Extraction of Cyst Nematodes from Organic Soils

The effects of extraction technique, sample size, soil moisture level, and overflow rate on recovery of Globodera rostochiensis and (or) Heterodera schachtii cysts from organic soils were investigated. A modified Fenwick can (MFC) and an underflow elutriator (UE) described in this paper were evaluated and compared for cyst recovery efficiency and amount of organic flotsam collected. The MFC and UE extracted similar numbers of cysts, but the UE collected 50% less flotsam than the MFC. Sample size was negatively correlated with cyst recovery and positively correlated with amount of flotsam. The amount of flotsam recovered with the MFC was correlated with overflow speed. Presoaking air dried samples for 30 minutes halved the amount of flotsam without affecting cyst recovery. Extracting cysts from wet soil without prior drying resulted in negligible recovery with both extraction techniques. There were no significant differences in cyst recovery of the two genera tested.     

Seasonal Fluctuations of Globodera tabacum solanacearum as Estimated by Two Soil Extraction Techniques

Two soil extraction methods were compared to determine their efficiency in recovering cysts and juveniles of a tobacco cyst nematode, Globodera tabacum solanacearum. The methods were equally efficient when extracting nematodes from soil samples seeded in the laboratory; however, there was a significant extraction method × month interaction when the methods were used to estimate field soil populations over 2 years. The centrifugal sugar flotation method recovered greater numbers of cysts when densities were near 400 cysts/100 cm³ soil and greater numbers of juveniles in all samples. The sugar flotation method recovered greater numbers of cysts during months when densities were less than 400 cysts/100 cm³ soil. Numbers of cysts and juveniles were lowest in June and July following land tillage in May. A soil freeze in January 1982 may have been responsible for unusually high numbers of recovered cysts in February and March 1982, a pattern that did not occur in 1983.     

Impact of Planting 'Bell', a Soybean Cultivar Resistant to Heterodera glycines, in Wisconsin

Although the soybean cyst nematode (SCN), Heterodera glycines, has been known to exist in Wisconsin for at least 14 years, relatively few growers sample for SCN or use host resistance as a means to manage this nematode. The benefit of planting the SCN-resistant cultivar Bell on a sandy soil in Wisconsin was evaluated in 1992 and 1993. A range of SCN population densities was achieved by planting 11 crops with varying degrees of susceptibility for 1 or 2 years before the evaluation. Averaged over nematode population densities, yield of ''Bell'' was 30 to 43% greater than that of the susceptible cultivars, ''Corsoy 79'' and ''BSR 101''. Counts of cysts collected the fall preceding soybean were more predictive of yield than counts taken at planting. Yields of all three cultivars were negatively related (P < 0.001) to cyst populations. Fewer (P < 0.01) eggs were produced on ''Bell'' than on the susceptible cultivars. The annual (fall to fall) change in cyst population densities was dependent on initial nematode density for all cultivars in 1992 and for the susceptible cultivars in 1993. Yield reductions induced by the SCN under the conditions of this study indicate that planting a SCN-resistant cultivar in Wisconsin can be beneficial if any cysts are detected.     

Serological identification and quantification of potato cyst nematodes from clean cysts and processed soil samples

Two monoclonal antibodies which specifically recognise each of the two species of potato cyst nematodes (PCN) and do not cross react with other species of soil nematodes, were used successfully in an immunoassay to identify and quantify PCN species using clean cysts and mixed populations. These antibodies show reactivity only towards antigens prepared from live eggs and they recognise antigens which are easily released from the nematodes. The results presented in this paper show that serological identification and quantification of PCN, not only from clean cysts but also from processed soil samples is achievable. A simple procedure to recover nematodes from soil samples and then to release nematode antigens was devised. The use of these procedures and the immunoassay for quantification of PCN was validated in tests with soil samples from Northern Portugal. The flotation method proved to be as efficient as the Fen wick Can for cyst recovery from soil samples. A significant correlation was obtained between results from immunoassay estimates and the traditional method of cyst picking and egg counting. The amount of organic matter (OM) present in the soil samples affected the sensitivity of the immunoassay but quantification of nematodes extracted from soil samples was possible with soils containing up to 14% of OM. The challenge remains to optimise the extraction procedures and the immunoassay, in practical conditions with highly organic soils containing cysts of different sizes and ages.     

Effect of Controlled Cold Storage on Recovery of Rotylenchulus reniformis from Naturally Infested Soil

Rotylenchulus reniformis is rapidly becoming the most economically important pest associated with cotton in the southeastern United States. Incentive programs have been implemented to support sampling of production fields to determine the presence and abundance of R. reniformis. These sampling programs have dramatically increased the number of soils samples submitted to nematology laboratories during autumn. The large numbers of samples overwhelm most labs and require placement in cold storage until extraction. Therefore, the objective of this study was to examine the length of time soils infested with R. reniformis can be stored before nematode extraction without compromising the accuracy of estimates of population densities. A sandy loam and a silty loam were the two cotton production soils used in this study. Rotylenchulus reniformis numbers decreased 61%during the first 180 days of storage in both soils. Rotylenchulus reniformis numbers from the initial sampling through 180 days decreased as a linear function. The decline of R. reniformis numbers during storage was estimated as 0.28% of the population lost daily from the maximum population through 180 days. The diminution of nematode numbers from 180 through 1,080 days in storage continued, but at a slower rate. Numbers of R. reniformis declined to less than 89%, 93%, and 99% of the initial population within 360, 720, and 1,080 days, respectively, of storage. The reduction of R. reniformis numbers over 180 days can be adjusted, allowing a more accurate estimation of R. reniformis levels in soil samples stored at 4 °C.     

Response of Soybean to Heterodera glycines Races 1 and 2 in Different Soil Types

Experiments were conducted for 3 years at four locations and 1 year with six soil types at a common location in North Carolina to determine damage and control-cost functions for Heterodera glycines races 1 and 2 on soybean. In the experiments on native loamy sand and sandy soils, tolerance limits for initial population densities were 0 or very low, whereas in a muck, the tolerance limit was 315 eggs/500 cm³ soil. The aggressive race 2 was more damaging than race 1 in Lakeland sand and Norfolk loamy sand. The crop response was not different between races in the Appling sandy clay loam and Belhaven muck. Soybean yield responses to H. glycines were linear in six soil types in microplots at a common site. The amount of damage varied among these soil types, with lowest yields in the muck because of severe drought stress in this soil. An exponential function adequately described soybean yield response relative to nematode control with increasing rates of aldicarb in Norfolk loamy sand. Treatment with aldicarb in the Lakeland sand decreased the effective egg population of H. glycines but had only a minor effect in the muck.     

Anhydrobiotic Coiling of Nematodes in Soil

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.     

Impact of Rotylenchulus reniformis on Cotton Yield as Affected by Soil Texture and Irrigation

The effects of soil type, irrigation, and population density of Rotylenchulus reniformis on cotton were evaluated in a two-year microplot experiment. Six soil types, Fuquay sand, Norfolk sandy loam, Portsmouth loamy sand, Muck, Cecil sandy loam, and Cecil sandy clay, were arranged in randomized complete blocks with five replications. Each block had numerous plots previously inoculated with R. reniformis and two or more noninoculated microplots per soil type, one half of which were irrigated in each replicate for a total of 240 plots. Greatest cotton lint yields were achieved in the Muck, Norfolk sandy loam, and Portsmouth loamy sand soils. Cotton yield in the Portsmouth loamy sand did not differ from the Muck soil which averaged the greatest lint yield per plot of all soil types. Cotton yield was negatively related to R. reniformis PI (initial population density) in all soil types except for the Cecil sandy clay which had the highest clay content. Supplemental irrigation increased yields in the higher yielding Muck, Norfolk sandy loam, and Portsmouth loamy sand soils compared to the lower yielding Cecil sandy clay, Cecil sandy loam, and Fuquay sand soils. The Portsmouth sandy loam was among the highest yielding soils, and also supported the greatest R. reniformis population density. Cotton lint yield was affected more by R. reniformis Pi with irrigation in the Portsmouth loamy sand soil with a greater influence of Pi on lint yield in irrigated plots than other soils. A significant first degree PI × irrigation interaction for this soil type confirms this observation.     

Development of Heterodera glycines on Soybean Damaged by Soybean Looper and Stem Canker

Short-term greenhouse studies with soybean (Glycine max cv. Bragg) were used to examine interactions between the soybean cyst nematode (Heterodera glycines) and two other common pests of soybean, the stem canker fungus (Diaporthe phaseolorum var. caulivora) and the soybean looper (Pseudoplusia includens), a lepidopterous defoliator. Numbers of cyst nematode juveniles in roots and numbers of cysts in soil and roots were reduced on plants with stem cankers. Defoliation by soybean looper larvae had the opposite effect; defoliation levels of 22 and 64% caused stepwise increases in numbers of juveniles and cysts in both roots and soil, whereas numbers of females in roots decreased. In two experiments, stem canker length was reduced 40 and 45% when root systems were colonized by the soybean cyst nematode. The absence of significant interactions among these pests indicates that the effects of soybean cyst nematode, stem canker, and soybean looper on plant growth and each other primarily were additive.     

Portable Cyst Extractor: Detecting Cyst Nematodes in the Field

For on-site detection of cysts, a portable cyst extraction kit was constructed from nine readily available items. The portable cyst extractor detected cysts in a range of 1-117 cysts/100 g soil from 42 fields. Samples processed by this kit in fields were clean and easy to examine, possibly because the kit is a compact version of the standard wet-sieving technique used in the laboratory. The portable cyst extractor has several advantages over traditional survey methods: i) diagnosis of cyst infestations in the field accurately and rapidly most of the year and ii) reduction in the labor of samplings and transportation of soil samples.     

Effect of Within-field Variation in Soil Texture on Heterodera glycines and Soybean Yield

The influence of soil texture on Soybean yield in the presence of Heterodera glycines was investigated by comparing yields of susceptible cultivars with a resistant cultivar for 2 years. Soybean yield was negatively correlated with increasing sand content (P = 0.05). Yields of susceptible cultivars were suppressed with increasing sand content. Final nematode population densities were lowest in plots with greatest sand content. Soybean infection by SCN, as determined by the number of cysts 30 days after planting, was not consistently related to soil texture over 2 years. Initial nematode population density was positively related to soybean yield the first year and negatively related to soybean yield the second, probably a result of greater yield suppression by H. glycines in plots with greater sand content.     

Soybean Photosynthesis and Yield as Influenced by Heterodera glycines,Soil Type and Irrigation

The effects of soil types and soil water matric pressure on the Heterodera glycines-Glycine max interaction were examined in microplots in 1988 and 1989. Reproduction of H. glycines was restricted in fine-textured soils as compared with coarse-textured ones. Final population densities of this pathogen in both years of the study were greater in nonirrigated soils than in irrigated soils. The net photosynthetic rate of soybean (per unit area of leaf) was suppressed only slightly or not at all in response to infection by H. glycines and other stresses. Relative soybean-yield suppression in response to H. glycines was not affected by water content in fine-textured soils, but slopes of the damage functions were steepest in sand, sandy loam, and muck soils at high water content (irrigated plots). Yield restriction of soybean in response to this pathogen under irrigation was equal to or greater than the yield suppression under dry conditions. Although yield potential may be elevated by irrigation when soil-water content is inadequate, supplemental irrigation cannot be used to circumvent nematode damage to soybean.     

Factors Influencing Emergence of Juveniles from Cysts of Heterodera zeae

Several factors were studied to determine their effects on hatch and emergence of second-stage juveniles (J2) from cysts of Heterodera zeae. The optimum temperature for emergence of J2 from cysts of H. zeae was 30 C. No juveniles emerged from cysts at 10 or 40 C. Immersion of cysts in 4 mM zinc chloride solution stimulated 10% greater emergence of J2 than occurred in tap water controls during 28 days. Fresh corn rhizosphere leachates from 25-day and older plants growing in sand or sandy field soil stimulated 22-24% greater emergence of J2 from cysts than occurred in tap water after 28 days. Rhizosphere leachates stored for 30 days at 4 C and leachates of sand, sandy field soil, and silty field soil inhibited emergence of J2 from cysts by 7-12% compared to tap water. Rhizosphere leachates from corn plants aged 20, 30, 40, 50, or 60 days growing in sandy field soil stimulated emergence of J2 from cysts. Similar numbers of J2 emerged from cysts regardless of whether the source of cysts was field microplot cultures, greenhouse cultures, or growth chamber cultures. Fertilizing growth chamber cultures of H. zeae on corn plants resulted in a doubling of the numbers of cysts produced in the cultures, and those cysts yielded 2-3 times as many emerged J2 in hatching tests compared to cysts from similar unfertilized cultures.     

Effects of Initial Nematode Density on Population Dynamics of Globodera rostochiensis on Resistant and Susceptible Potatoes

The influence of resistant and susceptible potato cultivars on Globodera rostochiensis population density changes was studied at different nematode inoculum levels (Pi) in the greenhouse and field. Soil in which one susceptible and two resistant cultivars were grown and fallow soil in pots was infested with cysts to result in densities of 0.04-75 eggs/cm³ soil. A resistant cultivar was grown in an infested field with Pi of 0.7-16.7 eggs/cm³ soil. Pi was positively correlated with decline of soil population densities due to hatch where resistant potatoes were grown in the greenhouse and in the field but not in fallow soil. However, Pi was not correlated with in vitro hatch of G. rostochiensis cysts in water or potato root diffusate. Under continuous culture o f a resistant cultivar, viable eggs per cyst declined 60-90% per plant growth cycle (4 weeks) and the number of cysts containing viable eggs had decreased by 77% after five cycles. The rate of G. rostochiensis reproduction on both resistant and susceptible cultivars was negatively correlated with Pi. These data were used to predict the effect of resistant and susceptible potato cultivars on G. rostochiensis soil population dynamics.     

Extraction of Cysts and Eggs of Heterodera schachtii from Soil with an Assessment of Extraction Efficiency

Techniques are described for the extraction and enumeration of cysts and contained eggs from soil samples. The average recovery of cysts from seeded soil samples of differing soil texture was 82.7 ± 2.1%. Recovery from sandy clay soil samples seeded at 1 cyst/100 g soil was 63.4 ± 5.5%; at 4.2 cysts/100 g soil recovery was 89.6 ± 1.8%. Recovery of cysts from naturally infested clay soil was 88.3 ± 2.05%. Egg extraction efficiency for seeded samples was calculated as 78%, and for naturally infested soil was estimated as 83%.     

Extraction Efficiency of Belonolaimus longicaudatus from Sandy Soil

Numbers of Belonolaimus longicaudatus extracted from sandy soils (91-92% sand) by sieving and centrifugation were only 40-55% of those extracted by sieving and incubation on a Baermann tray. Residues normally discarded at each step of the sieving plus Baermann tray extraction procedure were examined for nematodes to obtain estimates of extraction efficiencies. For third-stage and fourth-stage juveniles, males, and females, estimates of extraction efficiency ranged from 60 to 65% in one experiment and 73 to 82% in another. Estimated extraction efficiencies for second-stage juveniles were lower (33% in one experiment, 67% in another) due to losses during sieving. When sterilized soil was seeded with known numbers of B. longicaudatus, 60% of second-stage juveniles and 68-76% of other stages were recovered. Most stages of B. longicaudatus could be extracted from these soils by sieving plus Baermann incubation with an efficiency of 60-70%.     

Effects of Soil Type on the Damage Potential of Meloidogyne incognita on Soybean

Effects of soil type on the reproduction and damage potential of Meloidogyne incognita on soybean, Glycine max (L.) Merr., were determined at five locations in North Carolina, including one site where plots with six soil types were established. M. incognita reproduced readily on a susceptible soybean cultivar in most soil types, with somewhat limited reproduction in muck soils. The relationship between initial population densities and yield varied among soil types and nematode populations. Yield losses were greatest in sandy and muck soil types, with less nematode damage occurring in the clay soil types. A North Carolina and a Georgia population of M. incognita differed greatly in their ability to reproduce on soybean and suppress growth. The North Carolina population had a moderate effect on yield in 1981 and only a slight effect in 1982. In contrast, a Georgia population severely limited soybean growth and yield at lower initial population densities in 1983, Initial population densities of the nematodes and physical and chemical edaphic factors accounted for much of the variation of soybean yield and nematode reproduction.     

Influence of Population Densities of Heterodera schachtii on Sugar Beets Grown in Microplots

High initial population densities of Heterodera schachtii larvae (36 and 108/gm of soil) greatly retarded the seedling emergence of sugar beet ''Monogerm CSF 1971'' in Vineland fine sandy loam. In comparison with controls, initial population densities (P_i''s) of 1.7, 3.0, 6.2, and 14.4 larvae/gm of soil respectively reduced the weight of storage roots by 38, 56, 64, and 92%. Weights of tops also decreased with increases in P_i; weights of tap and small feeder roots tended to be higher at all P_i''s except the highest. Sucrose percentage was not affected by any initial nematode density. The populations were lower at midseason than at seeding, and at harvest had increased greatly, with respective populations of 339, 402, 222, and 140 larvae/gm of soil. At harvest, cysts/gm of soil and cysts/gm of root were respectively 4.4 and 72, 6.1 and 99, 6.1 and 191, and 5.8 and 140. The maximum rate of multiplication was 150-200. and maximum density was 400 larvae/gm of soil. The high pathogenicity and multiplication rate of the nematode was attributed to optimum temperature conditions and soil type.     

Suppression of Heterodera schachtii Populations by Dactylella oviparasitica in Four Soils

The effects of Dactylella oviparasitica strain 50 applications on sugarbeet cyst nematode (Heterodera schachtii) population densities and plant weights were assessed in four agricultural soils. The fungus was added to methyl iodide-fumigated and nonfumigated portions of each soil. The soils were seeded with Swiss chard. Four weeks later, soils were infested with H. schachtii second-stage juveniles (J2). Approximately 1,487 degree-days after infestation, H. schachtii cyst, egg and J2 numbers and plant weights were assessed. In all four fumigated soils, D. oviparasitica reduced all H. schachtii population densities and increased most of the plant weights compared to the nonamended control soils. In two of the nonfumigated soils (10 and SC), D. oviparasitica reduced H. schachtii population densities and increased most plant weight values compared to the nonamended control soils. For the other two nonfumigated soils (44 and 48), which exhibited pre-existing levels of H. schachtii suppressiveness, fungal applications had relatively little impact on H. schachtii population densities and plant weights. The results from this study combined with those from previous investigations suggest that D. oviparasitica strain 50 could be an effective biological control agent.     

The Effect of Soil Texture and Irrigation on Rotylenchulus reniformis and Cotton

The reniform nematode, Rotylenchulus reniformis, is the most damaging nematode pathogen of cotton in Alabama. Soil texture is currently being explored as a basis for the development of economic thresholds and management zones within a field. Trials to determine the reproductive potential of R. reniformis as influenced by soil type were conducted in microplot and greenhouse settings during 2008 to 2010. Population density of R. reniformis was significantly influenced by soil texture and exhibited a general decrease with increasing median soil particle size (MSPS). As the MSPS of a soil increased from 0.04 mm in clay soil to > 0.30 mm in very fine sandy loam and sandy loam soils, R. reniformis numbers decreased. The R. reniformis population densities on all soil types were also greater with irrigation. Early season cotton development was significantly affected by increasing R. reniformis Pi, with plant shoot-weight-to-root-weight ratios increasing at low R. reniformis Pi and declining with increasing R. reniformis Pi. Plant height was increased by irrigation throughout the growing season. The results suggests that R. reniformis will reach higher population densities in soils with smaller MSPS; however, the reduction in yield or plant growth very well may be no greater than in a soil that is less preferential to the nematode.