Parasitism of Heterodera schachtii and Meloidogyne javanica by Hirsutella rhossiliensis in Microplots over Two Growing Seasons

Numbers of cyst and root-knot nematodes and percentage parasitism by the nematophagous fungus Hirsutella rhossiliensis were quantified in microplots over 2 years. The microplots contained either sugarbeets in loam infested with Heterodera schachtii or tomatoes in sand infested with Meloidogyne javanica. The fungus was added to half of the microplots for each crop. Although H. rhossiliensis established in both microplot soils, the percentage of nematodes parasitized did not increase with nematode density and nematode numbers were not affected by the fungus. The results indicate that long-term interactions between populations of the fungus and cyst or root-knot nematodes will not result in biological control.     

Control of Ditylenchus dipsaci in Infected Garlic Seed Cloves by Nonfumigant Nematicides

Different rates of granular formulations ofaldicarb, carbofuran, ethoprop, fensulfothion, and phenamiphos were applied directly onto garlic seed cloves in the seed furrow in sandy clay loam, clay loam, and loam soils at planting to assess efficacy for control of Ditylenchus dipsaci in infected seed cloves. All treatments were compared to hotwater-formalin clove dip disinfection treatment and to nontreated infected controls. Aldicarb and phenamiphos at 2.52 and 5.04 kg a.i./ ha, but not at lower rates, effectively suppressed infection by D. dipsaci and increased yields. Although both nematicides slightly slowed the rate of plant emergence, normal stands were established. Trace levels of infection occurred in all treatments, including the hotwater-formalin dip. Carbofuran at 5.04 kg a.i./ha controlled the nematode but was phytotoxic. Ethoprop was phytotoxic. Fensulfothion did not control D. dipsaci even at the highest application rate, 8.90 kg a.i./ha. Single and multiple applications of oxamyl at 1.12-8.96 kg a.i./ha, applied as a surface spray or in furrow irrigation water, slowed the early progression of disease symptoms but failed to provide season-long nematode control.     

Comparative Disc-Electrophoretic Protein Analyses of Selected Meloidogyne,Ditylenchus, Heterodera and Aphelenchus spp.

Disc-electrophoretic separation of soluble proteins from whole nematode homogenates yielded band profiles useful for distinguishing selected species of Meloidogyne and Ditylenchus, and the genera Heterodera, and Aphelenchus. Certain protein bands were common to all the species of Meloidogyne, whereas other bands were specific. Meloidogyne spp. and Heterodera glycines shared some protein similarities, but other genera differed distinctly. Protein profiles of Meloidogyne spp. were not significantly altered by the host on which the nematode was cultured.     

Disinfection Alternatives for Control of Ditylenchus dipsaci in Garlic Seed Cloves

Hot-water dips with and without the additives abamectin and sodium hypochlorite were evaluated for control of Ditylenchus dipsaci infection of garlic seed cloves. All treatments were compared to hot water-formalin clove dip disinfection and to nontreated infected controls for garlic emergence, midseason infection, bulb damage, and yield at harvest in field plots in 12 experiments. Hot-water treatments without additives only partially controlled D. dipsaci when a warming presoak dip (38 C) of 30, 45, or 60 minutes'' duration was followed by a hot-water dip (49 C) of 15-30 minutes'' duration. Exposure to 49 C for 30 minutes caused slight retardation of garlic emergence, although normal stand was established. Abamectin at 10-20 ppm as the 20-minute hot dip (49 C) or as a 20-minute cool dip (18 C) following a 20-minute hot-water dip and sodium hypochlorite at 1.052-1.313% aqueous solution as the 20-minute hot dip were highly effective in controlling D. dipsaci and were noninjurious to garlic seed cloves. None of these treatments was as effective as a hot water-formalin dip and were noneradicative, but showed high efficacy on heavily infected seed cloves relative to nontreated controls. Abamectin was most effective as a cool dip. These abamectin cool-dip (following hot-water dip) and sodium hypochlorite hot-dip treatments can be considered as effective alternatives to replace formalin as a dip additive for control of clove-borne D. dipsaci. Sodium hypochlorite was less effective as the cool dip, and at concentrations of 1.75-2.63% was phytotoxic to garlic.     

Effect of Carbamate,Organophosphate, and Avermectin Nematicides on Oxygen Consumption by Three Meloidogyne spp.

Second-stage juveniles (I2) of Meloidogyne arenaria consumed more oxygen (P ≤ 0.05) than M. incognita J2, which in turn consumed more than M. javanica J2 (4,820, 4,530, and 3,970 μl per hour per g nematode dryweight, respectively). Decrease in oxygen consumption depended on the nematicide used. Except for aldicarb, there was no differential sensitivity among the three nematode species. Meloidogyne javanica had a greater percentage decrease (P ≤ 0.05) in oxygen uptake when treated with aldicarb, relative to the untreated control, than either M. arenaria or M. incognita. Meloidogyne javanica J2 had a greater degree of recovery from fenamiphos or aldicarb intoxication, after subsequent transfer to water, than did M. incognita. This finding may relate to differential sensitivity among Meloidogyne spp. in the field. Degree of respiratory inhibition and loss of nematode motility for M. javanica after exposure to the nematicides were positively correlated (P ≤ 0.05).     

Growth of Carrot and Tomato from Oxamyl-coated Seed and Control of Meloidogyne hapla

Oxamyl was coated on carrot (Daucus carota L. cv. Spartan Fancy-80) and tomato (Lycopersicon esculentum Mill. cv. Glamour) seeds with a polymer sticker for the control of Meloidogyne hapla. The sticker diluted in water 1:1 delayed carrot seedling emergence. Oxamyl at 40 mg/ml in a 1:5 dilution of sticker lowered the rate of carrot seedling emergence until day 13 and plant growth until day 28. Oxamyl at 20 or 40 mg/ml in a 1:5 dilution of sticker on carrot seeds planted in M. hapla-infested muck soil resulted in fewer galled tap roots and fewer galls per root system 4 weeks after planting. Tap root lengths were greater than those of the control. Tomato seedling emergence was delayed and top and root weights were reduced, relative to the control, at 25 days by the sticker diluted 1:1 to 1:3. Oxamyl at 20 or 40 mg/ml in a 1:5 diluted sticker delayed tomato seedling emergence. Top weights of tomato seedlings from seeds coated with 20 mg/ml of oxamyl in a 1:5 diluted sticker planted in a silt loam were greater than control top weights at 4 and 6 weeks. Root weights were greater than those of the control only at 4 weeks. There were fewer galls per gram of root on seedlings from oxamyl-coated seeds and fewer juveniles per pot of soil, relative to the controls, only at 4 weeks.     

Control of Meloidogyne javanica by Formulations of Inula viscosa Leaf Extracts

Inula viscosa is a perennial plant that is widely distributed in Mediterranean countries. Formulations of I. viscosa extracts were tested for their effectiveness in control of Meloidogyne javanica in laboratory, growth chamber, microplot, and field experiments. Oily pastes were obtained by extraction of dry leaves with a mixture of acetone and n-hexane or n-hexane alone, followed by evaporation of the solvents. Emulsifiable concentrate formulations of the pastes killed M. javanica juveniles in sand at a concentration of 0.01% (paste, w/w) or greater and reduced the galling index of cucumber seedlings as well as the galling index and numbers of nematode eggs on tomato plants in growth chamber experiments. In microplot experiments, the hexane-extract formulation at 26 g paste/m² reduced nematode infection on tomato plants in one of two experiments. In a field experiment, a reduction of 40% in root galling index by one of two formulations was observed on lettuce plants. The plant extracts have potential as a natural nematicide, although the formulations need improvement.     

Meloidogyne javanica Parasitic on Peanut

Peanut fields in four governorates of Egypt were surveyed to identify species of Meloidogyne present. Fourteen populations obtained from peanut roots were all identified as M. javanica based on perineal patterns, stylet and body lengths of second-stage juveniles, esterase phenotypes, and restriction fragment length polymorphisms of mtDNA. Three of 14 populations, all from contiguous fields in the Behara governorate, had individuals with a unique two-isozyme esterase phenotype. All populations of M. javanica tested on peanut had levels of reproduction on the M. arenaria-susceptible peanut cultivar Florunner that were not different from M. arenaria (P = 0.05), and had lower levels of reproduction on the M. arenaria-resistant genotype TxAG-7 than on Florunner (P = 0.05). Reproduction of the five Egyptian populations of M. javanica tested was lower on root-knot nematode resistant tomato cultivars Better Boy and Celebrity than on the root-knot nematode susceptible cultivar Rutgers (P = 0.05). These data are evidence that some populations of M. javanica are parasitic on peanut and that the peanut and tomato genotypes resistant to M. arenaria are also resistant to these populations of M. javanica.     

Resistance of Interspecific Arachis Breeding Lines to Meloidogyne javanica and an Undescribed Meloidogyne Species

Resistance to a peanut-parasitic population of Meloidogyne javanica and an undescribed Meloidogyne sp. in peanut breeding lines selected for resistance to Meloidogyne javanica was examined in greenhouse tests. The interspecific hybrid TxAG-7 was resistant to reproduction of Meloidogyne javanica, M. javanica, and Meloidogyne sp. An Meloidogyne javanica-resistant selection from the second backcross (BC) of TxAG-7 to the susceptible cultivar Florunner also was resistant to M. javanica but appeared to be segregating for resistance to the Meloidogyne sp. When reproduction of M. javanica and Meloidogyne javanica were compared on five BC4F3 peanut breeding lines, each derived from Meloidogyne javanica-susceptible BC4F2 individuals, all five lines segregated for resistance to M. javanica, whereas four of the lines appeared to be susceptible to Meloidogyne javanica. These data indicate that several peanut lines selected for resistance to Meloidogyne javanica also contain genes for resistance to populations of M. javanica and the undescribed Meloidogyne sp. that are parasitic on peanut. Further, differences in segregation patterns suggest that resistance to each Meloidogyne sp. is conditioned by different genes.     

Meloidogyne javanica on Peanut in Florida

A mixed population of Meloidogyne arenaria race 1 and M. javanica race 3 is reported on peanut from a field in Levy County, Florida. Confirmation of M. javanica on peanut is based on esterase and malate dehydrogenase isozyme patterns resolved on polyacrylamide slab gels following electrophoresis, and perineal patterns. Up to 29% of 290 individual females collected from peanut roots in the field in autumn 2002 showed a typical esterase J3 phenotype for M. javanica. This is the third report of M. javanica infecting peanut in the United States.     

Sensitive PCR Detection of Meloidogyne arenaria, M. incognita, and M. javanica Extracted from Soil

We have developed a simple PCR assay protocol for detection of the root-knot nematode (RKN) species Meloidogyne arenaria, M. incognita, and M. javanica extracted from soil. Nematodes are extracted from soil using Baermann funnels and centrifugal flotation. The nematode-containing fraction is then digested with proteinase K, and a PCR assay is carried out with primers specific for this group of RKN and with universal primers spanning the ITS of rRNA genes. The presence of RKN J2 can be detected among large numbers of other plant-parasitic and free-living nematodes. The procedure was tested with several soil types and crops from different locations and was found to be sensitive and accurate. Analysis of unknowns and spiked soil samples indicated that detection sensitivity was the same as or higher than by microscopic examination.     

Reproduction and Development of Meloidogyne incognita and M. javanica on Guardian Peach Rootstock

Guardian peach rootstock was evaluated for susceptibility to Meloidogyne incognita race 3 (Georgia-peach isolate) and M. javanica in the greenhouse. Both commercial Guardian seed sources produced plants that were poor hosts of M. incognita and M. javanica. Reproduction as measured by number of egg masses and eggs per plant, eggs per egg mass, and eggs per gram of root were a better measure of host resistance than number of root galls per plant. Penetration, development, and reproduction of M. incognita in Guardian (resistant) and Lovell (susceptible) peach were also studied in the greenhouse. Differences in susceptibility were not attributed to differential penetration by the infectivestage juveniles (J2) or the number of root galls per plant. Results indicated that M. incognita J2 penetrated Guardian roots and formed galls, but that the majority of the nematodes failed to mature and reproduce.     

Evidence for a Dosage Effect of the Mi gene on Partially Virulent Isolates of Meloidogyne javanica

The reproduction of single egg-mass isolates of Meloidogyne javanica from Crete that differed in virulence were compared on tomato (Lycopersicon esculentum) genotypes homozygous or heterozygous for the Mi gene. The reproduction of three isolates with partial virulence was much greater on tomato genotypes heterozygous for the Mi gene (cultivars Scala, Bermuda, and 7353) than on two homozygous genotypes (F8 inbred lines derived from Scala). The reproduction of a highly virulent isolate on the homozygous and heterozygous genotypes was similar to that on a susceptible cultivar. These results pose questions regarding the nature of partial virulence and indicate a quantitative effect of the Mi gene in relation to such virulence.     

Disc-electrophoretic Patterns of Enzymes and Soluble Proteins of Ditylenchus dipsaci and D. triformis

Soluble protein, esterase and oxidative enzyme patterns of the Waynesville, North Carolina, (WNC) and Raleigh, North Carolina, (RNC) populations of Ditylenchus dipsaci were compared. Polyacrylamide gel electrophoretic patterns of soluble protein extracts of nematodes of the two populations differed. Esterase and catalase patterns, however, were identical. Peroxidatic activity of the catalase isoenzymes from nematodes of the two populations differed when catechol was used as a cosubstrate. Distinct differences were demonstrated in soluble protein and enzyme patterns between D. dipsaci and D. triiormis.     

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.     

Temporal Efficacy of Selected Nematicides on Meloidogyne Species on Tobacco

Aldicarb, ethoprop, and fenamiphos were evaluated for their efficacy in controlling various species of root-knot nematodes on flue-cured tobacco and for their residual activity, as determined through periodic sampling and bioassays of soil taken from field plots. Field experiments were conducted at five locations over 2 years with flue-cured tobacco. Soil in plots treated with nematicides were formed into high, wide beds before transplanting with ''Coker 371-Gold'' or ''K 326'' tobacco. Residual control of Meloidogyne spp. was greatest (P ≤ 0.05) with fenamiphos (in some cases up to 10 weeks, as measured in tomato bioassays of infested soil and root fragments). Suppression of nematode reproduction by ethoprop was short-lived, and numbers of second-stage juveniles + eggs and numbers of galls in bioassays sometimes surpassed those of untreated plots within 4 weeks after treatment. Aldicarb gave intermediate control over time as compared to the other compounds. Although nematicidal efficacy of all compounds varied with site and season, fenamiphos and aldicarb generally produced the highest yields.     

Interaction of Ditylenchus dipsaci and Meloidogyne hapla on Resistant and Susceptible Plant Species

Numbers ofDitylenchus dipsaci or Meloidogyne hapla invading Ranger alfalfa, Tender crop bean, Stone Improved tomato, AH-14 sugarbeet, Yellow sweet clover, and Wasatch wheat from single inoculations were not significantly different from numbers by invasion of combined inoculations. D. dipsaci was recovered only from shoot and M. hapla only from root tissue. Combined inoculations did not affect reproduction of either D. dipsaci or M. hapla. D. dipsaci suppressed shoot growth of all species at 15-30 C, and M. hapla suppressed shoot growth of tomato, sugarbeet, and sweet clover at 20, 25, and 30 C. There was a positive correlation (P < 0.05) between shoot and root growth suppression by D. dipsaci on all cultivars except wheat at 20 C and tomato at 30 C. M. hapla suppressed (P < 0.05) root growth of sugarbeet at 20-50 C and wheat at 30 C. Growth suppression was synergistic in combined inoculations of sweet clover shoot growth at 15 C and root growth at 20-30 C, wheat root growth at 15 and 20 C, and tomato root growth at 15-30 C (P < 0.05) D. dipsaci invasions caused mortality of alfalfa and sweet clover at 15-30 C and sugarbeet at 20-30 C. Mortality rates of alfalfa and sweet clover increased synergistically (P < 0.05) from combined inoculations.     

A Nematicide Seed Treatment to Control Ditylenchus dipsaci on Seedling Alfalfa

Three nematicides were evaluated as seed treatments to control the alfalfa stem nematode (Ditylenchus dipsaci) on seedling alfalfa. Alfalfa seeds were soaked for 10 hours in a 0.5% (formulated by weight) concentration of either carbofuran, phenamiphos or oxamyl in acetone with no adverse effect on seed germination. All three treatments decreased nematode damage and increased survival of ''Ranger'' (susceptible) and ''Lahontan'' (resistant) alfalfa plants, when seeds were planted in soil infested with D. dipsaci. Mean live plant counts after 6 weeks in the untreated control, acetone alone, carbofuran, phenamiphos, and oxamyl treatments, respectively, were 4.3, 6.3, 19.0, 19.8, and 19.0 for Lahontan and 4.5, 1.5, 18.5, 19.3, and 18.0 for Ranger from 20 seeds/pot. Nematicide seed treatments resulted in significantly healthier Ranger alfalfa plants 4 months after planting. The combination of seed treatment and host resistance may provide a means of establishing alfalfa in an alfalfa monocropped system where soil populations of D. dipsaci are high.