Biochemical and nanotechnological approaches to combat phytoparasitic nematodes.

The foundation of most food production systems underpinning global food security is the careful management of soil resources. Embedded in the concept of soil health is the impact of diverse soil-borne pests and pathogens, and phytoparasitic nematodes represent a particular challenge. Root-knot nematodes and cyst nematodes are severe threats to agriculture, accounting for annual yield losses of US$157 billion. The control of soil-borne phytoparasitic nematodes conventionally relies on the use of chemical nematicides, which can have adverse effects on the environment and human health due to their persistence in soil, plants, and water. Nematode-resistant plants offer a promising alternative, but genetic resistance is species-dependent, limited to a few crops, and breeding and deploying resistant cultivars often takes years. Novel approaches for the control of phytoparasitic nematodes are therefore required, those that specifically target these parasites in the ground whilst minimizing the impact on the environment, agricultural ecosystems, and human health. In addition to the development of next-generation, environmentally safer nematicides, promising biochemical strategies include the combination of RNA interference (RNAi) with nanomaterials that ensure the targeted delivery and controlled release of double-stranded RNA. Genome sequencing has identified more than 75 genes in root knot and cyst nematodes that have been targeted with RNAi so far. But despite encouraging results, the delivery of dsRNA to nematodes in the soil remains inefficient. In this review article, we describe the state-of-the-art RNAi approaches targeting phytoparasitic nematodes and consider the potential benefits of nanotechnology to improve dsRNA delivery.

Impacts of the Ban on the Soil-Applied Fumigant Methyl Bromide.

The loss of the soil fumigant methyl bromide (MeBr) and adoption of soil fumigant alternatives has been challenging for farmers, particularly for those crops in which pathogens previously controlled by MeBr have emerged as significant problems, but it has resulted in some unanticipated benefits for the scientific community and the environment. Applauded as one of the most effective environmental agreements to date, the universally accepted Montreal Protocol on Ozone Depleting Substances has had a significant impact on the environment, reducing the release of halogenated compounds from anthropogenic sources enough to mitigate global warming by an estimated 1.1°C by 2021. The funding associated with various MeBr transition programs has increased collaboration across scientific disciplines, commodity groups, industry, and regulatory agencies. Chemical alternatives and improved application strategies, including the development of gas-retentive agricultural films, coupled with sound efficacy data and grower ingenuity have resulted in the sustained production of many of the impacted crops; although there has been some loss of acreage and value, particularly for Florida fumigated crops, for some, value has continued to increase, allowing production to continue. The loss of a single, broad-spectrum tool for pest control has led to a deeper understanding of the specific pest complexes impacting these at-risk crops, as well as the development of new, biologically based management tools for their control while increasing our understanding of the role of the soil microbiome in pest control and crop production.

Application of Thymol Vapors to Control Postharvest Decay Caused by Penicillium digitatum and Lasiodiplodia theobromae in Grapefruit.

Two of the major postharvest diseases impacting grapefruit shelf life and marketability in the state of Florida (USA) are stem-end rot (SER) caused by Lasiodiplodia theobromae and green mold (GM) caused by Penicillium digitatum. Here, we investigated the in vitro and in vivo efficacy of vapors of thymol, a natural compound found in the essential oil of various plants and the primary constituent of thyme (Thymus vulgaris) oil, as a potential solution for the management of GM and SER. Thymol vapors at concentrations lower than 10 mg L-1 significantly inhibited the mycelial growth of both pathogens, causing severe ultrastructural damage to P. digitatum conidia. In in vivo trials, the incidence and lesion area of GM and SER on inoculated grapefruit were significantly reduced after a 5 d exposure to 50 mg L-1 thymol vapors. In addition, the in vitro and in vivo sporulation of P. digitatum was suppressed by thymol. When applied in its vapor phase, thymol had no negative effect on the fruit, neither introducing perceivable off-flavor nor causing additional weight loss. Our findings support the pursuit of further studies on the use of thymol, recognized as safe for human health and the environment, as a promising strategy for grapefruit postharvest disease management.

Yield performance, mineral profile, and nitrate content in a selection of seventeen microgreen species.

Introduction: Originally regarded as garnish greens, microgreens are increasingly valued for their nutritional profile, including their mineral content. Methods: A study was conducted under controlled environmental conditions utilizing a selection of seventeen microgreen species belonging to seven different botanical families to investigate the genetic variation of macro- and micro-minerals and nitrate (NO3 -) content. Plants were grown in a soilless system using a natural fiber mat as the substrate. After germination, microgreens were fertigated with a modified half-strength Hoagland solution prepared using deionized water and without adding microelements. At harvest (10 to 19 days after sowing, based on the species), yield components were measured and dry tissue samples were analyzed for the concentration of total nitrogen (N), NO3 -, P, K, Ca, Mg, S, Na, Fe, Zn, Mn, Cu, and B. Results and discussion: Genotypic variations were observed for all of the examined parameters. Nitrogen and K were the principal macronutrients accounting for 38.4% and 33.8% of the total macro-minerals concentration, respectively, followed in order by Ca, P, S, and Mg. Except for sunflower (Helianthus annuus L.), all the tested species accumulated high (1,000-2,500 mg kg-1 FW) or very high (>2,500 mg kg-1 FW) NO3 - levels. Eight of the studied species had a K concentration above 300 mg 100 g-1 FW and could be considered as a good dietary source of K. On the other hand, scallion (Allium fistulosum L.), red cabbage (Brassica oleracea L. var. capitata), amaranth (Amaranthus tricolor L.), and Genovese basil (Ocinum basilicum L.) microgreens were a good source of Ca. Among micro-minerals, the most abundant was Fe followed by Zn, Mn, B, and Cu. Sunflower, scallion, and shiso (Perilla frutescens (L.) Britton) were a good source of Cu. Moreover, sunflower was a good source of Zn, whereas none of the other species examined could be considered a good source of Fe and Zn, suggesting that supplementary fertilization may be required to biofortify microgreens with essential microminerals. In conclusion, the tested microgreens can be a good source of minerals showing a high potential to address different dietary needs; however, their yield potential and mineral profile are largely determined by the genotype.

Soil Amendment and Storage Effect the Quality of Winter Melons (Benincasa hispida (Thunb) Cogn.) and Their Juice.

Winter melon fruits were grown in the field using anaerobic soil disinfestation (ASD) and conventional fertilizer alone as the control treatment. Fruits were harvested and stored at 20 °C for 120 d, the juice was processed on day one and day 120, and the effects of soil amendment and 120 d storage on the juice's physical and chemical (sugars, acids, volatile and nutritional compounds) properties were evaluated. Fruit juice extracted from ASD-grown fruit had greater magnitude of zeta potential than the control juice, indicating it was physically more stable than the juice obtained from the control conditions. ASD fruit juice had lower soluble solids content (SSC), and lower volatile compounds that contribute green, grass, and sulfur notes, and negatively influence flavor quality. ASD fruit juice had higher vitamin B5 and cytidine. Juice processed from 120 d stored fruit had less yield due to 12.4-15.6% weight loss. The non-soluble solids content was higher and particle size was larger, and the SSC and individual sugars decreased. However, titratable acidity (TA) increased primarily due to increased citric acid. Out of 16 free amino acids, 6 increased and only 1 decreased. However, three out of five nucleosides decreased; vitamins B1 and B6 increased; vitamins B2, B3 and C decreased. Overall, juice derived from fruit produced using ASD was physically more stable and had less SSC and off-odor volatiles than the control, while the fruit juice of those stored for 120 d had lower SSC and higher TA and nutritional profiles, comparable to freshly harvested fruit.

Quantifying the effects of anaerobic soil disinfestation and other biological soil management strategies on nitrous oxide emissions from raised bed plasticulture tomato production.

Soilborne pests are a major obstacle that must be overcome for the production of horticultural crops. Methyl bromide (MBr) was an effective preplanting soil broad-spectrum biocide, but its use has been banned due to its role in depleting the ozone layer. As a result, sustainable alternative methods for controlling soilborne pathogens and pests are needed. Nitrous oxide (N2 O) emissions are of concern in crop production due to the role of N2 O as a greenhouse gas. Agricultural lands are known sources for emission of N2 O into the atmosphere. Emissions are related to many environmental factors as well as fertilization and fumigation practices. This study evaluated the influence of different alternatives to MBr on N2 O emissions throughout a tomato production season in two locations representative of southern and northern Florida. We evaluated eight soil management practices, including (a) untreated controls; (b) chemical soil fumigation; (c) anaerobic soil disinfestation using molasses (M) + composted poultry litter and (d and e) M + composted yard waste (CYW, at two rates); (f) Soil Symphony Amendment (SSA), a commercially available mix of microbes and nutrients; (g) CYW alone; and (h) CYW + SSA. Nitrous oxide emissions were measured throughout the cropping season. Emissions were highest on the day of planting (Day 21), ranging from 213 to 1,878 µg m-2 h-1 , likely due to the release of N2 O that had accumulated under the totally impermeable film when it was punctured for planting. However, statistical significance varied between sites. Estimated cumulative emissions of N2 O throughout the production season ranged from 1.3 to 4.8 kg N2 O-N ha-1 .

Effects of Harvest Maturity, Refrigeration and Blanching Treatments on the Volatile Profiles of Ripe "Tasti-Lee" Tomatoes.

The interactive effects of six maturity stages and refrigerated storage (chilling)/blanching (heating) treatments on the volatile profiles of ripe tomatoes were studied. A total of 42 volatiles were identified, of which 19 compounds had odor activity values equal to or greater than 1. Of those, "green" and "leafy" aroma volatiles were most abundant. Chilling and heating treatments both suppressed overall volatile production, with chilling having the greater impact, regardless of harvest maturity. However, fruit harvested at the turning stage had the least volatile suppression by chilling and heating treatments in comparison with fruit harvested earlier or later, mostly in the fatty acid- and phenylalanine-derived volatiles. Volatiles derived from amino acids were promoted by heat treatment for fruit harvested at all maturities, and those derived from carotenoid and phenylalanine pathways and harvested at advanced harvest maturities were stimulated by chilling treatment. Volatile production is generally believed to be improved by delayed harvest, with vine-ripe being optimum. However, opposite results were observed possibly because the later-harvested fruit had longer exposure to open-field weather stress. The best harvest maturity recommendation is the turning stage where fruit developed abundant volatiles and were least impacted by chilling and heating treatments.

Anaerobic Soil Disinfestation Efficacy Against Fusarium oxysporum Is Affected by Soil Temperature, Amendment Type, Rate, and C:N Ratio.

A meta-analysis of anaerobic soil disinfestation (ASD) efficacy against Fusarium oxysporum and F. oxysporum f. sp. lycopersici was conducted emphasizing effects of environment and organic amendment characteristics and pot and field studies conducted on ASD amendment C:N ratio and soil temperature effects on F. oxysporum f. sp. lycopersici inoculum survival. In a pot study, two organic amendments, dry molasses-based or wheat bran-based, applied at 4 mg of C/g of soil, with 40:1, 30:1, 20:1, and 10:1 C:N ratios, were evaluated against F. oxysporum f. sp. lycopersici at 15 to 25°C. This study was followed by a pot study with temperature regimes of 15 to 25°C and 25 to 35°C and two C:N ratios (20:1 and 40:1), and a field study at 40:1, 30:1, 20:1, and 10:1 C:N ratios, a 30:1 C:N ratio at a lower C rate (2 mg of C/g of soil), and an anaerobic control. Soil temperature >25°C and more labile amendments increased ASD suppression of F. oxysporum/F. oxysporum f. sp. lycopersici in the meta-analysis. In pot studies, F. oxysporum f. sp. lycopersici survival was reduced for molasses-based mixtures at 20:1 and 30:1 C:N ratios compared with wheat bran-based mixtures but not compared with the anaerobic control. At 25 to 35°C, all ASD treatments suppressed F. oxysporum f. sp. lycopersici relative to controls. In the field, all ASD treatments reduced F. oxysporum f. sp. lycopersici survival compared with the anaerobic control, and 4 mg of C/g of soil amendment rates induced higher anaerobic conditions and higher F. oxysporum f. sp. lycopersici mortality compared with the 2 mg of C/g of soil rate. Although amendment C:N ratios from 10 to 40:1 were similarly suppressive of F. oxysporum, lower temperatures reduced ASD effectiveness against F. oxysporum/F. oxysporum f. sp. lycopersici and further work is warranted to enhance suppression at soil temperatures <25°C.

Organic Amendments for Pathogen and Nematode Control.

The loss of methyl bromide as a soil fumigant and minimal advances in the development and registration of new chemical fumigants has resulted in a resurgence of interest in the application of organic amendments (OAs) for soilborne plant pathogen and plant-parasitic nematode management. Significant progress has been made in the characterization of OAs, application of strategies for their use, and elucidation of mechanisms by which they suppress soilborne pests. Nonetheless, their utility is limited by the variability of disease control, expense, and the logistics of introducing them into crop production systems. Recent advances in molecular techniques have led to significant progress in the elucidation of the role of bacteria and fungi and their metabolic products on disease suppression with the addition of OAs. Biosolarization and anaerobic soil disinfestation, developed to manipulate systems and favor beneficial microorganisms to maximize their impact on plant pathogens, are built on a strong historical research foundation in OAs and the physical, chemical, and biological characteristics of disease-suppressive soils. This review focuses on recent applications of OAs and their potential for the management of soilborne plant pathogens and plant-parasitic nematodes, with emphasis primarily on annual fruit and vegetable production systems.

Dissipation of fomesafen in fumigated, anaerobic soil disinfestation-treated, and organic-amended soil in Florida tomato production systems.

BACKGROUND: Fumigated, anaerobic soil disinfestation-treated (ASD), and organic-amended soil management strategies have been investigated as potential methyl bromide (MBr) alternatives for controlling diseases, nematodes, and weeds in soil. Nutsedge and broadleaf weed control using fomesafen has been reported to be comparable to MBr in normal cropping systems. There is no information on the fate of fomesafen used in combination with alternative practices. In this study, the fate of fomesafen in these alternative systems was measured by liquid chromatography-tandem mass spectrometry (LC/MS-MS) following extraction using a modified Quick Easy Cheap Effective Safe (QuEChERS) method. RESULTS: The reported half-life (DT50 ) values for fomesafen in the top 15 cm of soil were from 62.9 to 107.3 days. The DT50 values in organic-amended soil were higher than in ASD-treated soil in the top 15 cm. For all treatments, reductions in concentrations were positively correlated with lower redox potentials and organic matter content. Some leaching of fomesafen into the 16-30 cm zone was observed in all treatments. CONCLUSIONS: The DT50 values in this study were generally higher than those reported in previous studies performed at different locations. Due to increased losses of the herbicide and subsequent reduction in weed control, fomesafen is likely not to be suitable for effective weed control in systems using ASD techniques employing composted poultry litter and molasses. Integration of fomesafen using composted yard waste 1 (CYW1) and Soil Symphony Amendment (SSA) may result in acceptable weed control. Given that the soil was very sandy and the pH was higher than the pKa, fomesafen might leach deeper than 30 cm, particularly with the use of chemical soil fumigants (CSFs). © 2019 Society of Chemical Industry.

Evaluation of Steam and Soil Solarization for Meloidogyne arenaria Control in Florida Floriculture Crops.

Steam and soil solarization were investigated for control of the root-knot nematode Meloidogyne arenaria in 2 yr of field trials on a commercial flower farm in Florida. The objective was to determine if preplant steam treatments in combination with solarization, or solarization alone effectively controlled nematodes compared to methyl bromide (MeBr). Trials were conducted in a field with naturally occurring populations of M. arenaria. Treatments were solarization alone, steam treatment after solarization using standard 7.6-cm-diameter perforated plastic drain tile (steam 1), steam treatment following solarization using custom-drilled plastic drain tile with 1.6-mm holes spaced every 3.8 cm (steam 2), and MeBr applied at 392 kg/ha 80:20 MeBr:chloropicrin. Drain tiles were buried approximately 35 cm deep with four tiles per 1.8 by 30 m plot. Steam application followed a 4-wk solarization period concluding in mid-October. All steam was generated using a Sioux propane boiler system. Plots were steamed for sufficient time to reach the target temperature of 70°C for 20 min. Solarization plastic was retained on the plots during steaming and plots were covered with a single layer of carpet padding to provide additional insulation. The floriculture crops larkspur (Delphinium elatum and Delphinium × belladonna), snapdragon (Antirrhinum majus), and sunflower (Helianthus annuus) were produced according to standard commercial practices. One month after treatment in both years of the study, soil populations of M. arenaria were lower in both steam treatments and in MeBr compared to solarization alone. At the end of the season in both years, galling on larkspur, snapdragon, and sunflowers was lower in both steam treatments than in solarization. Both steam treatments also provided control of M. arenaria in soil at the end of the season comparable to, or exceeding that provided by MeBr. Both steam treatments also reduced M. arenaria in snapdragon roots comparable to, or exceeding control with MeBr. Meloidogyne arenaria in soil increased in solarization alone. Solarization alone also had higher gall ratings on larkspur, snapdragon, and sunflower than all other treatments. Steam provided excellent control of M. arenaria in this study.

Grafting and Paladin Pic-21 for Nematode and Weed Management in Vegetable Production.

Two years of field trials conducted in a Meloidogyne incognita-infested field evaluated grafting and Paladin Pic-21 (dimethyl disulfide:chloropicrin [DMDS:Pic] 79:21) for root-knot nematode and weed control in tomato and melon. Tomato rootstocks evaluated were; 'TX301', 'Multifort', and 'Aloha'. 'Florida 47' was the scion and the nongrafted control. A double crop of melon was planted into existing beds following tomato harvest. Melon rootstocks, C. metulifer and 'Tetsukabuto', were evaluated with nongrafted 'Athena' in year 1. In year 2, watermelon followed tomato with scion variety 'Tri-X Palomar' as the control and also grafted onto 'Emphasis' and 'Strongtosa' rootstocks. Four soil treatments were applied in fall both years under Canslit metalized film; Paladin Pic-21, methyl bromide:chloropicrin (MeBr:C33, 67:33), Midas (iodomethane:chloropicrin 50:50), and a herbicide-treated control. M. incognita J2 in soil were highest in herbicide control plots and nongrafted tomato. All soil treatments produced similar tomato growth, which was greater than the herbicide control. All treatments reduced M. incognita J2 in roots compared to the herbicide control. 'Multifort' rootstock produced the largest and healthiest roots; however, the number of M. incognita isolated from roots did not differ among the tomato rootstocks tested. Galling on tomato was highest in herbicide control plots and nongrafted plants. In melon, M. incognita J2 in soil did not differ among melon rootstocks, but numbers isolated from melon rootstocks increased in 'Tetsukabuto' compared with C. metuliferus. 'Tetsukabuto' were larger root systems than nongrafted 'Athena'. All fumigants provided protection for all melon rootstocks against galling by M. incognita compared to the herbicide control. Galling on C. metuliferus rootstock was less in all fumigant treatments compared with nongrafted 'Athena' and 'Tetsukabuto'. In watermelon, M. incognita in soil and roots did not differ among soil treatments or watermelon rootstocks, and yield was lower in both grafted rootstocks compared with the nongrafted control. All soil treatments increased average fruit weight of watermelon compared with the herbicide control, and provided effective weed control, keeping the most predominant weed, purple nutsedge (Cyperus rotundus L.), density at or below 1/m row. Grafting commercial scions onto M. incognita-resistant rootstocks has potential for nematode management combined with soil treatments or as a stand-alone component in crop production systems.

Deep Characterization of the Microbiomes of Calophya spp. (Hemiptera: Calophyidae) Gall-Inducing Psyllids Reveals the Absence of Plant Pathogenic Bacteria and Three Dominant Endosymbionts.

Bacteria associated with sap-feeding insect herbivores include not only symbionts that may increase their hosts' fitness but also harmful plant pathogens. Calophya spp. gall-inducing psyllids (Hemiptera: Calophyidae) are being investigated for their potential as biological control agents of the noxious weed, Brazilian peppertree (Schinus terebinthifolia), in Florida. Although there are no examples of plant pathogen transmission by members of the family Calophyidae, several insects in the superfamily Psylloidea are known to transmit pathogenic bacteria in the genera Candidatus Liberibacter and Candidatus Phytoplasma. To determine whether Calophya spp. harbor potentially harmful plant pathogenic bacteria, we sequenced small subunit (SSU) ribosomal RNA (rRNA) gene amplicons generated from individuals from four Calophya spp. populations: All microbial SSU gene sequences fell into the bacterial domain, with 98-99% belonging to the Proteobacteria. The Calophya microbiomes contained a relatively simple community, with 49-79 operational taxonomic units (OTUs; 97%) detected, and only 5-8 OTUs with greater than 1% abundance. Candidatus Carsonella showed the highest relative abundance, with OTUs from this candidate genus representing between 51-65% of all recovered sequences. The next most abundant clade observed was an unclassified Enterobacteriacae group closely related to bacteria from the genera Buchnera and Blochmannia that ranged from 20-31% in relative abundance. Wolbachia populations were the third most abundant group and represented 7-27% of the diversity in microbial OTUs. No SSU rRNA gene sequences from putative pathogenic bacteria from the genera Ca. Liberibacter or Ca. Phytoplasma were detected in the microbiomes of the four Calophya populations. The probability that infected psyllids were present in our colonies, but were not sampled, was extremley low (1.39 x 10(-10)). As far as we are aware, our study is the first to characterize the microbiome of a candidate biological control agent, and coupled with previous work demonstrating a high degree of host specificity and absence of plant viruses, suggests that releasing Calophya spp. in United States poses minimal risk to non-target plants.

The effect of transitional organic production practices on soilborne pests of tomato in a simulated microplot study.

The perceived risk of pest resurgence upon transition from conventional to organic-based farming systems remains a critical obstacle to expanding organic vegetable production, particularly where chemical fumigants have provided soilborne pest and disease control. Microplots were used to study the effects of soil amendments and cropping sequences applied over a 2-year transitional period from conventional to organic tomato (Solanum lycopersicum) cultivation on the incidence of bacterial wilt caused by Ralstonia solanacearum, purple nutsedge (Cyperus rotundus) reproduction, root galling by Meloidogyne incognita, and soil nematode populations. A continuation of tomato monoculture during the transitional period resulted in a disease incidence of 33%, as compared with 9% in microplots that were rotated with sunn hemp (Crotalaria juncea) and Japanese millet (Echinochloa crusgalli var. frumentacea). The benefits of disease control from a crop rotation extended into to a second season of organic tomato cultivation season, where bacterial wilt declined from 40% in microplots with a tomato monoculture to 17% in plots with a crop rotation sequence. Combining applications of urban plant debris with a continued tomato monoculture increased the incidence of bacterial wilt to 60%. During the transition period, tomato plants following a cover crop regime also had significantly lower levels of root galling from root-knot nematode infection compared with plants in the continuous tomato monoculture. Nutsedge tuber production was significantly increased in plots amended with broiler litter but not urban plant debris. Compared with a continuous monoculture, the results illustrate the importance of a systems-based approach to implementing transitional organic practices that is cognizant of their interactive effects on resident soilborne disease, weed, and pest complexes.

Evaluation of Cover Crops with Potential for Use in Anaerobic Soil Disinfestation (ASD) for Susceptibility to Three Species of Meloidogyne.

Several cover crops with potential for use in tropical and subtropical regions were assessed for susceptibility to three common species of root-knot nematode, Meloidogyne arenaria, M. incognita, and M. javanica. Crops were selected based on potential use as organic amendments in anaerobic soil disinfestation (ASD) applications. Nematode juvenile (J2) numbers in soil and roots, egg production, and host plant root galling were evaluated on arugula (Eruca sativa, cv. Nemat), cowpea (Vigna unguiculata, cv. Iron & Clay), jack bean (Canavalia ensiformis, cv. Comum), two commercial mixtures of Indian mustard and white mustard (Brassica juncea & Sinapis alba, mixtures Caliente 61 and Caliente 99), pearl millet (Pennisetum glaucum, cv. Tifleaf III), sorghum-sudangrass hybrid (Sorghum bicolor × S. bicolor var. sudanense, cv. Sugar Grazer II), and three cultivars of sunflower (Helianthus annuus, cvs. 545A, Nusun 660CL, and Nusun 5672). Tomato (Solanum lycopersicum, cv. Rutgers) was included in all trials as a susceptible host to all three nematode species. The majority of cover crops tested were less susceptible than tomato to M. arenaria, with the exception of jack bean. Sunflower cv. Nusun 5672 had fewer M. arenaria J2 isolated from roots than the other sunflower cultivars, less galling than tomato, and fewer eggs than tomato and sunflower cv. 545A. Several cover crops did not support high populations of M. incognita in roots or exhibit significant galling, although high numbers of M. incognita J2 were isolated from the soil. Arugula, cowpea, and mustard mixture Caliente 99 did not support M. incognita in soil or roots. Jack bean and all three cultivars of sunflower were highly susceptible to M. javanica, and all sunflower cultivars had high numbers of eggs isolated from roots. Sunflower, jack bean, and both mustard mixtures exhibited significant galling in response to M. javanica. Arugula, cowpea, and sorghum-sudangrass consistently had low numbers of all three Meloidogyne species associated with roots and are good selections for use in ASD for root-knot nematode control. The remainder of crops tested had significant levels of galling, J2, and eggs associated with roots, which varied among the Meloidogyne species tested.

Susceptibility of Several Common Subtropical Weeds to Meloidogyne arenaria, M. incognita, and M. javanica.

Experiments were conducted in the greenhouse to assess root galling and egg production of three root-knot nematode species, Meloidogyne arenaria, M. incognita, and M. javanica, on several weeds common to Florida agricultural land. Weeds evaluated were Amaranthus retroflexus (redroot pigweed), Cyperus esculentus (yellow nutsedge), Eleusine indica (goosegrass), Portulaca oleracea (common purslane), and Solanum americanum (American black nightshade). Additionally, although it is recommended as a cover crop in southern regions of the U.S., Aeschynomene americana (American jointvetch) was evaluated as a weed following the detection of root galling in a heavy volunteer infestation of an experimental field in southeastern Florida. Weeds were propagated from seed and inoculated with 1000 nematode eggs when plants reached the two true-leaf stage. Tomato (Solanum lycopersicum 'Rutgers') was included as a positive control. Aeschynomene americana and P. oleracea roots supported the highest number of juveniles (J2) and had the highest number of eggs/g of root for all three species of Meloidogyne tested. However, though P. oleracea supported very high root levels of the three nematode species tested, its fleshy roots did not exhibit severe gall symptoms. Low levels of apparent galling, combined with high egg production, increase the potential for P. oleracea to support populations of these three species of root-knot nematodes to a degree that may not be appropriately recognized. This research quantifies the impact of P. oleracea as a host for M. arenaria, M. incognita, and M. javanica compared to several other important weeds commonly found in Florida agricultural production, and the potential for A. americana to serve as an important weed host of the three species of root-knot nematode tested in southern regions of Florida.

Differential performance of tropical soda apple and its biological control agent Gratiana boliviana (Coleoptera: Chrysomelidae) in open and shaded habitats.

The leaf feeding beetle Gratiana boliviana Spaeth has been released since 2003 in the southeastern United States for biological control of tropical soda apple, Solanum viarum Dunal. In Florida, G. boliviana can be found on tropical soda apple growing in open pastures as well as in shady wooded areas. The objectives of this study were to determine the effect of light intensity on the performance of tropical soda apple and G. boliviana under greenhouse conditions, and to determine the abundance and mortality of G. boliviana in open and shaded habitats. Leaves growing in the shade were less tough, had higher water and nitrogen content, lower soluble sugars, and less dense and smaller glandular trichomes compared with leaves growing in the open. Plants grew slightly taller and wider under shaded conditions but total biomass was significantly reduced compared with plants grown in the open. In the greenhouse, G. boliviana had higher immature survival, greater folivory, larger adult size, and higher fecundity when reared on shaded plants compared with open plants. Sampling of field populations revealed that the overall abundance of G. boliviana was lower but leaf feeding damage was higher in shaded habitats compared with the open habitats. The percentage of eggs surviving to adult was greater in shaded compared with open habitats. The abundance of predators was higher in the open pasture and was positively correlated with the abundance of G. boliviana. These results indicate that not only plant quality but also habitat structure are important to the performance of weed biological control agents.

Microplot Evaluation of Rootstocks for Control of Meloidogyne incognita on Grafted Tomato, Muskmelon, and Watermelon.

Microplot experiments were conducted over two years (four growing seasons) to evaluate Meloidogyne incognita resistance in rootstocks used for grafted tomato (Solanum lycopersicum), muskmelon (Cucumis melo), and watermelon (Citrullus lanatus). Three tomato rootstocks; 'TX301', 'Multifort', and 'Aloha', were tested in addition to the nongrafted scion, 'Florida-47'. Two muskmelon rootstocks; Cucumis metuliferus and 'Tetsukabuto' (Cucurbita maxima × Cucurbita moschata) were evaluated with the nongrafted scion 'Athena'. Two watermelon rootstocks included 'Emphasis', a lagenaria-type, and an interspecific squash hybrid 'StrongTosa', which were grafted to the scion 'TriX Palomar' and planted only in the second year. Microplots were infested with M. incognita eggs in September each year. Tomatoes were planted in September followed by melons in March. In both years of the study, M. incognita juveniles (J2) in soil were similar among all tomato rootstocks, but numbers in roots were higher in the nongrafted Florida 47 than in all grafted rootstocks. In muskmelon only C. metuliferus rootstock reduced galling in nematode infested soil. Tetsukabuto did not reduce numbers of M. incognita J2 in either soil or roots either year. There were no differences in nematode numbers, galling, or plant growth parameters among the watermelon rootstocks tested. The use of resistant rootstocks has great potential for improving nematode control in the absence of soil fumigants.

A Nylon Membrane Bag Assay for Determination of the Effect of Chemicals on Soilborne Plant Pathogens in Soil.

A new nylon membrane bag (NMB) assay was developed for studies to determine the effect of chemicals added to soil on survival of soilborne plant pathogens. The rapid and effective assay can be used to study organisms for which there are no selective media or for which a selective medium is expensive or difficult to prepare. This assay consists of placing pathogens inside a bag made of small-pore (0.22-µm) nylon filtration membrane, which is placed in soil and later retrieved to determine survival of the pathogens on nonselective media. Chemicals but not other microorganisms can enter the bag from the soil. Using this assay, Streptomyces scabies, Fusarium oxysporum f. sp. lycopersici race 3, and Ralstonia solanacearum were successfully recovered from soil after 72 h as demonstrated by growth on a semiselective Streptomyces medium (S. scabies) or nonselective potato dextrose agar medium (F. oxysporum f. sp. lycopersici race 3 and R. solanacearum) with minimal microbial contamination. Addition of acetic acid (200 mM) to soil killed 100% of S. scabies. SPK (a mixture of organic chemicals) at a concentration of 1,500 mg kg-1 of soil killed 83.3% of F. oxysporum f. sp. lycopersici race 3 culture plugs, 100% of F. oxysporum f. sp. lycopersici race 3 spores, and 97.2% of R. solanacearum cells. SPK at 1,000 mg kg-1 of soil killed 50% of F. oxysporum f. sp. lycopersici race 3 culture plugs, 68.2% of F. oxysporum f. sp. lycopersici race 3 spores, and 12% of R. solanacearum. Benlate (500 to 1,500 mg kg-1 of soil) did not kill the culture plugs of F. oxysporum f. sp. lycopersici race 3 but reduced the growth rate of F. oxysporum f. sp. lycopersici race 3. Benlate (500, 1,000, and 1,500 mg kg-1 of soil) reduced F. oxysporum f. sp. lycopersici race 3 spore germination by 39.4, 49.3, and 50.4%, respectively. Streptomycin sulfate (1,500, 800, 400, and 200 mg kg-1 of soil) caused 75.3, 21, 11.9, and 0.9% mortality, respectively, of R. solanacearum.

Effects of midas® on nematodes in commercial floriculture production in Florida.

Cut flower producers currently have limited options for nematode control. Four field trials were conducted in 2006 and 2007 to evaluate Midas® (iodomethane:chloropicrin 50:50) for control of root-knot nematodes (Meloidogyne arenaria) on Celosia argentea var. cristata in a commercial floriculture production field in southeastern Florida. Midas (224 kg/ha) was compared to methyl bromide:chloropicrin (98:2, 224 kg/ha), and an untreated control. Treatments were evaluated for effects on Meloidogyne arenaria J2 and free-living nematodes in soil through each season, and roots at the end of each season. Plant growth and root disease were also assessed. Population levels of nematodes isolated from soil were highly variable in all trials early in the season, and generally rebounded by harvest, sometimes to higher levels in fumigant treatments than in the untreated control. Although population levels of nematodes in soil were not significantly reduced during the growing season, nematodes in roots and galling at the end of the season were consistently reduced with both methyl bromide and Midas compared to the untreated control. Symptoms of phytotoxicity were observed in Midas treatments during the first year and were attributed to Fe toxicity. Fertilization was adjusted during the second year to investigate potential fumigant/fertilizer interactions. Interactions occurred at the end of the fourth trial between methyl bromide and fertilizers with respect to root-knot nematode J2 isolated from roots and galling. Fewer J2 were isolated from roots treated with a higher level of Fe (3.05%) in the form of Fe sucrate, and galling was reduced in methyl bromide treated plots treated with this fertilizer compared to Fe EDTA. Reduced galling was also seen with Midas in Fe sucrate fertilized plots compared to Fe EDTA. This research demonstrates the difficulty of reducing high root-knot nematode population levels in soil in subtropical conditions in production fields that have been repeatedly fumigated. Although soil population density may remain stable, root population density and disease can be reduced.