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

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

Glucuronidase Expression in Transgenic Tobacco Roots with a Parasponia Promoter on Infection with Meloidogyne javanica

The expression of a g-us reporter gene linked to a Parasponia andersonii hemoglobin promoter has been studied in transgenic tobacco plants after infection by Meloidogyne javanica. Transgenic roots were harvested at different times after nematode inoculation, and stained histochemically for expression of the gus gene. During the early stages of infection (0-2 weeks) there was little expression in giant cells, in contrast to other cells of the root. In later stages of infection (3-6 weeks) there was strong gus expression in giant cells, with virtually no expression in other cells of the root. The Parasponia hemoglobin promoter therefore appears to direct down-regulation of linked genes on induction of giant cells, but up-regulation in mature giant cells. This reflects different metabolic activities in the giant cells depending on their stage of development. The Parasponia hemoglobin promoter may respond to oxygen tension in giant cells. This suggests that oxygen tension may be limited in the metabolically active giant cells that are associated with egg-laying females.     

Evaluation of Paecilomyces lilacinus as a Biocontrol Agent of Meloidogyne javanica on Tobacco

The efficacy of the nematode parasite Paecilomyces lilacinus, alone and in combination with phenamiphos and ethoprop, for controlling the root-knot nematode Meloidogyne javanica on tobacco and the ability of this fungus to colonize in soil under field conditions were evaluated for 2 years in microplots. Combinations and individual treatments of the fungus grown on autoclaved wheat seed, M. javanica eggs (76,000 per plot), and nematicides were applied to specified microplots at the time of transplanting tobacco the first year. Vetch was planted as a winter cover crop, and the fungus and nematicides were applied again the second year to specified plots at transplanting time. The fungus did not control the nematode in either year of these experiments. The average root-gall index (0 = no visible galls and 5 = > 100 galls per root system) ranged from 2.7 to 3.9 the first year and from 4.3 to 5.0 the second in nematode-infested plots treated with nematicides. Plants with M. javanica alone or in combination with P. lilacinus had galling indices of 5.0 both years; the latter produced lower yields than all other treatments during both years of the study. Nevertheless, the average soil population densities of P. lilacinus remained high, ranging from 1.2 to 1.3 × 106 propagules/g soil 1 week after the initial inoculation and from 1.6 to 2.3 × 104 propagules/g soil at harvest the second year. At harvest the second year the density of fungal propagules was greatest at the depth of inoculation, 15 cm, and rapidly decreased below this level.     

Efficacy of Ethoprop on Meloidogyne hapla and M. chitwoodi and Enhanced Biodegradation in Soil

Responses of egg masses, free eggs, and second-stage juveniles (J2) ofMeloidogyne hapla and M. chitwoodi to ethoprop were evaluated. The results indicated that J2 were the most sensitive, followed by free eggs and egg masses. In general, M. chitwoodi was more susceptible to ethoprop than M. hapla. Ethoprop at 7.2 μg a.i./g soil protected tomato roots from upward migrating M. chitwoodi for 5 weeks. The zone of protection was extended to 10 and 20 cm below the root zone when 3.6 and 7.2 cm water were applied over 8 days. Ethoprop at 1.8, 3.6, and 7.2 μg a.i./g soil degraded faster and killed fewer M. chitwoodi J2 in potato field soil previously exposed to ethoprop than in unexposed soil or sterilized exposed soil. The enhanced biodegradation property of the exposed soil lasted 17 months after the last application of ethoprop. The limited downward movement of ethoprop in the soil, migration of M. chitwoodi J2 into the treated zone, presence of resistant life stage(s) at the time of application, and loss of efficacy due to enhanced biodegradation may have a significant effect on the performance of ethoprop.     

Suppression of Meloidogyne incognita and M. javanica by Pasteuria penetrans in Field Soil

The role of Pasteuria penetrans in suppressing numbers of root-knot nematodes was investigated in a 7-year monocuhure of tobacco in a field naturally infested with a mixed population of Meloidogyne incognita race 1 and M. javanica. The suppressiveness of the soil was tested using four treatments: autoclaving (AC), microwaving (MW), air drying (DR), and untreated. The treated soil bioassays consisted of tobacco cv. Northrup King 326 (resistant to M. incognita but susceptible to M. javanica) and cv. Coker 371 Gold (susceptible to M. incognita and M. javanica) in pots inoculated with 0 or 2,000 second-stage juveniles of M. incognita race 1. Endospores of P. penetrans were killed by AC but were only slightly affected by MW, whereas most fungal propagules were destroyed or inhibited in both treatments. Root galls, egg masses, and numbers of eggs were fewer on Coker 371 Gold in MW, DR, and untreated soil than in AC-treated soil. There were fewer egg masses than root galls on both tobacco cultivars in MW, DR, and untreated soil than in the AC treatment. Because both Meloidogyne spp. were suppressed in MW soil (with few fungi present) as well as in DR and untreated soil, the reduction in root galling, as well as numbers of egg masses and eggs appeared to have resulted from infection of both nematode species by P. penetrans.     

Yield-loss Models for Tobacco Infected with Meloidogyne incognita as Affected by Soil Moisture

Yield-loss models were developed for tobacco infected with Meloidogyne incognita grown in microplots under various irrigation regimes. The rate of relative yield loss per initial nematode density (Pi), where relative yield is a proportion of the value of the harvested leaves in uninfected plants with the same irrigation treatment, was greater under conditions of water stress or with high irrigation than at an intermediate level of soil moisture. The maximum rate of plant growth per degree-day (base 10 C) was reduced as nematode Pi increased when plots contained adequate water. When plants were under water stress, increasing Pi did not luther reduce the maximum rate of plant growth (water stress was the limiting factor). Cumulative soil matric potential values were calculated to describe the relationship between available water in the soil (matric potential) due to the irrigation treatments and subsequent plant growth.     

Pathogenicity of Meloidogyne hapla to Lettuce as Affected by Inoculum Level, Plant Age at Inoculation and Temperature

Pathogenicity of Meloidogyne hapla to lettuce was influenced by inoculum level, age of plant at inoculation and temperature. Top weight of ''Minetto'' lettuce was reduced 32% when 2-week-old lettuce plants were each inoculated with five egg masses. Higher inoculum levels did not further decrease top weight significantly. Inoculation at seeding reduced top growth more than inoculation of 1-, 2- or 3-week-old seedlings. M. hapla reduced growth more at the intermediate (21.1 C night and 26.7 C day), than at the low (15.5 C night and 21.1 C day) or high (26.7 C night and 32.2 C day), temperature regimes.     

Relationship between Meloidogyne incognita and Rotylenchulus reniformis as Influenced by Soybean Genotype

The effect of soybean genotype on competition between Meloidogyne incognita race 2 (Mi) and Rotylenchulus reniformis (Rr) was evaluated in greenhouse and microplot replacement series experiments. Soil in pots containing seedlings of ''Davis'' (susceptible to Mi) or ''Buckshot 66'' (resistant to Mi) was infested with 1,000 vermiform individuals in the following Mi:Rr ratios: 0:0, 100:0, 75:25, 50:50, 25:75, or 0:100. After 91 days, the relative nematode yields (number of nematodes in mixed culture divided by the number in nonmixed culture) of each species were calculated based on soil and root nematode populations expressed as nematodes per gram of dry root tissue. To define the relationship between the two species, calculated relative nematode yields were compared with a theoretical noncompetition model using lack-of-fit regression. In the greenhouse, Mi populations on ''Davis'' were stimulated in the presence of Rr. In microplots, low Mi and Rr population densities likely resulted from severe galling and destruction of feeder roots that probably occurred early in the season. Enhanced susceptibility to Mi was not observed on ''Buckshot 66'', which remained resistant to Mi even when colonized by Rr. Host resistance is a key factor in determining the nature of the relationship between Mi and Rr.     

Predisposition of Broadleaf Tobacco to Fusarium Wilt by Early Infection with Globodera tabacum tabacum or Meloidogyne hapla

In greenhouse experiments, broadleaf tobacco plants were inoculated with tobacco cyst (Globodera tabacum tabacum) or root-knot (Meloidogyne hapla) nematodes 3, 2, or 1 week before or at the same time as Fusarium oxysporum. Plants infected with nematodes prior to fungal inoculation had greater Fusarium wilt incidence and severity than those simultaneously inoculated. G. t. tabacum increased wilt incidence and severity more than did M. hapla. Mechanical root wounding within 1 week of F. oxysporum inoculation increased wilt severity. In field experiments, early-season G. t. tabacum control by preplant soil application of oxamyl indirectly limited the incidence and severity of wilt. Wilt incidence was 48%, 23%, and 8% in 1989 and 64%, 60%, and 19% in 1990 for 0.0, 2.2, and 6.7 kg oxamyl/ha, respectively. Early infection of tobacco by G. t. tabacum predisposed broadleaf tobacco to wilt by F. oxysporum.     

Population Development of Meloidogyne incognita on Soybean Defoliated by Pseudoplusia includens

Greenhouse studies examined population densities of Meloidogyne incognita race 4 on soybean (Glycine max ''Davis'') defoliated by larvae of soybean looper (Pseudoplusia indudens (Walker)). Plants were defoliated over a 2-week period beginning 5 weeks after seedlings were transplanted. Four groups of plants were infested with nematodes (5,000 eggs/pot) at 2-week intervals to allow harvesting of plants at 0, 2, 4, and 6 weeks postdefoliation (WPD). Plants in each group were harvested 4 weeks after nematode infestation. Root and nodule weights of defoliated plants were suppressed at 0 WPD, but differences were not detectable at 2, 4, and 6 WPD. Population densities of M. incognita were similar on defoliated and control plants at 0 WPD but were greater on defoliated plants at 4 and 6 WPD. Percentage hatching of eggs produced on the latter plants also was higher. Effects of insect-induced defoliation on development of M. incognita remained detectable even after soybean plant growth apparently returned to normal.     

Nicotine Content of Tobacco Roots and Toxicity to Meloidogyne incognita

The motility of Meloidogyne incognita second-stage juveniles (J2) and their ability to induce root galls in tomato were progressively decreased upon exposure to nicotine at concentrations of 1-100 μg/ml. EC₅₀ values ranged from 14.5 to 22.3 μg/ml, but J2 motility and root-gall induction were not eliminated at 100 μg/ml nicotine. Nicotine in both resistant NC 89 and susceptible NC 2326 tobacco roots was increased significantly 4 days after exposure to M. incognita. The increase was greater in resistant than in susceptible tobacco. Root nicotine concentrations were estimated to be 661.1-979.1 μg/g fresh weight. More M. incognita were detected in roots of susceptible than in roots of resistant tobacco. Numbers of nematodes within resistant roots decreased as duration of exposure to M. incognita was increased from 4 to 16 days. Concentrations of nicotine were apparently sufficient to affect M. incognita in both susceptible and resistant tobacco roots. Localization of nicotine at infection sites must be determined to ascertain its association with resistance.     

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.     

Effects of Concomitant Development on Reproduction of Meloidogyne incognita and Rotylenchulus reniformis on Sweet Potato

The influence of various factors on reproduction of concomitant Meloidogyne incognita (Mi) and Rotylenchulus reniformis (Rr) on sweet potato were studied in the greenhouse. Reproduction of Rr was reduced by Mi at all inoculum levels and experiment durations used, while Mi reproduction was not inhibited. Both species failed to affect each other when inoculated simultaneously onto root systems developed in separate pots from different nodes of the same plant. Reproduction of each species was not significantly greater when inoculation of the second species was delayed 1-2 weeks compared to simultaneous inoculation. After shoot excision, Rr increased in the soil but Mi decreased. Fibrous root weights of plants inoculated with Rr + Mi in some tests were higher than those inoculated with Mi alone, indicating an early suppression of Mi and/or root stintulation by Rr. Drought stress delayed Rr egg hatching and movement of larvae into the soil, but had little effect on Mi reproduction.     

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).     

Reproduction of Meloidogyne javanica on Plant Roots Genetically Transformed by Agrobacterium rhizogenes

Reproduction of Meloidogyne javanica was compared on several Agrobacterium rhizogenes-transformed root cultures under monoxenic conditions. M. javanica reproduced on all transformed roots tested; however, more females and eggs were obtained on potato and South Australian Early Dwarf Red tomato than on bindweed, Tropic tomato, lima bean, or carrot. Roots that grew at moderate rates into the agar and produced many secondary roots supported the highest reproduction. Numbers of females produced in cultures of transformed potato roots increased with increasing nematode inoculum levels, whether inoculum was dispersed eggs or juveniles. Females appeared smaller, produced fewer eggs, and were found in coalesced galls at the higher inoculum levels. The ratio between the final and initial population decreased sharply as the juvenile inoculum increased. The second-stage juvenile was preferred to dispersed eggs or egg masses for inoculation of tissue culture systems because quantity and viability of inoculum were easily assessed. Meloidogyne javanica reared on transformed root cultures were able to complete their life cycles on new transformed root cultures or greenhouse tomato plants.     

Host-Specific Pathogenicity and Genome Differences between Inbred Strains of Meloidogyne hapla

Five isolates of M. hapla originating from the Netherlands and California were inbred by sequential transfer of single egg masses to produce six strains. Cytological examination showed that oocytes of these strains underwent meiosis and had n = 16 chromosomes. Strains were tested for ability to infect and to develop on several hosts by in vitro assays. The two strains from California infected tomato roots at a higher rate than those from the Netherlands, but no difference among strains was seen for ability to develop on tomato with or without the resistance gene Mi-1. All strains developed on the common bean cultivar Kentucky Wonder, but strains differed in ability to develop on the nematode-resistant cultivar NemaSnap. Strain-specific differences were also seen in ability to infect and to develop on Solanum bulbocastanum clone SB-22. Strain VW13, derived from nematodes treated with the mutagen EMS, was defective in ability to infect tomato and potato roots in vitro. Comparison of DNA using AFLP markers showed an average of 4% of the bands were polymorphic across the six strains, but no correlation was observed between the geographical origin or virulence and DNA polymorphism pattern.     

Effects of Population Densities of Meloidogyne hapla on Growth and Yield of Tomato

Growth and yield of ''Veebrite'' tomato were studied in 20-cm (i.d.) clay-tile microplots containing initially 260, 1,840, 6,120, or 27,950 Meloidogyne hapla larvae/kg of soil. Low nematode numbers stimulated, and the highest nematode population suppressed, vegetative plant growth. More tomatoes, with a higher total weight, were harvested from plants infested with 260 and 1,840 nematode larvae at planting than from those with initial densities of 6,120 and 27,950 larvae. At the two highest densities, the cumulative fruit production (weight) was suppressed by 10% and 40%, respectively. The increase in growth and yield at the lower densities appeared to be due to an increase in the size of the root systent. However, at the higher densities, yield was no longer directly related to root weight. The reproduction factor of M. hapla was negatively correlated with initial density; for the lowest and highest initial densities, it was 96X and 7X at midseason, and 354X and 3X at harvest, respectively. The equilibrium density was 63,000 larvae/kg of soil; initial densities larger than 2,000 larvae/kg of soil may require control.     

Relationship Between Population Densities of Pratylenchus penetrans and Crop Losses in Flue-Cured Tobacco in Ontario

Flue-cured tobacco was grown in microplots consisting of concrete drainage tries, 40-cm (i.d.), infested with 0; 666 ; 2000; 6000 or 18,000 root-lesion nematodes, Pratylenchus penetrans/kg of soil. Yield and grade index decreased with preplant soil population densities in excess of 2000/kg of soil. At initial densities of 6000 and 18,000/kg of soil losses in crop returns were 11.0% and 27.5%, respectively. Decreases in the maturity index and in percentage dry stalk weight with increasing densities showed that the nematode delayed maturity. Increases in population densities of nematodes were correlated with decreases in weights of tops and roots and in plant height. All final population densities in soil were lower than the initial densities except at the lowest pre-plant density. All soil populations at midseason were lower than those at the beginning and end of the growing season. Populations of P. penetrans at harvest were in excess of half a million per root system with the 18,000/kg initial soil population density. The results suggest that fumigation, which costs $75/ha, or approximately 2% of the crop value, is economically warranted at preplant densities in excess of 2000/kg of soft.     

Influence of Initial Population Densities of Meloidogyne incognita on Three Chile Cultivars

The effects of Meloidogyne incognita on the Big Jim, Jalapeno, and New Mexico No. 6 chile (Capsicum annuum) cultivars were investigated in microplots for two growing seasons. All three cultivars were susceptible to M. incognita and reacted similarly to different initial populations of this nematode. Severe stunting and yield suppressions occurred at all initial M. incognita densities tested ranging from 385 to 4,230 eggs and larvae/500 cm³ soil. Regression analysis of the microplot data from a sandy loam soil showed yield losses of 31% for the 1978 season and 25% for the 1979 season for the three cultivars for each 10-fold increase in the initial population of M. incognita.