Behavioral Response of Nothanguina phyllobia to Selected Plant Species

The silver-leaf nightshade nenmtode, Nothanguina phyllobia, is a promising biological control agent for its only reported host, Solanum elaeagnifolium Cav. When infective larvae of N. phyllobia and stem tissue of 39 econmnically important plant species were suspended in 0.5% water agar, nematodes aggregated about S. elaeagnifolium, Solanum carolinense L., Solanum melongena L., Solanum tuberosum L., and Prunus caroliniana (Mill.) Ait. Nematodes responded to Solanum spp. via positive chemotaxis and/or klinokinesis, but aggregated near tissue of P. caroliniana as a result of orthokinetic effects. Nematodes aggregated away from tissue of Hibiscus esculentus L., Triticum aestivum L., Santolina sp., Rosa sp., and Kochia scoparia (L.) Schrad. in the absence of orthokinetic effects. Experiments that excluded light and maintained relative humidity at 100% showed N. phyllobia to ascend the stems of 35 plant species to a height of > 9 cm within 12 h. Differences in stem ascension were not attributable to stem surface characteristics.     

Distribution of Nothanguina phyllobia and Its Potential as a Biological Control Agent for Silver-leaf Nightshade

The nematode Nothanguina phyllobia Thorne was found within large foliar galls on the perennial weed Solanum elaeagnifolium Cav. in west Texas. A two-year survey of a 6400 sq-km area in west Texas showed extensive distribution of the nematode. No hosts other than S. elaeagnifolium were observed. Densities of juvenile nematodes in the soil were high. N. phyllobia spread rapidly after small numbers of infective juveniles were applied in a foliar spray to an S. elaeagnifolium population. The host plant declined in vigor and frequently died. Artificial inoculation of an S. elaeagnifolium population with large numbers of the nematodes by broadcasting infected plant tissue resulted in high infection incidence.     

Effects of Oxygen and Temperature on the Activity and. Survival of Nothanguina phyllobia

The effects of oxygen and temperature on the activity and survival of infective forth-stage juveniles of Nothanguina phyllobia Thorne were examined in aqueous suspension. Rate of movement was not affected by a wide range of O₂ concentration (0.8-8.6 ppm). Activity decreased below 0.8 ppm 0 2, and at 0.15 ppm O₂ nematodes became motionless. Activity increased as a linear function of temperature up to a thermal optimum of 24 C; beyond 24 C activity decreased. Survival was greatly prolonged at low temperature. At 23 C, 50% mortality occurred within 7 d, whereas at 4 C, 70% survived after 98 d.     

Histopathogenesis of Galls Induced by Meloidogyne naasi in Wheat Roots

Histopathogenesis of galls induced by Meloidogyne naasi in wheat roots was studied. Large numbers of larvae penetrated wheat root tips within 24 hr; larvae migrated both inter- and intracellularly, causing cortical hypertrophy. Giant cells were formed in the stele around the head of each nematode within 4 to 5 days. Initial pathological alterations in giant cell formation consisted of hypertrophy of protophloem and protoxylem cells, their nuclei and nucleoli. Giant ceils contained 2 to 8 agglomerated multinucleolate nuclei. Synchronous mitotic divisions were first observed 9 days after inoculation. After 21 days, giant cells became highly vacuolate. Observations 40 days after inoculation revealed a complete degeneration of cell contents in many giant cells but their thick walls remained intact. Abnormal xylem completely surrounded the degenerated or partially degenerated giant cells.     

Temperature Effects on the Parasitic Phase of Romanomermis culicivorax in Culex pipiens

The developmental period for the parasitic stage of Romanomermis culicivorax in Culex pipiens was determined at constant and fluctuating temperatures. The median developmental times at 15, 18, 20, 27, and 32 C were 27.6, 17.2, 11.5, 7.1, and 5.8 days, respectively. The optimum temperature range for development of the parasitic stage in C. pipiens was 20 to 32 C. The threshold for development was calculated as 10.4 C, and the heat units required for development were 122.2 ± 3.6 day-degrees. Development at fluctuating temperatures conformed to that predicted by the constant temperature data.     

Chloroplast Differentiation in Tomato Root Galls Induced by the Root Knot Nematode Meloidogyne incognita

Primary roots of tomato, Lycopersicon esculentum cv. Marglobe, were cultured aseptically on agar containing a standard nutrient formulation with or without kinetin. When secondary roots developed, cultures were inoculated with the root-knot nematode, Meloidogyne incognita. Following inoculation, the cultures were divided into two groups which were incubated either in total darkness or in 16-h light-8-h dark cycles. At 24 h, 1, 2, 3, and 4 wk after incubation, roots from all cultures were processed for transmission electron microscopy. Fine structural observation of the parenchyma tissue in galls from the inoculated cultures indicated that starch containing plastids or amyloplasts, which are usually present and remain undifferentiated in these root cells, developed into chloroplasts. These chloroplasts contained a membrane system indistinguishable from those found in leaves of intact plants. Although plastid development was not affected when uninoculated cultures were incubated in the light, differentiation of the amyloplast was induced when roots were cultured on the medium containing kinetin. These results suggest that plastid differentiation in the inoculated tissue may be influenced by an accumulation of kinetin in the gall, which is induced by the nematode and serves as the nutrient sink for its feeding.     

Antagonists of Plant-parasitic Nematodes in Florida Citrus

In a survey of antagonists of nematodes in 27 citrus groves, each with a history of Tylenchulus semipenetrans infestation, and 17 noncitrus habitats in Florida, approximately 24 species of microbial antagonists capable of attacking vermiform stages of Radopholus citrophilus were recovered. Eleven of these microbes and a species of Pasteuria also were observed attacking vermiform stages of T. semipenetrans. Verticillium chlamydosporium, Paecilomyces lilacinus, P. marquandii, Streptomyces sp., Arthrobotrys oligospora, and Dactylella ellipsospora were found infecting T. semipenetrans egg masses. Two species of nematophagous amoebae, five species of predatory nematodes, and 29 species of nematophagous arthropods also were detected. Nematode-trapping fungi and nematophagous arthropods were common inhabitants of citrus groves with a history of citrus nematode infestation; however, obligate parasites of nematodes were rare.     

Survey of Heterorhabditidae and Steinernematidae (Rhabditida,Nematoda) in Western Canada

A survey was done in the summer months along the Alaska Highway, in other parts of British Columbia, in northern Alberta, and in the Yukon Territory for steinernematid and heterorhabditid nematodes occurring in the top 10 cm of soil. Steinernema feltiae and Steinernema spp. were found at 18 and Heterorhabditis megidis at 7 sites of 125 sampled. Most nematodes were found where visible insect infestation occurred and where human influence on the habitat was substantial (e.g., agricultural, forested and bush-hedgerow habitats); none was found in grassland or virgin forests. Heterorhabditis megidis occurred in only the southern, warmer, drier region of British Columbia. In the laboratory some steinernematid isolates and H. megidis killed Galleria mellonella larvae at 13 and 22 C, whereas some isolates of Steinernema killed the larvae at only 13 C. Steinernema spp. from three high altitude sites with low, average July temperatures (13-14 C) are cold-active in that they produced infective juveniles at 13 C and killed G. mellonella at 6 C.     

Ultrastructural Changes Caused by Fusarium oxysporum f. sp. lycopersici in Meloidogyne javanica Induced Giant Cells in Fusarium Resistant and Susceptible Tomato Cultivars

Tomato (Lycopersicon esculentum Mill.) seedlings, susceptible (cv. Pearson A-I Improved) and resistant (cv. Pearson Improved) to race 1 Fusarium oxysporum f. sp. lycopersici (Sacc.) Snyd &Hans., were inoculated with Meloidogyne javanica (Trueb) Chitwood second-stage juveniles and 3 weeks later with race 1 F. oxysporum f. sp. lycopersici spores. One week after fungal inoculation, no fungus was visible in root tissue of the tomato cultivars and the giant cells were normal. Two weeks after fungal inoculation, abundant hyphae were visible in xylem tissues of Fusarium-susceptible but not of Fusarium-resistant plants. In susceptible plants, giant cell degeneration occurred, characterized by membrane and organelle disruption. In addition, where hyphae were in direct contact with the giant cell, dissolution of the giant cell wall occurred. Three weeks after fungal inoculation, fungal hyphae and spores were visible inside xylem tissues and giant cells in Fusarium-susceptible plants and in xylem tissue of the resistant plants. In susceptible and resistant plants, giant cell degeneration was apparent. Giant cell walls were completely broken down in Fusarium-susceptible tomato plants. In both cultivars infected by Fusarium, giant cell nuclei became spherical and dark inclusions occurred within the chromatin material which condensed adjacent to the fragmented nuclear membrane. No such ultrastructural changes were seen in the giant cells of control plants inoculated with nematode alone. Giant cell deterioration in both cultivars is probably caused by toxic fungal metabolites.     

Effects of Hydroxyurea on the Ultrastructure of Giant Cells in Galls Induced by Meloidogyne javanica

Hydroxyurea (HU) at concentrations of 10 or 20 mg/liter was included in a medium on which excised tomato roots infected with the root-knot nematode Meloidogyne javanica were grown. In the HU, treated roots, giant cells were small and contained large vacuoles. Giant cell nuclei were amoeboidal with relatively small nucleoli in treated roots, compared with giant cells of nontreated galls. In treated-root giant cells, the cytoplasm was diffuse and few organelles such as mitochondria, dictyosomes, and endoplasmic reticulum were detected; also, walls of giant cells were thin with less extensive ingrowths than in nontreated roots. We conclude that HU suppressed normal giant cell formation interfering with its function as a feeding cell.     

Morphological and Histochemical Changes Occurring during the Life-span of Root-tip Galls on Lolium perenne Induced by Longidorus elongatus

The RNA and protein content of perennial ryegrass root-tip galls induced by Longidorus elongatus were measured from transverse sections and the morphology described. Galls progressed through five distinct stages and were viable for only 10-12 days at 18 C, after which they collapsed and became necrotic. In the initial stage hypertrophy occurred and cells contained enlarged nuclei and nucleoli, a greater proportion of cytoplasm, and increased concentrations of protein. This was followed by hyperplasia; cells divided to give two or four daughter cells, accompanied by a proportionate reduction in volumes of cytoplasm, nuclei, and nucleoli and reduced concentrations of RNA and protein. The third stage was secondary hypertrophy with enlarged, amoeboid nuclei and nucleoli and a significant increase in concentration of RNA and protein. In the final two stages, as feeding by L. elongatus progressively removed cell contents, most cells were devoid of inclusions and galls collapsed and were invaded by soil bacteria. This ordered development and exploitation of galls suggests that L. elongatus may have two phases in its feeding.     

Development of Meloidogyne incognita Inhibited by Digtaria decumbens cv. Pangola

Population densities of the root-knot nematode, Meloidogyne incognita, were lower after 90 days in soil planted to digitgrass (Digitaria decumbens cv. Pangola) than in soil left fallow or planted to tomato. Roots of tomato seedlings interplanted with Pangola digitgrass were less galled than were roots of tomato seedlings planted alone. Fewer second stage larvae invaded roots of Pangola digitgrass than tomato and those that entered the grass roots failed to develop beyond the late second stage.     

Suppression of Meloidogyne incognita and M. javanica by Pasteuria penetrans in Field Soil

The role of Pasteuria penetrans in suppressing numbers of root-knot nematodes was investigated in a 7-year monocuhure of tobacco in a field naturally infested with a mixed population of Meloidogyne incognita race 1 and M. javanica. The suppressiveness of the soil was tested using four treatments: autoclaving (AC), microwaving (MW), air drying (DR), and untreated. The treated soil bioassays consisted of tobacco cv. Northrup King 326 (resistant to M. incognita but susceptible to M. javanica) and cv. Coker 371 Gold (susceptible to M. incognita and M. javanica) in pots inoculated with 0 or 2,000 second-stage juveniles of M. incognita race 1. Endospores of P. penetrans were killed by AC but were only slightly affected by MW, whereas most fungal propagules were destroyed or inhibited in both treatments. Root galls, egg masses, and numbers of eggs were fewer on Coker 371 Gold in MW, DR, and untreated soil than in AC-treated soil. There were fewer egg masses than root galls on both tobacco cultivars in MW, DR, and untreated soil than in the AC treatment. Because both Meloidogyne spp. were suppressed in MW soil (with few fungi present) as well as in DR and untreated soil, the reduction in root galling, as well as numbers of egg masses and eggs appeared to have resulted from infection of both nematode species by P. penetrans.     

Susceptibility of Various Species of Lepidopterous Pupae to the Entomogenous Nematode Neoaplectana carpocapsae

The susceptibility of certain species of lepidopterous pupae occurring in different ecological situations to the entomogenous nematode, Neoaplectana carpocapsae, was tested. Soil- or litter-pupating lepidopterous insects were not highly susceptible to N. carpocapsae. The most susceptible insect pupating in the soil was Spodoptera exigua with 63% pupal mortality, while Harrisinia brillians, which pupates in litter, had 55% mortality. Other soil- and litter-pupating insects had mortalities of less than 25%. Some insect species that pupate above ground were highly susceptible (> 84% mortality) to N. carpocapsae infection.     

Galls induced by Calophya latiforceps (Hemiptera: Calophyidae) reduce leaf performance and growth of Brazilian peppertree

Schinus terebinthifolia Raddi (Anacardiaceae) (Brazilian peppertree) is one of the most serious terrestrial invasive plants in Florida. The leaf galling psyllid, Calophya latiforceps Burckhardt (Hemiptera: Calophyidae), was discovered feeding on the Brazilian peppertree in Bahia, Brazil, in 2010, and is being evaluated for release as a biological control agent. In order to understand how this candidate biological control agent affects the growth of S. terebinthifolia, we measured the effect of the psyllid on photosynthesis, leaf chlorophyll content and plant growth. Infested plants had lower photosynthesis compared to uninfested plants 30 and 45 days after gall initiation, and chlorophyll content was lower over a 70-d period. Plant height was reduced 31% and biomass 11% after three months of infestation. Results of these studies suggest that C. latiforceps will negatively affect the growth of the Brazilian peppertree if released in Florida.     

The Effect of Arthrobotrys conoides on Meloidogyne incognita Population Densities in Corn as Influenced by Temperature,Fungus Inoculum Density,and Time of Fungus Introduction in the Soil

In greenhouse experiments, the effect of Arthrobotrys conoides on Meloidogyne incognita population densities as affected by soil temperature, inoculum density, and green alfalfa was determined. The effect on M. incognita population densities was greater at a soil temperature of 25 C than at 18 or 32 C. Nematode control by A. conoides was most effective when the fungus was introduced into the soil 2 wk prior to nematode inoculation and planting of corn. Inoculum density of A. conoides was positively correlated with plant shoot weight (r = 0.81) and negatively correlated with numbers of Meloidogyne juveniles (r = -0.96), eggs (-0.89) and galls per gram of root (-0.91). A. conoides was not isolated from green alfalfa, but was isolated from alfalfa-amended soil to which no fungus had been added.     

Root Penetration by Meloidogyne incognita Juveniles Infected with Bacillus Penetrans

Bacillus penetrans inhibited penetration by Meloidogyne incognita second-stage juveniles (J2) into tomato roots in the laboratory and greenhouse. Spores from this Florida population of B. penetrans attached to J2 of M. javanica, M. incognita, and M. arenaria. A greater proportion of J2 of M. javanica were infected than were J2 of either M. incognita or M. arenaria, and a greater number of spores attached to M. incognita than to M. arenaria.