Hirsutella rhossiliensisand Verticillium chlamydosporium as Biocontrol Agents of the Root-knot Nematode Meloidogyne hapla on Lettuce

Hirsutella rhossiliensis and Verticillium chlamydosporium infected second-stage juveniles (J2) and eggs of Meloidogyne hapla, respectively, in petri dishes and in organic soil in pots planted to lettuce in the greenhouse. In vitro, H. rhossiliensis produced 78 to 124 spores/infected J2 of M. hapla. The number of J2 in roots of lettuce seedlings decreased exponentially with increasing numbers of vegetative colonies of H. rhossiliensis in the soil. At an infestation of 8 M. hapla eggs/cm³ soil, 1.9 colonies of H. rhossiliensis/cm³ soil were needed for a 50% decrease in J2 penetration of lettuce roots. Egg-mass colonization with V. chlamydosporium varied from 16% to 43% when soil was infested with 8 M. hapla eggs and treated with 5,000 or 10,000 chlamydospores of V. chlamydosporium/cm³ soil. This treatment resulted in fewer J2 entering roots of bioassay lettuce seedlings planted in the infested soils after harvesting the first lettuce plants 7 weeks after infestation with M. hapla. Hirsutella rhossiliensis (0 to 4.3 colonies/cm3 soil), V. chlamydosporium (500 to 10,000 chlamydospores/cm3 soil), or their combination, added to organic soils with 8 M. hapla eggs/cm³ soil, generally did not affect lettuce weight, root galling, or egg production of M. hapla. However, when lettuce was replanted in a mix of infested and uninfested soil (1:3 and 1:7, v:v), egg production was lower in soils with V. chlamydosporium than in soils without the fungus. Both fungi have potential to reduce the M. hapla population, but at densities below 8 eggs/cm³ soil.     

Damage Functions for Meloidogyne arenaria on Peanut

Microplot experiments were conducted in 1989 and 1990 to determine the relationship between yield of peanut (Arachis hypogaea) and inoculum density ofMeloidogyne arenaria race 1. Nine inoculum densities were used, ranging from 0-200 eggs/100 cm³ soil (1989) or from 0-100 eggs/100 cm³ (1990), and each density was replicated 10 times. In 1989, higher final densities (mean of 1,171 juveniles [J2]/100 cm³ soil) were obtained in plots inoculated with 0.5 to 50 eggs/100 cm³ soil than in plots inoculated with 100 to 200 eggs/100 cm³ (313 J2/100 cm³ soil). In 1990, final densities of M. arenaria reached high levels (≥ 1,111 J2/100 cm³ soil) in all inoculated plots. Pod yield and dry weight of foliage at harvest were negatively correlated (P ≤ 0.05) with inoculum density in both seasons. In 1989, the relationship between pod weight (y) and initial density (x) was described by Seinhorst''s equation, with y = 0.088 + 0.91(0.90)⁽x⁻¹⁾ and r² = 0.826. In 1990, the relationship was y = 0.22 + 0.78(0.97)⁽x⁻¹⁾ and r² = 0.794. These equations suggest tolerance limits of approximately 1 egg/100 cm³ soil, which may require specialized methods, such as bioassay, for detection.     

Management of Globodera rostochiensis as Influenced by Nematode Population Densities and Soil Type

The effects of aldicarb, oxamyl, 1,3-D, and plastic mulch (solarization) on soil population densities of the golden nematode (GN) Globodera rostochiensis was assessed in field and microplot experiments with different soil types. Oxamyl was evaluated in both soil and foliar treatments, whereas aldicarb, 1,3-D, and solarization were applied only to soil. Soil applications of aldicarb and oxamyl resulted in reduced nematode populations after GN-susceptible potatoes in plots with initial population densities (Pi) of > 20 and 7.5 eggs/cm³ soil, respectively, but nematode populations increased in treated soil when Pi were less than 20 and 7.5 eggs/cm³soil. In clay loam field plots with Pi of 19-76 eggs/cm³ soil, nematode densities increased even with repeated foliar applications of oxamyl, whereas nematode populations at Pi greater than 76 eggs/cm³ soil were reduced by foliar oxamyl. Treatment with 1,3-D or solarization, singly or in combination, reduced GN soil population densities regardless of soil type or Pi. Temperatures lethal to GN were achieved 5 cm deep under clear plastic but not 10 or 15 cm deep.     

Crop Rotation and Races of Meloidogyne incognita in Cotton Root-knot Management

The influence o f various crop rotations and nematode inoculum levels on subsequent population densities of Meloidogyne incognita races 1 and 3 were studied in microplots. Ten different 3-year sequences o f cotton, corn, peanut, or soybean, all with cotton as the 3rd-year crop, were grown in microplots infested with each race. Cotton monoculture, two seasons o f corn, or cotton followed by corn resulted in high race 3 population densities and severe root galling on cotton the 3rd year. Peanut for 2 years preceding cotton most effectively decreased the race 3 population and root galls on cotton the 3rd year. Race 1 did not significantly influence cotton growth or yield at initial populations of up to 5,000 eggs/500 cm³ soil. At 5,000 eggs/500 cm³, cotton growth was suppressed by race 3 but yield was not affected.     

Probability of Globodera rostochiensis Spread on Equipment and Potato Tubers

The probability of spreading cysts of Globodera rostochiensis on farming equipment and potato tubers was investigated in naturally infested field plots. The number of cysts recovered from soil that adhered to equipment differed significantly between different pieces of equipment. These differences were related to initial nematode density and, in most cases, to the volume of soil that adhered to the equipment. At an initial density of 0.04 egg/cm³ of soil, significantly more cysts were recovered from a potato digger than from a potato hiller, cultivator, or plow. At an initial density of 0.90 egg/cm³ of soil, significantly more cysts were recovered from the plow than from the other equipment. Although the population density was 22 times greater, only 10 times more cysts adhered 3 to equipment used in soil with a density of 0.90 egg/cm³ of soil than when used in soil infested at 0.04 egg/cm³. The number of potato tuber samples (4.5 kg) that contained cysts with viable eggs was positively correlated with the initial densities of G. rostochiensis in soil in which they were produced. The percentage of tuber samples with cysts containing viable eggs was 10-12% for tubers harvested from soil with densities less than 1 egg/cm³ and 30-76% for tubers harvested from soil with densities greater than 4 eggs/cm³ of soil.     

Population Densities of Meloidogyne incognita and Yield of Capsicum annuum

Two microplot experiments in 1981 and 1983 provided information on the effect of different population densities of Meloidogyne incognita race 1 and yield of sweet pepper. Microplots were square concrete pipes (30 × 30 cm and 50 cm long) filled with 40 liters of soil infested with 0, 0.062, 0.125, 0.25, 0.5, 1, 2, 4, 8, 16, 32, 64, 128, 256, and 512 eggs and juveniles/cm³ soil. Tolerance limits of 2.2 and 0.165 eggs and juveniles/cm³ soil and minimum yields of 58% and 20% of the controls were obtained in 1981 and 1983, respectively. Maximum reproduction rates of the nematode were 274 and 1,498 at the lowest initial population density. The population of the nematode declined rapidly after harvest, and only 13% and 6.5% of eggs and juveniles were detected in the soil after 1 and 6 months, respectively.     

Comparative Response of Alfalfa to Pratylenchus penetrans Populations

Four populations of Pratylenchus penetrans did not differ (P > 0.05) in their virulence or reproductive capability on Lahontan alfalfa. There was a negative relationship (r = -0 .7 9 ) between plant survival and nematode inocula densities at 26 ± 3 C in the greenhouse. All plants survived at an inoculum level (Pi) of 1 nematode/cm³ soil, whereas survival rates were 50 to 55% at 20 nematodes/cm³ soil. Alfalfa shoot and root weights were negatively correlated (r = - 0.87; P < 0.05) with nematode inoculum densities. Plant shoot weight reductions ranged from 13 % at Pi 1 nematode/cm³ soil to 69% for Pi 20 nematodes/cm³ soil, whereas root weight reductions ranged from 17% for Pi 1 nematode/cm³ soil to 75% for Pi 20 nematodes/cm³ soil. Maximum and minimum nematode reproduction (Pf/Pi) for the P. penetrans populations were 26.7 and 6.2 for Pi 1 and 20 nematodes/cm³ soil, respectively. There were negative correlations between nematode inoculum densities and plant survival (r = 0.84), and soil temperature and plant survival (r = -0 .7 8 ). Nematode reproduction was positively correlated to root weight (r = 0.89).     

Factors Affecting the Biology and Pathogenicity of Heterodera schachtii on Sugarbeet

A direct relationship exists between soil temperature and Heterodera schachtii development. The average developmental period of two nematode populations from Lewiston, Utah, and Rupert, Idaho, from J2 to J3, J4, adult, and the next generation J2 at soil temperatures of 18-28 C were 100, 140,225, and 399 degree-days (base 8 C), respectively. There was a positive relationship (P < 0.05) between nematode Pi, nematode generations, and sugarbeet yields. The greatest sugarbeet growth inhibition (87%) occurred when sugarbeets were exposed to a Pi of 12 eggs/cm³ soil for five generations (1,995 degree-days), compared with a 47% inhibition when plants were exposed to the same Pi for two generations. There was a negative correlation (P < 0.05) between the Pi, Pf, and sugarbeet yield for each population threshold. The smaller the Pi, the greater the sugarbeet yields and the greater the Pf. Root yields were 80 and 29 t /ha and Pf were 8.4 and 3.6 eggs/cm³ soil when sugarbeet seeds were planted at Pi of 0.4 and 7.9 eggs/cm³. respectively, at a soil temperature of 8 C. The number of years rotation with a nonhost crop required to reduce the nematode population density below a damage threshold level of 2 eggs/cm³ depends on the Pi. A Pi of 33.8 eggs/cm³ soil required a 5-year crop rotation, whereas a Pi of 8.4 eggs/cm³ soil required a 2-year crop rotation.     

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.     

Effect of Initial Nematode Density on Managing Globodera rostochiensis with Resistant Cultivars and Nonhosts

Cropping systems in which resistant potato cultivars were grown at different frequencies in rotation with susceptible cultivars and a nonhost (oats) were evaluated at four initial nematode population densities (Pi) for their ability to maintain Globodera rostochiensis at a target level of <0.2 egg/cm³ of soil. At a Pi of 0.1 to 1 egg/cm³ of soil, cropping systems with 2 successive years of a resistant cultivar every 3 years of potato production reduced and maintained G. rostochiensis at <0.2 egg/cm³ of soil. At a Pi of 1 to 4 eggs/cm³ of soil, 2 successive years of a resistant cultivar followed by 1 year of oats for every 4 years of production were necessary to reduce and maintain G. rostochiensis populations at <0.2 egg/cm³ of soil. At a Pi greater than 4 eggs/cm³ of soil, 2 successive years of a resistant cultivar plus 1 year of oats reduced G. rostochiensis densities to <0.2 egg/cm³ of soil, but the population increased above that density after cropping 1 year to a susceptible cultivar. The numbers of cysts and eggs per cyst in the final population (Pf) of G. rostochiensis were influenced by initial density and the frequency of growing a susceptible cultivar in a cropping system. The lowest number of cysts and eggs per cyst in the final G. rostochiensis population occurred with a cropping system consisting of 2 successive years of a resistant cultivar followed by oats with a susceptible cultivar grown the fourth year of production.     

Effects of Interactions among Heterodera glycines,Meloidogyne incognita,and Host Genotype on Soybean Yield and Nematode Population Densities

The effects of host genotype and initial nematode population densities (Pi) on yield of soybean and soil population densities of Heterodera glycines (Hg) race 3 and Meloidogyne incognita (Mi) race 3 were studied in a greenhouse and field microplots in 1983 and 1984. Centennial (resistant to Hg and Mi), Braxton (resistant to Mi, susceptible to Hg), and Coker 237 (susceptible to Hg and Mi) were planted in soil infested with 0, 31, or 124 eggs of Hg and Mi, individually and in all combinations, per 100 cm³ soil. Yield responses of the soybean cultivars to individual and combined infestations of Hg and Mi were primarily dependent on soybean resistance or susceptibility to each species separately. Yield of Centennial was stimulated or unaffected by nematode treatments, yield of Braxton was suppressed by Hg only, and yield suppressions caused by Hg and Mi were additive and dependent on Pi for Coker 237. Other plant responses to nematodes were also dependent on host resistance or susceptibility. Population densities of Mi second-stage juveniles (J2) in soil were related to Mi Pi and remained constant in the presence of Hg for all three cultivars. Population densities of Hg J2 on the two Hg-susceptible Cultivars, Braxton and Coker 237, were suppressed in the presence of Mi at low Hg Pi.     

Yield Reduction and Root Damage to Cotton Induced by Belonolaimus longicaudatus

Sting nematode (Belonolaimus longicaudatus) is recognized as a pathogen of cotton (Gossypium hirsutum), but the expected damage from a given population density of this nematode has not been determined. The objective of this study was to quantify the effects of increasing initial population densities (Pi) of B. longicaudatus on cotton yield and root mass. In a field plot study, nematicide application and cropping history were used to obtain a wide range of Pi values. Cotton yields were regressed on Pi density of B. longicaudatus to quantify yield losses in the field. In controlled environmental chambers, cotton was grown in soil infested with increasing Pi''s of B. longicaudatus. After 40 days, root systems were collected, scanned on a desktop scanner, and root lengths were measured. Root lengths were regressed on inoculation density of B. longicaudatus to quantify reductions in the root systems. In the field, high Pi''s (>100 nematodes/130 cm³ of soil) reduced yields to near zero. In controlled environmental chamber studies, as few as 10 B. longicaudatus/130 cm³ of soil caused a 39% reduction in fine cotton roots, and 60 B. longicaudatus/130 cm³ of soil caused a 70% reduction. These results suggest that B. longicaudatus can cause significant damage to cotton at low population densities, whereas at higher densities crop failure can result.     

Shade Tobacco Yield Loss and Globodera tabacum tabacum Population Changes in Relation to Initial Nematode Density

Field microplot experiments were conducted from 1987 to 1992 to determine the relationship between fresh weight leaf yield of shade tobacco (Nicotiana tabacum) and initial density of Globodera tabacum tabacum (encysted J2 per cm³ soil). Initial nematode densities of 0.1 to 1,097 J2/cm³ soil were negatively correlated with leaf yield, total shoot weight, and normalized plant height 5 to 6 weeks after transplanting (r = -0.73, -0.73, and -0.52, respectively). Nonlinear damage functions were used to relate initial G. t. tabacum densities to the yield and shoot weight data. The model described leaf yield losses of < 5 % for initial nematode densities of less than 100 J2/cm³ soil. Densities above 100 J2 resulted in yields decreasing exponentially to a maximum yield loss of >40% at 500 to 1,000 J2/cm³ soil. A similar initial density tolerance threshold relationship was observed for total shoot weight. No threshold effect was evident for standardized plant height, which was a poor predictor of leaf yield. Globodera tabacum tabacum population increase over a growing season was described by a linear relation on a log/log plot (R² = 0.73).     

Broadleaf Tobacco Yield Loss in Relation to Initial Globodera tabacum tabacum Population Density

Field microplot experiments were conducted from 1995 to 1998 to determine the relationship between fresh shoot weight of stalk-cut broadleaf and shade-grown cigar wrapper tobacco types (Nicotiana tabacum L.) and initial density of Globodera tabacum tabacum second stage juveniles (J2) per cm³ soil. Total shoot weight was negatively correlated with initial nematode densities of 12.3 to 747.3 J2/cm³ soil (r = -0.53 and -0.70 for broadleaf and shade-grown tobacco, respectively). Nonlinear damage functions were used to relate initial G. t. tabacum densities to shoot weight. The models described shoot weight losses of less than 14% or 39% for broadleaf and shade tobacco, respectively, at G. t. tabacum densities below 50 J2/cm³ soil. Total shoot weights were reduced by 40% and 60% of uninfested plots as preplant nematode densities approached maximum levels (>600 J2/cm³ soil) for broadleaf and shade tobacco, respectively. Globodera t. tabacum population increase over a growing season was described by a linear relation on a log/log plot (R² = 0.07 and 0.61 for broadleaf and shade, respectively). These experiments demonstrate that G. t. tabacum can directly reduce shoot weight of stalk-cut broadleaf tobacco. Broadleaf is more tolerant to nematode infection than shade tobacco, as shade tobacco shoot weight reductions were greater at the same initial nematode densities in the same years.     

Influence of Meloidogyne incognita on Resistant and Susceptible Sweet Potato Cultivars

Effects of several population densities ofMeloidogyne incognita on the sweet potato cultivars Centennial (susceptible) and Jasper (moderately resistant) were studied. Field plots were infested with initial levels (Pi) of 0, 10, 100, 1,000, 5,000, and 10,000 eggs and juveniles/500 cm³ soil in 1980 and 0, 100, 1,000, 2,000, 3,000, 4,000, and 5,000 in 1981. M. incognita population development trends were similar on both cultivars; however, at high Pi, more eggs and juveniles were recovered from Centennial than from Jasper. The highest Pi did not result in the highest mid-season (Pm) counts. Pi was negatively correlated with the number of marketable roots and root weight but positively correlated with total cracked roots, percentage of cracked roots, and cracking severity. Jasper tolerated higher Pi with greater yields and better root quality than Centennial. Cracking of fleshy roots occurred with both cultivars at low Pi.     

Influence of Meloidogyne chitwoodi and M. hapla on Wheat Growth

Meloidogyne chitwoodi reduced the growth of winter wheat ''Nugaines'' directly in relation to nematode density in the greenhouse, The relationship between top dry weight and initial nematode density suggests a tolerance limit of Nugaines wheat to M. chitwoodi of between 0.03 and 0.18 eggs/cm³ of soil; the value for relative minimum plant top weight was 0.45 g and 0.75 g, respectively. Growth of wheat in field microplots containing four population densities (0.003, 0.05, 0.75 and 9 eggs/cm³ soil) was not affected significantly at any inoculum level compared to controls during September to July, However, suppression of head weights of ''Fielder'' spring wheat grown May-July occurred in microplots initially infested with 0.75 and 9 eggs/cm³ soil. Reproduction (Pf/Pi) was poorer at these two inoculum levels as compared to the lower densities. In another greenhouse experiment, roots of wheat cultivars Fielder, ''Fieldwin,'' ''Gaines,'' ''Hyslop,'' and Nugaines became infected by M. chitwoodi, but not by M. hapla. Reproduction of M. chitwoodi was less on Gaines and Nugaines than on Fielder, Fieldwin, or Hyslop.     

Yield Response and Injury Levels of Meloidogyne incognita and M. javanica on the Susceptible Tobacco 'McNair 944'

The effects of Meloidogyne incognita and M. javanica on a susceptible tobacco (Nicotiana tabacum L.) cv. McNair 944 were investigated in field microplots during 1978 and 1979. Three initial inoculum levels—4, 16, and 64 nematode eggs and/or second-stage larvae per 100 cm³ of soil—were used for each nematode species. Data obtained from the experiments included plant yield and the amount of reproduction of the two nematode species. At comparative inoculum levels, M. javanica was more aggressive than M. incognita on tobacco and caused approximately twofold more yield suppression than M. incognita. The calculated initial population of M. incognita, derived from the average for 2 yr, which produced a 7% suppression in plant yield was four eggs and/or second-stage larvae per 100 cm³ of soil; whereas less than one M. javanica egg and/or second-stage larvae per 100 cm³ of soil was needed to achieve similar suppression. Nematode reproduction varied in the 1978 and 1979 tests, but similar trends were observed. Early season M. javanica populations were greater than those of M. incognita, but late season populations of M. incognita were twice anti three times those of M. javanica.     

Response of Resistant Soybean Plant Introductions to Meloidogyne arenaria Races 1 and 2

Resistant plant introductions, PI 230977 and PI 200538, and partially resistant Jackson and susceptible CNS were evaluated for seed yield in response to races 1 and 2 of Meloidogyne arenaria. Initial soil population densities (Pi) of the nematode were 0, 31, 125, and 500 eggs/100 cm³ soil. At the highest Pi, yield suppressions of CNS, Jackson, PI 230977, and PI 200538 were 55, 28, 31, and 29%, and 99, 86, 66, and 58% for races 1 and 2 compared with uninfested controls. Numbers of second-stage juveniles (J2) present in roots 14 days after planting increased as Pi increased, but did not differ between the two races. At the highest Pi, fewer race 1 (40-57%) and race 2 (53-68%) J2 were present in roots of the plant introductions than in roots of Jackson. Soil population densities of race 1 J2 at 135 days after planting were 83-89% lower on the resistant genotypes than on CNS. These numbers did not differ for race 2. Reproductive factors were considerably higher for race 2 compared to race 1 for all genotype by Pi combinations, except for CNS at the highest Pi.     

Evaluation of Dry Ice as a Potential Cryonematicide for Meloidogyne incognita in Soil

Solid CO₂ (dry ice) was added to pots containing soil that was infested either with eggs of the root-knot nematode, Meloidogyne incognita, or with tomato (Lycopersicon esculentum ''Rutgers'') root fragments that were infected with various stages of the nematode. Two hours after dry ice was added, thermocouples in the soil recorded temperatures ranging from -15 °C to -59 °C. One day after treatment with the dry ice, the temperature of the soil was allowed to equilibrate with that of the greenhouse, and susceptible tomato seedlings were planted in pots containing infested soil treated or untreated (controls) with dry ice. After 5 weeks, roots were removed from the pots and nematode eggs were extracted and counted. Plants grown in soil infested with eggs and receiving dry ice treatment had less than 1% of the eggs found in the controls; plants from soil infested with root fragments and receiving dry ice treatment had less than 4% of the eggs found in controls. Dry ice used to lower soil temperature may have potential as a cryonematicide.     

A Novel Technique for Infesting Field Sites with Encapsulated Eggs of Meloidogyne spp.

Eggs of Meloidogyne arenaria race 1 were encapsulated in calcium alginate for use as inoculum to infest peanut field plots. Some eggs within the capsules remained viable up to 10 weeks after preparation. A field site was successfully infested at peanut planting and (or) 6 weeks later. Dual applications of nematode inoculum (at planting and 6 weeks later) were superior to single applications (at planting or 6 weeks after planting). Field-site infestation levels at the end of the first year were related to the amount of inoculum dispersed and timing of the infestation (P = 0.001). Peanut yield was only slightly affected in the first year, but significant (P = 0.02) yield suppression occurred during the second year after field infestations. The negative relationship between the numbers of M. arenaria eggs and juveniles per 500 cm³ soil in the fall and the percentage of peanut hull galled the second year was described by a quadratic model (P = 0.002, R² = 0.41).