Interaction of Population Levels of Fusarium oxysporum f. sp. vasinfectum and Meloidogyne incognita on Cotton

In autoclaved greenhouse soil without Fusarium oxysporum f. sp. vasinfectum, Meloidogyne incognita did not cause leaf or vascular discoloration of 59-day-old cotton plants. Plants had root galls with as few as 50 Meloidogyne larvae per plant. Root galling was directly proportional to the initial nematode population level. Fusarium wilt symptoms occurred without nematodes with 77,000 fungus propagules or more per gram of soil. As few as 50 Meloidogyne larvae accompanying 650 fungus propagules caused Fusarium wilt. With few exceptions, leaf symptoms appeared sooner as numbers of either or both organisms increased. In soils infested with both organisms, the extent of fungal invasion and colonization was well correlated with the extent of nematode galling and other indications of the Fusarium wilt syndrome.     

Histopathology of Selected cultivars of Tobacco infected with Meloidogyne species

Rates of nematode penetration and the histopathology of root infections in fluecured tobacco cultivars ''McNair-944,'' ''Speight G-28,'' and ''NC-89'' with either Meloidogyne arenaria, M. incognita, M. hapla, or M. javanica were investigated. Penetration of root tips by juveniles of all species into the M. incognita-resistant NC-89 and G-28 was much less than that on the susceptible McNair-944. Few juveniles of M. incognita were detected in resistant cultivars 7 and 14 days after inoculation. Infection sites exhibited some cavities and extensive necrotic tissue at 14 days; less necrotic tissue and no intact nematodes were observed 35 days after inoculation. Although some females of M. arenaria reached maturity and produced eggs, considerable necrosis was induced in the resistant cultivars. Meloidogyne hapla and M. javanica developed on all cultivars, but there was necrotic tissue at some infection sites in the resistant cultivars. The occurrence of single multistructured nuclei in the syncytia of most M. hapla infections differed from the numerous small nuclei found in syncytia caused by the other three species.     

Influence of Soil Fumigation on the Fusarium-Root-knot Nematode Disease Complex of Cotton in California

For control of the root-knot nematode, Meloidogyne incognita, and the pathogenic wilt fungus, Fusarium oxysporum, on cotton, soil fumigants were applied in the field at conventional and higher rates. Conventional rates suppressed Fusarium wilt but higher rates gave quicker early growth, better stands, less stand loss over the season, a lower percentage of plants infected with wilt, fewer plants with vascular discoloration, and fewer nematodes. The best treatment about doubled the yields of untreated controls in one experiment and quadrupled them in another.     

Interaction of Fusarium oxysporum f. sp. ciceri and Meloidogyne javanica on Cicer arietinum

Interaction of Meloidogyne javanica and Fusarium oxysporum f. sp. ciceri was studied on Fusarium wilt-susceptible (JG 62 and K 850) and resistant (JG 74 and Avrodhi) chickpea cultivars. In greenhouse experiments, inoculation of M. javanica juveniles prior to F. oxysporum f. sp. ciceri caused greater wilt incidence in susceptible cultivars and induced vascular discoloration in roots of resistant cultivars. Nematode reproduction was greatest (P = 0.05) at 25 °C. Number of galls and percentage of root area galled increased when the temperature was increased from 15 °C to 25 °C. Wilt incidence was greater at 20 °C than at 25 °C. Chlorosis of leaves and vascular discoloration of plants did not occur at 15 °C. The nematode enhanced the wilt incidence in wilt-susceptible cultivars only at 25 °C. Interaction between the two pathogens on shoot and root weights was significant only at 20 °C, and F. o. ciceri suppressed the nematode density at this temperature. Wilt incidence was greater in clayey (48% clay) than in loamy sand (85% sand) soils. The nematode caused greater plant damage on loamy sand than on clayey soil. Fusarium wilt resistance in Avrodhi and JG 74 was stable in the presence of M. javanica across temperatures and soil types.     

Role of Nematodes,Nematicides, and Crop Rotation on the Productivity and Quality of Potato,Sweet Potato,Peanut, and Grain Sorghum

The objective of this experiment was to determine the effects of fenamiphos 15G and short-cycle potato (PO)-sweet potato (SP) grown continuously and in rotation with peanut (PE)-grain sorghum (GS) on yield, crop quality, and mixed nematode population densities of Meloidogyne arenaria, M. hapla, M. incognita, and Mesocriconema ornatum. Greater root-gall indices and damage by M. hapla and M. incognita occurred on potato than other crops. Most crop yields were higher and root-gall indices lower from fenamiphos-treated plots than untreated plots. The total yield of potato in the PO-SP and PO-SP-PE-GS sequences increased from 1983 to 1985 in plots infested with M. hapla or M. arenaria and M. incognita in combination and decreased in 1986 to 1987 when root-knot nematode populations shifted to M. incognita. The total yields of sweet potato in the PO-SP-PE-GS sequence were similar in 1983 and 1985, and declined each year in the PO-SP sequence as a consequence of M. incognita population density increase in the soil. Yield of peanut from soil infested with M. hapla increased 82% in fenamiphos-treated plots compared to untreated plots. Fenamiphos treatment increased yield of grain sorghum from 5% to 45% over untreated controls. The declining yields of potato and sweet potato observed with both the PO-SP and PO-SP-PE-GS sequences indicate that these crop systems should not be used longer than 3 years in soil infested with M. incognita, M. arenaria, or M. hapla. Under these conditions, these two cropping systems promote a population shift in favor of M. incognita, which is more damaging to potato and sweet potato than M. arenaria and M. hapla.     

Effects of Meloidogyne spp. and Rhizoctonia solani on the Growth of Grapevine Rootings

A disease complex involving Meloidogyne incognita and Rhizoctonia solani was associated with stunting of grapevines in a field nursery. Nematode reproduction was occurring on both susceptible and resistant cultivars, and pot experiments were conducted to determine the virulence of this M. incognita population, and of M. javanica and M. hapla populations, to V. vinifera cv. Colombard (susceptible) and to V. champinii cv. Ramsey (regarded locally as highly resistant). The virulence of R. solani isolates obtained from roots of diseased grapevines also was determined both alone and in combination with M. incognita. Ramsey was susceptible to M. incognita (reproduction ratio 9.8 to 18.4 in a shadehouse and heated glasshouse, respectively) but was resistant to M. javanica and M. hapla. Colombard was susceptible to M. incognita (reproduction ratio 24.3 and 41.3, respectively) and M. javanica. Shoot growth was suppressed (by 35%) by M. incognita and, to a lesser extent, by M. hapla. Colombard roots were more severely galled than Ramsey roots by all three species, and nematode reproduction was higher on Colombard. Isolates of R. solani assigned to putative anastomosis groups 2-1 and 4, and an unidentified isolate, colonized and induced rotting of grapevine roots. Ramsey was more susceptible to root rotting than Colombard. Shoot growth was inhibited by up to 15% by several AG 4 isolates and by 20% by the AG 2-1 isolate. AG 4 isolates varied in their virulence. Root rotting was higher when grapevines were inoculated with both M. incognita and R. solani and was highest when nematode inoculation preceded the fungus. Shoot weights were lower when vines were inoculated with the nematode 13 days before the fungus compared with inoculation with both the nematode and the fungus on the same day. It was concluded that both the M. incognita population and some R. solani isolates were virulent against both Colombard and Ramsey, and that measures to prevent spread in nursery stock were therefore important.     

Effect of Fusarium oxysporum f. sp. glycines and Sclerotium rolfsii on the pathogenicity of Meloidogyne incognita race 2 to soybean

Screenhouse studies were conducted to investigate the effects of Fusarium oxysporum f. sp. glycines and Sclerotium rolfsii on the pathogenicity of Meloidogyne incognita race 2 on soybean and the influence of the nematode on wilt incidence and growth of soybean. The interaction of each fungus with the nematode resulted in reduced shoot and root growth. Final nematode population was also reduced with concomitant inoculation of nematode and fungus or inoculation of fungus before nematode. While M. incognita suppressed wilt incidence in two nematode-susceptible cultivars of soybean (TGX 1485-2D and TGX 1440-IE), it had limited effect on wilt incidence in the nematode resistant cultivar of soybean (TGX 1448-2E). When F. oxysporumwas inoculated with the nematode, the mean number of nematodes that penetrated soybean roots decreased by 75% in TGX 1448-2E, 68% in TGX 1485-1D and 65% in TGX 1440-1E. Similarly when the soil was treated with S. rolfsii, the number decreased by 78% in TGX 1448-2E, 77% in TGX 1485-1D and 68% in TGX 1440-1E. The nematode did not develop beyond second-stage juvenile in TGX-1448-2E.     

The effect of root-knot nematode on vascular resistance to Fusarium oxysporum f. sp. vasinfectum in the stems of cotton plants

The effect of root-knot nematode (Meloidogyne incognita) on external wilt symptoms and on the cotton plant's vascular response to stem-inoculation with Fusarium oxysporum f. sp. vasinfectum was investigated. Wilt symptoms were more severe in all plants inoculated with both organisms than with the fungus alone but relative wilt resistance of the cultivars was maintained. Greater symptom severity was associated with greater fungal proliferation in the stele and this was related to the ability of the nematode to reduce the efficiency of vascular occlusion. The nematode had no effect on the accumulation of infection-induced terpenoid aldehyde compounds.     

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.     

Interactions of Concomitant Species of Nematodes and Fusarium oxysporum f. sp. vasinfectum on Cotton

Meloidogyne incognita, Hoplolaintus galeatus, and North Carolina and Georgia populations of Belonolaimus longicaudatus were introduced singly and in various combinations with Fusarium oxysporum f. sp. vasinfectum on wilt-susceptible ''Rowden'' cotton. Of all the nematodes, the combination of the N. C. population of B. longicaudatus with Fusarium promoted greatest wilt development. H. galeatus had no effect on wilt. With Fusarium plus M. incognito or B. longicaudatus, high nematode levels promoted greater wilt than low levels. The combination of either population of B. longicaudatus with M. incognita and Fusarium induced greater wilt development than comparable inoculum densities of either nematode alone or where H. galeatus was substituted for either of these nematodes. Nematode reproduction was inversely related to wilt development. Without Fusarium, however, the high inoculum level resulted in greater reproduction of all nematode species on cotton. Combining M. incognita with B. longicaudatus or H. galeatus gave mutually depressive effects on final nematode populations. The interactions of H. gateatus with B. longicaudatus varied with two populations of the latter.     

Interaction between Meloidogyne incognita and Fusarium oxysporum f. sp. phaseoli on Selected Bean Genotypes

Four bean genotypes (IPA-1, A-107, A-211, and Calima), representing all possible combinations of resistance and susceptibility to Fusarium oxysporum f. sp. phaseoli (Fop) and Meloidogyne incognita, were each inoculated with three population densities of these pathogens. Calima and A-107 were resistant to Fop; A-107 and A-211 were resistant to M. incognita; and IPA-1 was susceptible to both pathogens. In Fop-susceptible lines (IPA-1 and A-211), the presence of M. incognita contributed to an earlier onset and increased severity of Fusarium wilt symptoms and plant stunting. However, the Fop-resistant Calima developed symptoms of Fusarium wilt only in the presence of M. incognita. Genotype A-107 (resistant to both M. incognita and Fop) exhibited Fusarium wilt symptoms and a moderately susceptible reaction to Fop only after the breakdown of its M. incognita resistance by elevated incubation temperatures (27 C). Root galling and reproduction of M. incognita was generally increased as inoculum density of M. incognita was increased on the M. incognita susceptible cultivars. However, these factors were decreased as the inoculum density of Fop was increased. It was concluded that severe infections of bean roots by M. incognita increase the severity of Fusarium wilt on Fop-susceptible genotypes and may modify the resistant reaction to Fop.     

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.     

Relationships of Initial Population Densities of Meloidogyne incognita and M. hapla to Yield of Tomato

Microplots 80 × 100 cm, infested with varying initial population densities (P_i) of Meloidogyne incognita or M. hapla, were planted to tomato at two locations. Experiments were conducted in a sandy loam soil at Fletcher, N. C. (mountains) where the mean temperature for May to September is ca 20.7 C, and in a loamy saml at Clayton, N. C. (coastal plain) where the mean temperature for May to Septemher is ca 24.8 C. In these experimentally infested plots, M. incognita and M. hapla caused maximunt yield losses of 20-30%, at lhe mountain site with Pi of 0-12,500 eggs and larvae/500 cm³ of soil. In the coaslal plain, M. incognita suppressed yields up to 85%, and M. hapla suppressed yields up to 50% in comparison with the noninfested control. A part of the high losses at this site apparently was due to M. incognita predisposing tomato to the early blight fungus. In a second experintent, in which a nematicide was used to obtain a range of P_is (with P_i as high as 25,000/50 cm³ of soil) at Fletcher, losses due to M. incognita were as great as 50%, but similar densities of M. hapla suppressed yields by only 10-25%. Approximate threshold densities for both species ranged from 500 to 1,000 larvae and eggs (higher for surviving larvae) for the mountain site, whereas nutnbers as low as 20 larvae/500 cm³ of soil of either species caused signiticant damage in the coastal plain. Chemical soil treatments proved useful in obtaining various initial population densities; however, problems were encountered in measuring effective inoculum after such treatments, especially in the heavier soil.     

Identification of Single Meloidogyne Juveniles by Polymerase Chain Reaction Amplification of Mitochondrial DNA

Polymerase chain reaction (PCR) was used to amplify a specific 1.8-kb sequence of mitochondrial DNA from single juveniles and eggs from 17 populations of Meloidogyne incognita, M. hapla, M. javanica, and M. arenaria. Approximately 2 μg amplified product were produced per reaction. Restriction digestion of the amplified product with HinfI permitted discrimination of clonal lineages of the four species. Meloidogyne javanica, however, could not be separated from M. hapla by the enzymes used in these experiments. Various amplification conditions and nematode lysis procedures were examined in order to optimize the speed and quality of identifications.     

The Development and Influence of Meloidogyne incognita and M. javanica on Wheat

The effects of soil temperature and initial inoculum density (Pi) of Meloidogyne incognito and M. javanica on growth of wheat (Triticum aestivum cv. Anza) and nematode reproduction were studied in controlled temperature baths in the glasshouse. Nematode reproduction was directly proportional to temperature between 14 and 30 C for M. incognita and between 18 and 26 C for M. javanica. Reproduction rates (Pf/Pi, where Pf = final number of eggs) for Pi''s of 3,000, 9,000, and 30,000 eggs/plant were greatest at each temperature when Pi = 3,000. Maximum M. incognita reproduction rate (Pf/Pi = 51.12) was at 30 C. At 26 C, M. javanica reproduction (Pf/Pi = 14.82, 9.02, and 4.23 for Pi = 3,000, 9,000, and 30,000, respectively) was about half that of M. incognita when Pi = 3,000 or 9,000 but similar when Pi = 30,000. Reproduction of both species was depressed between 14 and 18 C. Shoot and root growth and head numbers were inversely related to soil temperature between 14 and 30 C but were not affected by the Pi of M. incognita when 7 d old seedlings were inoculated. When newly germinated seedlings were inoculated with M. incognita or M. javanica, the Pi did not affect shoot and root fresh weights, shoot/root ratio, and tillering, but it did reduce root dry weight (M. javanica at 26 C) and increase shoot dry weight (M. incognita at 18-22 C). The optimum temperature range is lower for wheat growth than for nematode reproduction. Wheat cv. Anza is a good host for M. incognita and M. javanica, but it is tolerant to both species.     

Pathogenicity of Pythium aphanidermatum to Chrysanthemum in Combined Inoculations with Belonolaimus longicaudatus or Meloidogyne incognita

Rooted cuttings of ''Iceberg'' chrysanthemum in steamed soil were inoculated with the nematodes Belonolaimus longicaudatus, and Meloidogyne incognita, alone and combined with Pythium aphanidermatum, a fungus pathogen of chrysanthemum. B. longicaudatus alone severely restricted the root system; with P. aphanidermatum also present, plant weight and height were further reduced and onset of symptoms was earlier. M. incognita + fungus interaction was similar but less intense. The fungus suppressed egg production of M. incognita but not the reproduction of B. Iongicaudatus. However, all three pathogens combined significantly suppressed reproduction of both nematodes and caused greatest inhibition of plant growth.     

Application of Isoelectric Focusing to the Taxonomic Identification of Meloidogyne spp.

Meloidogyne incognita, M. arenaria, M. hapla, and M. javanica were distinguishable from each other by isoelectric focusing (IEF) of nematode egg proteins. Proteins extracted from larvae and adults of Hoplolaimus columbus and from eggs of Heterodera glycines had distinctive profiles, also. Protein profiles from eggs, preparasitic larvae and egg-laying adults of M. incognita showed differences. It was necessary to compare samples run at the same time to ensure reliability.     

A Polymerase Chain Reaction Method for Identification of Five Major Meloidogyne Species

A polymerase chain reaction (PCR) method for discriminating Meloidogyne incognita, M. arenaria, M. javanica, M. hapla, and M. chitwoodi was developed. Single juveniles were ruptured in a drop of water and added directly to a PCR reaction mixture in a microcentrifuge tube. Primer annealing sites were located in the 3'' portion of the mitochondrial gene coding for cytochrome oxidase subunit II and in the 16S rRNA gene. Following PCR amplification, fragments of three sizes were detected. The M. incognita and M. javanica reactions produced a 1.7-kb fragment; the M. arenaria reaction, a 1.1-kb fragment; and the M. hapla and M. chitwoodi reactions resulted in a 0.52-kb fragment. Digestion of the amplified product with restriction endonucleases allowed discrimination among species with identically sized amplification products. Dra I digestions of the 0.52-kb amplification product produced a characteristic three-banded pattern in M. chitwoodi, versus a two-banded pattern in M. hapla. Hinf I digestion of the 1.7-kb fragment produced a two-banded pattern in M. javanica, versus a three-banded pattern in M. incognita. Amplification and digestion of DNA from juveniles from single isolates of M. marylandi, M. naasi, and M. nataliei indicated that the diagnostic application of this primer set may extend to less frequently encountered Meloidogyne species.     

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.     

Effect of Carbamate,Organophosphate, and Avermectin Nematicides on Oxygen Consumption by Three Meloidogyne spp.

Second-stage juveniles (I2) of Meloidogyne arenaria consumed more oxygen (P ≤ 0.05) than M. incognita J2, which in turn consumed more than M. javanica J2 (4,820, 4,530, and 3,970 μl per hour per g nematode dryweight, respectively). Decrease in oxygen consumption depended on the nematicide used. Except for aldicarb, there was no differential sensitivity among the three nematode species. Meloidogyne javanica had a greater percentage decrease (P ≤ 0.05) in oxygen uptake when treated with aldicarb, relative to the untreated control, than either M. arenaria or M. incognita. Meloidogyne javanica J2 had a greater degree of recovery from fenamiphos or aldicarb intoxication, after subsequent transfer to water, than did M. incognita. This finding may relate to differential sensitivity among Meloidogyne spp. in the field. Degree of respiratory inhibition and loss of nematode motility for M. javanica after exposure to the nematicides were positively correlated (P ≤ 0.05).