Infection of Seedlings of Alfalfa and Red Clover by Concomitant Populations of Meloidogyne incognita and Pratylenchus penetrans

Invasion of 2-day-old seedlings of ''Buffalo'' alfalfa and ''Kenland'' red clover by larvae of M. incognita and adults of P. penetrans, during 1-3 day periods of incubation at 24 C, was investigated in 50-mm petri dishes on 1% agar. Penetration by both nematodes increased arithmetically with increased numbers in inocula. P. penetrans invaded alfalfa more readily than red cover, but M. incognita invaded red clover more readily than alfalfa. Both nematodes inhibited root-elongation of alfalfa more than that of red clover. In combinations of 10 and 50 of both nematodes, invasion of both plants by both nematodes was the same as for each nematode alone. Penetration by M. incognita into alfalfa, but not into red clover, was significantly reduced when combinations of 50 M. incognita and 200 P. penetrans were inoculated simultaneously. In the presence of large numbers of entrant P. penetrans in both plants, penetration by M. incognita was highly significantly reduced. Penetration by P. penetrans was unaffected in the reciprocal situations.     

Infectivity of Pratylenchus penetrans on Alfalfa

The infectivity of Pratylenchus penetrans on alfalfa seedlings cv. Du Pulls was studied. The dense root-hair zone was the preferred zone of penetration by females, males, and third-stage larvae. A lesion initially appeared as a water-soaked area at the root surface, becoming yellow and elliptical as the nematode entered the cortex, with dark-brown cells later appearing in the centre as the nematode fed. At 20 C, females penetrated roots earlier, faster, and in greater numbers than either males or third-stage larvae. Females penetrated roots at temperatures from 5 to 35 C, with maximum penetration between 10 and 30 C, while males and third-stage larvae penetrated roots only between 10 and 30 C with maximum penetration a t 20 C. Penetration of roots by females, males, and third-stage larvae increased after storage of 5 C for 35 days, but decreased after storage of 140 days or more. Combinations of the three life stages in pairs neither enhanced nor inhibited penetration of roots by individual life stages; males were not attracted to females. Increasing inoculum density up to 20 nematodes/seedling did not affect penetration.     

Effect of Age of Alfalfa Root on Penetration by Pratylenchus penetrans

Penetration by all migratory life stages of Pratylenchus penetrans into roots of alfalfa (Medicago sativa L. cv. Du Puits) was inversely proportional to tissue age. Two-day-old tissue in the root hair zone was penetrated twice as much as 10- or 20-day-old sections of the tap root. Age-related differences were also observed in branch roots; these differences were not affected by increasing the number of nematodes from 1 to 10 per inoculation site, nor by increasing the length of the incubation period from 6 to 96 h. Age-related differences were only significant with 3-wk-old plants, not with 2- and 1-wk-old seedlings. Nematodes entered roots at temperatures from 5 to 30 C with maximum entry at 20 C and minimum at 5 C. At all temperatures, except 5 C, penetration into young tissue (2 days) was significantly greater than into medium (10 days) and old (20 days) tissue. Females and third-stage larvae entered the different-aged root sections 122% and 83%, respectively, more than did males. Two-day-old seedlings of the alfalfa cultivars Vernal, Saranac, and Du Puits were penetrated equally by P. penetrans. Perhaps the inverse relationship between penetration and age of root is, in part, responsible for the increasing resistance or tolerance of plants to nematode damage as they grow older.     

Nematode Response to Carbofuran

Higher populations of Meloidogyne incognita larvae and Pratylenchus penetrans were recovered from soil treated with carbofuran 10 and 15 days after treatment, respectively, than were recovered from untreated control soil. The number of P. penetrans, however, was lower 50 days after treatment, and symptoms developed only occasionally on the root systems of host plants. Populations of Tylenchorhynchus claytoni inoculated at different distances from the base of corn seedlings growing in carbofuran-treated soil did not move toward the plant, whereas they were attracted in untreated soil from a distance of 12 cm. P. penetrans moved at random in treated agar medium when inoculations occurred 4 cm away from the root tips of tomato seedlings under aseptic conditions. Those nematodes that reached the roots were never observed feeding during a 20-day observation period. Specimens of P. penetrans placed on the developing roots moved at random and never penetrated. In contrast, numerous P. penetrans penetrated roots of seedlings growing in untreated medium.     

Penetration and Development of Meloidogyne arenaria on Two New Grape Rootstocks

Penetration, development, and reproduction of a virulent ''Harmony'' population of Meloidogyne arenaria was studied on two nematode-resistant grape rootstocks 10-17A and 6-19B. ''Cabernet Sauvignon'' was used as a susceptible control for comparison. Plants were inoculated with 100 freshly hatched second-stage juveniles (J2) of M. arenaria. Greater numbers of J2 penetrated roots of ''Cabernet'' than 10-17A, and none penetrated roots of 6-19B 4 days after inoculation (DAI). At 7 DAI, vermiform J2 advanced to sausage-shaped J2 in roots of ''Cabernet,'' penetrated roots of 6-19B, and had egressed from roots of 10-17A. Resistant rootstocks expressed hypersensitive responses to penetrating J2 along the root epidermis, among the cortical cells, and along the differentiating vascular bundles. At 13 DAI, 68% of the J2 had attained globose stage in roots of ''Cabernet,'' whereas there was no development of vermiform J2 in roots of the other two rootstocks. The nematodes reproduced only in roots of ''Cabernet.'' Lack of development of J2 in roots of the two resistant grape rootstocks might be the result of a hypersensitive response to J2 feeding.     

Penetration of Celery and Alfalfa Roots by Pratylenchus penetrans as Affected by Temperature

A greater percentage of females than juveniles or males of P. penetrans penetrated celery roots grown in infested soil at 5, 18, or 30 C; the difference was greatest at 5 C. The time of initial penetration of alfalfa seedlings inoculated with single nematodes on water agar varied with temperature. Females penetrated the seedlings earlier and over a wider range of temperatures than did males or juveniles. The rate of penetration was highest for females. After initial penetration, the penetration rate decreased with time. At 13-28 C, approximately 80% of roots were penetrated by females and only 25-30% by males and juveniles by the end of the experiment.     

Nature of Sweet Potato Resistance to Meloidogyne incognita and the Effects of Temperature on Parasitism

Penetration, rate of development, and total population of Meloidogyne incognita in roots of susceptible ''Allgold'' and resistant ''Nemagold'' sweet potatoes increased with temperature 24-32 C. Rate of larval penetration in ''Allgold'' was significantly higher than in ''Nemagold'' after 48 hr of root exposure at 24, 28, and 32 C. At 24, 28, and 32 C (16 hr) day and 20 C (8 hr) night temperature the life cycle of M. incognita required 42, 32, and 28 days in ''Allgold'', and 44, 33, and 31 days in ''Nemagold''; mature females in the first generation were 40, 40, 40, and 10, 22, 20 respectively. The correlation between the length of time roots were allowed to grow in the soil prior to inoculation and number of larvae recovered from the roots after inoculation was positive for ''Allgold'' and negative for ''Nemagold''. Therefore, a root exudate repellent to M. incognita larvae is proposed as a hypothetical basis for resistance to M. incognita in sweet potatoes.     

Anisohydric behaviour in grapevines results in better performance under moderate water stress and recovery than isohydric behaviour

Aims

Arbuscular mycorrhizal fungi (AMF) can control root-knot nematode infection, but the mode of action is still unknown. We investigated the effects of AMF and mycorrhizal root exudates on the initial steps of Meloidogyne incognita infection, namely movement towards and penetration of tomato roots.

Methods

M. incognita soil migration and root penetration were evaluated in a twin-chamber set-up consisting of a control and mycorrhizal (Glomus mosseae) plant compartment (Solanum lycopersicum cv. Marmande) connected by a bridge. Penetration into control and mycorrhizal roots was also assessed when non-mycorrhizal or mycorrhizal root exudates were applied and nematode motility in the presence of the root exudates was tested in vitro.

Results

M. incognita penetration was significantly reduced in mycorrhizal roots compared to control roots. In the twin-chamber set-up, equal numbers of nematodes moved to both compartments, but the majority accumulated in the soil of the mycorrhizal plant compartment, while for the control plants the majority penetrated the roots. Application of mycorrhizal root exudates further reduced nematode penetration in mycorrhizal plants and temporarily paralyzed nematodes, compared with application of water or non-mycorrhizal root exudates.

Conclusions

Nematode penetration was reduced in mycorrhizal tomato roots and mycorrhizal root exudates probably contributed at least partially by affecting nematode motility.     

Effects of Cycloate on Development of Heterodera schachtii and Growth of Three Beta Species

Greenhouse tests were set up to evaluate the effects of the herbicide, cycloate (S-ethyl cydohexylethylthiocarbamate), oil development of Heterodera schachtii and growth of three Beta species. Cycloate added to infested soil enhanced cyst development/gm root on B. vulgaris and larvae/gm of root in B. patellaris and B. procumbens at 4, 16, and 16 μg(a.i.)/gm of soil, respectively. Total numbers of nematodes/individual root system decreased because of poor root growth of seedlings in cycloate-amended soil. Penetration and larval development through stage three did occur in the wild Beta species in any treatment. Thus, resistance of B. patellaris and B. pocumbens to development of H. schachtii was not altered by cycloate. Cycloate also retarded growth (P = 0.05) of the sugarbeet cultivars and B. patellaris at 4 μg(a.i.)/gm and B. procumbens at 16 μg(a.i.)/gm of soil. Higher concentrations of nematodes/gm root in plants growing in cycloate-amended soil may be attributed to factors such as fewer roots available for penetration, possible effects of cycloate on egg hatch, greater attraction of nematodes to roots, and increased susceptibility of roots to larval penetration. Suppression of seedling growth in cycloate-amended soil may be attributed in part to higher nematode density and in part to direct root damage from cycloate.     

Pathogenicity of the Lesion Nematode,Pratylenchus brachyurus,on Six Soybean Cultivars

Pathogenicity tests of Pratylenchus brachyurus on selected greenhouse-grown soybean cultivars indicated the nematode reduced seed yield of ''Hood'' but not that of ''Custer'', ''Bragg'', ''Dyer'', or ''Pickett''. Root weights of all cultivars were reduced. Damage and numbers of nematodes within soybean roots growing at 13, 21, and 29 C were greater at higher temperatures. At 29 C, root pruning was prominent in ''Hood'' and ''Pickett'' but limited in ''Custer'' and ''Hill''. Root pruning was not observed at 13 C and only ''Pickett'' showed pruning at 21 C. Plant height and foliage weight were not affected. P. brachyurus had no effect on the emergence of ''Pickett'' or ''Bragg'' soybeans. Nematode counts from roots of ''Pickett'' at intervals after inoculation indicated that hatching of second generation second-stage larvae occurred about 15 days after egg laying. An average of 68% of the initial inoculum penetrated the roots within five days of inoculation, the highest observed was 81% in five days. Details of structural damage in penetrated tissues were studied in sectioned roots. In soybean roots infected by P. brachyurus and/or Rhizoctonia solani greater damage occurred with nematode and fungus combined than with either acting alone.     

Interaction of Meloidogyne naasi,Pratylenchus penetrans,and Tylenchorhynchus agri on Creeping Beentgrass

The pathogenicity and interactions of Meloidogyne naasi, Pratylenchus penetrans, and Tylenchorhynchus agri on ''Toronto C-15'' creeping bentgrass, Agrostis palustris, was studied in a long-term greenhouse experiment. Based on dry weights of roots and clippings, M. naasi alone and in all combinations with P. penetrans and T. agri was highly pathogenic to creeping bentgrass. P. penetrans and T. agri alone and in combination inhibited root growth but adversely affected top growth only when the two were co-inoculated. In combination, the effects of each species on top growth were additive, with M. naasi the dominant pathogen. Creeping bentgrass was an excellent host for M. naasi and T. agri, but a poor host for P. penetrans. T. agri inhibited population increase of M. naasi, indicating nematode-nematode competition, but neither T. agr/ nor P. penetrans was affected by any of the combinations.     

Pathogenicity of Pratylenchus penetrans,Heterodera glycines,and Meloidogyne incognita on Soybean Genotypes

The pathogenicity of Heterodera glycines, Meloidogyne incognita, and Pratylenchus penetrans on H. glycines-resistant ''Bryan,'' tolerant-susceptible ''G88-20092,'' and intolerant-susceptible ''Tracy M'' soybean cultivars was tested using plants grown in 800 cm³ of soil in 15-cm-diam. clay pots in three greenhouse experiments. Plants were inoculated with 0, 1,000, 3,000, or 9,000 H. glycines race 3 or M. incognita eggs, or vermiform stages of P. penetrans/pot. Forty days after inoculation, nmnbers of all three nematodes, except H. glycines on Bryan, generally increased with increasing inoculum levels in Experiment I. Heterodera glycines and M. incognita significantly decreased growth only of Tracy M. At 45 and 57 days after inoculation with 6,000 individuals/pot in experiments II and III, respectively, significantly more P. penetrans and M. incognita than H. glycines were found on Bryan. However, H. glycines and M. incognita population densities were greater than P. penetrans on G88-20092 and Tracy M. Growth of Tracy M infected by H. glycines and M. incognita and growth of G88-20092 infected by M. incognita decreased in Experiment III. Pratylenchus penetrans did not affect plant growth. Reduction in plant growth differed according to the particular nematode species and cultivar, indicating that nematodes other than the species for which resistance is targeted can have different effects on cultivars of the same crop species.     

Symptomless Resistance of Alfalfa to Meloidogyne incognita acrita

Penetration, development and migration of the cotton root-knot nematode, Meloidogyne incognita acrita, in resistant and susceptible alfalfa varieties was compared. Larvae entered both resistant and susceptible plants in approximately the same numbers. After 3 to 4 days, the number of larvae in resistant roots decreased sharply until at 7 days fewer than 5 larvae/seedling and no nematode development could be found. In susceptible roots, larvae became sedentary and developed normally; egg production began as early as 18 days after penetration of the host.     

Effect of Hirsutella rhossiliensis on Infection of Potato by Pratylenchus penetrans

We evaluated the ability of the nematode-pathogenic fungus Hirsutella rhossiliensis (Deuteromycotina: Hyphomycetes) to reduce root penetration and population increase of Pratylenchus penetrans on potato. Experiments were conducted at 24 C in a growth chamber. When nematodes were placed on the soil surface 8 cm from a 14-day-old potato cutting, the fungus decreased the number entering roots by 25%. To determine the effect of the fungus on population increase after the nematodes entered roots, we transplanted potato cuttings infected with P. penetrans into Hirsutella-infested and uninfested soil. After 60 days, the total number of nematodes (roots and soil) was 20 ± 4% lower in Hirsutella-infested than in uninfested soil.     

Penetration and Development of Meloidogyne hapla in Resistant and Susceptible Alfalfa Under Differing Temperatures

Studies were conducted to examine under differing temperatures (12, 16, 20, 24, 28, and 32 C) the penetration anti development of Meloidogyne hapla in resistant lines ''298'' and ''Nev. Syn XX'', and susceptible ''Lahontan'' and ''Ranger'' hardy-type alfalfas. The results indicated that resistance to M. hapla was similar to that previously described for M. incognita in nonhardy alfalfa. Although initial penetration in resistant seedlings was similar to that of susceptible seedlings, nematode larvae failed to establish and develop in root tissues and nematode numbers subsequently declined. In susceptible seedlings, nematode development proceeded rapidly, and egg production began after 5 weeks. Temperature had little influence on the nematode development except to slow the response at the lower temperatures. Other studies were conducted to verify a previously reported immune (no penetration) reaction to M. hapla by the ''Vernal'' selection ''M-4''. When compared to the resistant (penetration without nematode development) Vernal selection ''M-9'' under differing temperatures (20, 24, 28, and 32 C), each selection was equally penetrated by M. hapla but at a lower level than in susceptible Ranger cuttings. Generally, no root galling was observed in either M-4 or M-9; however, very slight galling was found 35 days after inoculation on about 50% of these cuttings when grown at 32 C.     

Growth of Potato and Control of Pratylenchus penetrans with Oxamyl-treated Seed Pieces in Greenhouse Studies

Oxamyl was applied to both uncut and cut potato tubers in aqueous solutions of 1,000 to 32,000 μg/ml. Emergence in greenhouse pots was delayed for a day or more after soaking cut tuber pieces in 32,000 μg/ml. After 10 weeks plant growth was greater, relative to the control, when Pratylenchus penetrans-infested soil was planted with cut tubers soaked for 20 minutes in 32,000 μg/ml. Soaking for 40 minutes did not increase nematode control nor affect plant growth. Oxamyl applied to tubers at 1,000 μg/ml reduced the numbers of P. penetrans in the soil by 20% and in the roots by 35%; at 32,000 μg/ml, the numbers of P. penetrans in the soil were reduced by 73-86% and in the roots by 86-97%. The numbers of P. penetrans did not increase in the roots of plants developed from cut tubers soaked in 32,000 μg/ml over a period of 10 weeks, but numbers of lesion nematodes had begun to increase in the soil.     

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.     

Reproductive Variability of Field Populations of Meloidogyne spp. on Grape Rootstocks

Variability in penetration, development, and reproduction of two resistance-breaking field pathotypes (pt.) of Meloidogyne arenaria, M. incognita, and a population of mixed Meloidogyne spp. virulent to grape hosts were compared on two resistant Vitis rootstocks ''Freedom'' and ''Harmony'' in separate tests. ''Cabernet Sauvignon'' was included as a susceptible host to all four nematode populations. Secondstage juveniles (J2) of the mixed population failed to penetrate Freedom roots. By contrast, 6% of J2 in the M. incognita population penetrated Freedom roots but did not develop beyond the swollen J2 stage. The two resistance-breaking populations of M. arenaria differed in their virulence except on susceptible roots of Cabernet Sauvignon. More J2 of M. arenaria pt. Freedom penetrated Freedom roots and reached adult stage than did M. arenaria pt. Harmony. Later life stages of M. arenaria pt. Freedom occurred earlier and in greater numbers in Harmony roots than did M. arenaria pt. Harmony. Reproduction of M. arenaria pt. Freedom was greater in Freedom and Harmony roots than M. arenaria pt. Harmony. Thus, one population of M. arenaria is highly virulent and the other is moderately virulent.     

Effect of Single and Interplantings on Pathogenicity of Pratyenchus penetrans and P. neglectus to Alfalfa and Crested Wheatgrass

Alfalfa is a host of Pratylenchus penetrans and P. neglectus, whereas crested wheatgrass is a host of P. neglectus but not of P. penetrans. In a 120-day greenhouse experiment at 24 ñ 3 C, P. neglectus inhibited the growth of ''Lahontan'' alfalfa and ''Fairway'' crested wheatgrass. There were no differences in persistence and plant growth of alfalfa and crested wheatgrass, or reproduction of P. neglectus, in single plantings of alfalfa (AO) or crested wheatgrass (CWO), or in interplanted alfalfa and crested wheatgrass (ACW) treatments. On alfalfa, P. penetrans inhibited growth and reproduced more than did P. neglectus. Inhibition of plant growth and reproduction of P. penetrans was greater on alfalfa in AO than in ACW treatments. Pratylenchus penetrans did not reproduce on crested wheatgrass, but inhibited growth of crested wheatgrass in interplanted treatments and was avirulent in single planted treatments. Results were similar in a controlled growth chamber experiment at 15, 20, 25, and 30 C. Both nematode species inhibited alfalfa growth at all temperatures, and P. penetrans was more virulent than was P. neglectus to alfalfa at all temperatures and treatments. Plant growth inhibition and reproduction of P. penetrans on alfalfa in single and interplanted treatments were similar at 15-20 C, but were greater in single than in interplanted treatments at 25-30 C. Pratylenchus penetrans was avirulent to crested wheatgrass in the single planted treatments at all temperatures, but inhibited growth of crested wheatgrass in interplanted treatments at 20-30 C. Plant growth and reproduction of P. neglectus on crested wheatgrass was similar in single and interplanted treatments at 20-30 C and 15-30 C, respectively.     

Development of Meloidogyne chitwoodi on Wheat

Postinfection development of Meloidogyne chitwoodi from second-stage juveniles (J2) to mature females and egg deposition on ''Nugaines'' winter wheat required 105, 51, 36, and 21 days at 10, 15, 20, and 25 C. At 25 C, the J2 induced cavities and hyperplasia in the cortex and apical meristem of root tips with hypertrophy of cortical and apical meristem cell nuclei, 2 and 5 days after inoculation. Giant cells induced by late J2 were observed in the stele 10 days after inoculation. Clusters of egg-laying females were common on wheat root galls 25 days after inoculation. Juveniles penetrated wheat roots at 4 C and above, but not at 2 C, when inoculum was obtained from cultures grown at 20 C, but no penetration occurred at 4 C when inoculum was stored for 12 hours at 4 C before inoculation. In northern Utah, J2 penetrated Nugaines wheat roots in the field in mid-May, about 5 months after seedling emergence. M. chitwoodi eggs were first observed on wheat roots in mid-July when plants were in blossom. Only 40% of overwintered M. chitwoodi eggs hatched at 25 C.