Vertical Distribution of Rotylenchulus reniformis in Cotton Fields

The possible impact of Rotylenchulus reniformis below plow depth was evaluated by measuring the vertical distribution of R. reniformis and soil texture in 20 symptomatic fields on 17 farms across six states. The mean nematode population density per field, 0 to 122 cm deep, ranged from 0.4 to 63 nematodes/g soil, and in 15 fields more than half of the R. reniformis present were below 30.5 cm, which is the greatest depth usually plowed by farmers or sampled by consultants. In 11 fields measured, root density was greatest in the top 15 cm of soil; however, roots consistently penetrated 92 to 122 cm deep by midseason, and in five fields in Texas and Louisiana the ratio of nematodes to root-length density within soil increased with depth. Repeated sampling during the year in Texas indicated that up to 20% of the nematodes in soil below 60 cm in the fall survived the winter. Differences between Baermann funnel and sugar flotation extraction methods were not important when compared with field-to-field differences in nematode populations and field-specific vertical distribution patterns. The results support the interpretation that R. reniformis below plow depth can significantly impact diagnosis and treatment of cotton fields infested with R. reniformis.     

Histological Observations of Rotylenchulus reniformis on Gossypium longicalyx and Interspecific Cotton Hybrids

Observations on the development of reniform nematode (Rotylenchulus reniformis) on roots of Gossypium longicalyx, G. hirsutum, and two interspecific hybrids derived from them were made by light microscopy. Gossypium longicalyx is reported to be immune to reniform nematode, but the mechanism(s) for resistance are unknown. Penetration of G. longicalyx roots by female nematodes was confirmed, and incipient swelling of the females, indicating initiation of maturation of the reproductive system, was observed. Female maturation occurred up to the formation of a single embryo inside the female body but not beyond this point. In both hybrids, development was inhibited but progressed further than in the immune parent. Reactions ranged from highly compatible, with the formation of active syncytia and full development of females, to incompatible with little or no development of the female. Compatible plants showed characteristic hypertrophied cells, enlarged nuclei, dense cytoplasm, and partial dissolution of cell walls, whereas incompatible plant reactions included lignification of the cells adjacent to the nematode head, or the complete collapse and necrosis of the cells involved. The need to characterize reactions and to carefully select among the plants descended from the hybrids during the introgression process, as well as the importance of combining the results of reproduction tests with histological observation of the plant-nematode interactions, is discussed.     

Suppression of Rotylenchulus reniformis on Cotton by the Nematophagous Fungus ARF

The reniform nematode, Rotylenchulus reniformis Linford &Oliveira, has become a serious threat to cotton (Gossypium hirsutum L.) production in the United States during the past decade. The objective of this study is to isolate fungi from eggs of R. reniformis and select potential biological control agents for R. reniformis on cotton. Soil samples were collected from cotton fields located in Jefferson County, Arkansas. Eight genera of fungi were included in the 128 fungal isolates obtained, and among them were five strains of the nematophagous fungus ARF. The mtDNA RFLP pattern, colony growth characteristics, and pathogenicity indicate the five ARF isolates represent one described strain and one new strain. Light and electron microscopic observations suggest ARF is an active parasite of R. reniformis, with parasitism ranging from 48% to 79% in in vitro tests. Three greenhouse experiments demonstrated ARF successfully suppressed the number of reniform nematodes during the first and second generation of the nematode. Reductions in numbers of R. reniformis on the roots for the seven application rates of 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, and 0.5% ARF were 87%, 92%, 94%, 96%, 97%, 98%, and and 98%, respectively.     

Impact of Rotylenchulus reniformis on Cotton Yield as Affected by Soil Texture and Irrigation

The effects of soil type, irrigation, and population density of Rotylenchulus reniformis on cotton were evaluated in a two-year microplot experiment. Six soil types, Fuquay sand, Norfolk sandy loam, Portsmouth loamy sand, Muck, Cecil sandy loam, and Cecil sandy clay, were arranged in randomized complete blocks with five replications. Each block had numerous plots previously inoculated with R. reniformis and two or more noninoculated microplots per soil type, one half of which were irrigated in each replicate for a total of 240 plots. Greatest cotton lint yields were achieved in the Muck, Norfolk sandy loam, and Portsmouth loamy sand soils. Cotton yield in the Portsmouth loamy sand did not differ from the Muck soil which averaged the greatest lint yield per plot of all soil types. Cotton yield was negatively related to R. reniformis PI (initial population density) in all soil types except for the Cecil sandy clay which had the highest clay content. Supplemental irrigation increased yields in the higher yielding Muck, Norfolk sandy loam, and Portsmouth loamy sand soils compared to the lower yielding Cecil sandy clay, Cecil sandy loam, and Fuquay sand soils. The Portsmouth sandy loam was among the highest yielding soils, and also supported the greatest R. reniformis population density. Cotton lint yield was affected more by R. reniformis Pi with irrigation in the Portsmouth loamy sand soil with a greater influence of Pi on lint yield in irrigated plots than other soils. A significant first degree PI × irrigation interaction for this soil type confirms this observation.     

Interrelationships of Rotylenchulus reniformis with Rhizoctonia solani on Cotton

The interrelationships between reniform nematode (Rotylenchulus reniformis) and the cotton (Gossypium hirsutum) seedling blight fungus (Rhizoctonia solani) were studied using three isolates of R. solani, two populations of R. reniformis at multiple inoculum levels, and the cotton cultivars Dehapine 90 (DP 90) and Dehapine 41 (DP 41). Colonization of cotton hypocotyl tissue by R. solani resulted in increases (P ≤ 0.05) in nematode population densities in soil and in eggs recovered from the root systems in both 40- and 90-day-duration experiments. Increases in soil population densities resulted mainly from increases in juveniles. Enhanced reproduction of R. reniformis in the presence of R. solani was consistent across isolates (1, 2, and 3) of R. solani and populations (1 and 2) and inoculum levels (0.5, 2, 4, and 8 individuals/g of soil) of R. reniformis, regardless of cotton cultivar (DP 90 or DP 41). Severity of seedling blight was not influenced by the nematode. Rhizoctonia solani caused reductions (P ≤ 0.05) in cotton growth in 40- and 90-day periods. Rotylenchulus reniformis reduced cotton growth at 90 days. The relationship between nematode inoculum levels and plant growth reductions was linear. At 90 days, the combined effects of these pathogens were antagonistic to plant growth.     

Rotylenchulus reniformis below Plow Depth Suppresses Cotton Yield and Root Growth

Damage to cotton by Rotylenchulus reniformis below plow depth was evaluated in a sandy clay loam soil at Weslaco, Texas. In December 1999, 14 holes on 51-cm centers were dug 91 cm deep along the planting bed and adjacent furrow and 2 ml of 1,3-dichloropropene was placed 91, 61, and 30 cm deep as each hole was refilled and packed. This technique eliminated 96%, 81%, and 74% of R. reniformis down to 107 cm at distances 0, 25, and 51 cm laterally from the point of application (P ≤ 0.05), whereas chisel fumigation at 168 liters/ha 43 cm deep reduced nematode numbers only in the top 61 cm (P ≤ 0.001). Manual placement of fumigant increased yield 92%; chisel fumigation increased yield 88% (P ≤ 0.005). A second experiment in February 2001 placed fumigant 43 or 81 cm deep, or at both 43 and 81 cm. Holes alone had no significant effect on nematode density at planting, midseason or harvest, on root length density at midseason, or on cotton lint yield. Fumigant at 43 cm reduced nematode numbers above fumigant application depth at planting 94% (P ≤ 0.02), at midseason 37% (P ≤ 0.09), and at harvest 0%, increasing yield 57% (P ≤ 0.002). Fumigant at 81 cm reduced nematode numbers above fumigant application depth at planting 86% (P ≤ 0.02), at midseason 74% (P ≤ 0.02), and at harvest 48% (P ≤ 0.01), increasing yield 53% (P ≤ 0.002). Fumigating at both 43 and 81 cm reduced nematode numbers above 90 cm 94% at planting and 79% at midseason, increased midseason root-length density 14-fold below 76 cm, and doubled yield (P ≤ 0.02 in all cases).     

Postinfection Development of Rotylenchulus reniformis on Resistant Gossypium barbadense Accessions

The reniform nematode (Rotylenchulus reniformis) causes significant cotton (Gossypium hirsutum) losses in the southeastern United States. The research objective was to describe the effects of two resistant G. barbadense lines (cultivar TX 110 and accession GB 713) on development and fecundity of reniform nematode. Nematode development and fecundity were evaluated on the resistant lines and susceptible G. hirsutum cultivar Deltapine 16 in three repeated growth chamber experiments. Nematode development on roots early and late in the infection cycle was measured at set intervals from 1 to 25 d after inoculation (DAI) and genotypes were compared based on the number of nematodes in four developmental stages (vermiform, swelling, reniform, and gravid). At 15, 20, and 25 DAI, egg production by individual females parasitizing each genotype was measured. Unique reniform nematode developmental patterns were noted on each of the cotton genotypes. During the early stages of infection, infection and development occurred 1 d faster on susceptible cotton than on the resistant genotypes. Later, progression to the reniform and gravid stages of development occurred first on the susceptible genotype, followed by G. barbadense cultivar TX 110, and finally G. barbadense accession GB 713. Egg production by individual nematodes infecting the three genotypes was similar. This study corroborates delayed development previously reported on G. barbadense cultivar TX 110 and is the first report of delayed infection and development associated with G. barbadense accession GB 713. The different developmental patterns in the resistant genotypes suggest that unique or additional loci may confer resistance in these two lines.     

Comparisons of Female and Egg Assays to Identify Rotylenchulus reniformis Resistance in Cotton

More plants can be screened for reniform nematode resistance each year if the time involved can be shortened. In this study, the hypothesis that female counts are as efficient as egg counts in identifying resistant genotypes was tested. In two greenhouse experiments Gossypium genotypes which varied from resistant to susceptible to reniform nematode (Rotylenchulus reniformis) were compared to a susceptible control cultivar. Infested field soil served as the inoculum source for the first experiment, and vermiform stages extracted from greenhouse cultures were used to infest soil in the second experiment. Six replicates of each genotype were harvested 25 d after planting and swollen females were counted. The remaining plants were harvested 35 d after planting and eggs extracted from the roots were counted. Processing and counting times recorded in the first experiment were similar for both assessment methods, but 10 additional days were required for egg-based assessment. Contrast analyses showed that assessments based on females per gram of root were equivalent to assessments based on eggs per gram of root for the five genotypes tested in the first experiment and for an expanded set of 13 genotypes tested in the second experiment. The results indicated that either life stage can be used to screen for resistance.     

Rotylenchulus reniformis Management in Cotton with Crop Rotation

One-year crop rotations with corn or highly resistant soybean were evaluated at four locations for their effect on Rotylenchulus reniformis population levels and yield of a subsequent cotton crop. Four nematicide (aldicarb) regimes were included at two of the locations, and rotation with reniform-susceptible soybean was included at the other two locations. One-year rotations to corn or resistant soybean resulted in lower R. reniformis population levels (P ≤ 0.05) than those found in cotton at three test sites. However, the effect of rotation on nematode populations was undetectable by mid-season when cotton was grown the following year. Cotton yield following a one-year rotation to resistant soybean increased at all test locations compared to continuous cotton, and yield following corn increased at three locations. The optimum application rate for aldicarb in this study was 0.84 kg a.i./ha in furrow. Side-dress applications of aldicarb resulted in yield increases that were insufficient to cover the cost of application in 3 of the 4 years.     

Impact of Soil Texture on the Reproductive and Damage Potentials of Rotylenchulus reniformis and Meloidogyne incognita on Cotton

The effects of soil type and initial inoculum density (Pi) on the reproductive and damage potentials of Meloidogyne incognita and Rotylenchulus reniformis on cotton were evaluated in microplot experiments from 1991 to 1993. The equilibrium nematode population density for R. reniformis on cotton was much greater than that of M. incognita, indicating that cotton is a better host for R. reniformis than M. incognita. Reproduction of M. incognita was greater in coarse-textured soils than in fine-textured soils, whereas R. reniformis reproduction was greatest in a Portsmouth loamy sand with intermediate percentages of clay plus silt. Population densities of M. incognita were inversely related to the percentage of silt and clay, but R. reniformis was favored by moderate levels of clay plus silt (ca. 28%). Both M. incognita races 3 and 4 and R. reniformis effected suppression of seed-cotton yield in all soil types evaluated. Cotton-yield suppression was greatest in response to R. reniformis at high Pi. Cotton maturity, measured as percentage of open bolls at different dates, was affected by the presence of nematodes in all 3 years.     

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.     

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.     

Identification of Rotylenchulus reniformis Resistant Glycine Lines

Identification of resistance to reniform nematode (Rotylenchulus reniformis) is the first step in developing resistant soybean (Glycine max) cultivars that will benefit growers in the mid-South region of the United States. This study was conducted to identify soybean (G. max and G. soja) lines with resistance to this pathogen. Sixty-one wild and domestic soybean lines were evaluated in replicated growth chamber tests. Six previously untested soybean lines with useful levels of resistance to reniform nematode were identified in both initial screening and subsequent confirmation tests: released germplasm lines DS4-SCN05 (PI 656647) and DS-880 (PI 659348); accession PI 567516 C; and breeding lines DS97-84-1, 02011-126-1-1-2-1 and 02011-126-1-1-5-1. Eleven previously untested moderately susceptible or susceptible lines were also identified: released germplasm lines D68-0099 (PI 573285) and LG01-5087-5; accessions PI 200538, PI 416937, PI 423941, PI 437697, PI 467312, PI 468916, PI 594692, and PI 603751 A; and cultivar Stafford (PI 508269). Results of previously tested lines evaluated in the current study agreed with published reports 69.6% of the time for resistant lines and 87.5% of the time for susceptible lines. Soybean breeders may benefit from incorporating the newly identified resistant lines into their breeding programs.     

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.     

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.     

Influence of Rhizoctonia solani on Egg Hatching and Infectivity of Rotylenchulus reniformis

The effects of culture filtrates of Rhizoctonia solani and root exudates of R. solani-infected cotton (Gossypium hirsutum) seedlings on hatching of eggs and infectivity of females of Rotylenchulus reniformis were evaluated in an attempt to account for the enhanced nematode reproduction observed in the presence of this fungus. Crude filtrates of R. solani cultures growing over sterile, deionized distilled water did not affect egg hatching. Exudates from roots of cotton seedlings increased hatching of R. reniformis eggs over that observed in water controls. Exudates from cotton seedling roots not infected or infected with R. solani did not differ in their effect on egg hatching. However, infection of cotton seedlings by reniform females was increased in the presence of R. solani, resulting in the augmented egg production and juvenile population densities in soil observed in greenhouse studies.     

Natural Migration of Rotylenchulus reniformis In a No-Till Cotton System

Rotylenchulus reniformis is the most damaging nematode pathogen of cotton in Alabama. It is easily introduced into cotton fields via contaminated equipment and, when present, is difficult and costly to control. A trial to monitor the natural migration of R. reniformis from an initial point of origin was established in 2007 and studied over two growing seasons in both irrigated and non-irrigated no-till cotton production systems. Vermiform females, juveniles and males reached a horizontal distance of 200 cm from the initial inoculation point, and a depth of 91 cm in the first season in both systems. Irrigation had no effect on the migration of vermiform females and juveniles, but males migrated faster in the irrigated trial than in the non-irrigated trial. Population density increased steadily in the irrigated trial during both years, exceeding the economic threshold of 1,000 per 150 cm(3), but was highly correlated with rainfall in the non-irrigated trial. The average speed of migration ranged from 0- to 3.3-cm per day over 150 days. R. reniformis was able to establish in both the irrigated and non-irrigated trials in one season and to increase population density significantly.     

Effect of Temperature on the Embryogenesis of Geographic Populations of Rotylenchulus reniformis

The effect of temperature on the embryonic development of three populations of reniform nematode (Rotylenchulus reniformis) from the southeastern United States was studied. The development of eggs from single-cell stage to eclosion of second-stage juvenile was monitored at 20, 25, 30, and 35°C. All populations completed embryogenesis in 7 days at 25°C. The greatest differences among populations in time to completion of embryogenesis were observed at 20 and 35°C. Results at the intermediate temperatures (25 and 30°C) were similar for the three populations. The optimal temperature for embryogenesis was calculated to be 31.4°C for the population from Alabama, 28.4°C for the one from Mississippi, and 37.5°C for the one from South Carolina.     

Suppression of Rotylenchulus reniformis 122-cm Deep Endorses Resistance Introgression in Gossypium

Nine sources of resistance to Rotylenchulus reniformis in Gossypium (cotton) were tested by measuring population density (Pf) and root-length density 0 to 122 cm deep. A Pf in the plow layer less than the autumn sample treatment threshold used by consultants was considered the minimum criterion for acceptable resistance, regardless of population density at planting (Pi). Other criteria were ample roots and a Pf lower than on the susceptible control, as in pot studies. In a Texas field in 2001 and 2002, no resistant accessions had Pf less than the control but all did in microplots into which nematodes from Louisiana were introduced. An environmental chamber experiment ruled out nematode genetic variance and implicated unknown soil factors. Pf in field experiments in Louisiana, Mississippi, and Alabama were below threshold for zero, six and four of the accessions and above threshold in the control. Gossypium arboreum A2–87 and G. barbadense GB-713 were the most resistant accessions. Results indicate that cultivars developed from these sources will suppress R. reniformis populations but less than in pots in a single season.     

Classification of Rotylenchulus reniformis Numbers in Cotton Using Remotely Sensed Hyperspectral Data on Self-Organizing Maps

Rotylenchulus reniformis is one of the major nematode pests capable of reducing cotton yields by more than 60%, causing estimated losses that may exceed millions of dollars U.S. Therefore, early detection of nematode numbers is necessary to reduce these losses. This study investigates the feasibility of using remotely sensed hyperspectral data (reflectances) of cotton plants affected with different nematode population numbers with self-organizing maps (SOM) in correlating and classifying nematode population numbers extant in a plant's rhizosphere. The hyperspectral reflectances were classified into three classes based on R. renifomis population numbers present in plant's rhizosphere. Hyperspectral data (350-2500 nm) were also sub-divided into Visible, Red Edge + Near Infrared (NIR) and Mid-IR region to determine the sub-region most effective in spectrally classifying the nematode population numbers. Various combinations of different feature extraction and dimensionality reduction methods were applied in different regions to extract reduced sets of features. These features were then classified using a supervised-SOM classification method. Our results suggest that the overall classification accuracies, in general, for most methods in most regions (except visible region) varied from 60% to 80%, thereby, indicating a positive correlation between the nematode numbers present in plant's rhizosphere and the corresponding plant's hyperspectral signatures. Results showed that classification accuracies in the Mid-IR region were comparable to the accuracies obtained in other sub-regions. Finally, based on our findings, the use of remotely-sensed hyperspectral data with SOM could prove to be extremely time efficient in detecting nematode numbers present in the soil.