Sensitivity of Meloidogyne incognita and Rotylenchulus reniformis to Abamectin

Avermectins are macrocyclic lactones produced by Streptomyces avermitilis. Abamectin is a blend of B_1a and B_1b avermectins that is being used as a seed treatment to control plant-parasitic nematodes on cotton and some vegetable crops. No LD₅₀ values, data on nematode recovery following brief exposure, or effects of sublethal concentrations on infectivity of the plant-parasitic nematodes Meloidogyne incognita or Rotylenchulus reniformis are available. Using an assay of nematode mobility, LD₅₀ values of 1.56 μg/ml and 32.9 μg/ml were calculated based on 2 hr exposure for M. incognita and R. reniformis, respectively. There was no recovery of either nematode after exposure for 1 hr. Mortality of M. incognita continued to increase following a 1 hr exposure, whereas R. reniformis mortality remained unchanged at 24 hr after the nematodes were removed from the abamectin solution. Sublethal concentrations of 1.56 to 0.39 μg/ml for M. incognita and 32.9 to 8.2 μg/ml for R. reniformis reduced infectivity of each nematode on tomato roots. The toxicity of abamectin to these nematodes was comparable to that of aldicarb.     

Evaluating the predatory potential of carnivorous nematodes against Rotylenchulus reniformis and Meloidogyne incognita

Predatory behavior of a nematode is usually determined through gut content observation or prey delimitation counts. In this experiment, Mononchus and Neoactinolaimus predation of Rotylenchulus reniformis or Meloidogyne incognita was determined using a PCR-based nematode gut content analysis. Soil samples naturally infested with Mononchus were placed in tubes and potential prey nematodes R. reniformis, M. incognita, or a mixture of both were introduced. The gut contents of Mononchus were assayed for the DNA from R. reniformis or M. incognita using PCR specific primers. A higher % of Mononchus tested positive for DNA of R. reniformis than for M. incognita when the prey were added alone. However, when provided with both prey species, Mononchus was tested positive for DNA of M. incognita more frequently than for R. reniformis. Percent Mononchus testing positive for DNA of R. reniformis correlated positively with the abundance of R. reniformis, but this relationship was not observed between Mononchus and M. incognita. Neoactinolaimus was added to aqueous solution containing a mixture of free-living nematodes and R. reniformis. More Neoactinolaimus tested positive for DNA of R. reniformis than other predatory or omnivorous nematodes in the same samples. Based on regression analysis, the presence of fungivorous and other predatory nematodes in the soil could distract Neoactinolaimus from predation on R. reniformis. Our results suggested that Prismatolaimus, Mesodiplogasteroides and Eudorylaimus could also prey on R. reniformis. Although less than 40% of the predatory or omnivorous nematodes tested preyed on R. reniformis, this level of predation could contribute to reducing the population densities of plant-parasitic nematodes in the soil.     

Sensitivity of Meloidogyne incognita and Rotylenchulus reniformis to Fluopyram

Fluopyram is a succinate dehydrogenase inhibitor (SDHI) fungicide that is being evaluated as a seed treatment and in-furrow spray at planting on row crops for management of fungal diseases and its effect on plant-parasitic nematodes. Currently, there are no data on nematode toxicity, nematode recovery, or effects on nematode infection for Meloidogyne incognita or Rotylenchulus reniformis after exposure to low concentrations of fluopyram. Nematode toxicity and recovery experiments were conducted in aqueous solutions of fluopyram, while root infection assays were conducted on tomato. Nematode paralysis was observed after 2 hr of exposure at 1.0 µg/ml fluopyram for both nematode species. Using an assay of nematode motility, 2-hr EC₅₀ values of 5.18 and 12.99 µg/ml fluopyram were calculated for M. incognita and R. reniformis, respectively. Nematode recovery in motility was greater than 50% for M. incognita and R. reniformis 24 hr after nematodes were rinsed and removed from a 1-hr treatment of 5.18 and 12.99 µg/ml fluopyram, respectively. Nematode infection of tomato roots was reduced and inversely proportional to 1-hr treatments with water solutions of fluopyram at low concentrations, which ranged from 1.3 to 5.2 µg/ml for M. incognita and 3.3 to 13.0 µg/ml for R. reniformis. Though fluopyram is nematistatic, low concentrations of the fungicide were effective at reducing the ability of both nematode species to infect tomato roots.     

Pathogenicity of Pratylenchus penetrans,Heterodera glycines,and Meloidogyne incognita on Soybean Genotypes

The pathogenicity of Heterodera glycines, Meloidogyne incognita, and Pratylenchus penetrans on H. glycines-resistant ''Bryan,'' tolerant-susceptible ''G88-20092,'' and intolerant-susceptible ''Tracy M'' soybean cultivars was tested using plants grown in 800 cm³ of soil in 15-cm-diam. clay pots in three greenhouse experiments. Plants were inoculated with 0, 1,000, 3,000, or 9,000 H. glycines race 3 or M. incognita eggs, or vermiform stages of P. penetrans/pot. Forty days after inoculation, nmnbers of all three nematodes, except H. glycines on Bryan, generally increased with increasing inoculum levels in Experiment I. Heterodera glycines and M. incognita significantly decreased growth only of Tracy M. At 45 and 57 days after inoculation with 6,000 individuals/pot in experiments II and III, respectively, significantly more P. penetrans and M. incognita than H. glycines were found on Bryan. However, H. glycines and M. incognita population densities were greater than P. penetrans on G88-20092 and Tracy M. Growth of Tracy M infected by H. glycines and M. incognita and growth of G88-20092 infected by M. incognita decreased in Experiment III. Pratylenchus penetrans did not affect plant growth. Reduction in plant growth differed according to the particular nematode species and cultivar, indicating that nematodes other than the species for which resistance is targeted can have different effects on cultivars of the same crop species.     

SSR marker and ITS cleaved amplified polymorphic sequence analysis of soybean × Glycine tomentella intersubgeneric derived lines

Wild perennial Glycine species are an invaluable gene resource for the cultivated soybean [Glycine max (L.) Merr., 2n=40]. However, these wild species have been largely unexplored in soybean breeding programs because of their extremely low crossability with soybean and the need to employ in vitro embryo rescue methods to produce F₁ hybrids. The objective of this study was to develop molecular markers to identify gene introgression from G. tomentella, a wild perennial Glycine species, to soybean. A selection of 96 soybean simple sequence repeat (SSR) markers was evaluated for cross-specific amplification and polymorphism in G. tomentella. Thirty-two SSR markers (33%) revealed specific alleles for G. tomentella PI 483218 (2n=78). These SSR markers were further examined with an amphidiploid line (2n=118) and monosomic alien addition lines (MAALs), each with 2n=40 chromosomes from soybean and one from G. tomentella. The results show that the use of SSR markers is a rapid and reliable method to detect G. tomentella chromosomes in MAALs. We also developed a cleaved amplification polymorphism sequence (CAPS) marker according to the sequences of internal transcribed spacer (ITS) regions in soybean and G. tomentella. Four MAALs that carry the ITS (rDNA) locus from G. tomentella were identified. The SSR and ITS-CAPS markers will greatly facilitate the introgression and characterization of gene transfer from G. tomentella to soybean.Communicated by F.J. Muehlbauer     

Extraction of Root-associated Meloidogyne incognita and Rotylenchulus reniformis

A technique based on physical maceration of root tissue was developed to extract vermiform and swollen stages of Meloidogyne incognita and Rotylenchulus reniformis. Experiments conducted on soybean and tomato evaluated the efficiency of method (stir, grind), NaOC1 concentration (0%, 0.5%), and duration (lx, 2x) on extraction of nematodes and eggs from 60-day-old populations. Root-associated populations of R. reniformis were considerably lower than those of M. incognita, so development of the method focused on the latter. Grinding liberated more nematodes than stirring, but the reverse was true for egg extraction. Among grinding treatments, a duration of 10 seconds in 0.5% NaOCl provided the most efficient extraction of nematodes and eggs. Among stirring treatments, a duration of 10 minutes in 0.5% NaOCl provided the most efficient extraction of eggs. These techniques were compared on soybean roots 30 days older than those on which the procedures were first evaluated, with consistent results.     

Response of Resistant Soybean Plant Introductions to Meloidogyne incognita in Field Microplots

The response of two soybean plant introductions, PI 96354 and PI 417444, highly resistant to Meloidogyne incognita, to increasing initial soil population densities (Pi) (0, 31, 125, and 500 eggs/100 cm³ soil) of M. incognita was studied in field microplots for 2 years. The plant introductions were compared to the cultivars Forrest, moderately resistant, and Bossier, susceptible to M. incognita. Averaged across years, the yield suppressions of Bossier, Forrest, PI 417444, and PI 96354 were 97, 12, 18, and < 1%, respectively, at the highest Pi when compared with uninfested control plots. Penetration of roots by second-stage juveniles (J2) increased linearly with increasing Pi at 14 days after planting. At the highest Pi, 62% fewer J2 were present in roots of PI 96354 than in roots of the other resistant genotypes. Soil population densities of M. incognita were lower on both plant introductions than on Forrest. At 75 and 140 days after planting, PI 96354 had the lowest number of J2 in the soil, with 49% and 56% fewer than Forrest at the highest Pi. The resistance genes in PI 96354 should be useful in a breeding program to improve the level of resistance to M. incognita in soybean cultivars.     

Impact of Soil Texture on the Reproductive and Damage Potentials of Rotylenchulus reniformis and Meloidogyne incognita on Cotton

The effects of soil type and initial inoculum density (Pi) on the reproductive and damage potentials of Meloidogyne incognita and Rotylenchulus reniformis on cotton were evaluated in microplot experiments from 1991 to 1993. The equilibrium nematode population density for R. reniformis on cotton was much greater than that of M. incognita, indicating that cotton is a better host for R. reniformis than M. incognita. Reproduction of M. incognita was greater in coarse-textured soils than in fine-textured soils, whereas R. reniformis reproduction was greatest in a Portsmouth loamy sand with intermediate percentages of clay plus silt. Population densities of M. incognita were inversely related to the percentage of silt and clay, but R. reniformis was favored by moderate levels of clay plus silt (ca. 28%). Both M. incognita races 3 and 4 and R. reniformis effected suppression of seed-cotton yield in all soil types evaluated. Cotton-yield suppression was greatest in response to R. reniformis at high Pi. Cotton maturity, measured as percentage of open bolls at different dates, was affected by the presence of nematodes in all 3 years.     

Competition of Meloidogyne incognita and Rotylenchulus reniformis on Cotton Following Separate and Concomitant Inoculations

It has been hypothesized Rotylenchulus reniformis (Rr) has a competitive advantage over Meloidogyne incognita (Mi) in the southeastern cotton production region of the United States. This study examines the reproduction and development of Meloidogyne incognita (Mi) and Rotylenchulus reniformis (Rr) in separate and concomitant infections on cotton. Under greenhouse conditions, cotton seedlings were inoculated simultaneously with juveniles (J2) of M. incognita and vermiform adults of R. reniformis in the following ratios (Mi:Rr): 0:0, 100:0, 75:25, 50:50, 25:75, and 0:100. Soil populations of M. incognita and R. reniformis were recorded at 3, 6, 9, 14, 19, 25, 35, 45, and 60 days after inoculations. At each date, samples were taken to determine the life stage of development, number of egg masses, eggs per egg mass, galls, and giant cells or syncytia produced by the nematodes. Meloidogyne incognita and R. reniformis were capable of initially inhibiting each other when the inoculum ratio of one species was higher than the other. In concomitant infections, M. incognita was susceptible to the antagonistic effect of R. reniformis. Rotylenchulus reniformis affected hatching of M. incognita eggs, delayed secondary infection of M. incognita J2, reduced the number of egg masses produced by M. incognita, and reduced J2 of M. incognita 60 days after inoculations. In contrast, M. incognita reduced R. reniformis soil populations only when its proportion in the inoculum ratio was higher than that of R. reniformis. Meloidogyne incognita reduced egg masses produced by R. reniformis, but not production of eggs and secondary infection.     

Effects of Acibenzolar-S-Methyl Application to Rotylenchulus reniformis and Meloidogyne javanica

Effects of acibenzolar-s-methyl, an inducer of systemic acquired resistance in plants, on Rotylenchulus reniformis and Meloidogyne javanica in vitro and in vivo were determined. A single foliar application of acibenzolar at 50 mg/liter (5 ml of solution per plant) to 7-day-old cowpea or soybean seedlings decreased R. reniformis and M. javanica egg production by 50% 30 days after inoculation. The mechanism of acibenzolar on plant-parasitic nematodes was then investigated. Acibenzolar at 50 to 200 mg/liter did not affect movement of R. reniformis and M. javanica or penetration of second-stage juveniles (J2) of M. javanica on cowpea. However, M. javanica development was slowed and fecundity was reduced in plants treated with acibenzolar. On average, 50% of J2 that penetrated acibenzolar-treated cowpeas developed into mature females with eggs, whereas the other 50% exhibited arrested development. The number of eggs per egg mass was 450 in water-treated cowpeas, whereas the number declined to 250 in acibenzolar-treated plants. Acibenzolar may be responsible for stimulating the plants to express some resistance to the nematodes.     

Effect of Meloidogyne incognita on Reproduction of Pratylenchus penetrans in Red Clover and Alfalfa

Roots of seedlings of red clover and alfalfa growing on 10⁻¹ Hoagland and Arnon solution agar were inoculated with various combinations of Meloidogyne incognita and Pratylenchus penetrans. Egg-laying by P. penetrans decreased as the number of nematodes, the ratio of entrant M. incognita to entrant P. penetrans, and the priority of invasion of roots by M. incognita increased. Embryogeny and hatching of eggs of P. penetrans, and development of larvae of M. incognita, were not affected. In red clover, the greatest red uction occurred when there were 65 entrant nematodes, the ratio of M. incognita:P. penetrans was 4:1 and M. incognita was inoculated four days prior to P. penetrans. In alfalfa, the less-favorable host for both nematodes, the greatest reduction occurred when there were 45 entrant nematodes, the ratio of M. incognita:P. penetrans was 2:1, and M. incognita was inoculated 4 days prior to P. penetrans.     

Toxicity of Tioxazafen to Meloidogyne Incognita and Rotylenchulus Reniformis

Tioxazafen is a seed-applied nematicide used in row crops. Currently, there are no data on nematode toxicity, nematode recovery, or effects of low concentrations of tioxazafen on nematode infection of a host root for Meloidogyne incognita or Rotylenchulus reniformis. Nematode toxicity and recovery experiments were conducted in water solutions of tioxazafen, while root infection assays were conducted on tomato. Nematode paralysis was observed after 24 hr of exposure at 27.0 µg/ml tioxazafen for both the nematode species. Based on an assay of nematode motility, 24-hr EC₅₀ values of 57.69 µg/ml and 59.64 µg/ml tioxazafen were calculated for M. incognita and R. reniformis, respectively. Tioxazafen rates of 2.7 µg/ml and 27.0 µg/ml reduced the nematode hatch after 3 d of exposure for both the nematode species. There was no recovery in nematode motility after the 24-hr exposure of M. incognita and R. reniformis to their corresponding 48-hr EC₅₀ values of 47.15 µg/ml and 47.25 µg/ml tioxazafen, respectively. Mortality of M. incognita continued to increase after 24 hr exposure, whereas R. reniformis mortality remain unchanged after nematodes were rinsed and removed for 48 hr from the tioxazafen solution. A 24-hr exposure to low concentrations of 0.38 to 47.15 µg/ml for M. incognita and 47.25 µg/ml for R. reniformis reduced the infectivity of each nematode species on tomato roots. The toxicity of tioxazafen was similar between nematode species; however, a greater rate of tioxazafen was needed to suppress R. reniformis infection of tomato than for M. incognita.     

Pepper Rootstock Graft Compatibility and Response to Meloidogyne javanica and M. incognita

Resistance of pepper species (Capsicum annuum, C. baccatum, C. chinense, C. chacoense, and C. frutescens), cultivars and accessions to the root-knot nematodes Meloidogyne incognita race 2 and M. javanica, and their graft compatibility with commercial pepper varieties as rootstocks were evaluated in growth chamber and greenhouse experiments. Most of the plants tested were highly resistant to M. javanica but susceptible to M. incognita. Capsicum annuum AR-96023 and C. frutescens accessions as rootstocks showed moderate and relatively high resistance to M. incognita, respectively. In M. incognita-infested soil in a greenhouse, AR-96023 supported approximately 6-fold less nematode eggs per gram root and produced about 2-fold greater yield compared to a nongrafted commercial variety. The commercial variety grafted on AR-96023 produced a yield as great as the non-grafted variety in the root-knot nematode-free greenhouse. Some resistant varieties and accessions used as rootstocks produced lower yields (P < 0.01) than that of the non-grafted variety in the noninfested greenhouse. Use of rootstocks with nematode-resistance and graft compatibility may be effective for control of root-knot nematodes on susceptible pepper.     

Optimal Release Rates for Attracting Meloidogyne incognita,Rotylenchulus reniformis,and Other Nematodes to Carbon Dioxide in Sand

Movement of vermiform stages of Meloidogyne incognita, Rotylenchulus reniformis, Ditylenchus phyllobius, Steinernema glaseri, and Caenorhabditis elegans in response to carbon dioxide was studied in 40- and 72-mm-long cylinders of moist sand inside 38-mm-d acrylic tubes. Meloidogyne incognita, R. reniformis, and S. glaseri were attracted to CO₂ when placed on a linear gradient of 0.2%/cm at a mean CO₂ concentration of 1.2%. When CO₂ was delivered into the sand through a syringe needle at flow rates between 2 and 130 μl/minute, the optimal flow rate for attracting M. incognita and R. reniformis was 15 μl/minute, and maximal attraction of the two species from a distance of 52 mm was achieved after 29 and 40 hours, respectively. After 24 hours, a total CO₂ volume of 20 cm³ was sufficient to induce 96% of all M. incognita introduced to move into the half of the cylinder into which CO₂ was delivered and more than 75 % to accumulate in the 9 cm³ of sand volume nearest the source. Results indicate it may be possible to use a chemical or biological source of CO₂ to attract nematodes to nematicide granules or biocontrol agents.     

Damage and Reproductive Potentials of Pratylenchus brachyurus and P. penetrans on soybean

Damage and reproductive potentials of Pratylenchus brachyurus and P. penetrans on soybean, Glycine max, cvs. Essex, Forrest, and Lee 68, were determined in microplot tests. Cultivar Essex was generally tolerant to P. brachyurus. Yield of Forrest was suppressed linearly with increasing P_i''s in the sandy soil (r = -0.92) and loamy sand soil (r = -0.99). Low to moderate P_i''s in the sandy clay loam gave an increase in yields as compared to plants without nematodes. Yield was not affected by this nematode in muck. Lee 68 was very sensitive to P. penetrans in microplots. Yield vs. P_i was fitted by a quadratic model (r = 0.82) with yield decreasing sharply as P_i''s were increased. The reproduction of both species decreased with increases in P_i. Lee 68 was a good host for P. penetrans, whereas Essex and Forrest were fair to poor hosts for P. brachyurus.     

Suppression of Rotylenchulus reniformis 122-cm Deep Endorses Resistance Introgression in Gossypium

Nine sources of resistance to Rotylenchulus reniformis in Gossypium (cotton) were tested by measuring population density (Pf) and root-length density 0 to 122 cm deep. A Pf in the plow layer less than the autumn sample treatment threshold used by consultants was considered the minimum criterion for acceptable resistance, regardless of population density at planting (Pi). Other criteria were ample roots and a Pf lower than on the susceptible control, as in pot studies. In a Texas field in 2001 and 2002, no resistant accessions had Pf less than the control but all did in microplots into which nematodes from Louisiana were introduced. An environmental chamber experiment ruled out nematode genetic variance and implicated unknown soil factors. Pf in field experiments in Louisiana, Mississippi, and Alabama were below threshold for zero, six and four of the accessions and above threshold in the control. Gossypium arboreum A2–87 and G. barbadense GB-713 were the most resistant accessions. Results indicate that cultivars developed from these sources will suppress R. reniformis populations but less than in pots in a single season.     

Possible utilization of weeds for the management of plant parasitic nematodes infesting some vegetable crops

Leaf extracts of noxious weeds such as Solanum xanthocarpum and Argemone maxicana were used as bare-root dip treatment for the management of three important plant-parasitic nematodes, Meloidogyne incognita, Rotylenchulus reniformis and Tylenchorhynchus brassicae infesting tomato (Lycopersicon esculantum ) and chilli (Capsicum annuum) plants. Significant reduction was observed in the root-knot development caused by M. incognita, multiplication of nematode populations of R. reniformis and T. brassicae on both the test plants. Larval penetration of second stage juveniles of M. incognita was also inhibited at various concentrations of leaf extracts and dip durations. Leaf extract of S. xanthocarpum caused relatively more inhibition in root-knot development in case of root-knot nematode, nematode multiplication of reniform and stunt nematodes than that of A. maxicana. Because of dip treatment in leaf extracts of Argemone maxicana and Solanum xanthocarpum, the plants show better growth and at the same time the populations of nematodes such as M. incognita, R. reniformis and T. brassicae significantly decreased, which naturally improved plant growth. The efficacy of root-dip treatment with respect to improvement in plant weight and reduction in root-knot development and nematode populations, increased with increasing the concentration of leaf extracts and dip durations.     

Effects of Meloidogyne incognita on Nitrogen Fixation in Soybean

The effects of a North Carolina population of Meloidogyne incognita on N₂ fixation on root-knot-susceptible ''Lee 68'' and moderately resistant ''Forrest'' soybean were evaluated 50, 75, I00, and 135 days after inoculation with nematodes. Nematodes stimulated N₂ fixation in Lee 68 by 50 days and in Forrest by 75 days. At all other intervals, N₂ fixation was either depressed or unaffected by nematodes. Additional observations indicate that the susceptibility of Lee 68 is associated with greater rates of penetration by larvae and more favorable responses of host tissues to nematodes than occur in Forrest. With time, however, the histological reactions of both hosts became less favorable for nematode development. Resistant or hypersensitive responses became common in Forrest by 75 days but not in Lee 68 until 90 days after inoculation. This population of M. incognita may stimulate N₂ fixation at a specific time interval and depress it at others; therefore, disease of susceptible soybeans caused by this nematode is probably not primarily due to a net loss of fixed nitrogen but to pathogenicity similar to that which occurs on nonlegume hosts.     

Infection of Seedlings of Alfalfa and Red Clover by Concomitant Populations of Meloidogyne incognita and Pratylenchus penetrans

Invasion of 2-day-old seedlings of ''Buffalo'' alfalfa and ''Kenland'' red clover by larvae of M. incognita and adults of P. penetrans, during 1-3 day periods of incubation at 24 C, was investigated in 50-mm petri dishes on 1% agar. Penetration by both nematodes increased arithmetically with increased numbers in inocula. P. penetrans invaded alfalfa more readily than red cover, but M. incognita invaded red clover more readily than alfalfa. Both nematodes inhibited root-elongation of alfalfa more than that of red clover. In combinations of 10 and 50 of both nematodes, invasion of both plants by both nematodes was the same as for each nematode alone. Penetration by M. incognita into alfalfa, but not into red clover, was significantly reduced when combinations of 50 M. incognita and 200 P. penetrans were inoculated simultaneously. In the presence of large numbers of entrant P. penetrans in both plants, penetration by M. incognita was highly significantly reduced. Penetration by P. penetrans was unaffected in the reciprocal situations.     

Evaluation of Asteraceae Plants for Control of Meloidogyne incognita

Of the 56 species and 43 genera of Asteraceae tested, 9 were highly resistant or immune to Meloidogyne incognita and did not form root galls. Twenty-six species and six cultivars had 25% or fewer roots galled and were considered moderately resistant to M. incognita. Pre-planting Cosmos bipinnatus (F190), Gaillardia pulchella, Tagetes erecta, Tithonia diversifolia, or Zinnia elegans (F645) reduced root galling and M. incognita J2 in and around Ipomoea reptans. Amendment of soils with roots, stems, or leaves of G. pulchella was effective in controlling M. incognita on I. reptans. Tissue extracts of G. pulchella were lethal to various plant-parasitic nematodes but were innocuous to free-living nematodes. Root exudates of G. pulchella were lethal to J2 of M. incognita and were inhibitory to the hatch of eggs at the concentration of 250 ppm or higher. Gaillardia pulchella could be used to manage M. incognita as a rotation crop, a co-planted crop, or a soil amendment for control of root-knot nematode.