Development of Four Populations of Meloidogyne hapla on Two Cultivars,of Cucumber at Different Temperatures

The infectivity and development of four populations of Meloidogyne hapla were compared, at three temperatures, on tomato and two varieties of cucumber. A population from Canada produced few root-galls on cucumber and, except at 24 C, no larvae developed into adult females and produced egg masses. In contrast, a population with 45 chromosomes from America produced many galls on cucumber and small proportions of larvae became females and produced egg masses at 20 and 24 C. At 18 C this population produced no egg masses on cucumber, but a population from Britain and one from America with 17 chromosomes produced more egg masses at this temperature than at 20 or 24 C. Dissection of the galls showed that on cucumber many larvae died or their growth and development was slowed.     

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.     

Sex Differentiation in Meloidogyne incognita and Anatomical Evidence of Sex Reversal

Sex differentiation was studied by examining the cellular structure of gonad primordia extracted from second-stage juveniles developing under different environmental conditions. In female jnveniles, divisions of the two somatic cells of the primordium occurred in mid-sccond stage and resulted in 12 cells. Two of them were differentiated as cap cells, two occupied the anterior central and eight the posterior central part of the V-shaped primordium. The two germinal cells divided at the 6-8 somatic-cell stage of the primordium; i.e., earlier than in any other plant-parasitic nematode. In male juveniles of similar developmental stage, divisions of somatic cells resulted in 10 cells: one cap cell at the posterior tip and nine cells at the anterior part of the rod-shaped primordium. Germinal cells divided at the 6-8 sontatic-cell stage. On the basis of gonad anatomy it was concluded that some female juveniles undergo sex reversal and proceed with further development as males. The degree of expression of intersexual features depends on the period at which sex reversal occurs. Sex reversal at an early period gives rise to males with one testis, almost indistinguishable front true males. Sex reversal at mid-second stage involves degeneration of the nucleus of one of the cap cells resulting in males with an atrophied testis and a well-developed testis. More delayed sex reversal results in males with two testes of approximately equal size. To explain these patterns of development, it is assumed that sex differentiation is hormonally controlled and that the environment influences hormonal balance by affecting gene expression.     

Parasitic Variability of Meloidogyne incognita Populations on Susceptible and Resistant Cotton

Root gall induction and egg production by the four recognized host races and two cytological races of Meloidogyne incognita were compared on cotton Gossypium hirsutum cvs. Deltapine 16 (root-knot susceptible) and Auburn 634 (highly resistant). The 12 nematode populations included in the study were from various parts of the world. No population increases occurred on the highly resistant cultivar. After 45 days, populations of host races 1 and 2 induced slight root galling on both cuhivars with only limited reproduction. Host race 4 populations induced moderate root galling with higher reproduction on Deltapine 16 than that of race 1 or race 2 populations. Host race 3 populations induced severe root galling with population density increases of 7-30-fold. In a complementary study, 24 cotton cultivars or breeding lines were compared for suitability as hosts for a typical population of M. incognita race 3. The poorest hosts, ''Aubnru 623,'' ''Auburn 634,'' and ''McNair 220,'' yielded fewer eggs after 45 days than were added initially. The best hosts - ''M-8.'' ''DES 24-8,'' ''McNair 235,'' and ''Coker 20l'' - yielded > 5 times as many eggs as were added initially.     

Early Root Response to Meloidogyne incognita in Resistant and Susceptible Alfalfa Cultivars

The early events of Meloidogyne incognita behavior and associated host responses following root penetration were studied in resistant (cv. Moapa 69) and susceptible (cv. Lahontan) alfalfa. Ten-day-old seedlings of alfalfa cultivars were inoculated with second-stage juveniles (J2) and harvested 12, 24, 48, and 72 hours and 7, 14, and 21 days later. Both cultivars supported similar root penetration and initial J2 migration. By 72 hours after inoculation the majority of J2 were amassed inside the vascular cylinder in roots of susceptible Lahontan, while J2 had not entered the vascular cylinder of resistant Moapa 69 and remained clumped at the root apex. Nematode development progressed normally in Lahontan, but J2 were not observed in Moapa 69 after day 7. The greatest differences between RNA translation products isolated from inoculated and uninoculated roots of Lahanton occurred 72 hours after inoculation. Only minor differences in gene expression were observed between inoculated and uninoculated Moapa 69 roots at 72 hours. Comparison of translation products from inoculated versus mechanically wounded Lahontan roots revealed products that were specific to or enhanced in nematode-infected plants. Moapa 69 appears to possess a type of resistance to M. incognita that does not depend on a conventional hypersensitive response.     

Dynamics of Winter Survival of Eggs and Juveniles of Meloidogyne incognita and M. arenaria

Winter survival dynamics of Meloidogyne incognita and M. arenaria were studied at nine sites in Texas for 2 years. Population survival from October until April was variable among sites, ranging from 0.1% to 33%. A negative correlation (r = -0.86, P = 0.01) was observed between initial population, densities in October and survival percentage until the following April. Total population (eggs + J2) and population of eggs declined continuously during the survival period. Populations of juveniles (J2) increased initially, then declined. The total populations were 82% eggs in October; hatch of these eggs was believed responsible for the observed increase in the population density of J2. Viable eggs were recovered from the soil until March. Eggs are as important as J2 in winter survival of M. incognita and M. arenaria in Texas. Survival data were analyzed by a simple mathematical model.     

Dynamics of Concomitant Populations of Hoplolaimus columbus,Scutellonema brachyurum,and Meloidogyne incognita on Cotton

Cotton seedlings grown in a greenhouse and a growth chamber were inoculated with Scutellonema brachyurum, Hoplolaimus columbus, and Meloidogyne incognita, singly and in all possible combinations, at two initial population (Pi) levels (100 and 300/100 cm³). S. brachyurum alone was not pathogenic to cotton at these population levels. It fed primarily as an ectoparasite but matured and reproduced within the root when it penetrated. Populations of S. brachyurum increased in the presence of H. columbus but were suppressed by M. incognita. H. columbus suppressed dry shoot weights of cotton (P = 0.05) at a Pi of 300/100 cm³ soil. Simultaneous inoculation of H. columbus with either M. incognita or S. brachyurum increased H. columbus populations over treatments with H. columbus alone, both at 60 and 90 d after inoculation. M. incognita suppressed cotton shoot weights significantly (P = 0.05) at both Pi levels. Inoculation with S. brachyurum increased M. incognita populations 60 d after inoculation, while H. columbus suppressed populations of M. incognita. Most larvae of M. incognita did not develop to maturity in the presence of H. columbus. Giant cells aborted and were necrotic 20-25 d after inoculation. Since M. incognita and H. columbus feed on different tissues, the inhibition of M. incognita may have resulted from a physiological effect of H. columbus on the host.     

Relationship Between Morphology and Parasitism in Two Populations of Meloidogyne incognita

The reliability of morphological characters and host differential plants for distinguishing between two populations of Meloidogyne incognita was studied. Population A (originally from North Carolina) had incognita-type perineal patterns. A single egg mass subpopulation of population A had a mixture of incognita and acrita perineal patterns with 33% of the patterns atypical for either species. Population B (from Georgia) had predominantly acrita-type patterns with only about 5% atypical patterns. The head shapes of males from both populations were mainly M. incognita. On the basis of stylet length, both populations conformed to M. incognita acrita. Both populations were identified as M. incognita race 1 by reaction on the North Carolina differential hosts. Reactions on azalea and pepper gave no clear identification of the populations. We concluded that there is no relation between perineal pattern, male head shape, and parasitism of host differentials with the two populations studied.     

Influence of Soil Temperature on Meloidogyne incognita Resistant and Susceptible Cotton,Gossypium hirsutum

The degree of resistance by a cotton plant to Meloidogyne incognita is affected by soil temperature, particularly in moderately resistant cultivars, The total number of nematodes in the resistant and moderately resistant rools at 35 C was equal to, or greater than, the number in susceptible roots at 20, 25, or 30 C. A shift in numbers to developing and egg-bearing forms of nematodes in the susceptible cultivar as tentperature increased indicates development was affected by temperature rather than by genetic resistance mechanisms. However, the nematode resistant cultivar did not support maturation of nematodes until a soil tempurature of 35 C was attained. This indicated that resistance mechanisms are partially repressed at 35 C and differences in nematode development cannot be explained in terms of accumulated heat units. The moderately resistant cultivar was significantly more sensitive to the effects of high temperature than was the resistant cultivar.     

Free Amino Acids in Roots of Infected Cotton Seedlings Resistant and Susceptible to Meloidogyne incognita

Quantities of free amino acids in segments of cotton roots resistant and susceptible to Meloidogyne incognita were compared. Following infection, the root-knot susceptible cultivar, M8, had greater percentage increases of certain individual free amino acids than the resistant cultivar, Clevewilt, but the sum total of free amino acids was greatest in the resistant cultivar. More free amino acids were present in infected than in noninfected plants of both cultivars. The overall concn of glycine declined over the I 0-day period following inoculation. The concns of the aromatic amino acids, tyrosine and phenylalanine, varied as functions of infection, cultivar, and time of harvest. Proline in susceptible M8 increased nearly 2000-fold 10 days after infection, when considerable thickening of syncytial walls is occurring.     

Ultrastructural Cytochemistry of Secretory Granules of Esophageal Glands of Meloidogyne incognita

Ultrastructural cytochemical tests for several enzymes, proteins, carbohydrates, and nucleic acids were conducted on secretory granules o£ dorsal and subventral esophageal glands of preparasitic second-stage juveniles and the dorsal gland of adult females of Meloidogyne incognita. Secretory granules in the subventral glands of juveniles stained positive for acid phosphatase. Peroxidase, DNase, RNase, cellulase, and nucleic acids were not detected in these granules. Secretory granules in the dorsal gland of adult females stained positive for peroxidase (pH 7.6) in < 50% of the tests, Acid phosphatase, β-glucuronidase, DNase, RNase, polyphenoloxidase, cellulase, and carbohydrates were not detected in dorsal gland granules in adult females. Positive staining with cobalt thiocyanate, a stain for amino groups of basic proteins, occurred in secretory granules in the dorsal gland, ribosomes, and chromatin in adult females. Ribosomes, nuclei, and secretory granules of the dorsal gland of adult females intensely stained when incubated in three reagents specific for nucleic acid.     

Retention of Resistance to Meloidogyne incognita in Lycopersicon Genotypes at High Soil Temperature

Lycopersicon glandulosum and L. peruvianum clones and L. esculentum cultivars ''VFN8'' (resistant) and ''Rutgers'' (susceptible) were tested for their resistance to Meloidogyne incognita (race l) at soil temperatures of 25 and 32 C. L. esculentum cv. VFN8 and L. peruvianum Acc. No. 128657, both of which possess the Mi gene, were resistant at 25 C but were susceptible at 32 C. L. glandulosum Acc. No. 126443 and L. peruvianum Acc. No. 270435, with combined resistance to M. hapla and M. incognita, and L. peruvianum Acc. Nos. 129152 and LA2157, with resistance to M. incognita, were highly resistant at both temperatures. In a second experiment three of these accessions under heat stress simulated by 32 C ambient and soil temperature retained a high level of resistance. Two clones of L. glandulosum Acc. No. 126440, with resistance to M. hapla, were moderately susceptible to M. incognita at 25 and highly susceptible at 32 C. M. incognita produced significantly (P = 0.01) more eggs on L. esculentum cv. Rutgers at 32 than at 25 C. This study supports the existence of genes other than the Mi gene that confer resistance to M. incognita and are functional at high soil temperatures.     

Tolerance to Rotylenchulus reniformis and Resistance to Meloidogyne incognita Race 3 in High-Yielding Breeding Lines of Upland Cotton

Field experiments in 1992 and 1994 were conducted to determine the effect of Rotylenchulus reniformis, reniform nematode, on lint yield and fiber quality of 10 experimental breeding lines of cotton (Gossypium hirsutum) in untreated plots or plots fumigated with 1,3-dichloropropene. Controls were La. RN 1032, a germplasm line possessing some resistance to R. reniformis, and Stoneville 453, a cultivar that is susceptible to reniform nematode. Several breeding lines produced greater lint yields than Stoneville 453 or La. RN 1032 in both fumigated and untreated plots. Average lint yield suppression due to R. reniformis for six of the 10 breeding lines was less than half of the 52% yield reduction sustained by Stoneville 453. In growth chamber experiments, R. reniformis multiplication factors for La. RN 1032 and breeding lines N222-1-91, N320-2-91, and N419-1-91 were significantly lower than on Deltapine 16 and Stoneville 453 at 6 weeks after inoculation. R. reniformis populations increased by more than 50-fold on all entries within 10 weeks. In growth chambers, the breeding lines N220-1-92, N222-1-91, and N320-2-91 were resistant to Meloidoglyne incognita race 3; multiplication factors were ≤1.0 at both 6 weeks and 10 weeks after inoculation compared with 25.8 and 26.5 for Deltapine 16 at 6 and 10 weeks after inoculation, respectively, and 9.1 and 2.6 for Stoneville 453. Thus, the results indicate that significant advances have been made in developing improved cotton germplasm lines with the potential to produce higher yields in soils infested with R. reniformis or M. incogaita. In addition to good yield potential, germplasm lines N222-1-91 and N320-2-91 appear to possess low levels of resistance to R. reniformis and a high level of resistance to M. incognita. This germplasm combines high yield potential with significant levels of resistance to both R. reniformis and M. incognita.     

Nematode Temperature Responses: a Niche Dimension In Populations of Bacterial-Feeding Nematodes

The optimum temperatures for population development were determined for six species of bacterial-feeding nematodes from among eight temperatures, ranging from 5 to 40 C. Four of the species are cohabiting species. The range of temperatures over which population development occurs (temperature niche breadth) is different for the cohabiting species. This difference may be a means of reducing competition between species, thus increasing temperatures over which habitats can be exploited.     

Nematicidal Principles from Two Species of Lamiaceae

Aqueous extracts of Ocimum sanctum and O. basilicum leaves contained compounds that killed Meloidogyne incognita larvae in 160 min. Thin layer and gas-liquid chromatography, and infrared spectrophotometry indicated that the essential oils eugenol and linalool were the active nematicidal compounds.     

Relationships Between the Population Density of Meloidogyne incognita and Growth of Tobacco

Seedlings of tobacco cultivars resistant (NC95) and susceptible (McNair 30) to Meloidogyne incognita were grown in 15-cm diameter clay pots containing steamed soil infested with 0, l, 2, 4, 8, 16, 32, and 64 eggs of M. incognita per 1.5 cm³ soil. Plants were maintained in the greenhouse for 3 weeks, and then transferred to the field for 12 weeks. Growth of tobacco was expressed separately as dry weight of leaves and as plant height. Least squares regression analysis showed that tobacco growth-nematode density interactions are in agreement with Seinhorst''s exponential model Y = m + (l-m) czp. Tobacco growth was not affected significantly as nematode density was increased from 0 to tolerance levels, which were approximately 2 and 1 eggs per 1.5 cm³ soil for the resistant and susceptible cultivars, respectively. As nematode density was increased beyond tolerance level, tobacco growth decreased sharply until a minimum yield was approached. The minimum leaf weights and plant heights of the resistant cultivar at the highest nematode density were greater than those of the susceptible cultivar.     

Energetics of Meloidogyne incognita on Resistant and Susceptible Alyceclover Genotypes

To determine the energy cost of a population of Meloidogyne incognita on the roots of alyceclover, nematode biomass was estimated and equations in the literature were used to calculate energy budgets. Amounts of energy consumed, respired, or used in production of nematode biomass were calculated. Results suggested that severe infestations of root-knot nematodes can remove significant quantities of energy from their hosts. Over a 36-day period, a population of 2.6 females of M. incognita per root system removed less than 0.4 calories of energy from a resistant alyceclover plant but over 11 calories were removed by 28 females from a susceptible alyceclover. The calculations indicate that on the resistant alyceclover line, 53% of the energy assimilated by the root-knot population was allocated to respiration, with only 47% allocated to production, whereas on the susceptible line, 65% of the assimilated energy was allocated to production. Such energy demands by the parasite could result in significant reductions in yield quantity and quality at a field production level.     

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.