Effects of Tobacco Cyst Nematode on Growth of Flue-cured Tobacco

The effects of infection by tobacco cyst nematode (Globodera tabacum solanacearum) on growth of flue-cured tobacco cultivars NC 567 (resistant) and K 326 (susceptible) were evaluated in the field in 1993 and 1994. Infection by G. t. solanacearum suppressed number of leaves, plant height, and fresh weight of leaves and feeder roots. Correlations between weekly egg densities of G. t. solanacearum collected from soil and host growth during 11 weeks after transplanting (WAT) were often inconsistent between cultivars and years. However, consistent correlations were obtained between root weight and egg densities collected 9 WAT, as well as between leaf weight from susceptible K 326 and nematode egg densities 6 WAT. Leaf and feeder root weights were significantly correlated with the area under the curve for all nematodes per gram of feeder root for K 326 in 1993 and for both cultivars in 1994. Reduction in feeder root weight by G. t. solanacearum was similar for the resistant and susceptible cultivars. Reduction in fresh leaf weight by G. t. solanacearum was twice as great (P ≤ 0.07) for K 326 as for NC 567 in 1994. Incorporating nematode resistance into germplasm possessing improved yield and quality traits should produce cultivars more acceptable to growers.     

Genetics of Burley and Flue-Cured Tobacco Resistance to Globodera tabacum tabacum

Genotypes of burley (cultivars B-21 and B-49), flue-cured (line VA-81 and cultivar PD-4), and Connecticut broadleaf (cultivar C9) tobacco (Nicotiana tabacum) resistant (R) or susceptible (S) to the tobacco cyst nematode Globodera tabacum tabacum were crossed. F1 progeny of burley and susceptible broadleaf were selfed and backcrossed to produce additional progeny for evaluation of resistance in greenhouse experiments. Plants without adult female nematodes visible (×10 magnification) on the root surface 6 weeks after inoculation were classified as resistant, whereas those plants in which one or more females were evident were classified as susceptible. Segregation ratios for progeny of resistant and susceptible plants were not different from 3:1 and 1:1 for F2 (F1 × F1) and BC1 (F1 × S) lines, respectively, indicating that resistance in burley to G. t. tabacum is conferred by a single, dominant gene. Segregation ratios for resistance in crosses between nematode-resistant burley and flue-cured tobacco (F1 and F2 progeny) and between burley-flue-cured hybrids and broadleaf BC1 (F1 × S) and BC2 (BC1 × S) progeny were consistent with the assumption that resistance to G. t. tabacum in burley and flue-cured tobacco is conferred by the same or closely linked single, dominant gene(s).     

Enhanced Hatching of Globodera tabacum solanacearum Juveniles by Root Exudates of Flue-cured Tobacco

Stimulation of hatching of a tobacco cyst nematode (Globodera tabacum solanacearum) by root exudates from resistant NC 567 and susceptible K 326 cultivars of flue-cured tobacco, Nicotiana tabacum, was investigated. Root exudates were collected by soaking seedlings in deionized water for 2 hours at 22 °C in the dark. Fifteen mature and uniformly sized cysts were exposed at 15, 20, or 25 °C to undiluted root exudate, root exudate diluted 1:1 or 1:3 with deionized water, or deionized water alone. Hatched juveniles were counted and removed at weekly intervals during 42 and 53 days of exposure in experiments conducted in 1994 and 1995, respectively. Root exudates from both susceptible cultivar K 326 and resistant cultivar NC 567 stimulated more hatching than deionized water at 25 °C in 1994, and at all three tested temperatures in 1995. In 1994, dilution of root exudates 1:3 reduced stimulation of hatching at 25 °C compared to undiluted exudate. Hatching at 25 °C was similarly stimulated by exposure to undiluted root exudate and exudate diluted 1:1. In 1995, both dilutions reduced stimulation of hatching by root exudates at all the temperatures.     

Identification of Sources of Resistance to Four Species of Root-knot Nematodes in Tobacco

Resistance to the southern root-knot nematode, Meloidogyne incognita races 1 and 3, has been identified, incorporated, and deployed into commercial cultivars of tobacco, Nicotiana tabacum. Cultivars with resistance to other economically important root-knot nematode species attacking tobacco, M. arenaria, M. hapla, M. javanica, and other host-specific races of M. incognita, are not available in the United States. Twenty-eight tobacco genotypes of diverse origin and two standard cultivars, NC 2326 (susceptible) and Speight G 28 (resistant to M. incognita races 1 and 3), were screened for resistance to eight root-knot nematode populations of North Carolina origin. Based on root gall indices at 8 to 12 weeks after inoculation, all genotypes except NC 2326 and Okinawa were resistant to M. arenaria race 1, and races 1 and 3 of M. incognita. Except for slight root galling, genotypes resistant to M. arenaria race 1 responded similarly to races 1 and 3 of M. incognita. All genotypes except NC 2326, Okinawa, and Speight G 28 showed resistance to M. javanica. Okinawa, while supporting lower reproduction of M. javanica than NC 2326, was rated as moderately susceptible. Tobacco breeding lines 81-R-617A, 81-RL- 2K, SA 1213, SA 1214, SA 1223, and SA 1224 were resistant to M. arenaria race 2, and thus may be used as sources of resistance to this pathogen. No resistance to M. hapla and only moderate resistance to races 2 and 4 of M. incognita were found in any of the tobacco genotypes. Under natural field infestations of M. arenaria race 2, nematode development on resistant tobacco breeding lines 81-RL-2K, SA 1214, and SA 1215 was similar to a susceptible cultivar with some nematicide treatments; however, quantity and quality of yield were inferior compared to K 326 plus nematicides.     

Control of Globodera tabacum solanacearum by Alternating Host Resistance and Nematicide

The feasibility of alternating use of resistant vs. susceptible flue-cured tobacco cultivars to improve control of Globodera tabacum subsp, solanacearum (TCN) was investigated at two Virginia locations in 1984-86. Post-harvest TCN population densities were reduced in each year of the study when fenamiphos was used with a TCN-resistant cultivar (NC 567), relative to susceptible cultivars (K 326 or Mc 944). Using NC 567 with fenamipbos also reduced preplant TCN population densities in the next growing season. Egg population densities before planting in 1986 were significantly lower in plots planted with NC 567 in 1984, even when a susceptible cultivar had been planted in 1985. Use of fenamiphos with NC 567 in 1984 and 1985 further reduced preplant egg population densities in 1986. Economic returns were significantly greater in 1984 when NC 567 was used with fenamiphos, rather than a susceptible cultivar. Treatments involving fenamiphos and (or) NC 567 in 1984 and 1985 resulted in higher economic returns in 1986 than did treatments using a susceptible cultivar without fenamiphos in both previous years. Economic returns were highest in 1986 when fenamiphos and NC 567 were used in 1984 and 1985 and a susceptible cultivar was planted in 1986.     

Effects of Continuous Cropping of Resistant and Susceptible Cultivars on Reproduction Potentials of Heterodera glycines and Globodera tabacum solanacearum

The reproductive potentials of Heterodera glycines (mixture of races 3 and 4 and unidentified races) and a tobacco cyst nematode Globodera tabacum solanacearum were studied in the field. The experiments involved four cultivars of soybean Glycine max and four cultivars of Nicotiana tabacum. The reproductive potential of the H. glycines population was high on Essex and Lee 74 soybean but low on Forrest and Bedford over the 3 years (1982-84) of continuous cropping. The reproductive potential of H. glycines was 12% on Forrest and 6% on Bedford in 1982 but increased to 37 and 35% in 1983 and to 71 and 41% in 1984, respectively, on these two cultivars. The reproductive potential of G. tabacum solanacearum was high on McNair 944 and Coker 319 tobacco cultivars and low on VA 81 and PD 4 over the 3 years of cropping. The reproductive potential of G. tabacum solanacearum on VA 81 and PD 4 was 18 and 17% in 1982, 7 and 16% in 1983, and 5 and 5% in 1984, respectively. The changes in reproductive potentials of H. glycines and G. tabacum solanacearum may be related to inherent genetic variability in the systems that control reproduction of the two cyst nematodes and nature of resistance incorporated in the soybean and tobacco cultivars.     

Intraspecific Variability within Globodera tabacum solanacearum Using Random Amplified Polymorphic DNA

Random amplified polymorphic DNA (RAPDs) were used to investigate the intraspecific variability among 19 geographic isolates of Globodera tabacum solanacearum from eight counties in Virginia and one county in North Carolina. Globodera tabacum tabacum, G. t. virginiae, and the Mexican cyst nematode (MCN) were included as outgroups. Six primers were used and 119 amplification products were observed. Each primer yielded reproducible differences in fragment patterns that differentiated the isolates and species. Hierarchical cluster analysis was performed to illustrate the relatedness among isolates and species. The average Jaccard''s similarity index among isolates of G. t. solanacearum was 74%, possibly representing greater variation than that reported in the literature across different pathotypes of the potato cyst nematode, Globodera pallida, in studies where RAPD were also employed. The RAPD markers described here may be useful for the development of specific primers or probes that could improve the identification of TCN populations. Such improvements in the characterization of TCN genotypes would facilitate the effective deployment of existing and future resistant cultivars to control these economically important pests.     

Variability among Populations of Meloidogyne arenaria

Variability in reproduction and pathogenicity of 12 populations of Meloidogyne arenaria race 1 was evaluated on Florunner peanut, Centennial soybean, Rutgers tomato, G70, K326, and Mc944 tobacco, and Carolina Cayenne, Mississippi Nemaheart, and Santanka pepper. Differences among M. arenaria populations in rates of egg production 45 days after inoculation were observed for all cultivars except Santanka pepper. Differences among populations in dry top weights or fresh root weights were recorded on all cultivars. Numbers of nematode eggs produced on Florunner peanut varied from 3,419 to 11,593/g fresh root weight. On resistant tobacco cultivars (G70 and K326), one nematode population produced high numbers of eggs (12,042 and 6,499/g fresh root weight on G70 and K326, respectively), whereas the other populations produced low numbers of eggs (less than 500 eggs/g fresh root weight on both cultivars). Two variant M. arenaria race 1 populations were identified by factor analysis of reproductive rates on all nine cultivars. Differences m reproduction and pathogenicity observed among populations would affect the design of sustainable management systems for M. arenaria.     

Trap Crops and Population Management of Globodera tabacum tabacum

Tobacco, eastern black nightshade, and tomato were grown for 3 to 13 weeks to assess differences in invasion, development, and soil density of Globodera tabacum tabacum (tobacco cyst nematode) in field plots and microplots over three seasons. Tobacco cyst nematodes invaded roots of resistant and susceptible tobacco, nightshade, and tomato. Nematode development was fastest in nightshade and slowest in tomato, and few adults developed in roots of nematode-resistant tobacco. Soil populations of tobacco cyst nematodes were reduced up to 80% by destroying nightshade or susceptible tobacco grown for 3 to 6 weeks. Nematode populations were reduced up to 96% by destroying tomato or resistant tobacco grown for 3 to 6 weeks. Timing of crop destruction was less critical with tomato and resistant tobacco, as nematode populations did not increase after 13 weeks of growth. These studies demonstrate that trap cropping, through crop destruction, can significantly reduce G. t. tabacum populations.     

Temporal Efficacy of Selected Nematicides on Meloidogyne Species on Tobacco

Aldicarb, ethoprop, and fenamiphos were evaluated for their efficacy in controlling various species of root-knot nematodes on flue-cured tobacco and for their residual activity, as determined through periodic sampling and bioassays of soil taken from field plots. Field experiments were conducted at five locations over 2 years with flue-cured tobacco. Soil in plots treated with nematicides were formed into high, wide beds before transplanting with ''Coker 371-Gold'' or ''K 326'' tobacco. Residual control of Meloidogyne spp. was greatest (P ≤ 0.05) with fenamiphos (in some cases up to 10 weeks, as measured in tomato bioassays of infested soil and root fragments). Suppression of nematode reproduction by ethoprop was short-lived, and numbers of second-stage juveniles + eggs and numbers of galls in bioassays sometimes surpassed those of untreated plots within 4 weeks after treatment. Aldicarb gave intermediate control over time as compared to the other compounds. Although nematicidal efficacy of all compounds varied with site and season, fenamiphos and aldicarb generally produced the highest yields.     

Broadleaf Tobacco Yield Loss in Relation to Initial Globodera tabacum tabacum Population Density

Field microplot experiments were conducted from 1995 to 1998 to determine the relationship between fresh shoot weight of stalk-cut broadleaf and shade-grown cigar wrapper tobacco types (Nicotiana tabacum L.) and initial density of Globodera tabacum tabacum second stage juveniles (J2) per cm³ soil. Total shoot weight was negatively correlated with initial nematode densities of 12.3 to 747.3 J2/cm³ soil (r = -0.53 and -0.70 for broadleaf and shade-grown tobacco, respectively). Nonlinear damage functions were used to relate initial G. t. tabacum densities to shoot weight. The models described shoot weight losses of less than 14% or 39% for broadleaf and shade tobacco, respectively, at G. t. tabacum densities below 50 J2/cm³ soil. Total shoot weights were reduced by 40% and 60% of uninfested plots as preplant nematode densities approached maximum levels (>600 J2/cm³ soil) for broadleaf and shade tobacco, respectively. Globodera t. tabacum population increase over a growing season was described by a linear relation on a log/log plot (R² = 0.07 and 0.61 for broadleaf and shade, respectively). These experiments demonstrate that G. t. tabacum can directly reduce shoot weight of stalk-cut broadleaf tobacco. Broadleaf is more tolerant to nematode infection than shade tobacco, as shade tobacco shoot weight reductions were greater at the same initial nematode densities in the same years.     

Early Crop Root Destruction for Management of Tobacco Cyst Nematodes

Prompt tillage after crop harvest was investigated as a cultural control for the tobacco cyst nematode, Globodera tabacum tabacum, on stalk-cut broadleaf cigar wrapper tobacco. Stalk stumps and roots remaining after harvest were destroyed by tilling immediately or from 2 to 6 wk after harvest in field experiments over 4 yr. Cyst nematode Pf/Pi ratios ranged from 0.65 to 1.62 when plants were tilled immediately after harvest and 1.13 to 5.88 when tillage was delayed. Nematode population development was monitored by inoculating plants in pots placed in fields with J2 in eggs and sampling over time (8 to 18 wk). Three generations per year were observed, and G. t. tabacum generation time was as short as 6 wk for each generation. Destroying stalks and root systems remaining after harvesting stalk-cut broadleaf cigar wrapper tobacco removes the host to preclude development of nematodes at the end of the second and entire third generation. Early tillage resulted in consistently lower tobacco cyst nematode populations than allowing viable roots to remain in fields for an additional 8 to 18 wk. This management tactic reduces the need for nematicide application to slow nematode population increases over time and can reduce losses due to infection by G. t. tabacum.     

Morphometrics of Globodera tabacum tabacum,G. t. virginiae,and G. t. solanacearum (Nemata: Heteroderinae)

A morphometric evaluation of second-stage juveniles (J2), males, females, cysts, and eggs of several isolates of the tobacco cyst nematode (TCN) complex, Globodera tabacum tabacum (GTT), G. t. virginiae (GTV), and G. t. solanacearum (GTS) is presented. Morphometrics of eggs, J2, and males are considerably less variable than of females and cysts. No measurements of eggs and J2 are useful for identification of the three subspecies. Distance from the median bulb and excretory pore to the head end in J2 and males is quite stable. Stylet knob width of males is useful for identifying GTV isolates and tail length in separating males of GTT isolates from GTV and GTS. Body length/width (L/W) ratio of females and cysts discriminates GTT from GTV and GTS; stylet knob width is an auxiliary character for identifying GTV. This subspecies complex has a continuum of values for the other characters. Data suggest a close relationship between GTV and GTS, which also occur in close proximity in Virginia.     

Effects of Initial Nematode Density on Population Dynamics of Globodera rostochiensis on Resistant and Susceptible Potatoes

The influence of resistant and susceptible potato cultivars on Globodera rostochiensis population density changes was studied at different nematode inoculum levels (Pi) in the greenhouse and field. Soil in which one susceptible and two resistant cultivars were grown and fallow soil in pots was infested with cysts to result in densities of 0.04-75 eggs/cm³ soil. A resistant cultivar was grown in an infested field with Pi of 0.7-16.7 eggs/cm³ soil. Pi was positively correlated with decline of soil population densities due to hatch where resistant potatoes were grown in the greenhouse and in the field but not in fallow soil. However, Pi was not correlated with in vitro hatch of G. rostochiensis cysts in water or potato root diffusate. Under continuous culture o f a resistant cultivar, viable eggs per cyst declined 60-90% per plant growth cycle (4 weeks) and the number of cysts containing viable eggs had decreased by 77% after five cycles. The rate of G. rostochiensis reproduction on both resistant and susceptible cultivars was negatively correlated with Pi. These data were used to predict the effect of resistant and susceptible potato cultivars on G. rostochiensis soil population dynamics.     

Shade Tobacco Yield Loss and Globodera tabacum tabacum Population Changes in Relation to Initial Nematode Density

Field microplot experiments were conducted from 1987 to 1992 to determine the relationship between fresh weight leaf yield of shade tobacco (Nicotiana tabacum) and initial density of Globodera tabacum tabacum (encysted J2 per cm³ soil). Initial nematode densities of 0.1 to 1,097 J2/cm³ soil were negatively correlated with leaf yield, total shoot weight, and normalized plant height 5 to 6 weeks after transplanting (r = -0.73, -0.73, and -0.52, respectively). Nonlinear damage functions were used to relate initial G. t. tabacum densities to the yield and shoot weight data. The model described leaf yield losses of < 5 % for initial nematode densities of less than 100 J2/cm³ soil. Densities above 100 J2 resulted in yields decreasing exponentially to a maximum yield loss of >40% at 500 to 1,000 J2/cm³ soil. A similar initial density tolerance threshold relationship was observed for total shoot weight. No threshold effect was evident for standardized plant height, which was a poor predictor of leaf yield. Globodera tabacum tabacum population increase over a growing season was described by a linear relation on a log/log plot (R² = 0.73).     

Impact of Herbicides on Heterodera glycines Susceptible and Resistant Soybean Cultivars

Several abiotic and biotic stresses can affect soybean in a growing season. Heterodera glycines, soybean cyst nematode, reduces yield of soybean more than any other pathogen in the United States. Field and greenhouse studies were conducted to determine whether preemergence and postemergence herbicides modified the reproduction of H. glycines, and to determine the effects of possible interactive stresses caused by herbicides and H. glycines on soybean growth and yield. Heterodera glycines reproduction factor (Rf) generally was less on resistant than susceptible cultivars, resulting in a yield advantage for resistant cultivars. The yield advantage of resistant cultivars was due to more pods per plant on resistant than susceptible cultivars. Pendimethalin reduced H. glycines Rf on the susceptible cultivars in 1998 at Champaign, Illinois, and in greenhouse studies reduced dry root weight of H. glycines-resistant and susceptible cultivars, therefore reducing Rf on the susceptible cultivars. The interactive stresses from acifluorfen or imazethapyr and H. glycines reduced the dry shoot weight of the resistant cultivar Jack in a greenhouse study. Herbicides did not affect resistant cultivars'' ability to suppress H. glycines Rf; therefore, growers planting resistant cultivars should make herbicide decisions based on weeds present and cultivar tolerance to the herbicide.     

Hatch and Reproduction of Globodera tabacum tabacum in Response to Tobacco,Tomato, or Black Nightshade

The effects of broadleaf tobacco, tomato, and black nightshade on juvenile hatch and reproduction of Globodera tabacum tabacum were determined in laboratory and greenhouse experiments. Root exudates from nightshade stimulated greater egg hatch than those from either ''Rutgers'' tomato or ''86-4'' tobacco. Hatch was greater at higher proportions of root exudates for all three plant species. Root exudates from plants greater than 3 weeks old stimulated more hatch than younger plants. No regression relationships existed between plant age and nematode batch. In other experiments, hatch from eggs in cysts was higher for tomato and nightshade after 10 weeks in greenhouse pots compared to tobacco and bare soil. Numbers of second-stage juveniles in eggs in cysts produced from a previous generation on the same host were highest on nightshade and less on tomato and tobacco. Cysts of variable age recovered from field soil had increased hatch in both root exudates or water compared to recently produced cysts from plants in growth chambers. Globodera t. tabacum may be subject to both host and environmentally mediated diapause.     

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.     

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.