Do biological control agents adapt to local pest genotypes? A multiyear test across geographic scales.

Parasite local adaptation has been a major focus of (co)evolutionary research on host-parasite interactions. Studies of wild host-parasite systems frequently find that parasites paired with local, sympatric host genotypes perform better than parasites paired with allopatric host genotypes. In contrast, there are few such tests in biological control systems to establish whether biological control parasites commonly perform better on sympatric pest genotypes. This knowledge gap prevents the optimal design of biological control programs: strong local adaptation could argue for the use of sympatric parasites to achieve consistent pest control. To address this gap, we tested for local adaptation of the biological control bacterium Pasteuria penetrans to the root-knot nematode Meloidogyne arenaria, a global threat to a wide range of crops. We measured the probability and intensity of P. penetrans infection on sympatric and allopatric M. arenaria over the course of 4 years. Our design accounted for variation in adaptation across scales by conducting tests within and across fields, and we isolated the signature of parasite adaptation by comparing parasites collected over the course of the growing season. Our results are largely inconsistent with local adaptation of P. penetrans to M. arenaria: in 3 of 4 years, parasites performed similarly well in sympatric and allopatric combinations. In 1 year, however, infection probability was 28% higher for parasites paired with hosts from their sympatric plot, relative to parasites paired with hosts from other plots within the same field. These mixed results argue for population genetic data to characterize the scale of gene flow and genetic divergence in this system. Overall, our findings do not provide strong support for using P. penetrans from local fields to enhance biological control of Meloidogyne.

Development and Genetic Characterization of Peanut Advanced Backcross Lines That Incorporate Root-Knot Nematode Resistance From Arachis stenosperma.

Crop wild species are increasingly important for crop improvement. Peanut (Arachis hypogaea L.) wild relatives comprise a diverse genetic pool that is being used to broaden its narrow genetic base. Peanut is an allotetraploid species extremely susceptible to peanut root-knot nematode (PRKN) Meloidogyne arenaria. Current resistant cultivars rely on a single introgression for PRKN resistance incorporated from the wild relative Arachis cardenasii, which could be overcome as a result of the emergence of virulent nematode populations. Therefore, new sources of resistance may be needed. Near-immunity has been found in the peanut wild relative Arachis stenosperma. The two loci controlling the resistance, present on chromosomes A02 and A09, have been validated in tetraploid lines and have been shown to reduce nematode reproduction by up to 98%. To incorporate these new resistance QTL into cultivated peanut, we used a marker-assisted backcrossing approach, using PRKN A. stenosperma-derived resistant lines as donor parents. Four cycles of backcrossing were completed, and SNP assays linked to the QTL were used for foreground selection. In each backcross generation seed weight, length, and width were measured, and based on a statistical analysis we observed that only one generation of backcrossing was required to recover the elite peanut's seed size. A populating of 271 BC3F1 lines was genome-wide genotyped to characterize the introgressions across the genome. Phenotypic information for leaf spot incidence and domestication traits (seed size, fertility, plant architecture, and flower color) were recorded. Correlations between the wild introgressions in different chromosomes and the phenotypic data allowed us to identify candidate regions controlling these domestication traits. Finally, PRKN resistance was validated in BC3F3 lines. We observed that the QTL in A02 and/or large introgression in A09 are needed for resistance. This present work represents an important step toward the development of new high-yielding and nematode-resistant peanut cultivars.

Festulolium and fungal endophyte associations: host status for Meloidogyne incognita and nematotoxic plant extracts.

Festulolium hybrids are forage grasses used worldwide in temperate climates. They are associated with the fungal endophyte Epichloë uncinata, which aids in nutrient uptake, drought tolerance, and production of metabolites that protect against parasites and herbivores. Epichloë uncinata produces loline alkaloids, which can deter insect pests. Festulolium has not been widely studied for susceptibility to plant-parasitic nematodes, so Festulolium lines, with and without fungal endophytes, were tested in the greenhouse for host status to the root-knot nematode Meloidogyne incognita. All were poor hosts, regardless of line or endophyte status. Pepper seedlings planted into soil following removal of the Festulolium plants were infected by nematodes, likely because of surviving nematodes from the original inoculation combined with some reproduction on Festulolium. Lolines were found in shoots and roots of all endophyte-associated lines, and some types of lolines in roots increased after nematode infection. Methanolic extracts from roots and shoots of a tested Festulolium line did not inhibit egg hatch, but killed nearly a third of second-stage juveniles whether an endophyte was present or not. Further studies would indicate whether these Festulolium lines aid in suppressing field populations of M. incognita.Festulolium hybrids are forage grasses used worldwide in temperate climates. They are associated with the fungal endophyte Epichloë uncinata, which aids in nutrient uptake, drought tolerance, and production of metabolites that protect against parasites and herbivores. Epichloë uncinata produces loline alkaloids, which can deter insect pests. Festulolium has not been widely studied for susceptibility to plant-parasitic nematodes, so Festulolium lines, with and without fungal endophytes, were tested in the greenhouse for host status to the root-knot nematode Meloidogyne incognita. All were poor hosts, regardless of line or endophyte status. Pepper seedlings planted into soil following removal of the Festulolium plants were infected by nematodes, likely because of surviving nematodes from the original inoculation combined with some reproduction on Festulolium. Lolines were found in shoots and roots of all endophyte-associated lines, and some types of lolines in roots increased after nematode infection. Methanolic extracts from roots and shoots of a tested Festulolium line did not inhibit egg hatch, but killed nearly a third of second-stage juveniles whether an endophyte was present or not. Further studies would indicate whether these Festulolium lines aid in suppressing field populations of M. incognita.

A new source of root-knot nematode resistance from Arachis stenosperma incorporated into allotetraploid peanut (Arachis hypogaea).

Root-knot nematode is a very destructive pathogen, to which most peanut cultivars are highly susceptible. Strong resistance is present in the wild diploid peanut relatives. Previously, QTLs controlling nematode resistance were identified on chromosomes A02, A04 and A09 of Arachis stenosperma. Here, to study the inheritance of these resistance alleles within the genetic background of tetraploid peanut, an F2 population was developed from a cross between peanut and an induced allotetraploid that incorporated A. stenosperma, [Arachis batizocoi x A. stenosperma]4×. This population was genotyped using a SNP array and phenotyped for nematode resistance. QTL analysis allowed us to verify the major-effect QTL on chromosome A02 and a secondary QTL on A09, each contributing to a percentage reduction in nematode multiplication up to 98.2%. These were validated in selected F2:3 lines. The genome location of the large-effect QTL on A02 is rich in genes encoding TIR-NBS-LRR protein domains that are involved in plant defenses. We conclude that the strong resistance to RKN, derived from the diploid A. stenosperma, is transferrable and expressed in tetraploid peanut. Currently it is being used in breeding programs for introgressing a new source of nematode resistance and to widen the genetic basis of agronomically adapted peanut lines.

Evaluation of Selected Nonfumigant Nematicides on Increasing Inoculation Densities of Meloidogyne incognita on Cucumber.

The southern root-knot nematode (RKN), Meloidogyne incognita, is particularly difficult to manage because of high susceptibility of all commercial cucumber (Cucumis sativus) cultivars to this nematode. Growers have conventionally relied on nematicide applications to control RKN. Two microplot experiments were conducted in which four nonfumigant nematicides, oxamyl, fluopyram, fluensulfone, and fluazaindolizine, were examined for their efficacy in reducing gall severity and postharvest soil nematode numbers in microplots inoculated with increasing inoculation densities (1,000, 5,000, 10,000, and 20,000 nematodes/microplot), and improving growth and yield of cucumber. Nematicides were applied 1 day prior to transplanting cucumber seedlings, except fluensulfone, which was applied 7 days before transplanting. At harvest, root gall indices differed significantly (P < 0.0001) among nematode inoculation densities and nematicides. All four nematicides were effective in reducing the root gall index when compared with the untreated control on a consistent basis at all M. incognita inoculation densities. At the lowest inoculation density, no significant difference in gall index or final population density was observed among nematicides; however, gall index increased with increasing nematode inoculation densities in nematicide-treated microplots. Correlations between gall index and inoculation density clearly showed that soil treatment with fluensulfone, fluazaindolizine, or fluopyram was more effective in reducing gall severity than treatment with oxamyl. Regression analysis also indicated no significant effect of nematode inoculation densities on yield of cucumber treated with these nematicides. Results of this study will provide guidance for improving nematicide efficiencies in soil with varying inoculation densities of RKN.

Rapid change in host specificity in a field population of the biological control organism Pasteuria penetrans.

In biological control, populations of both the biological control agent and the pest have the potential to evolve and even to coevolve. This feature marks the most powerful and unpredictable aspect of biological control strategies. In particular, evolutionary change in host specificity of the biological control agent could increase or decrease its efficacy. Here, we tested for change in host specificity in a field population of the biological control organism Pasteuria penetrans. Pasteuria penetrans is an obligate parasite of the plant parasitic nematodes Meloidogyne spp., which are major agricultural pests. From 2013 through 2016, we collected yearly samples of P. penetrans from eight plots in a field infested with M. arenaria. Plots were planted either with peanut (Arachis hypogaea) or with a rotation of peanut and soybean (Glycine max). To detect temporal change in host specificity, we tested P. penetrans samples annually for their ability to attach to (and thereby infect) four clonal lines of M. arenaria. After controlling for temporal variation in parasite abundance, we found that P. penetrans from each of the eight plots showed temporal variation in their attachment specificity to the clonal host lines. The trajectories of change in host specificity were largely unique to each plot. This result suggests that local forces, at the level of individual plots, drive change in specificity. We hypothesize that coevolution with local M. arenaria hosts may be one such force. Lastly, we observed an overall reduction in attachment rate with samples from rotation plots relative to samples from peanut plots. This result may reflect lower abundance of P. penetrans under crop rotation, potentially due to suppressed density of host nematodes. As a whole, the results show local change in specificity on a yearly basis, consistent with evolution of a biological control organism in its ability to infect and suppress its target pest.

Maternal Stress Reduces the Susceptibility of Root-Knot Nematodes to Pasteuria Penetrans.

Pasteuria penetrans is an obligate parasite of root-knot nematodes (Meloidogyne spp.). Endospores of P. penetrans attach to the cuticle of second-stage juveniles (J2) and complete their life cycle within the nematode female body. Infected females will be filled with spores and will be sterilized. Studies with Daphnia magna and its parasite Pasteuria ramosa showed that a poor maternal environment can lead to offspring resistant to P. ramosa. Therefore, we hypothesized that Meloidogyne arenaria females raised under a stressed environment would produce offspring that were more resistant to P. penetrans. Females were exposed to a stressed environment created by crowding and low-food supply, or a non-stressed environment and their offspring evaluated for endospore attachment and infection by P. penetrans. No difference in spore attachment was observed between the two treatments. However, infection rate of P. penetrans in the stressed treatment was significantly lower than that in the non-stressed treatment (8 vs 18%). Mothers raised under stressed conditions appeared to produce more resistant offspring than did mothers raised under favorable conditions. Under stressful conditions, M. arenaria mothers may provide their progeny with enhanced survival traits. In the field, when nematode populations are not managed, they often reach the carrying capacity of their host plant by the end of the season. This study suggests that the next generation of inoculum may be more resistant to infection by P. penetrans.

Fluensulfone sorption and mobility as affected by soil type.

BACKGROUND: Fluensulfone is a fluoroalkenyl chemical with activity against multiple genera of plant-parasitic nematodes. The adsorption, desorption, and mobility of fluensulfone were evaluated on multiple soils from the USA in laboratory and column experiments. RESULTS: Adsorption data regressed to the logarithmic Freundlich equation resulted in isotherm values of 1.24 to 3.28. Soil adsorption of fluensulfone correlated positively with organic matter (0.67) and clay (0.34), but negatively with sand (-0.54). Fluensulfone soil desorption correlated to pH (0.38) and cation exchange capacity (0.44). Fluensulfone desorption from Arredondo sand soil was 26%, and from other soils ranged from 43 to 70%. In mobility experiments, fluensulfone in the leachate peaked at 3 h, gradually declining and becoming undetectable after 9 h. Recovery from leachate was 45% of the initial fluensulfone applied to the soil surface. In separate experiments, 30-cm-long soil columns were saturated with 1 L of water, and then segregated into three 10-cm sections. Fluensulfone recovery was 41, 34, 29, and 13% in Chualar sandy loam, Arredondo sand, Greenville sandy clay loam, and Tifton loamy sand, respectively, in the top 10-cm section. CONCLUSION: Data indicated that soil organic matter and clay contents will affect sorption, mobility, and dissipation of fluensulfone. © 2017 Society of Chemical Industry.

Influence of Root Exudates and Soil on Attachment of Pasteuria penetrans to Meloidogyne arenaria.

The bacterium Pasteuria penetrans is a parasite of root-knot nematodes (Meloidogyne spp.). Endospores of P. penetrans attach to the cuticle of second-stage juveniles (J2) and subsequently sterilize infected females. When encumbered by large numbers of spores, juveniles are less mobile and their ability to infect roots is reduced. This study looked at different factors that influence spore attachment of P. penetrans to the root-knot nematode Meloidogyne arenaria. Pretreatment of J2 with root exudates of eggplant (Solanum melongena cv. Black beauty) reduced spore attachment compared with pretreatment with phosphate-buffered saline (PBS), suggesting that the nematode surface coat was altered or the spore recognition domains on the nematode surface were blocked. Spore attachment was equally reduced following exposure to root exudates from both host and nonhost plants for M. arenaria, indicating a common signal that affects spore attachment. Although phytohormones have been shown to influence the lipophilicity of the nematode surface coat, auxins and kinetins did not affect spore attachment compared with PBS. Root exudates reduced spore attachment more in sterilized soil than in natural soil. Sterilization may have eliminated microbes that consume root exudates, or altered the chemical components of the soil solution or root exudates. Root exudates caused a greater decrease in spore attachment in loamy sand than in a sandy loam soil. The sandy loam had higher clay content than the loamy sand, which may have resulted in more adsorption of compounds in the root exudates that affect spore attachment. The components of the root exudates could have also been modified by soil type. The results of this study demonstrate that root exudates can decrease the attachment of P. penetrans endospores to root-knot nematodes, indicating that when these nematodes enter the root zone their susceptibility to spore attachment may decrease.

Gene expression profiling describes the genetic regulation of Meloidogyne arenaria resistance in Arachis hypogaea and reveals a candidate gene for resistance.

Resistance to root-knot nematode was introgressed into cultivated peanut Arachis hypogaea from a wild peanut relative, A. cardenasii and previously mapped to chromosome A09. The highly resistant recombinant inbred RIL 46 and moderately resistant RIL 48 were selected from a population with cv. Gregory (susceptible) and Tifguard (resistant) as female and male parents, respectively. RNA-seq analysis was performed on these four genotypes using root tissue harvested from root-knot nematode infected plants at 0, 3, 7 days after inoculation. Differential gene expression analysis provides evidence that root-knot nematodes modulate biological pathways involved in plant hormone, defense, cell signaling, cytoskeleton and cell wall metabolism in a susceptible reaction. Corresponding to resistance reaction, an effector-induced-immune response mediated by an R-gene was identified in Tifguard. Mapping of the introgressed region indicated that 92% of linkage group A09 was of A. cardenasii origin in Tifguard. RIL46 and RIL 48 possessed 3.6% and 83.5% of the introgression on A09, respectively. Within the small introgressed region carried by RIL 46, a constitutively expressed TIR-NBS-LRR gene was identified as the candidate for nematode resistance. Potential defense responsive pathways include effector endocytosis through clathrin-coated vesicle trafficking, defense signaling through membrane lipid metabolism and mucilage production.

Efficacy of Various Application Methods of Fluensulfone for Managing Root-knot Nematodes in Vegetables.

Fluensulfone is a new nematicide in the flouroalkenyl chemical group. A field experiment was conducted in 2012 and 2013 to evaluate the efficacy of various application methods of fluensulfone for control of Meloidogyne spp. in cucumber (Cucumis sativus). Treatments of fluensulfone (3.0 kg a.i./ha) were applied either as preplant incorporation (PPI) or via different drip irrigation methods: drip without pulse irrigation (Drip NP), pulse irrigation 1 hr after treatment (Drip +1P), and treatment at the same time as pulse irrigation (Drip =P). The experiment had eight replications per treatment and also included a PPI treatment of oxamyl (22.5 kg a.i./ha) and a nontreated control. Compared to the control, neither the oxamyl nor the fluensulfone PPI treatments reduced root galling by Meloidogyne spp. in cucumber. Among the drip treatments, Drip NP and Drip +1P reduced root galling compared to the control. Cucumber yield was greater in all fluensulfone treatments than in the control. In a growth-chamber experiment, the systemic activity and phytotoxicity of fluensulfone were also evaluated on tomato (Solanum lycopersicum), eggplant (Solanum melongena), cucumber, and squash (Curcurbita pepo). At the seedling stage, foliage of each crop was sprayed with fluensulfone at 3, 6, and 12 g a.i./liter, oxamyl at 4.8 g a.i./liter, or water (nontreated control). Each plant was inoculated with Meloidogyne incognita juveniles 2 d after treatment. There were six replications per treatment and the experiment was conducted twice. Foliar applications of fluensulfone reduced plant vigor and dry weight of eggplant and tomato, but not cucumber or squash; application of oxamyl had no effect on the vigor or weight of any of the crops. Typically, only the highest rate of fluensulfone was phytotoxic to eggplant and tomato. Tomato was the only crop tested in which there was a reduction in the number of nematodes or galls when fluensulfone or oxamyl was applied to the foliage compared to the nontreated control. This study demonstrates that control of Meloidogyne spp. may be obtained by drip and foliar applications of fluensulfone; however, the systemic activity of fluensulfone is crop specific and there is a risk of phytotoxicity with foliar applications.

Efficacy of Fluensulfone in a Tomato-Cucumber Double Cropping System.

Vegetable crops in the southeastern United States are commonly grown on plastic mulch with two crop cycles produced on a single mulch application. Field trials were conducted in 2013 and 2014 in two locations to evaluate the efficacy of fluensulfone for controlling Meloidogyne spp. when applied through drip irrigation to cucumber in a tomato-cucumber double-cropping system. In the spring tomato crop, 1,3-dichloropropene (1,3-D), fluensulfone, and a resistant cultivar significantly decreased root galling by 91%, 73%, and 97%, respectively, compared to the untreated control. Tomato plots from the spring were divided into split plots for the fall where the main plots were the spring treatment and the subplots were cucumber either treated with fluensulfone (3.0 kg a.i./ha. via drip irrigation) or left untreated. The fall application of fluensulfone improved cucumber vigor and reduced gall ratings compared to untreated subplots. Fluensulfone reduced damage from root-knot nematodes when applied to the first crop as well as provided additional protection to the second crop when it was applied through a drip system.

Conserving and enhancing biological control of nematodes.

Conservation biological control is the modification of the environment or existing practices to protect and enhance antagonistic organisms to reduce damage from pests. This approach to biological control has received insufficient attention compared with inundative applications of microbial antagonists to control nematodes. This review provides examples of how production practices can enhance or diminish biological control of plant-parasitic nematodes and other soilborne pests. Antagonists of nematodes can be enhanced by providing supplementary food sources such as occurs when organic amendments are applied to soil. However, some organic amendments (e.g., manures and plants containing allelopathic compounds) can also be detrimental to nematode antagonists. Plant species and genotype can strongly influence the outcome of biological control. For instance, the susceptibility of the plant to the nematode can determine the effectiveness of control; good hosts will require greater levels of suppression than poor hosts. Plant genotype can also influence the degree of rhizosphere colonization and antibiotic production by antagonists, as well the expression of induced resistance by plants. Production practices such as crop rotation, fallow periods, tillage, and pesticide applications can directly disrupt populations of antagonistic organisms. These practices can also indirectly affect antagonists by reducing their primary nematode host. One of the challenges of conservation biological control is that practices intended to protect or enhance suppression of nematodes may not be effective in all field sites because they are dependent on indigenous antagonists. Ultimately, indicators will need to be identified, such as the presence of particular antagonists, which can guide decisions on where it is practical to use conservation biological control. Antagonists can also be applied to field sites in conjunction with conservation practices to improve the consistency, efficacy, and duration of biological control. In future research, greater use should be made of bioassays that measure nematode suppression because changes in abundance of particular antagonists may not affect biological control of plant parasites.

Population Dynamics of Dactylella oviparasitica and Heterodera schachtii: Toward a Decision Model for Sugar Beet Planting.

A series of experiments were performed to examine the population dynamics of the sugarbeet cyst nematode, Heterodera schachtii, and the nematophagus fungus Dactylella oviparasitica. After two nematode generations, the population densities of H. schachtii were measured in relation to various initial infestation densities of both D. oviparasitica and H. schachtii. In general, higher initial population densities of D. oviparasitica were associated with lower final population densities of H. schachtii. Regression models showed that the initial densities of D. oviparasitica were only significant when predicting the final densities of H. schachtii J2 and eggs as well as fungal egg parasitism, while the initial densities of J2 were significant for all final H. schachtii population density measurements. We also showed that the densities of H. schachtii-associated D. oviparasitica fluctuate greatly, with rRNA gene numbers going from zero in most field-soil-collected cysts to an average of 4.24 x 10(8) in mature females isolated directly from root surfaces. Finally, phylogenetic analysis of rRNA genes suggested that D. oviparasitica belongs to a clade of nematophagous fungi that includes Arkansas Fungus strain L (ARF-L) and that these fungi are widely distributed. We anticipate that these findings will provide foundational data facilitating the development of more effective decision models for sugar beet planting.

Induction of Systemic Acquired Resistance by Rotylenchulus reniformis and Meloidogyne incognita in Cotton Following Separate and Concomitant Inoculations.

Systemic acquired resistance (SAR) can be elicited by virulent and avirulent pathogenic strains and SAR against plant-parasitic nematodes has been documented. Our objective was to determine whether co-infection of cotton by Meloidogyne incognita and Rotylenchulus reniformis affects the population level of either nematode compared to infection by each species individually. Split-root trials were conducted in which plants were inoculated with i) R. reniformis only, ii) M. incognita only, iii) both R. reniformis and M. incognita, or iv) no nematodes. Half of the root system was inoculated with R. reniformis or M. incognita on day 0 and the other half with M. incognita or R. reniformis on day 0 or day 14 depending on the experiment. Experiments were conducted on cotton cultivar DP 0935 B2RF (susceptible to both nematodes), LONREN-1 (germplasm line resistant to R. reniformis), and M-120 RNR (germplasm line resistant to M. incognita), and tests were terminated 8 wk after the last inoculation. Both soil (vermiform) and roots (egg) extracted from each half of the root system to determine the total nematode population levels, and root galling was rated on a 0 to 10 scale. Mixed models analysis and comparison of least squares means indicated no differences in root galling (except on LONREN-1) or population levels when the two nematode species were introduced on the same day. When M. incognita was introduced 14 d after R. reniformis, reduction in galling (36% on DP 0935 and 33% on LONREN-1) and M. incognita population levels (35% on DP 0935 and 45% on LONREN-1) were significant (P ≤ 0.05). When R. reniformis was inoculated 14 d after M. incognita, reduction in R. reniformis population levels (18% on DP 0935 and 26% on M-120) were significant. This study documents for the first time that infection of cotton by a nematode can elicit SAR to another nematode species.

Influence of infection of cotton by rotylenchulus reniformis and meloidogyne incognita on the production of enzymes involved in systemic acquired resistance.

Systemic acquired resistance (SAR), which results in enhanced defense mechanisms in plants, can be elicited by virulent and avirulent strains of pathogens including nematodes. Recent studies of nematode reproduction strongly suggest that Meloidogyne incognita and Rotylenchulus reniformis induce SAR in cotton, but biochemical evidence of SAR was lacking. Our objective was to determine whether infection of cotton by M. incognita and R. reniformis increases the levels of P-peroxidase, G-peroxidase, and catalase enzymes which are involved in induced resistance. A series of greenhouse trials was conducted; each trial included six replications of four treatments applied to one of three cotton genotypes in a randomized complete block design. The four treatments were cotton plants inoculated with i) R. reniformis, ii) M. incognita, iii) BTH (Actigard), and iv) a nontreated control. Experiments were conducted on cotton genotypes DP 0935 B2RF (susceptible to both nematodes), LONREN-1 (resistant to R. reniformis), and M-120 RNR (resistant to M. incognita), and the level of P-peroxidase, G-peroxidase, and catalase activity was measured before and 2, 4, 6, 10, and 14 d after treatment application. In all cotton genotypes, activities of all three enzymes were higher (P ≤ 0.05) in leaves of plants infected with M. incognita and R. reniformis than in the leaves of control plants, except that M. incognita did not increase catalase activity on LONREN-1. Increased enzyme activity was usually apparent 6 d after treatment. This study documents that infection of cotton by M. incognita or R. reniformis increases the activity of the enzymes involved in systemic acquired resistance; thereby providing biochemical evidence to substantiate previous reports of nematode-induced SAR in cotton.

Evaluation of an antibiotic-producing strain of Pseudomonas fluorescens for suppression of plant-parasitic nematodes.

The antibiotic 2,4-diacetylphloroglucinol (DAPG), produced by some strains of Pseudomonas spp., is involved in suppression of several fungal root pathogens as well as plant-parasitic nematodes. The primary objective of this study was to determine whether Wood1R, a D-genotype strain of DAPG-producing P. fluorescens, suppresses numbers of both sedentary and migratory plant-parasitic nematodes. An experiment was conducted in steam-heated soil and included two seed treatments (with Wood1R and a control without the bacterium) and six plant-nematode combinations which were Meloidogyne incognita on cotton, corn, and soybean; M. arenaria on peanut; Heterodera glycines on soybean; and Paratrichodorus minor on corn. Wood 1R had no effect on final numbers of M. arenaria, P. minor, or H. glycines; however, final numbers of M. incognita were lower when seeds were treated with Wood1R than left untreated, and this reduction was consistent among host plants. Population densities of Wood1R were greater on the roots of corn than on the other crops, and the bacterium was most effective in suppressing M. incognita on corn, with an average reduction of 41%. Despite high population densities of Wood1R on corn, the bacterium was not able to suppress numbers of P. minor. When comparing the suppression of M. incognita on corn in natural and steam-heated soil, egg production by the nematode was suppressed in natural compared to steamed soil, but the presence of Wood1R did not result in additional suppression of the nematodes in the natural soil. These data indicate that P. fluorescens strain Wood1R has the capacity to inhibit some populations of plant-parasitic nematodes. However, consistent suppression of nematodes in natural soils seems unlikely.

Population dynamics of Meloidogyne arenaria and Pasteuria penetrans in a long-term crop rotation study.

The endospore-forming bacterium Pasteuria penetrans is an obligate parasite of root-knot nematodes (Meloidogyne spp.). The primary objective of this study was to determine the effect of crop sequence on abundance of P. penetrans. The experiment was conducted from 2000 to 2008 at a field site naturally infested with both the bacterium and its host Meloidogyne arenaria and included the following crop sequences: continuous peanut (Arachis hypogaea) (P-P-P) and peanut rotated with either 2 years of corn (Zea mays) (C-C-P), 1 year each of cotton (Gossypium hirsutum) and corn (Ct-C-P), or 1 year each of corn and a vegetable (V-C-P). The vegetable was a double crop of sweet corn and eggplant (Solanum melongena). A bioassay with second-stage juveniles (J2) of M. arenaria from a greenhouse (GH) population was used to estimate endospore abundance under the different crop sequences. A greater numerical increase in endospore densities was expected in the P-P-P and V-C-P sequences than in the other sequences because both peanut and eggplant are good hosts for M. arenaria. However, endospore densities, as determined by bioassay, did not substantially increase in any of the sequences during the 9-year experiment. To determine whether the nematode population had developed resistance to the resident P. penetrans, five single egg-mass (SEM) lines from the field population of M. arenaria were tested alongside the GH population for acquisition of endospores from the field soil. Four of the five SEM lines acquired 9 to 14 spores/J2 whereas the GH population and one of the SEM lines acquired 3.5 and 1.8 spores/J2, respectively. Endospore densities estimated with the four receptive SEM lines were highest in the P-P-P plots (14-20 spores/J2), intermediate in the V-C-P plots (6-7 spores/J2), and lowest in the Ct-C-P plots (< 1 spore/J2). These results indicate that the field population of M. arenaria is heterogeneous for attachment of P. penetrans endospores. Moreover, spore densities increased under intensive cropping of hosts for M. arenaria, but the GH population of the nematode was not receptive to spore attachment. However, previously, the GH population was very receptive to spore acquisition from this field site. One explanation for this inconsistency is that the M. arenaria population in the field became resistant to the dominant subpopulation of P. penetrans that had been present, and this led to the selection of a different subpopulation of the bacterium that is incompatible with the GH population.

Comparison of Methods for Assessing Resistance to Meloidogyne arenaria in Peanut.

Use of resistant cultivars is a desirable approach to manage the peanut root-knot nematode (Meloidogyne arenaria). To incorporate resistance into commercially acceptable cultivars requires reliable, efficient screening methods. To optimize the resistance screening protocol, a series of greenhouse tests were done using seven genotypes with three levels of resistance to M. arenaria. The three resistance levels could be separated based on gall indices as early as two weeks after inoculation (WAI) using 8,000 eggs of M. arenaria per plant, while four or more weeks were needed when 1,000-6,000 eggs/plant were used. High inoculum densities (over 8,000 eggs/plant) were needed to separate the three resistance levels based on eggs per gram of root within eight WAI. A gall index based on percentage of galled roots could separate the three resistance levels at lower inoculum levels and earlier harvest dates than other assessment methods. The use of eggs vs. second-stage juveniles (J2) as inoculum provided similar results; however, it took three to five more days to collect J2 than to collect eggs from roots. Plant age affected gall index and nematode reproduction on peanut, especially on the susceptible genotypes AT201 and D098. The genotypes were separated into their correct resistance classes when inoculated 10 to 30 days after planting, but were not separated correctly when inoculated on day 40.

Reproduction of Meloidogyne incognita on Winter Cover Crops Used in Cotton Production.

Substantial reproduction of Meloidogyne incognita on winter cover crops may lead to damaging populations in a subsequent cotton (Gossypium hirsutum) crop. The amount of population increase during the winter depends on soil temperature and the host status of the cover crop. Our objectives were to quantify M. incognita race 3 reproduction on rye (Secale cereale) and several leguminous cover crops and to determine if these cover crops increase population densities of M. incognita and subsequent damage to cotton. The cover crops tested were 'Bigbee' berseem clover (Trifolium alexandrinum), 'Paradana' balansa clover (T. balansae), 'AU Sunrise' and 'Dixie' crimson clover (T. incarnatum), 'Cherokee' red clover (T. pratense), common and 'AU Early Cover' hairy vetch (Vicia villosa), 'Cahaba White' vetch (V. sativa), and 'Wrens Abruzzi' rye. In the greenhouse tests, egg production was greatest on berseem clover, Dixie crimson clover, AU Early Cover hairy vetch, and common hairy vetch; intermediate on Balansa clover and AU Sunrise crimson clover; and least on rye, Cahaba White vetch, and Cherokee red clover. In both 2002 and 2003 field tests, enough heat units were accumulated between 1 January and 20 May for the nematode to complete two generations. Both AU Early Cover and common hairy vetch led to greater root galling than fallow in the subsequent cotton crop; they also supported high reproduction of M. incognita in the greenhouse. Rye and Cahaba White vetch did not increase root galling on cotton and were relatively poor hosts for M. incognita. Only those legumes that increased populations of M. incognita reduced cotton yield. In the southern US, M. incognita can complete one to two generations on a susceptible winter cover crop, so cover crops that support high nematode reproduction may lead to damage and yield losses in the following cotton crop. Planting rye or Meloidogyne-resistant legumes as winter cover crops will lower the risk of increased nematode populations compared to most vetches and clovers.