Meloidogyne incognita and Rotylenchulus reniformis and Associated Soil Textures from Some Cotton Production Areas of Texas.

The incidence of Meloidogyne incognita and Rotylenchulus reniformis on cotton was determined in 1989-92 from 1,089 soil samples collected from 31 counties that account for nearly 60% of the 2.2 million hectares planted to cotton in Texas. Meloidogyne incognita was commonly found in the Southern High Plains and Brazos River Valley regions of Texas (57% and 34%, respectively, of samples) but was found in less than 8% of samples from the Central Blacklands, Coastal Bend, Low Plains, or the Upper Gulf Coast regions. Rotylenchulus reniformis was widely distributed in the Brazos River Valley (24% of samples) and found occasionally in the Upper Gulf Coast (8% of samples). Meloidogyne incognita was found only rarely in soils with greater than 40% clay content, whereas Rotylenchulus reniformis was frequently found in finely textured soils but was less common in soils with greater than 40% sand content. In samples infested with M. incognita or R. reniformis, population densities of these species were at least 10-fold greater than population densities of other plant-parasitic species present in the sample. Root-knot and reniform nematodes were not found together in high population densities (>100 individuals/500 cm(3)) in the same sample.

Counting nematodes with a microplate reader.

The feasibility of counting plant-parasitic nematodes in aqueous suspensions by measuring light transmittance through aqueous suspensions with an ELISA microplate reader was explored. Absorbance readings for eggs or vermiform stages of three species were linearly related (R(2) > 0.99) to concentrations between 0 and 10,000 nematodes/ml. Coefficients of variation ranged from 12-23%, depending on the species and developmental stage used. The method, therefore, was at least as accurate as direct counts of nematodes in aliquots on a microscope and more than 100 times as fast. The method should have direct application in research programs on plant resistance to nematodes, nematode population dynamics, and nematode behavior.

Carbon Dioxide and Temperature Gradielits in Baermann Funnel Extraction of Rotylenchulus reniformis.

Vermiform Rotylenchulus reniformis were anesthetized in water by 10-40% CO but were fully motile for 24 hours in water below 5% CO. When air containing 2.5% CO was blown onto agar, nematodes accumulated at the point of highest CO concentration. Nematodes also accumulated when chilling (0.2-1 C) of agar by the gas flow at the accumulation point was offset with heat from a fiber optic. In Baermann funnels containing R. reniformis in silt loam and sandy clay loam soils, CO in funnel water increased during 24 hours from 0 to ca. 1%; more CO accumulated below the soil layer than above. Bubbling air with 2.5% CO into water below soil in covered funnels increased the CO gradient and increased nematode extraction, whereas bubbling air without CO below soil purged CO from the water and decreased nematode extraction. Manipulation of CO within funnels usually increased extraction by only 30% and never by more than 3-fold. Controlling temperature gradients consistently increased extraction by 2-30-fold.

Survey of Current Distribution of Rotylenchulus reniformis in the United States.

The reniform nematode, Rotylenchulus reniformis, has been reported from all Gulf Coast states, Arkansas, Hawaii, North Carolina, and South Carolina. Experts in 11 states identified the counties or parishes where the nematode is found and provided information regarding associated soil parameters, climate, crops, and crop management. Rotylenchulus reniformis has been detected in 187 counties and parishes of the southeastern United States and is most widespread in Louisiana, Mississippi, Alabama, Florida, and Georgia. In every state except Florida and Hawaii, economically damaging soil populations were associated with continuous cotton production. Other crops considered to be damaged by R. reniformis were soybean, tobacco, several vegetables, and pineapple (Hawaii). There was no consistent relationship between the nematode's presence and soil texture, soil pH, rainfall, or irrigation regime. However, certain respondents associated damage from the nematode primarily with silty or clay soils (Texas, Hawaii, Florida, and Georgia) or with silty soils with exceptionally tow pH (Hawaii and Louisiana).

Influence of Glomus intraradices and Soil Phosphorus on Meloidogyne incognita Infecting Cucumis melo.

The interaction among Glomus intraradices, Meloidogyne incognita, and cantaloupe was studied at three soil phosphorus (P) levels in a greenhouse. All plants grew poorly in soil not amended with P, regardless of mycorrhizal or nematode status. In soil amended with 50 mug P /g soil, M. incognita suppressed the growth of nonmycorrhizal plants by 84%. In contrast, growth of mycorrhizal plants inoculated with M. incognita was retarded by only 21%. A similar trend occurred in plants grown in soil with 100 mug P /g soil. Mycorrhizal infection had no effect on the degree of root-knot gall formation and did not affect the number of nematode eggs per egg mass. Mineral levels in plant shoots generally declined as soil P levels increased and were not significantly influenced by G. intraradices or M. incognita.

Accelerated movement of nematodes from soil in baermann funnels with temperature gradients.

Baermann funnels were modified to eliminate or reverse the small temperature gradient (1-2 C/cm) across the soil layer that normally results from water evaporation. Effects of modifications on extraction efficiency were examined at various ambient temperatures and after overnight adaptation of three nematode species at 20 and 30 C. Extraction of Meloidogyne incognita from sandy loam, Tylenchulus semipenetrans from sandy clay loam, and Rotylenchulus reniformis from silt was greatly accelerated simply by covering funnels to prevent evaporation. In most cases, covering increased the nematodes extracted by 10-100 times after 5.5-48 hours. Faster and more efficient extraction of R. reniformis occurred over a wide range of ambient temperature (18-29 C). Effects of ambient temperature and temperature gradient direction on Baermann funnel extraction of R. reniformis were partly inconsistent with the behavior of R. reniformis in agar. Nematodes in agar moved toward cold at some ambient temperatures and toward heat at other temperatures. They always appeared to move toward cold on Baermann funnels. Differences were not attributable to blockage of gas exchange by covers. In agar and in funnels, the patterns of response to ambient temperature were shifted in the direction of the storage temperature.

Effect of Temperature on Infection and Survival of Rotylenchulus reniformis.

From infestation of lettuce with preinfective females to egg deposition, populations of Rotylenchulus reniformis from Baton Rouge, Louisiana; Lubbock and Weslaco, Texas; and Mayaguez, Puerto Rico, required 41, 13, 7, and 7 days at 15, 20, 25, and 34 C, respectively. No nematode infection occurred at 10 C with any R. reniformis population, and the population from Puerto Rico did not reproduce at 15 C. Nematode survival was not influenced by temperature, since populations from Texas and Louisiana survived for 6 months without a host at - 5 , - 1 , 4, and 25 C. Survival of R. reniformis was substantially influenced by soil moisture. Soil moistures greater than 7% (< 1 bar) aided nematode survival at storage temperature of 25 C, whereas moisture adversely affected nematode survival below freezing. Soil moisture below 4% (> 15 bars) favored nematode survival below freezing but adversely affected nematodes in soils stored at 25 C. Soil moisture effects on nematode survival were less accentuated at 4 and 0 C.

Effects of Soil Solarization on Rotylenchulus reniformis in the Lower Rio Grande Valley of Texas.

Soil solarization was evaluated for control of Rotylenchulus reniformis in the lower Rio Grande Valley of Texas. In field experiments, solarization significantly reduced soil nematode population densities 0-15 cm deep and increased yields of lettuce and cowpea. The length of time required for 90% mortality of nematodes in soil heated under controlled conditions in the laboratory varied from 25 hours to less than 1 hour between 41 and 47 C. Daily exposures of nematode-infested soil to lethal temperatures for sublethal time periods had a cumulative lethal effect. In water, vermiform stages required up to 10 days to recover from sublethal thermal stress. Eggs were similar to juveniles in their sensitivity to high temperatures. Lethal time-temperatures under controlled conditions were in general agreement with field results.

Geographical Distributions of Rotylenchulus reniformis, Meloidogyne incognita, and Tylenchulus semipenetrans in the Lower Rio Grande Valley as Related to Soil Texture and Land Use.

A survey was conducted over a 22-year period to evaluate the influence of soil texture and land use on the geographical distributions of Rotylenchulus reniformis, Meloidogyne incognita, and Tylenchulus semipenetrans in the lower Rio Grande valley. The distributions of R. reniformis and M. incognita were related to soil texture, whereas T. semipenetrans occurred wherever host plants were present regardless of soil texture. The incidence of M. incognita was greatest in elevated sandy loams and moderately well-drained silts of modern flood terraces of the Rio Grande river. Rotylenchulus reniformis occurred predominantly in clay silts and clays of ancient flood terraces. Clay loams and sandy clay loams of the central, irrigated portion of the lower Rio Grande valley appeared favorable for M. incognita and R. reniformis. Differences between the geographical distributions of these two species could not be attributed to host crops.

Histopathology of Okra and Ridgeseed Spurge Infected with Meloidodera charis.

Histological observations of okra Abelomoschus esculentus 'Clemson Spineless' and ridgeseed spurge Euphorbia glyptosperma (a common weed) infected with Meloidodera charis Hopper, indicated that the juvenile nematode penetrated the roots intercellularly. Within 5 days after plant emergence the nematode positioned its body in the cortical tissue parallel to the vascular system. By 10 days after plant emergence the juvenile had extended its head into the vascular system and initiated giant cell formation, generally in protophloem tissue. Giant cells were one celled and usually multi-nucleate. Eggs were observed in the female body 30 days after plants emerged and juveniles were found within the female body by 40 days. Nematode development progressed equally in the root system of either host plant. Generally, throughout the nematode's life cycle its entire body remained inside the cortical tissue of okra. In ridgeseed spurge, however, the posterior portion of the female erupted through the host epidermis as early as 15 days after plant emergence; only the head and neck remained embedded in the host. The nematode caused extensive tissue disruption in the cortical and vascular system of both plant species. Corn, Zea mays, was another host of the nematode.

Pathogenicity and Histopathology of Rotylenchulus reniformis Infecting Cantaloup.

Rotylenchulus reniformis was pathogenic to cantaloup (Cucumis melo 'Perlita') under greenhouse conditions. These findings confirm field symptoms of cantaloup infected with R. reniformis. Histopathological studies show that the nematode penetrates the cortex perpendicular to the vascular system and comes to rest with the head against the endodermis in young roots. Feeding stimulated the pericycle to either side of the endodermal feeding cell and caused cell hypertrophy with enlargement of the nucleoli and granular thickening of the cytoplasm. In older roots where the endodermis had collapsed, the nematode fed directly into the pericycle and caused similar symptoms. Nematode development was more rapid at 27 C than at 21 C.

Factors Affecting the Control of Rotylenchulus reniformis with Electromagnetic Energy.

The reniform nematode Rotylenchulus reniformis was reduced in the upper 10 cm of soil with application of UHF electromagnetic energy. Bioassay of treated soil indicated no delayed effect on the population from the treatment. The population was significantly reduced by hot water treatments at 40 C for 10 min, and at 45 C for 5 and 10 min, 50 C and above killed all nematodes. Data were inconclusive as to whether the effect of UHF electromagnetic energy was thermal or nonthermal.

Effect of RotyIenchulus reniformis on reflectance of cotton plant leaves.

Differences between light reflectance from leaves of cotton (Gossypiurn hirsutum) plants grown with a low- or no-nematode (Rotylenchulus reniformis) population (nonstressed), and from leaves grown with a high nematode population (stressed) were measured in field and greenhouse experiments. Reflectance was measured spectrophotometrically in the laboratory on single leaves and spectroradiometrically in the field on plant canopies. Nematode-stressed cotton plants were stunted with fewer, smaller, and darker-green leaves than nonstressed plants. Over the 0.5- to 2.5-/microm waveband, stressed leaves had lower reflectance than nonstressed leaves of the same chronological age for both field- and greenhouse-grown plants. Reflectance differences between stressed and nonstressed leaves in the visible (0.5 to 0.75 microm), near-infrared (0.75 to 1.35 mum) and infrared water absorption (1.35 to 2.5 microm) regions were primarily caused by differences in leaf chlorophyll concentration, mesophyll structure, and water content, respectively. Results indicate the potential for remotely sensing nematode-infested plants to distinguish them from normal plants.

Interaction of Rotylenchulus reniformis, Soil Salinity, and Cotton.

Rotylenchulus reni]ormis occurred equally in relatively non-saline (4.0 mmhos/cm) and highly-saline (16.5 mmhos/cm) soils in sampling transects across zones of depressed plant growth in six Texas cotton fields.Results from greenhouse pot experiments indicated progressive positive interaction of salinity and R. reni[ormis pathogenicity in the range 6-18 mmhos/cm.

Pathogenicity and Histopathology of Meloidogyne graminis Infecting 'Tifdwarf' Bermudagrass Roots.

In a greenhouse experiment Meloidogyne graminis was pathogenic to 'Tifdwarf' bermudagrass, causing significant reduction in plant weight. Roots and tops of inoculated grass weighed 28.4% less than non-inoculated grass 8 months after inoculation. Clipping weight of nematode-infected turf weighed 68.9% less than clippings from non-infected turf. Histopathological studies showed that the head of the female nematode penetrated the vascular system and resulted in giant cell formation in the feeding area. The nematode body remained in the cortex parallel to the vascular system. Eggs were deposited at the posterior of the nematode in a gelatinous matrix in the cortex. M. graminis fed with its anterior end oriented toward the growing root tip. M. incognita had no set body orientation pattern when feeding on bermudagrass.