Replacement Series: A Tool for Characterizing Competition between Phytoparasitic Nematodes

The replacement series approach was used to detect and define competition between Meloidogyne incognita (Mi) and Rotylenchulus reniformis (Rr) on soybean. In three greenhouse tests, soybean cv. Davis seedlings were inoculated with 1,000 vermiform nematodes in the following Mi:Rr ratios: 0:0, 100:0, 75:25, 50:50, 25:75, and 0:100. After 86 days, relative nematode-yield values (number of each species in mixed culture divided by number in nonmixed culture) were calculated based on nematodes in soil per gram of dry root tissue. Calculated values were plotted and the resulting line compared with a reference line representing equal inter- and intraspecific competition predicted by the replacement series. Relative yields for Mi were higher than predicted at all ratios where Mi and Rr occurred together (lack-of-fit regression, F= 5.9401, P = 0.0008), indicating increased reproduction in the presence of Rr. Relative yields for Rr did not differ from predicted yields (lack-of-fit regression, F= 0.7565, P = 0.5203), indicating no effect of Mi on Rr. These relationships were not detected using analysis of variance. The relationship between Mi and Rr was independent of host colonization by Diaporthe phaseolorum var. caulivora, the stem canker fungus.     

Effects of Heterodera glycines and Meloidogyne incognita on Early Growth of Soybean

Greenhouse and field microplot studies were conducted to compare soybean shoot and root growth responses to root penetration by Heterodera glycines (Hg) and Meloidogyne incognita (Mi) individually and in combination. Soybean cultivars Centennial (resistant to Hg and Mi), Braxton (resistant to Mi, susceptible to Hg), and Coker 237 (susceptible to Hg and Mi) were selected for study. In the greenhouse, pot size and number of plants per pot had no effect on Hg or Mi penetration of Coker 237 roots; root weight was higher in the presence of either nematode species compared with the noninoculated controls. In greenhouse studies using a sand or soil medium, and in field microplot studies, each cultivar was grown with increasing initial population densities (Pi) of Hg or Mi. Interactions between Hg and Mi did not affect early plant growth or number of nematodes penetrating roots. Root penetration was the only response related to Pi. Mi penetration was higher in sand than in soil, and higher in the greenhouse than in the field, whereas Hg penetration was similar under all conditions. At 14 days after planting, more second-stage juveniles were present in roots of susceptible than in roots of resistant plants. Roots continued to lengthen in the greenhouse in the presence of either Mi or Hg regardless of host genotype, but only in the presence of Mi in microplots; otherwise, responses in field and greenhouse studies were similar and differed only in magnitude and variability.     

Response of Resistant and Susceptible Bell Pepper (Capsicum annuum) to a Southern California Meloidogyne incognita Population from a Commercial Bell Pepper Field

To determine the presence and level of root-knot nematode (Meloidogyne spp.) infestation in Southern California bell pepper (Capsicum annuum) fields, soil and root samples were collected in April and May 2012 and analyzed for the presence of root-knot nematodes. The earlier samples were virtually free of root-knot nematodes, but the later samples all contained, sometimes very high numbers, of root-knot nematodes. Nematodes were all identified as M. incognita. A nematode population from one of these fields was multiplied in a greenhouse and used as inoculum for two repeated pot experiments with three susceptible and two resistant bell pepper varieties. Fruit yields of the resistant peppers were not affected by the nematodes, whereas yields of two of the three susceptible pepper cultivars decreased as a result of nematode inoculation. Nematode-induced root galling and nematode multiplication was low but different between the two resistant cultivars. Root galling and nematode reproduction was much higher on the three susceptible cultivars. One of these susceptible cultivars exhibited tolerance, as yields were not affected by the nematodes, but nematode multiplication was high. It is concluded that M. incognita is common in Southern California bell pepper production, and that resistant cultivars may provide a useful tool in a nonchemical management strategy.     

Population Dynamics of Meloidogyne incognita and Rotylenchulus reniformis Alone and in Combination,and Their Effects on Sweet Potato

Meloidogyne incognita (Mi) and Rotylenchulus reniformis (Rr) interactions on sweet potato were studied in naturally and artificially infested field plots for 3 years. In a naturally infested field, early season counts of Mi or Rr were positively correlated with later counts of the same nematode, but negative correlations were found between early Mi and subsequent Rr, and early Rr and subsequent Mi counts. In field plots fumigated with methyl bromide and then infested with low levels of Rr, Mi, and Rr + Mi, final population densities of Mi juveniles were reduced by Rr, but Rr was not affected by Mi. In field plots with a high natural population density of Rr, artificial infestation with high levels of Mi in both fumigated and nonfumigated treatments inhibited Rr, while the final Mi juvenile population density was not affected. Results indicate that a competitive interaction exists with each species capable of inhibiting the other and becoming the dominant population. The nematodes had no apparent effect on yield at the inoculum densities used, either alone or mixed. Both nematodes increased cracking of sweet potatoes, but mixed populations did not differ in incidence of cracking from either Rr or Mi alone.     

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.     

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.     

Competition of Meloidogyne incognita and Rotylenchulus reniformis on Cotton Following Separate and Concomitant Inoculations

It has been hypothesized Rotylenchulus reniformis (Rr) has a competitive advantage over Meloidogyne incognita (Mi) in the southeastern cotton production region of the United States. This study examines the reproduction and development of Meloidogyne incognita (Mi) and Rotylenchulus reniformis (Rr) in separate and concomitant infections on cotton. Under greenhouse conditions, cotton seedlings were inoculated simultaneously with juveniles (J2) of M. incognita and vermiform adults of R. reniformis in the following ratios (Mi:Rr): 0:0, 100:0, 75:25, 50:50, 25:75, and 0:100. Soil populations of M. incognita and R. reniformis were recorded at 3, 6, 9, 14, 19, 25, 35, 45, and 60 days after inoculations. At each date, samples were taken to determine the life stage of development, number of egg masses, eggs per egg mass, galls, and giant cells or syncytia produced by the nematodes. Meloidogyne incognita and R. reniformis were capable of initially inhibiting each other when the inoculum ratio of one species was higher than the other. In concomitant infections, M. incognita was susceptible to the antagonistic effect of R. reniformis. Rotylenchulus reniformis affected hatching of M. incognita eggs, delayed secondary infection of M. incognita J2, reduced the number of egg masses produced by M. incognita, and reduced J2 of M. incognita 60 days after inoculations. In contrast, M. incognita reduced R. reniformis soil populations only when its proportion in the inoculum ratio was higher than that of R. reniformis. Meloidogyne incognita reduced egg masses produced by R. reniformis, but not production of eggs and secondary infection.     

In Vitro Culture and Feeding Behavior of Belonolaimus longicaudatus on Excised Zea mays Roots

A greenhouse population of the sting nematode, Belonolaimus longicaudatus, obtained from an infested golf course in California''s Coachella Valley, was surface-decontaminated and cuhured on excised roots of Zea mays supported by Gamborg''s B5 medium. At 26-27 °C the females laid eggs, and newly emerged juveniles of the second generation completed three molts within 29 days after egg deposition. Sixty days after inoculation with 60 females and 40 males, an average of 529 nematodes and 83 eggs were recovered from the culture. The feeding process consisted of probing, stylet penetration, ingestion, and stylet retraction. Feeding seemed to be necessary before egg deposition or molting occurred. The sting nematode was observed feeding exclusively as an ectoparasite and preferably at the region of cell division and elongation. Vigorous feeding by many nematodes usually caused discoloration of root tips and termination of growth.     

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.     

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).     

Interactions between Meloidogyne incognita,M. hapla,and Pratylenchus brachyurus in Tobacco

In a greenhouse pot experiment on the pathogenicity and interactions of Meloidogyne incognita, M. hapla and Pratylenchus brachyurus on four cultivars o f tobacco the cultivars ''Hicks'' and ''NC 2326'' were susceptible to each nematode and "NC 95'' and ''NC 2512'' resistant only to M. incognita.Mean heights of susceptible plants were depressed but fresh weight of tops did not differ significantly. Meloidogyne spp. increased fresh weight of susceptible (but not the resistant) roots.Reproduction of M. incognita was decreased in the presence of P. brachyurus in one case. M. hapla reproduction was less with either of the other nematodes in five out of eight cases. In 12 combinations involving P. brachyurus, reproduction of this species was depressed in seven, not affected in four and increased in one.Mechanisms involved in associative interactions were not identified but appeared to be indirect and to involve individual host-nematode responses.     

Competition between Heterodera glycines and Meloidogyne incognita or Pratylenchus penetrans: Independent Infection Rate Measurements

Competition on soybean between Heterodera glycines (race 3) and Meloidogyne incognita or H. glycines and Pratylenchus penetrans were investigated in greenhouse experiments. Each pair of nematode species was mixed in 3-ml suspensions at ratios of 1,000:0, 750:250, 500:500, 250:750, and 0:1,000 second-stage juveniles or mixed stages for P. penetrans. Nematodes from a whole root system were counted and infection rates standardized per 1,000 nematodes (per replication) prior to testing the null hypothesis through a lack-of-fit F-test. Although the effect of increasing H. glycines proportions on the infection rate of M. incognita was generally adverse, the rate deviated significantly from a trend of linear decline at the 75% H. glycines level in one of two experiments. All lack-of-fit F-tests for the H. glycines and P. penetrans mix were significant, indicating that infection rates for both nematodes varied considerably across inocula. The infection rate of H. glycines decreased with increasing P. penetrans proportions. The rate of P. penetrans infection increased with increasing H. glycines proportions up to the 50% level, but declined at the 75% level. Competition had no effect on nematode development. The general adverse relationships between M. incognita and H. glycines and those between P. penetrans and H. glycines showed a linear trend. The relationship between H. glycines and P. penetrans indicates that the former may be competitive when present at higher proportions than the latter. In this study we have evaluated nematode competition under controlled conditions and provide results that can form a basis for understanding the physical and physiological trends of multiple nematode interactions. Methods critical to data analyses also are outlined.     

Early Growth of Soybean as Altered by Heterodera glycines,Phenamiphos and/or Alachlor

Greenhouse and field experiments were conducted to determine the effects of phenamiphos and/or alachlor on early growth of soybean, root morphology, and infection and resurgence of Heterodera glycines (race 1). All tests were planted to ''Ransom'' soybeans. In greenhouse experiments without nematodes, root growth was inhibited at 5 days by alachlor treatments and at 10 days by phenamiphos treatments; with nematodes, phenamiphos treatments enhanced root growth. Phenamiphos also suppressed early penetration of soybean roots by H. glycines in the greenhouse. Early soybean growth parameters among treatments were generally similar in the field. Nematode penetration was limited with treatments containing phenamiphos at one location. Plants treated with only alachlor had less nematode infection than did the control; however, plants treated with herbicide/nematicide combinations had more nematode penetration than did plants treated with phenamiphos alone. Alterations of root growth and interference with the efficacy of phenamiphos are two processes by which alachlor may enhance soybean susceptibility or suitability to H. glycines.     

Effects and Dynamics of a Nematode Community on Soybean

The relationships between densities of all members of a plant-parasitic nematode community and yield of ''Davis'' soybean and between final and preplant population levels were examined in small plots on sandy soils in north-central Florida. Plant-parasitic nematodes present in the community included Belonolaimus longicaudatus, Criconemella sphaerocephala, Meloidogyne incognita, Paratrichodorus minor, Pratylenchus brachyurus, and Xiphinema sp. Plant growth, including stand count, soybean yield (kg/ha), and size of young plants, was occasionally inversely correlated (P ≤ 0.05) with densities of B. longicaudatus or P. brachyurus, but not with densities of other species or with a range of soil variables. The nature of this relationship varied with season, with more severe stand losses noted during 1987 than in 1988. Final population densities (Pf) of most nematode species showed significant (P ≤ 0.05) linear relationships to densities measured at planting or earlier (Pi). These relationships were stronger (higher r²) with the ectoparasite B. longicaudatus than with the endoparasites M. incognita and P. brachyurus. Criconemella sphaerocephala declined under soybean cultivation, reaching levels near zero after two seasons. A quadratic model showed an improvement (P ≤ 0.05) over the linear model in describing the relationship between Pf and Pi measured at planting for B. longicaudatus, and gave a better indication of the leveling off of Pf at high values of Pi.     

Infection,Reproduction Potential,and Root Galling by Root-knot Nematode Species and Concomitant Populations on Peanut and Tobacco

Single populations of Meloidogyne arenaria races 1 (MA1) and 2 (MA2) and M. hapla (MH), and mixed populations of MA1 + MA2 and MA1 + MH with four inoculum levels of eggs were tested on peanut cv. ''Florigiant'' and M. incognita-resistant tobacco cv. ''McNair 373'' in a greenhouse experiment. Root infection, female development, and reproduction of MA2 on peanut and MA1 on resistant tobacco were limited at 2 and 6 weeks. MA1, MH, and MA1 + MH on peanut had similar root infection (total parasitic forms per root unit) at both 2 and 6 weeks, and similar female development and reproduction potentials at 6 weeks. MA2 tended to depress root infection, female development, and reproduction of MA1 on peanut. MH had little effect on MA1 on this crop. On tobacco, MA2 population had greater incidence of root infection than did MH at 2 weeks. The two nematode species had similar development in roots at 6 weeks. All of these processes were restricted when either MA2 or MH was present together with MA1. As initial inoculum level of parasitically fit populations increased, relative infection ratio on both peanut and tobacco, and reproduction factor on peanut decreased. Populations that had high infection incidence and reproduction rates induced greater root galling than did other populations. Root galling was suppressed in the presence of antagonistic response between nematode populations.     

Impact of Plant Nutrition on Pratylenchus penetrans Infection of Prunus avium Rootstocks

A hypothesis that cherry rootstocks grown under optimal nutrient conditions are affected less by Pratylenchus penetrans infection than those grown under deficient nutrient conditions was tested by growing four Prunus avium L. rootstocks (''Mazzard'', ''Mahaleb'', ''GI148-1'', and ''GI148-8'') at a soil pH of 7.0 over a period of 3 months under greenhouse conditions (25 ñ 2 °C). Pratylenchus penetrans was inoculated at 0 (control) or 1,500 nematodes per g fresh root weight for a total of 3,600, 4,200, 10,500, and 11,400 per plant on Mazzard, Mahaleb, GI148-1, and GI148-8, respectively, with nutrients (commercial fertilizer) applied once at planting (deficient) or twice weekly (optimal). The experiment was repeated once. The optimum nutrient regime resulted in greater soil nutrient levels and plant growth; higher leaf concentrations of N, P, K, and Mg; and fewer P. penetrans than under the deficient nutrient regime. The addition of fertilizer either may increase nematode mortality in the soil or improve rootstock resistance to nematode infection. Increases in Ca in leaves from the nutrient-deficient and nematode-infected treatments suggested the plants were physiologically stressed. The Pf/Pi ratios indicated that these rootstocks may have had resistance to P. penetrans; however, because of the dominant role of nutrition in the experimental design, the question of resistance could not be properly addressed.     

Pepper Rootstock Graft Compatibility and Response to Meloidogyne javanica and M. incognita

Resistance of pepper species (Capsicum annuum, C. baccatum, C. chinense, C. chacoense, and C. frutescens), cultivars and accessions to the root-knot nematodes Meloidogyne incognita race 2 and M. javanica, and their graft compatibility with commercial pepper varieties as rootstocks were evaluated in growth chamber and greenhouse experiments. Most of the plants tested were highly resistant to M. javanica but susceptible to M. incognita. Capsicum annuum AR-96023 and C. frutescens accessions as rootstocks showed moderate and relatively high resistance to M. incognita, respectively. In M. incognita-infested soil in a greenhouse, AR-96023 supported approximately 6-fold less nematode eggs per gram root and produced about 2-fold greater yield compared to a nongrafted commercial variety. The commercial variety grafted on AR-96023 produced a yield as great as the non-grafted variety in the root-knot nematode-free greenhouse. Some resistant varieties and accessions used as rootstocks produced lower yields (P < 0.01) than that of the non-grafted variety in the noninfested greenhouse. Use of rootstocks with nematode-resistance and graft compatibility may be effective for control of root-knot nematodes on susceptible pepper.     

Pathological Effects of Pratylenchus neglectus on Wheatgrasses

In controlled greenhouse and growth chamber studies, Pratylenchus neglectus reduced dry shoot and dry root weight of rangeland grasses. Greenar intermediate wheatgrass and Secar Snake River wheatgrass were more susceptible to P. neglectus than Hycrest crested wheatgrass, Fairway crested wheatgrass, and Nordan crested wheatgrass at a greenhouse bench temperature of 26 ± 3 C. Hycrest was the most tolerant to parasitism by P. neglectus. An initial nematode inoculum density of four nematodes/cm³ soil reduced dry shoot weights of Hycrest, Fairway, Nordan, Greenar, and Secar by 22%, 33%, 36%, 47%, and 49%, and reduced dry root weights by 26%, 31%, 32%, 38%, and 42%. There was a positive relationship between dry root weight, the nematode inoculum density, and the nematode reproduction index (final nematode population/initial nematode inoculum). However, there were more nematodes/g root tissue on Secar than on the crested wheatgrasses, and significantly more nematodes/g root tissue on Greenar, Fairway, and Nordan than on Hycrest. Pratylenchus neglectus was most pathogenic at four nematodes/cm³ soil at 30 C and least pathogenic at one nematode/cm³ soil at 15 C. Greenar and Secar were more susceptible to the nematode than Hycrest, Fairway, and Nordan at two and four nematodes/cm³ soil at 20 to 30 C. The nematode reproductive indices were greatest at 30 C and were positively correlated with dry root weight. Secar supported the most and Hycrest had the fewest nematodes/g root.     

Temporal Efficacy of Selected Nematicides on Meloidogyne Species on Tobacco

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

Effects of Meloidogyne incognita on Nitrogen Fixation in Soybean

The effects of a North Carolina population of Meloidogyne incognita on N₂ fixation on root-knot-susceptible ''Lee 68'' and moderately resistant ''Forrest'' soybean were evaluated 50, 75, I00, and 135 days after inoculation with nematodes. Nematodes stimulated N₂ fixation in Lee 68 by 50 days and in Forrest by 75 days. At all other intervals, N₂ fixation was either depressed or unaffected by nematodes. Additional observations indicate that the susceptibility of Lee 68 is associated with greater rates of penetration by larvae and more favorable responses of host tissues to nematodes than occur in Forrest. With time, however, the histological reactions of both hosts became less favorable for nematode development. Resistant or hypersensitive responses became common in Forrest by 75 days but not in Lee 68 until 90 days after inoculation. This population of M. incognita may stimulate N₂ fixation at a specific time interval and depress it at others; therefore, disease of susceptible soybeans caused by this nematode is probably not primarily due to a net loss of fixed nitrogen but to pathogenicity similar to that which occurs on nonlegume hosts.