wheat-growing region, and may be a problem each growing season. Nematicides controlled the nematode and increased yields, but they were not economical. No resistance was found in existing commercial wheat cultivars. A pest management approach using variety selection, nitrogen fertilizer, planting in cool soil (15 C) and a crop rotation avoiding wheat after wheat was the most practical solution to this problem on a commercial scale.
lesion nematode; resistance; crop rotation; chemical control; fertilizer; temperature; nematicides
In microaerobic and anaerobic environments the principal glycolytic end-product of A. avenae and Caenorhabditis sp. was lactic acid during the first 12-16 hr, after which it was ethanol. Upon return to aerobiosis, ¹⁴C-labeled ethanol in the medium was utilized by the nematodes; ¹⁴CO₂ and some ¹⁴C-labeled glycogen was detected. Total dry weight loss of non-feeding nematodes was 25% greater in the absence of alcohol than in the presence of ethanol or n-propanol. Physical movement and respiration increased and reproduction was extended by alcohol in the bathing solution.
Lactic acid; Acetaldehyde; Succinic acid; Aerobic; Anaerobic; Microaerobic; Glycolysis; Alcohol dehydrogenase; Lactic dehydrogenase
Starving Aphelenchus avenae survived 3-4 weeks in microaerobic and anaerobic environments, but Caenorhabditis sp. survived less than 80 hr. Aerobically, both nematodes metabolize neutral lipid reserves: there was no microaerobic ( <5% O₂) or anaerobic neutral lipid catabolism. Early in anaerobiosis both nematodes utilized endogenous glycogen. Caenorhabditis sp. depleted the glycogen and died. A. avenae under oxygen stress longer than 120 hr entered cryptobiosis, during which there was neither measurable O₂ uptake nor glycogen or neutral lipid utilization, Only when re-aerated, did A. avenae recover and resume "'normal" metabolism.
Aeration; Glycogen metabolism; Neutral lipid catabolism; Aphelenchus avenae; Caenorhabditis sp.; Cryptobiosis
Reproduction of Aphelenchus avenae, reared on Rhizoctonia solani growing on steamed wheat seeds and Caenorhabditis sp., reared on a mixed bacterial culture grown on oatmeal, was significantly reduced at 5% oxygen and inhibited at 4% oxygen and below. Aeration ranging from atmospheric air (21%) to 10% oxygen had no effect on reproduction. Close interval (5 days or less) fluctuations, between high and low oxygen concentrations, inhibited population buildup of Hemicycliophora arenaria on tomato in soil, and of A. avenae and Caenorhabditis sp. in vitro. In soil tests with H. arenaria exposed to 12 hr of nitrogen every three days (in air) inhibited the rate of buildup compared to controls maintained in continuous air. With the in vitro studies, as little as 4 hr nitrogen every 3 days (stored in air) significantly influenced the population numbers.
Hemicycliophora arenaria; Aphelenchus avenae; Caenorhabditis sp.; Oxygen; Nitrogen; Aeration; Reproduction; Population
The onset of molting in all stages of Hemicycliophora arenaria was preceded by the appearance of numerous, discrete globular structures which were termed "molting bodies" because they were present in the hypodermis only during the production of the new cuticle. In all parasitic stages the molt commenced with the separation of the cuticle from the hypodermis from which the new sheath and cuticle were differentiated. Following completion of the new sheath and cuticle most of the old outer covering was apparently absorbed before ecdysis. Electronmicrographs of body wall cross sections in molting L4 male specimens revealed the final molt to be a double molt in which an additional sixth cuticle was produced. Since both a new sheath and cuticle were produced during the molt of each stage, the sheath must be considered as an integral part of the cuticle and not as a residual cuticle or the result of an incomplete additional molt. Molting in Aphelenchus avenae and Hirschmanniella gracilis was less complex and "molting bodies" were not observed. After cuticle separation the hypodermis gave rise to a new trilaminate zone, the future cortex, and (later) the matrix and striated basal layers.
Cuticle; Molting; Ultrastructure; Hemicycliophora arenaria; Aphelenchus avenae; Hirschmanniella gracilis
Ultrathin sections of all parasitic stages of Hemicycliophora arenaria revealed two major divisions in the body covering. The outermost was a seven-layered sheath and the innermost a five-layered cuticle comprising three zones; an outer trilaminate cortex, a fibrillar matrix and a striated basal layer. The body covering of the nonparasitic males also exhibited two major divisions: the outer, a relatively thin four-layered sheath and the inner, a six-layered cuticle consisting of three zones; an outer trilaminate cortex, a two-layered matrix and a striated basal layer. The cuticles of all stages of Aphelenchus avenae were similar, consisting of five layers divisible into three zones; an outer trilaminate cortex, a fibrillar matrix and a striated basal layer. Hirschmanniella gracilis and H. belli cuticles were also similar in all stages examined, consisting of six layers divisible into three zones; an outer trilaminate cortex, a two-layered matrix and a striated basal layer.
Cuticle; Ultrastructure; Hemicycliophora arenaria; Aphelenchus avenae; Hirschmanniella gracilis; Hirschmanniella belli
In the present study, evaluations of the nominal species and descriptions (H. parvus, H. intermedius, H. insignis, H. microdoratus and H. brevicaudatus) are made of five new species. H. squamosus is proposed as a species inquirenda; H. mangiferae is judged a valid species. H. litchi and H. birchfieldi are synonymized with H. mangiferae and H. ureshinoensis with H. kanayaensis. H. strictathecatus is considered a valid species. A key to the species of the genus is given. The limitation of use of male diagnostic characters in a mixture of populations of Hemicriconemoides with some other criconematids is discussed. The life cycle of H. chitwoodi has four complete molts; one in the egg, three in soil, two of which have spines. The adult female has a sheath cuticle from an incomplete fifth molt.