Meloidogyne platani n. sp. (Meloidogynidae), a Root-knot Nematode Parasitizing American Sycamore

Meloidogyne platani n. sp. is described and illustrated from specimens obtained from roots of American sycamore, Platanus occidentalis, in Virginia. This new species shows certain similarities with M. arenaria but differs from it by a number of distinctive characters. The perineal pattern of females is rounded with fine, wavy to zig-zag striae and raised, convoluted striae in the inner lateral line regions. The stylet of females is 16.5 μm long with large, rounded stylet knobs set off from the shaft. Males have a low head cap and smooth head region. The styler length is 22.0 μm, and the stylet knobs are rounded and set off from the shaft. Mean second-stage juvenile length is 443.0 μm, and stylet length is 12.2 μm. The head region of juveniles is not annulated, and the tail has a definite terminus. This nematode causes severe galling and reproduces well on sycamore. Other good hosts include white ash and tobacco cv. NC 95. M. platani n. sp. reproduces by mitotic parthenogenesis and has a somatic chromosome number of approximately 45 (2n).     

Effect of Temperature on Expression of Resistance to Meloidogyne spp. In Common Bean (Phaseolus vulgaris)

The effect of soil temperature on the expression of resistance in several common bean lines carrying resistance to root-knot nematodes (Meloidogyne spp.) was studied under controlled temperatures in temperature tank and growth chamber conditions. Resistance to M. javanica and M. incognita race 1 in bean lines A315, A328, A445, G1805, and G2618 was stable at 24-30 C. However, there was a significant increase in reproduction of M. javanica on A315, A328, and A445 when temperature was increased from 26 to 30 C. This increase did not reflect a change from a resistant to a susceptible reaction or classification. Resistance in A315 is derived from G1805, whereas resistance in A328 and A445 is derived from G2618. Alabama No. 1, PI 165426, and PI 165435, with resistance to M. incognita race 2, were heat stressed at temperatures above 27 C. Resistance to M. incognita race 2 in Alabama No. 1 and PI 165435 was lost at 30 C, but PI 165426 supported low reproduction of M. incognita race 2 at all temperatures. Poor root development at 30 C may have been responsible, in part, for the poor development of M. incognita race 2 on PI 165426.     

Optimization of Mitochondrial DNA-based Hybridization Assays to Diagnostics in Soil

Nucleic acid hybridization among root-knot nematode mitochondrial DNAs can be used to identify several Meloidogyne species. Research was initiated to optimize mitochondrial DNA-based molecular diagnostics for the demanding environments likely to be encountered in field isolates. DNA hybridization using reconstituted DNA-soil mixtures revealed a loss of assay sensitivity ranging from 34% to 92% with four agronomic soils tested. This problem was alleviated by the addition of exogenously added DNA. Variation in nematode egg lysis procedures also affected hybridization efficiency, with NaOC1 treatment most effective at disrupting Meloidogyne eggs. These optimized conditions permit detection of mtDNA released from one to five Meloidogyne eggs using standard nucleic acid hybridization procedures.     

Development of Meloidogyne chitwoodi on Wheat

Postinfection development of Meloidogyne chitwoodi from second-stage juveniles (J2) to mature females and egg deposition on ''Nugaines'' winter wheat required 105, 51, 36, and 21 days at 10, 15, 20, and 25 C. At 25 C, the J2 induced cavities and hyperplasia in the cortex and apical meristem of root tips with hypertrophy of cortical and apical meristem cell nuclei, 2 and 5 days after inoculation. Giant cells induced by late J2 were observed in the stele 10 days after inoculation. Clusters of egg-laying females were common on wheat root galls 25 days after inoculation. Juveniles penetrated wheat roots at 4 C and above, but not at 2 C, when inoculum was obtained from cultures grown at 20 C, but no penetration occurred at 4 C when inoculum was stored for 12 hours at 4 C before inoculation. In northern Utah, J2 penetrated Nugaines wheat roots in the field in mid-May, about 5 months after seedling emergence. M. chitwoodi eggs were first observed on wheat roots in mid-July when plants were in blossom. Only 40% of overwintered M. chitwoodi eggs hatched at 25 C.     

Occurrence,Parasitism, and Pathogenicity of Nematodes Associated with Sycamore (Platanus occidentalis L.)

Ten species of stylet-bearing nematodes were recovered in a survey of sycamore (Platanus occidentalis L. ) stands in Georgia. Helicotylenchus, Xiphinema, and Criconemoides were the genera found most frequently. Populations of Hoplolaimus galeatus, Scutellonema brachyurum, Helicotylenchus dihystera and H. pseudorobustus increased on greenhouse-grown sycamore, but Trichodorus christiei, Xiphinema americanum, Meloidogyne hapla, M. arenaria and M. incognita did not. Hoplolaimus galeatus and S. brachyurum are semi-endoparasites; H. dihystera and H. pseudorobustus are migratory endoparasites. Hoplolaimus galeatus caused extensive root necrosis and marked decrease of fresh weights of seedling roots and tops. Helicotylenchus dihystera and S. brachyurum produced only qualitatively different sparse and unhealthy root growth. Helicotylenchus pseudorobustus caused only a reduction in root surface area.     

Host Suitability and Response of Asparagus Cultivars to Meloidogyne Species and Races

The host-parasite relationships of asparagus and Meloidogyne spp. were examined under greenhouse and microplot conditions. Meloidogyne species and races differed greatly in their ability to reproduce on asparagus seedlings. Meloidogyne hapla generally failed to reproduce, and M. javanica, M. arenaria race 1, and M. incognita race 3 reproduced poorly, with a reproduction factor (Rf = final population/initial population) usually < 1.0. Only M. arenaria race 2 and M. incognita races 1 and 4 reproduced consistently on all asparagus cultivars tested (Rf typically 1-11). No effect of M. incognita race 4 on host growth was detected. Meloidogyne arenaria race 2 and M. incognita race 1 had slight negative effects (5-10%) on plant and root growth.     

Molecular Diagnosis of Meloidogyne Species

Genetic variation within nuclear and mitochondrial DNA of Meloidogyne species and host races has been evaluated for the development of root-knot nematode molecular diagnostics. This review summarizes the distinctive features of several useful DNA-based assays for plant-parasitic nematodes, focusing upon the direct application of these procedures for Meloidogyne detection, identification, and systematics.     

Differential Response of Thor Alfalfa to Meloidogyne chitwoodi Races and M. hapla

Second-stage juveniles (J2) of races 1 and 2 of Meloidogyne chiiwoodi and M. hapla readily penetrated roots of Thor alfalfa and Columbian tomato seedlings; however, few individuals of M. chitwoodi race 1 were able to establish feeding sites and mature on alfalfa. Histopathological studies indicate that J2 of race 1 either failed to initiate feeding sites or they caused cell enlargement without typical cell wall thickening. The protoplasm of these cells coagulated, and juveniles of race 1 did not develop beyond the swollen J2 stage. A few females of race 1 fed on small giant cells and deposited a few eggs at least 20 and 30 days later than M. chitwoodi race 2 and M. hapla, respectively. Failure of race 1 to establish feeding sites was related to egression of J2 from the roots. The M. chitwoodi race 1 J2 egression from alfalfa roots was higher than egression of race 2 and M. hapla. Egression of J2 of M. chitwoodi races 1 and 2 from tomato roots was similar and higher than that of M. hapla. Thus egression plays an important role in the host-parasite relationship of M. chitwoodi and alfalfa.     

Influence of Aldicarb on the Growth and Yield of Tobacco

Microplot and field experiments were used to examine the plant-growth stimulation frequently associated with the use of aldicarb on tobacco in the absence of major pests. Aldicarb rates of 1.5-4.5 kg a.i./ha enhanced tobacco growth and yield in most experiments, but higher rates (≥ 4.5 kg) usually resulted in a neutral to negative effect. Tobacco cultivars NC 82 and Speight G-28 were more responsive than McNair 944 to the pesticide in microplots. Supplemental irrigation enhanced the responsiveness of Speight G-70 tobacco and McNair 944 to aldicarb, but excessive moisture (ca. 7-8 cm/week) limited cured-leaf yields. Aldicarb also resulted in the greatest mean tobacco yields in 35 field experiments involving Meloidogyne spp. over 3 years, relative to ethoprop, 1,3-dichloropropene (1,3-D), 1,3-D + chloropicrin, and nontreated controls. Thus, aldicarb generally enhanced tobacco growth and yield in the presence or absence of nematodes, but its impact is dependent on other variables, including cultivar, soil moisture, and soil type.     

Damage Functions for Three Meloidogyne Species on Arachis hypogaea in Texas

The yield response of Florunner peanut to different initial population (Pi) densities of Meloidogyne arenaria, M. javanica, and an undescribed Meloidogyne species (isolate 93-13a) was determined in microplots in 1995 and 1996. Seven Pi''s (0, 0.5, 1, 5, 10, 50, and 100 eggs and J2/500 cm³ soil) were used for each Meloidogyne species in both years. The three species reproduced abundantly on Florunner in both years. In 1995, mean reproduction differed among the three species; mean Rf values were 10,253 for isolate 93-13, 4,256 for M. arenaria, and 513 for M. javanica. In 1996, the reproduction of M. arenaria (mean Rf = 7,820) and isolate 93-13a (mean Rf = 7,506) were similar, and both had greater reproduction on peanut than did M. javanica (mean Rf = 2,325). All three nematode species caused root and pod galling, and a positive relationship was observed between Pi and the percentage of pods galled. Meloidogyne arenaria caused a higher percentage of pod galling than did M. javanica or isolate 93-13a. A negative linear relationship between log₁₀ (Pi + 1) and pod yield was observed for all three nematode species each year. The yield response slopes were similar except for that of M. javanica, which was less negative than that of isolate 93-13a in 1995, and less negative than that of M. arenaria and isolate 93-13a in 1996.     

Reproduction of Mi-Virulent Meloidogyne incognita Isolates on Lycopersicon spp.

Selection of detectable numbers of Mi-virulent root-knot nematodes has necessitated a greater understanding of nematode responses to new sources of resistance. During the course of this research, we compared the reproduction of four geographically distinct Mi-virulent root-knot nematode isolates on three resistant accessions of Lycopersicon peruvianum. Each accession carried a different resistant gene, Mi-3, Mi-7, or Mi-8. All nematode isolates were verified as Meloidogyne incognita using diagnostic markers in the mitochondrial genome of the nematode. Reproduction of Mi-virulent isolates W1, 133 and HM, measured as eggs per g of root, was greatest on the Mi-7 carrying accession and least on the Mi-8 carrying accession. In general, Mi-3 behaved similar to the Mi-8 carrying accession. Reproduction of the four nematode isolates was also compared on both Mi and non-Mi-carrying L. esculentum cultivars and a susceptible L. peruvianum accession. Resistance mediated by Mi in L. esculentum still impacted the Mi-virulent nematodes with fewer eggs per g of root on the resistant cultivar (P ≤ 0.05). Preliminary histological studies suggests that Mi-8 resistance is mediated by a hypersensitive response, similar to Mi.     

Effect of Temperature on Growth, Development and Reproduction of Meloidogyne hapla in Lettuce

Temperature was an important factor in growth, development and reproduction of Meloidogyne hapla in lettuce. Growth, as measured by increase in diameter of females, was not appreciably different at the intermediate (21.1 C night and 26.7 C day) and high (26.7 C night and 32.2 C day) temperature regimes, but was considerably less at the low temperature regime (15.5 C night and 21.1 C day) than at the two higher temperature regimes. Second-stage female larvae developed into adults 14 days after inoculation at the high, 18 days at the intermediate and 34 days at the low temperature regime. Eggs were observed 20 days after inoculation at the high, 26 days at the intermediate and 54 days at the low temperature regime. Number of eggs and larvae after 6 weeks was greater at the high than at the intermediate temperature regime and no eggs or larvae occurred at the low temperature regime during the observed 6 weeks.     

Description of Meloidogyne nataliei n. sp. (Nematoda: Meloidogynidae) from Grape (Vitis labrusca) in Michigan,with SEM Observations

Meloidogyne nataliei n. sp. is described and illustrated from grape (Vitis labrusca) in a declining vineyard at Mattawan, Michigan, USA. Infected grape roots exhibit no hyperplastic symptoms. Females protrude from the roots and are surrounded by a massive egg sac containing many eggs. This new species is distinguishable from other species of the genus especially by its large, striking perineal pattern with, usually, two ropelike separated striae on each side extending laterally from the vulval and anal areas. Among other diagnostic characters are the location of the female excretory pore adjacent to or near the base of the head and the heavy larval stylet averaging about 22 μm in length. Examination of males, females, and larvae with the scanning electron microscope confirmed observations made by optical microscopy and revealed diagnostic and other structures in greater detail. Of particular significance was the nature of the male head, with a massive circular labial disc on which is located a rectangular structure surrounding the oral opening, and the six distinct lips appearing as a rosette in en face view. The known distribution of this new species is presently limited to its original location in Michigan.     

Extremely Sensitive Thermotaxis of the Nematode Meloidogyne incognita

Eggs of the root-knot nematode Meloidogyne incognita were acclimated to 23 C. Newly hatched second-stage juveniles migrated toward higher temperatures when placed in shallow thermal gradients averaging 23 C. The threshold gradient for this response was below 0.001 C/cm, with a best estimate of 4 x 10⁻⁴ C/cm. Calculations of physical limitations on thermotaxis indicate that this sensitivity is well within the limits of what is physically possible.     

Correlation of Tobacco Yield,Value, and Root-Knot Index with Early-to-Midseason,and Postharvest Meloidogyne Population Densities

Certain nematicidal treatments for the control of root knot of tobacco in six field experiments in North Carolina were used to determine early, midseason, and postharvest densities of eggs and larvae of Meloidogyne spp., postharvest root-knot indices, and crop yield and value. Various statistical correlations showed that population densities determined 6-8 wk after transplanting were more indicative of treatment effectiveness than postharvest densities. Logarithmic transformation of early and midseason population data stabilized the variance.     

Variability in Reproduction of Four Races of Meloidogyne incognita on Two Cultivars of Soybean

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.     

Species-Specific Restriction Site Polymorphism in Root-knot Nematode Mitochondrial DNA

Research was initiated to physically characterize the mitochondrial genomes of several Meloidogyne spp. and host-races, to address questions regarding their systematics and dispersal, and to assess the possibility of developing molecular diagnostics for these nematodes. Techniques were developed for purification and rapid detection of mitochondrial DNA from root-knot nematodes. Mitochondrial DNAs among Meloidogyne spp. were demonstrated to exhibit extensive divergence. The potential for using the rapidly diverging mitochondrial genomes as a diagnostic assay for M. incognita, M. hapla, M. arenaria, and M. javanica is discussed.     

Response of Soybean Cultivars to Nematicidal Treatments of Soil Infested with Meloidogyne incognita

A comparison of untreated and nematicide-treated soil for soybean production revealed that Meloidogyne incognita hastened crop maturity and reduced plant ht, seed wt, and yield. Reductions of yield varied from 32-90% depending on cultivar susceptibility. DBCP was more consistent in increasing crop performance than organo-phosphale or oxime carbamate nematicides. Greatest yield increases were produced by nematicidal treatment of soils planted to soybean cultivars with the lowest susceptibility.     

Characterization of Carbohydrates on the Surface of Second-stage Juveniles of Meloidogyne spp.

Fluorescent conjugates of the lectins soybean agglutinin (SBA), Concanavalin A (Con A), wheat germ agglutinin (WGA), Lotus tetragonolobus agglutinin (LOT), and Limulus polyphemus agglutinin (LPA) bound primarily to amphidial openings and amphidial secretions of viable, preinfective second-stage juveniles (J2) of Meloidogyne incognita races 1 and 3 (Mil, Mi3) and M. javanica (Mj). No substantial difference in fluorescent lectin binding was observed among the populations examined. Binding of only LOT and LPA were inhibited in the presence of 0.1 M competitive sugar. Structural differences in amphidial carbohydrate complexes among populations of Mi 1, Mi3, and Mj were revealed by glycohydrolase treatment of preinfective J2 and subsequent labeling with fluorescent lectins. A quantitative microfiltration enzyme-linked lectin assay revealed previously undetected differences in lectin binding to nonglycohydrolase-treated J2. Freinfective J2 of Mj bound the greatest amount of SBA, LOT, and WGA, whereas J2 of Mil bound the most LPA.