Effects of Eight Herbicides on In Vitro Hatching of Heterodera glycines

Laboratory studies were conducted to evaluate effects of selected herbicides on hatching of free eggs of the soybean cyst nematode, Heterodera glycines. The herbicides used were Atrazine (atrazine), Basagran (bentazon), Bladex (cyanazine), Blazer (acifluorfen), Command (clomazone), Lasso (alachlor), Sonalan (ethalfluralin), and Treflan (trifluralin). Treatments comprised two concentrations of commercial herbicide formulations and deionized water and 3.14 mM zinc sulfate as negative and positive controls, respectively. Eggs were extracted from females and cysts, surface disinfested, and incubated in herbicide or control solutions at 25 ± 2 C in darkness. Hatched second-stage juveniles were counted every other day for 24 days. Hatching of H. glycines eggs in 50 and 500 μg/ml Blazer was 42 to 67% less than that in deionized water and 6l to 78% less than that in zinc sulfate solution. Zinc sulfate significantly increased hatching activity in 50 μg/ml but not 500 μg/ml Blazer. The other herbicides tested at various concentrations had no significant effect on egg hatching. The specific component of Blazer inhibiting egg hatching is unknown. Suppression of hatching by Blazer indicates that this postemergence soybean herbicide may have a potential role in managing H. glycines.     

Evaluation of Pea and Soybean as Trap Crops for Managing Heterodera glycines

Trap crops that stimulate nematode egg hatching but not reproduction have been reported as an effective means for managing certain nematodes. Studies were carried out at two field sites each year in 1998 and 1999 to evaluate the potential of trapping the soybean cyst nematode (Heterodera glycines) with soybean and pea in the corn year to manage the nematode in Minnesota. The trap crops were planted on the same day as corn at each site and later killed with the herbicide glyphosate. Nematode egg densities were determined at planting, 1 and 2 months after planting, and at harvest. Treatments included four seeding rates (0, 124,000, 247,000, and 494,000 seeds/ha) of resistant soybean as a trap crop and four kill dates (3, 4, 5, and 6 weeks after planting). No effects of the trap-crop and kill-date treatments on H. glycines population density, corn yield, and the followingyear soybean yield were observed at the two locations. In a second study, the experiment included four trap-crop comparisons (resistant soybean at 494,000 seeds/ha, susceptible soybean at 494,000 seeds/ha, pea at 1,482,000 seeds/ha, and no trap crop) and five kill dates (3, 4, 5, 6 weeks after planting, and no-kill). At the Waseca site, egg density at harvest was lower where resistant soybean was grown for 6 weeks and where pea was grown for 5 and 6 weeks compared with where no trap crop was grown. Maintaining pea plants for more than 5 weeks, however, reduced corn yield by 20% at the Waseca site. At the Lamberton site, egg density at harvest was lower where the susceptible soybean was grown for 5 weeks compared with where no trap crop was grown. Even with significant reduction of eggs in some treatments, use of soybean and pea as trap crops in the corn year was not an effective means for managing H. glycines.     

Influence of Selected Plant Species on Hatching of Eggs and Development of Juveniles of Heterodera glycines

The influence of selected plant species on egg hatch and subsequent development of Heterodera glycines race 3 was investigated. Plants tested included four soybean cultivars, red clover, alfalfa, hairy vetch, field corn, sweet corn, cabbage, tobacco, cotton, and wheat. Soybean stimulated egg hatching more than any of the other plant species, with H. glycines-resistant cultivars being more stimulating than susceptible ones. Hairy vetch also increased hatch. Roots of cabbage, red clover, alfalfa, and hairy vetch were readily penetrated by juveniles of H. glycines. Maturation to adult occurred only on soybean and hairy vetch.     

Heterodera glycines Cyst Components and Surface Disinfestants Affect H. glycines Hatching

We investigated the effects of Heterodera glycines cyst components and surface disinfestants on hatching of H. glycines eggs in vitro. Eggs were incubated in either H. glycines cyst wall fragments, cyst wall and egg rinsate, egg homogenate, or control solutions of soybean root diffusate, sterile distilled water, or zinc sulfate. Hatch in cyst wall and egg rinsate, and egg homogenate, was greater (α = 0.05) than hatch in sterile distilled water; however, it was not different from hatch in zinc sulfate according to Dunnett''s test. Hatch in cyst wall fragments was similar to hatch in sterile distilled water. To determine whether surface disinfestants affected hatch, eggs were treated first with chlorhexidine diacetate, mercuric chloride, sodium hypochlorite, or streptomycin sulfate and then incubated in H. glycines egg homogenate, soybean root diffusate, sterile distilled water, or zinc sulfate. Hatch of eggs treated with chlorhexidine diacetate, mercuric chloride, and streptomycin sulfate was reduced (α = 0.05), and hatch of eggs treated with sodium hypochlorite was increased (α = 0.05) relative to hatch of nontreated eggs in all incubation solutions except zinc sulfate according to Dunnett''s Test. Hatch in zinc sulfate was similar among all surface disinfestants except mercuric chloride, where hatch was reduced relative to hatch of nontreated and other surface disinfestant-treated eggs.     

Plant-Induced Hatching of Eggs of the Soybean Cyst Nematode Heterodera glycines

Root diffusate from soybean plants caused greater hatching of Heterodera glycines eggs during vegetative growth of the host, but the activity declined with plant senescence. Chelation of the root diffusate with ethylenediamine tetraacetic acid (EDTA) significantly increased hatching activity for H. glycines eggs. Diffusate from leafless plants caused little hatching, whereas treatment of intact plants with the growth regulators gibberellin and kinetin had no effect on the hatching activity of root diffusate. Treating H. glycines eggs with zinc chloride and root diffusate reduced egg hatching from zinc chloride alone. Levels of zinc in the root diffusate were insufficient to induce egg hatch, based on analysis by atomic absorption spectrophotometry. The enzymatic activity of leucine aminopeptidase in H. glycines eggs was not altered by treatment with chelated or nonchelated root diffusate.     

Influence of Herbicide Application to Soybeans on Soybean Cyst Nematode Egg Hatching

The hatching of Heterodera glycines eggs in soybean root exudates collected after postemergence application of three herbicides, and the hatching potential of H. glycines eggs from females feeding on herbicide-treated plants, were measured in vitro. Hatching in all root exudate solutions (RES) was greater than in deionized water but less than in 0.003 M ZnSO₄ solution. Filtering RES with a 0.22-μm-filter increased H. glycines hatching in RES. Application of acifluorfen, bentazon, and lactofen to foliage of soybean plants inhibited hatching of H. glycines eggs from the same plants. Hatching in RES from the different herbicide-treated soybeans was similar. Application of crop oil concentrate and non-ionic surfactant adjuvant to foliage did not affect hatching of H. glycines eggs from soybean plants.     

Population Dynamics of Heterodera glycines in Conventional Tillage and No-Tillage Soybean/Corn Cropping Systems

The effects of no-tillage (NT), conventional tillage (CT), and crop rotation on soybean yield and population dynamics of Heterodera glycines were compared during a 7-year study in a silty clay loam soil with 6% organic matter. Either H. glycines-resistant ''Linford'' soybean or susceptible ''Williams 82'' soybean was rotated with corn and grown on 76-cm-wide rows in both tillage systems. Soybean was planted in 1994, 1996, 1998, 1999, and 2000. Yield of Linford was significantly greater than Williams 82 in all years. Soybean yield was affected by tillage in 1999 and 2000. No-tillage production tended to support more reproduction (R = number of eggs at harvest/number of eggs at planting) on both cultivars. The largest R for Williams 82 were in 1998: 58.35 for NT plots and 11.78 for CT plots. For Linford, the largest R were 12.09 for NT plots in 1996, and 3.71 for CT in 1999. When corn was planted, R decreased more in NT. When soybean was planted in years subsequent to 1994, numbers of eggs at harvest (Pf) were greater for Williams 82 NT than for Williams 82 CT or Linford in both tillage systems; however, crop rotation with corn negated these population increases. The soil became suppressive to H. glycines in 1999 and was suppressive in 2000. After the 3 years of continuous soybean, Pf per 250 cm[sup3] soil were 2,870 for Williams 82 NT, 791 for Williams 82 CT, 544 for Linford NT, and 990 for Linford CT in 2000, compared with Pf of 13,100 for Williams 82 NT, 15,000 for Williams CT, 2,360 for Linford NT, and 2,050 for Linford CT in 1994. Describing population dynamics solely on the basis of R was not adequate, but also required independent examination of initial populations following overwintering and Pf after the growing season. Planting soybean either NT or CT in rotation with corn did not result in long-term increases in numbers of H. glycines eggs.     

Dormancy of Heterodera glycines in Missouri

A 2-year study was conducted in field microplots to determine the relative importance of soybean phenology and soil temperature on induction of dormancy in Heterodera glycines in Missouri. Four near-isogenic soybean lines differing for maturity date were planted in microplots infested with a race 5 isolate of H. glycines. Soil temperature was monitored at a depth of 15 cm. Eggs of H. glycines, extracted from cysts collected monthly from each microplot, were used in hatching tests and bioassays to determine dormancy. Egg hatching and second-stage juvenile (J2) infectivity rates decreased sharply from their highest levels in midsummer (July-August) to a low level by October of each year and remained low (< 10% hatching and < 0.2 J2/cm root) until May or June of the following year. The patterns of numbers of females and eggs in the bioassays were similar. The decreases were not related to soil temperature and did not differ consistently among soybean isolines. The monophasic changes in all nematode responses with peak midsummer rates suggest that H. glycines produces one primary generation per year in central Missouri. Changes in hatching rates and the timing of minimum and maximum rates suggested that H. glycines eggs exhibit more than one type of dormancy.     

Responses of Heterodera glycines Populations to a Postemergence Herbicide Mixture and Simulated Insect Defoliation

Effects of a mixture of the postemergence herbicides acifluorfen and bentazon, and simulated defoliation expected from green cloverworm on population densities of Heterodera glycines were determined in field plots in Iowa. The herbicide mixture and defoliation each suppressed soybean growth. Population densities of H. glycines were generally lower in herbicide-treated than untreated plots. Population densities of the nematode were unaffected by defoliation in 1988 and 1990-91, but were increased by the treatment in 1989.     

An Alternative Method for Evaluating Soybean Tolerance to Heterodera glycines in Field Plots

Alternate planting dates and periodic destruction of the previous year''s soybean crop as well as 1-year bare fallow were used to establish a range of population densities ofHeterodera glycines for the subsequent year. Soybean cultivar Coker 156 (susceptible, moderately tolerant) was compared to cultivars Essex (susceptible, intolerant) and Bedford (resistant) to evaluate tolerance at different H. glycines population densities established through the previous year''s treatments. Yield of Coker 156 was consistently intermediate between yields of Bedford and Essex in 1986 and 1987. Yield of Essex was negatively correlated (P = 0.05) with preplant egg numbers of H. glycines in 1987, whereas yield of Bedford and Coker 156 were not related to nematode density. Reproduction of H. glycines was greater (P = 0.05) on the moderately tolerant Coker 156 than on either of the other cultivars.     

Effect of Rotation Crops on Heterodera glycines Population Density in a Greenhouse Screening Study

Crop rotation is a common means of reducing pathogen populations in soil. Several rotation crops have been shown to reduce soybean cyst nematode (Heterodera glycines) populations, but a comprehensive study of the optimal crops is needed. A greenhouse study was conducted to determine the effect of growth and decomposition of 46 crops on population density of H. glycines. Crops were sown in soil infested with H. glycines. Plants were maintained until 75 days after planting, when the soil was mixed, a sample of the soil removed to determine egg density, and shoots and roots chopped and mixed into the soil. After 56 days, soil samples were again taken for egg counts, and a susceptible soybean (‘Sturdy’) was planted in the soil as a bioassay to determine egg viability. Sunn hemp (Crotalaria juncea), forage pea (Pisum sativum), lab-lab bean (Lablab purpureus), Illinois bundleflower (Desman-thus illinoensis), and alfalfa (Medicago sativa) generally resulted in smaller egg population density in soil or number of cysts formed on soybean in the bioassay than the fallow control. Sunn hemp most consistently showed the lowest numbers of eggs and cysts. As a group, legumes resulted in lower egg population densities than monocots, Brassica species, and other dicots.     

Differences in Hatching of Heterodera glycines Egg-mass and Encysted Eggs in vitro

Hatching studies with Heterodera glycines typically have been conducted with a mixture of egg-mass and encysted eggs. Laboratory research was conducted to compare hatching of H. glycines eggs from external egg masses with that of eggs extracted from within females and cysts (encysted eggs). Egg-mass eggs were collected by soaking infected soybean roots in 0.5% sodium hypochlorite, and encysted eggs were collected from females and cysts dislodged from the same roots with a stream of water. Eggs were incubated at 25 °C in deionized water, 3.0 mM ZnSO₄solution, or one of three synthetic H. glycines hatch inhibitors, mad hatched juveniles were counted every other day for 22 days. Samples of eggs collected at the beginning and end of all experiments were analyzed to determine extent of embryo development. Egg-mass eggs hatched more rapidly than encysted eggs during the first 16 days, but not thereafter. Throughout the experiments, hatch of egg-mass eggs in deionized water was greater than that of encysted eggs. From day 8 to day 22, egg-mass eggs were less sensitive than encysted eggs to the hatch inhibitor 2-(2''-carboxyethyl)-5-[carboxy(hydroxy)methylidenyl]cyclopentanone. A greater proportion of egg-mass eggs contained vermiform juveniles than did encysted eggs at the beginning of the experiments, but not at the end. Results indicated that H. glycines egg-mass and encysted eggs have different hatching behaviors that cannot be explained entirely by differences in embryological development.     

Influence of Rhizoctonia solani on Egg Hatching and Infectivity of Rotylenchulus reniformis

The effects of culture filtrates of Rhizoctonia solani and root exudates of R. solani-infected cotton (Gossypium hirsutum) seedlings on hatching of eggs and infectivity of females of Rotylenchulus reniformis were evaluated in an attempt to account for the enhanced nematode reproduction observed in the presence of this fungus. Crude filtrates of R. solani cultures growing over sterile, deionized distilled water did not affect egg hatching. Exudates from roots of cotton seedlings increased hatching of R. reniformis eggs over that observed in water controls. Exudates from cotton seedling roots not infected or infected with R. solani did not differ in their effect on egg hatching. However, infection of cotton seedlings by reniform females was increased in the presence of R. solani, resulting in the augmented egg production and juvenile population densities in soil observed in greenhouse studies.     

Interrelationship of Heterodera glycines and Fusarium solani in Sudden Death Syndrome of Soybean

Experiments were established in field microplots to examine the association between Heterodera glycines and the blue form of Fusarium solani in sudden death syndrome of soybean (SDS). Foliar disease symptoms occurred on more plants per plot, appeared 3 to 7 days earlier, and were more severe on plants grown in plots infested with F. solani + H. glycines than on those inoculated with F. solani only. Yields were suppressed only in treatments that included the nematode. Numbers of H. glycines cysts and second-stage juveniles were significantly lower in plots infested with F. solani + H. glycines than with the nematode alone. Fusarium solani was able to infect cysts and eggs.     

Population Dynamics of Heterodera glycines and Soybean Response in Soils Treated with Selected Nematicides and Herbicides

Two field experiments were conducted in two locations to determine the effects of the nematicides aldicarb, phenamiphos, and ethoprop and/or the herbicides alachlor, linuron, or metribuzin on the population dynamics of Heterodera glycines and soybean growth and yield. Population densities of H. glycines were greater, at some time during the growing season, in several treatments with alachlor alone and in combination with nematicides. Numbers of H. glycines at harvest were greater in plots treated with aldicarb than in those treated with ethoprop or phenamiphos. The numbers in aldicarb treated plots were generally reduced when plots also received a herbicide. Soybean yields were negatively correlated with numbers of H. glycines eggs and juveniles in early to mid season but positively correlated with late season population densities.     

Interaction of Fensulfothion and Phorate with Preemergence Herbicides on Soybean Parasitic Nematodes

The herbicides alachlor, linuron, vernolate, and metribuzin were applied to plots treated with the nematicide fensulfothion or the insecticide phorate and planted to soybean in two locations in North Carolina. In 1976 treatment with fensulfothion + alachlor or vernolate, phorate + alachlor or metribuzin resulted in greater nematode population densities than no treatment, or treatment with fensulfothion alone, or phorate alone. In 1977 fensulfothion and phorate alone and in combination with the preemergence herbicides effectively controlled Tylenchorhynchus cIaytoni. Late season population resurgence of Heterodera glycines occurred in fensulfothion + alachlor treated plots. Correlation coefficients for H. glycines vs. yield were -0.48 (P = 0.05) and -0.46 (P = 0.05) for 30 and 68 d after planting, respectively.     

Control of the Soybean Cyst Nematode by Crop Rotation in Combination with a Nematicide

An experiment to evaluate the control of soybean cyst nematodes compared 1-year, 2-year, and 3-year nonhost rotations with continuous soybeans (Glycine max) in 0.2-ha plots. In a second 1-year rotation, the plots were planted to soybean or corn (Zea mays) after fumigation in the spring with a split application of 1,3-dichloropropene (748.2 liters/ha). The effects of the nematicide were apparent the first year. Soybean yield was 1,482 kg/ha compared to 233 kg/ha in the untreated plots. In the second year, the highest yielding plants (2,035 kg/ha) were those following 1 year of corn that had been treated the previous year; plants in untreated plots yielded 288 kg/ha. Average yield of soybean following 1 year of corn was 957 kg/ha compared to 288 kg/ha for continuous soybean. In the third year, the effects of the nematicide were still evident. Soybean plants in plots treated the first year, followed by corn, then soybean, yielded 1,044 kg/ha compared to 761 kg/ ha for soybean following 1 year of corn and 991 kg/ha for soybean following 2 years of corn. Plots planted to soybean for 3 consecutive years yielded 337 kg/ha. Nematicidal effects were no longer evident during the fourth year. Yields were most improved by the greatest number of years in the nonhost crop; highest yields in descending order were from plants following 3 years of corn, 2 years of corn, and 1 year of corn. Plots planted to soybean for 4 consecutive years yielded 130 kg/ha. Highly significant negative correlations occurred each year between initial nematode population densities and seed yield.     

Density-Dependent Multiplication and Survival Rates in Heterodera glycines

Seasonal multiplication and overwinter survival are density-dependent in Heterodera glycines. At low to moderate population densities, the nematode is capable of large population increases on susceptible soybean cultivars and high rates of oversummer or overwinter survival in the absence of a host. To improve estimates of H. glycines multiplication and survival rates, egg densities were monitored for 12 cropping sequences across 10 years. Log-linear regression analysis was used to describe and compare density-dependent relationships. Growing-season change in H. glycines egg densities was density-dependent for all crops (susceptible soybean, resistant soybean, and nonhost), with slope estimates for the density-dependent relationship greater for susceptible soybean compared with a non-host crop. Overwinter population change also was density-dependent, with similar declines in survival rates observed for all crops as population densities increased. Survival was greater following susceptible soybean compared with resistant soybean, with an intermediate rate of survival associated with non-host crops. Survival estimates greater than 100% frequently were obtained at low population densities, despite attempts to account for sampling error. Rates of growing-season multiplication and survival, when standardized for population density, declined with year of the study. Standardized overwinter survival rates were inversely related to average daily minimum temperature and monthly snow cover.     

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.     

Population Development of Heterodera glycines and Soybean Yield in Soybean-maize Rotations Following Introduction into a Noninfested Field

An 11-year field study was initiated in 1979 to monitor population development of Heterodera glycines. Fifty cysts of a race 5 population were introduced into plots in a field with no history of soybean production and that had been in sod for 20 years. Soybean cultivars either susceptible or resistant to H. glycines were grown either in monoculture or rotated with maize in a 2-year rotation. During the first 5 years, resistant cultivars with the Peking source of resistance were planted. After year 5, monocuhure of Peking resistance resulted in 18 cysts/250 cm³ of soil, whereas populations resulting from the continuous cropping of susceptible soybean resulted in 45 cysts/250 cm³. Some plots in all treatments, including control plots, were contaminated at the end of year 5. Crop rotation delayed population development of H. glycines. During years 6 through 11 cv. Fayette (PI88.788 source of resistance) was planted. In year 6 numbers of cysts declined to 1/250 cm³ of soil in the treatment consisting of monocultured Fayette. At the end of year 10, cysts were below the detection level in all treatments in which Fayette was planted. Yield of susceptible soybean in monoculture with or without H. glycines infestation was lower beginning in year 6 when compared to yield of soybean grown in rotation and remained lower throughout the duration of the experiment except for 1987 (year 9). Yields of susceptible and resistant soybean were different each year except for drought years in 1980 and 1988. From 1979 to 1982 differences in yield were due to lower yield potential of resistant cultivars. Except for the drought year, yield of cv. Fayette was greater than susceptible Williams 82 during years 6 through 11.