Spatial Distribution of Plant-Parasitic Nematodes in Semi-Arid Vitis vinifera Vineyards in Washington

The most commonly encountered plant-parasitic nematodes in eastern Washington Vitis vinifera vineyards are Meloidogyne hapla, Mesocriconema xenoplax, Pratylenchus spp., Xiphinema americanum, and Paratylenchus sp.; however, little is known about their distribution in the soil profile. The vertical and horizontal spatial distribution of plant-parasitic nematodes was determined in two Washington V. vinifera vineyards. Others variables measured in these vineyards included soil moisture content, fine root biomass, and root colonization by arbuscular mycorhizal fungi (AMF). Meloidogyne hapla and M. xenoplax were aggregated under irrigation emitters within the vine row and decreased with soil depth. Conversely, Pratylenchus spp. populations were primarily concentrated in vineyard alleyways and decreased with depth. Paratylenchus sp. and X. americanum were randomly distributed within the vineyards. Soil water content played a dominant role in the distribution of fine roots and plant-parasitic nematodes. Colonization of fine roots by AMF decreased directly under irrigation emitters; in addition, galled roots had lower levels of AMF colonization compared with healthy roots. These findings will help facilitate sampling and management decisions for plant-parasitic nematodes in Washington semi-arid vineyards.     

Influence of Criconemella xenoplax and Pruning Time on Short Life of Peach Trees

Influences of Criconemella xenoplax and pruning dates were studied in field microplots with ''Nemaguard'' peach cuttings on a site not previously planted to peaches. Trees with or without C. xenoplax were pruned beginning in December 1984 or March 1985. Peach tree short life (PTSL) did not occur in the absence of C. xenoplax. PTSL occurred earlier in December-pruned than in March-pruned inoculated trees. Results confirm that "old" peach sites are not required for PTSL to occur. Pruning Nemaguard and ''Lovell'' greenhouse-grown seedlings reduced the root mass of both stocks and stimulated Nemaguard, but not Lovell, shoot regrowth. Numbers of C. xenoplax per gram of dry root were greater on pruned than on unpruned seedlings.     

Pathogenicity of Macroposthonia xenoplax to Walnut

Preplanting treatment of soil naturally infested with Macroposthonia xenoplax with 1,2-dibromoethane (ethylene dibromide) significantly increased the growth rate of Juglans hindsii seedlings. When seedlings of J. hindsii, J. regia CV "Serf" and J. regia CV Eureka were inoculated with M. xenoplax, their growth was signilicantly less than that of nematode-free controls. This retarded growth rate was accompanied hy feeder root necrosis, longitudinal cracks in the older roots, and distinct lesions in the secondary phloem.     

Hirsutella rhossiliensisand Verticillium chlamydosporium as Biocontrol Agents of the Root-knot Nematode Meloidogyne hapla on Lettuce

Hirsutella rhossiliensis and Verticillium chlamydosporium infected second-stage juveniles (J2) and eggs of Meloidogyne hapla, respectively, in petri dishes and in organic soil in pots planted to lettuce in the greenhouse. In vitro, H. rhossiliensis produced 78 to 124 spores/infected J2 of M. hapla. The number of J2 in roots of lettuce seedlings decreased exponentially with increasing numbers of vegetative colonies of H. rhossiliensis in the soil. At an infestation of 8 M. hapla eggs/cm³ soil, 1.9 colonies of H. rhossiliensis/cm³ soil were needed for a 50% decrease in J2 penetration of lettuce roots. Egg-mass colonization with V. chlamydosporium varied from 16% to 43% when soil was infested with 8 M. hapla eggs and treated with 5,000 or 10,000 chlamydospores of V. chlamydosporium/cm³ soil. This treatment resulted in fewer J2 entering roots of bioassay lettuce seedlings planted in the infested soils after harvesting the first lettuce plants 7 weeks after infestation with M. hapla. Hirsutella rhossiliensis (0 to 4.3 colonies/cm3 soil), V. chlamydosporium (500 to 10,000 chlamydospores/cm3 soil), or their combination, added to organic soils with 8 M. hapla eggs/cm³ soil, generally did not affect lettuce weight, root galling, or egg production of M. hapla. However, when lettuce was replanted in a mix of infested and uninfested soil (1:3 and 1:7, v:v), egg production was lower in soils with V. chlamydosporium than in soils without the fungus. Both fungi have potential to reduce the M. hapla population, but at densities below 8 eggs/cm³ soil.     

Competition Between Tylenchorhynchus annulatus and Mesocriconema xenoplax on Grain Sorghum as Influenced by Macrophomina phaseolina

Greenhouse experiments were conducted to examine competition between Tylenchorhynchus annulatus and Mesocriconema xenoplax on grain sorghum roots that were colonized by the fungus Macrophomina phaseolina or free from fungus colonization. An incomplete factorial treatment design consisted of two levels of M. phaseolina (0 or 10 colony-forming units/g soil) and 12 T. annulatus:M. xenoplax ratios: 1,000:0; 750:0; 500:0; 250:0; 0:0; 0:250; 0:500; 0:750; 0:1,000; 750:250; 500:500; and 250:750. Plants were harvested after 105 days. Despite similar feeding habits, competition between these ectoparasitic nematode species was limited. Tylenchorhynchus annulatus was more susceptible to antagonism by M. xenoplax than the reverse, but susceptibility depended on initial inoculum ratio. Root colonization by M. phaseolina reduced competitive effects of T. annulatus on M. xenoplax but not the reverse. Both nematode species reduced shoot dry weight but only T. annulatus reduced root dry weight. Both plant weight parameters were reduced by M. phaseolina.     

Effect of Entomopathogenic Nematodes on Mesocriconema xenoplax Populations in Peach and Pecan

The effect of Steinernema riobrave and Heterorhabditis bacteriophora on population density of Mesocriconema xenoplax in peach was studied in the greenhouse. Twenty-one days after adding 112 M. xenoplax adults and juveniles/1,500 cm³ soil to the soil surface of each pot, 50 infective juveniles/cm² soil surface of either S. riobrave or H. bacteriophora were applied. Another entomopathogenic nematode application of the same density was administered 3 months later. The experiment was repeated once. Mesocriconema xenoplax populations were not suppressed (P ≤ 0.05) in the presence of either S. riobrave or H. bacteriophora 180 days following ring nematode inoculation. On pecan, 200 S. riobrave infective-stage juveniles/cm² were applied to the soil surface of 2-year-old established M. xenoplax populations in field microplots. Additional applications of S. riobrave were administered 2 and 4 months later. This study was terminated 150 days following the initial application of S. riobrave. Populations of M. xenoplax were not suppressed in the presence of S. riobrave.     

Control of Meloidogyne javanica and M. arenaria on kenaf and roselle with genetic resistance and nematicides

Kenaf (Hibiscus cannabinus) and roselle (H. sabdarifla) were evaluated in nematicide-treated and untreated field soil naturally infested with either Meloidogyne javanica or M. arenaria. Root-knot indices indicated that the kenaf breeding line j-l-113 had moderate resistance to M. javanica and low resistance to M. arenaria. Kenaf cv Everglades 71 was highly susceptible to both M. javanica and M. arenaria, and roselle breeding line A59-56 was highly resistant. Both nematode species reproduced on all plant entries, but more larvae were recovered from the soil in plots planted to Everglades 71 than in plots planted to j-l-l13 or A59-56. In untreated soil infested with M. javanica, dry-matter yields were greater (P = 0.05) for j-l-l13 and A59-56 than for Everglades 71. The percentages of live plants at harvest were: j-l-l13, 88; A59-56, 93; and Everglades 71, 9. Ethylene dibromide (1,2-dibromoethane) at 73.9 kg a.i./ha and DBCP (1,2-dibromo-3-chloropropane) at 17.6 kg a.i./ha increased dry-matter yields significantly for all entries planted in soil infested with M. arenaria. Carbofuran (2.3-dihydro-2,2-dimethyl-7-benzofuranyl methylcarbamate) at 5.9 kg a.i./ha did not increase the dry-matter yields of any entry. None of the nematicides increased the growth of any entry significantly in soil infested with M. javanica.     

Effects of Macroposthonia xenoplax on the Growth of Concord Grape

Concord grape (Vitis labrusca) plants were inoculated with Macroposthonia xenoplax at levels of 100, 1,000, and 10,000 nematodes. After 4 months, plants inoculated with 10,000 M. xenoplax were stunted, and root systems were darker and had fewer feeder roots than those in other treatments. The lower nematode inoculation levels suppressed top growth but did not affect root growth. M. xenoplax reproduced well on Concord grapes.     

Effect of Initial Population Density of Criconemella xenoplax on Reducing Sugars,Free Amino Acids,and Survival of Peach Seedlings over Time

Percentage of mortality, growth suppression, and changes in free amino acid and reducing sugar content in root and (or) stem tissue of Nemaguard peach seedlings were studied in the greenhouse in relation to time and eight different initial population densities (Pi) of Criconemella xenoplax. After 90 and 180 days, free amino acid content in root tissue significantly increased with increasing nematode numbers. Suppression of root volume, dry root and stem weight, height increase, plant survival, and content of reducing sugars in root tissue were detected at 180 and 270 days and following pruning. All criteria were negatively correlated with nematode Pi. Changes in growth, metabolic parameters, and survival percentage were attributed to Pi density of C. xenoplax, duration of the experiment, and nematode reproduction rate.     

Evaluation of Host Suitability in Prunus for Criconemella xenoplax

Methods were developed for screening Prunus selections for host suitability to Criconemella xenoplax. The relative host suitability of selections was based upon a doubling accumulation value (β) that was defined as the number of degree-days (base 9 C) required for doubling of an increment of the initial nematode population. The β value characteristic for C. xenoplax (139 ± 8 degree-days) on suitable hosts was similar to the average β value determined for several peach rootstocks known to be suitable hosts. The β values were 144 ± 21 for Halford, 141 ± 16 for Lovell, and 138 ± 10 for Nemaguard. A higher value for β could indicate poorer host suitability or resistance of a selection to C. xenoplax. All of 369 Prunus accessions tested, including eight accessions that had survived well on a field site infested with C. xenoplax, were suitable hosts. Apparently, resistance to C. xenoplax was not a factor in survival of the accessions planted in the field. Seedlings from P. besseyi, P. pumila ''Mando'', and two interspecific hybrids, Redcoat and Sapalta IR 549-1, failed to support nematode population increase in 44-81% of tests conducted, but all selections supported population increase in some tests. These accessions may have resistance mechanisms that are active only under specific conditions.     

Tests for Transmission of Prunus Necrotic Ringspot and Two Nepoviruses by Criconemella xenoplax

In two of three trials, detectable color reactions in ELISA for Prunus necrotic ringspot virus (PNRSV) were observed for Criconemella xenoplax handpicked from the root zone of infected peach trees. Criconemella xenoplax (500/pot) handpicked from root zones of peach trees infected with PNRSV failed to transmit the virus to cucumber or peach seedlings. The nematode also failed to transmit tomato ringspot (TomRSV) or tobacco ringspot viruses between cucumbers, although Xiphinema americanum transmitted TomRSV under the same conditions. Plants of peach, cucumber, Chenopodium quinoa, and Catharanthus roseus were not infected by PNRSV when grown in soil containing C. xenoplax collected from root zones of PNRSV-infected trees. Shirofugen cherry scions budded on Mazzard cherry seedling rootstocks remained symptomless when transplanted into root zones of PNRSV-infected trees. Virus transmission was not detected by ELISA when C. xenoplax individuals were observed to feed on cucumber root explants that were infected with PNRSV and subsequently fed on roots of Prunus besseyi in agar cultures. Even if virus transmission by C. xenoplax occurs via contamination rather than by a specific mechanism, it must be rare.     

Population Dynamics of Pratylenchus penetrans,Paratylenchus sp., and Criconemella xenoplax on Western Oregon Peppermint

Endoparasitic nematode populations are usually measured separately for soil and roots without a determination of the quantitative relation between soil and root population components. In this study, Pratylenchus penetrans populations in peppermint soil, roots, and rhizomes were expressed as the density within a standardized core consisting of 500 g dry soil plus the roots and rhizomes contained therein. Populations of Paratylenchus sp. and Criconemella xenoplax in 500 g dry soil were also determined, thus measuring the total plant-parasitic nematode population associated with the plant. Mean wet root weight per standard core peaked in spring and again in late summer and was lowest early in the growing season and in early fall. Pratylenchus penetrans populations peaked 4 to 6 weeks after root weight peaks. The percentage of the total population in roots reached 70% to 90% in early April, decreased to 20% to 40% in August, and returned to higher percentages during the winter. Rhizomes never contained more than a minor proportion of the population. Mean Paratylenchus sp. populations increased through spring and peaked in late August. Mean C. xenoplax populations fluctuated, peaking in August or September. Populations of all parasitic species were lowest during winter. Evaluation using the standard core method permits assessment of the total P. penetrans population associated with the plant and of changes in root weight as well as the seasonal distribution of P. penetrans.     

Biological Control of Meloidogyne hapla on Alfalfa and Tomato with the Fungus Meria coniospora

This study was to determine whether Arthrobotrys flagrans, A. oligospora, and Meria coniospora would control the root-knot nematode Meloidogyne hapla on alfalfa and tomato. Alfalfa seeds were coated with a fungus-rye powder in 2% cellulose and were planted in infested soil. Three-week-old seedlings from seed treated with M. coniospora had 60% and 58% fewer galls in two experiments than did seedlings from untreated seeds. Numbers of J2 in the soil were not reduced. Plant growth did not improve. When seed of tomato were coated with M. coniospora and planted in M. hapla-infested soil, roots had 34% fewer galls and 47% fewer J2 in the soil at 28 days. After 56 days there was no reduction in J2 numbers. Plant growth did not improve. When roots of tomato transplants were dusted with M. coniospora fungus-rye powder or sprayed with a spore suspension before planting in M. hapla-infested soil, 42% and 35%, respectively, fewer galls developed in 28 days on treated roots than on roots not treated with fungus. The numbers of J2 extracted from roots or recovered from soil were not reduced, however, and plant growth did not improve.     

Biological Control of the Phytoparasitic Nematode Mesocriconema xenoplax on Peach Trees

Seven fluorescent Pseudomonas spp. capable of inhibiting reproduction of Mesocriconema xenoplax have been isolated from soil sites that suppress both nematode multiplication and Peach Tree Short Life (PTSL). One of these seven strains, Pseudomonas sp. BG33R, inhibits M. xenoplax multiplication in vivo and egg hatch in vitro. Mesocriconema xenoplax populations on peach seedlings inoculated with BG33R and planted into soil-solarized field plots remained at or below the economic threshold for nematicide treatment in South Carolina for nearly 18 months. Soil solarization alone induced a shift toward a microbial community that was suppressive to nematode multiplication. Additionally, five Tn5 mutants of BG33R, lacking the ability to kill eggs, have been generated. The Tn5 insertion site in each mutant has been cloned and sequenced. DNA sequence analysis has revealed a high degree of homology to several genes of interest because of their potential involvement in the production of the egg-kill factor. These Tn5 egg-kill negative mutants also no longer produce protease or salicylic acid while producing nearly twice the amount of fluorescent siderophore as the wild type parent.     

Dynamics of Concomitant Populations of Meloidogyne incognita and Criconemella xenoplax on Peach

The interaction between Meloidogyne incognita and Criconemella xenoplax on nematode reproduction and growth of Lovell peach was studied in field microlots and the greenhouse. Meloidogyne incognita suppressed reproduction of C. xenoplax in both field and greenhouse experiments. Tree growth, as measured by trunk diameter, was reduced (P ≤ 0.05) in the presence of M. incognita as compared with C. xenoplax of the uninoculated control trees 26 months following inoculation. A similar response regarding dry root weight was also detected in greenhouse-grown seedlings after 5 months. The presence of C. xenoplax did not affect Lovell tree growth. A synergistic effect causing a reduction (P ≤ 0.05) in tree growth was recorded 26 and 38 months following inoculation. The presence of M. incognita increased levels of malonyl-1-aminocyclopropane-1-carboxylic acid content in leaves of trees grown in field microplots 19 months after inoculaoon. Meloidogyne incognita appears to be a more dominant parasite than C. xenoplax on Lovell peach.     

Effects of Soil Temperature and Planting Date of Wheat on Meloidogyne incognita Reproduction,Soil Populations,and Grain Yield

Wheat cultivars Anza and Produra grown in winter in California were planted in Meloidogyne incognita infested and noninfested sandy loam plots in October (soil temperature 21 C) and November (soil temperature 16 C) of 1979. Meloidogyne incognita penetrated roots of mid-October planted Ataza (427 juveniles/g root), developed into adult females by January, and produced 75 eggs/g root by harvest in April. Penetration and development did not occur in late plantings. Anza seedlings grown in infested soil in pots buried in field soil in early spring were not invaded until soil temperature exceeded 18 C. Meloidogyne incognita juveniles can migrate through soil and penetrate roots at temperatures above 18 C (activity threshold), however development can occur at lower temperatures. Grain yields were not significantly different between nematode infested (3,390 kg/ha) and noninfested (2,988 kg/ha) plots. Winter decline of eggs and juveniles in two late plantings anti in fallow soil were 69, 72, and 77%, respectively, but egg and juvenile decline was only 40% in the early Anza plots that supported nematode reproduction in the spring. Delay of planting date until soil temperature is below 18 C is suggested to maximize the use of wheat in rotation as a nematode pest management cultural tactic for suppressing root-knot nematodes.     

Pathogenicity of Criconemoides xenoplax to Prune and Plum Rootstocks

Elimination of Criconemoides xenoplax from a prune orchard soil by fumigation with ethylene dibromide at the rate of 42 μliter/liter of soil (equivalent to about 13 gal/acre) improved the growth of Myrobalan plum, Addition of this nematode to Myrobalan seedlings or young ''Marianna 2624'' plants propagated from cuttings resulted in destruction of cortical root tissue, darkening of roots, alteration of water stress, lowering of nutrient levels in leaves, and reduction in plant weight. C. xenoplax increased on all nine Prunus cerasifera varieties and hybrids tested, including those used commonly as rootstocks for prunes and plums. Rhizoctonia solani isolated from Myrobalan seedlings infected with C. xenoplax caused lesions on the hypocotyls of young Myrobalan seedlings in the laboratory, but had no effect on older seedlings in the greenhouse, and did not alter the effect of C. xenoplax.     

Detection and Investigation of Soil Biological Activity against Meloidogyne incognita

Greenhouse experiments with two susceptible hosts of Meloidogyne incognita, a dwarf tomato and wheat, led to the identification of a soil in which the root-knot nematode population was reduced 5- to 16-fold compared to identical but pasteurized soil two months after infestation with 280 M. incognita J2/100 cm³ soil. This suppressive soil was subjected to various temperature, fumigation and dilution treatments, planted with tomato, and infested with 1,000 eggs of M. incognita/100 cm³ soil. Eight weeks after nematode infestation, distinct differences in nematode population densities were observed among the soil treatments, suggesting the suppressiveness had a biological nature. A fungal rRNA gene analysis (OFRG) performed on M. incognita egg masses collected at the end of the greenhouse experiments identified 11 fungal phylotypes, several of which exhibited associations with one or more of the nematode population density measurements (egg masses, eggs or J2). The phylotype containing rRNA genes with high sequence identity to Pochonia chlamydosporia exhibited the strongest negative associations. The negative correlation between the densities of the P. chlamydosporia genes and the nematodes was corroborated by an analysis using a P. chlamydosporia-selective qPCR assay.     

Suppression of Meloidogyne arenaria Race 1 by Soil Application of Endospores of Pasteuria penetrans

The potential of Pasteuria penetrans for suppressing Meloidogyne arenaria race 1 on peanut (Arachis hypogaea) was tested over a 2-year period in a field microplot experiment. Endospores of P. penetrans were mass-produced on M. arenaria race 1 infecting tomato plants. Endospores were inoculated in the first year only at rates of 0, 1,000, 3,000, 10,000, and 100,000 endospores/g of soil, respectively, into the top 20 cm of microplots that were previously infested with M. arenaria race 1. One peanut seedling was planted in each microplot. In the first year, root gall indices and pod galls per microplot were significantly reduced by 60% and 95% for 100,000 endospores/g of soil, and 20% and 65% for 10,000 endospores/g of soil, respectively. Final densities of second-stage juveniles (J2) in soil were not significantly different among the treatments. The number of endospores attached to J2 and percentage of J2 with attached endospores significantly increased with increasing endospore inoculation levels. Pasteuria penetrans significantly reduced the densities of J2 that overwintered. In the second year, root and pod gall indices, respectively, were significantly reduced by 81% and 90% for 100,000 endospores/g of soil, and by 61% and 82% of 10,000 endospores/g of soil. Pod yields were significantly increased by 94% for 100,000 and by 57% for 10,000 endospores/g of soil, respectively. The effect of P. penetrans on final densities of J2 in soil was not significant. Regression analyses verified the role of P. penetrans in the suppression of M. arenaria. The minimum number of endospores required for significantly suppressing M. arenaria race 1 on peanut was 10,000 endospores/g of soil.     

Relationship of Grapevine Yield and Growth to Nematode Densities

Yield, growth, and vigor of individual grape vines were correlated with nematode population densities in a series of California vineyards. In a Hanford sandy loam soil, Xiphinema americanum densities showed negative correlations with yield, growth, and vigor of vines. When vines were categorized according to vigor, X. americanurn densities had little relationship to yield of high-vigor vines, but were negatively correlated with yield of low-vigor vines. Densities of Paratylenchus harnatus were positively correlated with yield, growth, and vigor of vines. Correlations between Meloidogyne spp. densities and vine performance were variable, even when the vines were separated according to soil type and plant vigor. Densities of Meloidogyne spp. populations were generally higher on coarser-textured, sandy soils and the vines were less vigorous there. Densities of P. hamatus were greater in fine-textured soils.