Nuclear magnetic resonance: a tool for imaging belowground damage caused by Heterodera schachtii and Rhizoctonia solani on sugar beet.

Belowground symptoms of sugar beet caused by the beet cyst nematode (BCN) Heterodera schachtii include the development of compensatory secondary roots and beet deformity, which, thus far, could only be assessed by destructively removing the entire root systems from the soil. Similarly, the symptoms of Rhizoctonia crown and root rot (RCRR) caused by infections of the soil-borne basidiomycete Rhizoctonia solani require the same invasive approach for identification. Here nuclear magnetic resonance imaging (MRI) was used for the non-invasive detection of belowground symptoms caused by BCN and/or RCRR on sugar beet. Excessive lateral root development and beet deformation of plants infected by BCN was obvious 28 days after inoculation (dai) on MRI images when compared with non-infected plants. Three-dimensional images recorded at 56 dai showed BCN cysts attached to the roots in the soil. RCRR was visualized by a lower intensity of the MRI signal at sites where rotting occurred. The disease complex of both organisms together resulted in RCRR development at the site of nematode penetration. Damage analysis of sugar beet plants inoculated with both pathogens indicated a synergistic relationship, which may result from direct and indirect interactions. Nuclear MRI of plants may provide valuable, new insight into the development of pathogens infecting plants below- and aboveground because of its non-destructive nature and the sufficiently high spatial resolution of the method.

Systemically induced resistance and microbial competitive exclusion: implications on biological control.

The root-knot nematode, Meloidogyne incognita, is among the most damaging agricultural pests, particularly to tomato. The mutualistic endophytes Fusarium oxysporum strain Fo162 (Fo162) and Rhizobium etli strain G12 (G12) have been shown to systemically induce resistance toward M. incognita. By using triple-split-root tomato plants, spatially separated but simultaneous inoculation of both endophytes did not lead to additive reductions in M. incognita infection. More importantly, spatially separated inoculation of Fo162 and G12 led to a reduction in Fo162 root colonization of 35 and 39% when G12 was inoculated on a separate root section of the same plant in two independent experiments. In an additional split-root experiment, spatial separation of Fo162 and G12 resulted in a reduction of Fo162 root colonization of approximately 50% over the water controls in two independent experiments. The results suggested that the suppressive activity of G12 on Fo162 and M. incognita is possibly related to the induction of specific plant defense mechanisms. Thus, although Fo162 and G12 have the ability to systemically repress M. incognita infection in tomato, they can be considered incompatible biocontrol agents when both organisms are present simultaneously on the same root system.

Effect of Meloidogyne incognita inoculum density and application rate of Paecilomyces lilacinus strain 251 on biocontrol efficacy and colonization of egg masses analyzed by real-time quantitative PCR.

The fungal biocontrol agent, Paecilomyces lilacinus strain 251 (PL251), was evaluated for its potential to control the root-knot nematode Meloidogyne incognita on tomato at varying application rates and inoculum densities. Conversely to previous studies, significant dose-response relationships could not be established. However, we demonstrated that a preplanting soil treatment with the lowest dose of commercially formulated PL251 (2 × 10(5) CFU/g soil) was already sufficient to reduce root galling by 45% and number of egg masses by 69% when averaged over inoculum densities of 100 to 1,600 eggs and infective juveniles per 100 cm(3) of soil. To determine the role of colonization of M. incognita egg masses by PL251 for biocontrol efficacy, a real-time quantitative polymerase chain reaction (PCR) assay with a detection limit of 10 CFU/egg mass was used. Real-time PCR revealed a significant relationship between egg mass colonization by PL251 and the dose of product applied to soil but no correlation was found between fungal density and biocontrol efficacy or nematode inoculum level. These results demonstrate that rhizosphere competence is not the key mode of action for PL251 in controlling M. incognita on tomato.

MICAL-L1 is required for cargo protein delivery to the cell surface.

Secreted proteins are transported along intracellular route from the endoplasmic reticulum through the Golgi before reaching the plasma membrane. Small GTPase Rab and their effectors play a key role in membrane trafficking. Using confocal microscopy, we showed that MICAL-L1 was associated with tubulo-vesicular structures and exhibited a significant colocalization with markers of the Golgi apparatus and recycling endosomes. Super resolution STORM microscopy suggested at the molecular level, a very close association of MICAL-L1 and microdomains in the Golgi cisternae. Using a synchronized secretion assay, we report that the shRNA-mediated depletion of MICAL-L1 impaired the delivery of a subset of cargo proteins to the cell surface. The process of membrane tubulation was monitored in vitro, and we observe that recombinant MICAL-L1-RBD domain may contribute to promote PACSINs-mediated membrane tubulation. Interestingly, two hydrophobic residues at the C-terminus of MICAL-L1 appeared to be important for phosphatidic acid binding, and for association with membrane tubules. Our results reveal a new role for MICAL-L1 in cargo delivery to the plasma membrane.

Effect of different potting systems; inoculation time; nematode density and sources of cereal cyst nematode (Heterodera filipjevi) on juvenile penetration into wheat root system.

Investigations were designed to optimize testing systems for screening wheat breeding lines for resistance to Heterodera filipjevi. The effects of: 1) plant potting systems 2) inoculum level and time of inoculation 3) and type of inoculum of H. filipjevi on detection accuracy were examined in growth chamber experiments in Turkey. The rate of nematode penetration in the highly susceptible variety Bezostaya was used as the base measurement of efficacy. The results showed that the highest level of penetration coupled with high level of germination was obtained in plastic tubes (13 cm long x 3 cm in diam.) when compared to both small flower pots (400 cm3) and smaller plastic tubes (10.2 cm long x 0.8 cm in diam.). The highest rate of nematode penetration into wheat root system was obtained by inoculating the seedlings with 1000 J2 per plant. However, inoculation with 200 J2 at sowing or 200 J2 at sowing plus an additional 200 J2 after germination improved percent penetration when compared to inoculation with 600 or 1000 J2/plant at sowing. The test on the optimum form of inoculum showed that inoculating the seedling with J2's gave the highest rate of nematode penetration over inoculum with eggs or cysts. The results of these experiments demonstrated that screening wheat for resistance can be optimized by raising the seedlings in plastic tubes and inoculating them with 400 J2.

Comparison of Muscodor albus Volatiles with a Biorational Mixture for Control of Seedling Diseases of Sugar Beet and Root-Knot Nematode on Tomato.

A biorational synthetic mixture of organic components mimicking key antimicrobial gases produced by Muscodor albus was equivalent to the use of live M. albus for control of seedling diseases of sugar beet (Beta vulgaris) caused by Pythium ultimum, Rhizoctonia solani AG 2-2, and Aphanomyces cochlioides. The biorational mixture provided better control than the live M. albus formulation for control of root-knot nematode, Meloidogyne incognita, on tomato (Lycopersicon esculentum). The biorational mixture provided control of damping-off equal to a starch-based formulation of the live fungus for all three sugar beet pathogens, and significantly reduced the number of root-knot galls on tomato roots compared with a barley-based formulation. Rate studies with the biorational mixture showed that 2 and 0.75 µl/cm3 of soil were required to provide optimal control of Rhizoctonia and Pythium damping-off of sugar beet, respectively. Five microliters of biorational mixture per milliliter of water was required for 100% mortality in 24 h for Meloidogyne incognita in in vitro studies. In in vivo studies, 1.67 µl of the biorational mixture/cm3 of sand resulted in fewer root-knot galls than a Muscodor albus infested ground barley formulation applied at 5 g/liter of sand.

Suitability of Pueraria phaseoloides, Chromolaena odorata and Tithonia diversifolia as in-situ mulch for nematode management in musa cropping systems.

Mulching with plant organic matter has been shown to reduce nematode population densities in various cropping systems. The level of nematode control is increased when such mulches are incorporated into the soil as organic amendments. Chromolaena odorata, Tithonia diversifolia and Pueraria phaseoloides are common cover crops in West and Central Africa that produce large quantities of nutrient rich biomass. The aim of this study was to determine, if in-situ mulching of C. odorata, T. diversifolia and P. phaseoloides is suitable for nematode control in Musa production. In a pot trial, the susceptibility of these plants to spiral nematodes was investigated. The effects of different quantities of surface mulch on nematode population densities in the soil and in banana roots also were determined. All mulch types and all quantities led to a reduction in nematode population densities in the soil. The strongest nematode reductions were observed in the Pueraria treatments. In treatments containing banana plants mulching improved plant growth compared to the clean-fallowed soil and induced lower root infestation rates. However, nematode soil populations were higher in mulched than in non-mulched banana treatments. Plant parasitic nematodes also were isolated from roots of all three cover crop species and all three plants caused an increase in nematode numbers in the soil. Therefore, the tested cover crops proved unsuitable for nematode control in a system with the highly susceptible bananas. Further examinations are needed to determine whether or not the positive effects of surface mulching on plantain plant growth and root infestation rates also have positive effects on yield in an in-situ mulching system in the presence of nematodes.

Use of Trichoderma harzianum and Trichoderma viride for the biological control of Meloidogyne incognita on tomato.

Trichoderma harzianum and Trichoderma viride were tested for their capacity to reduce the incidence of the root-knot nematode Meloidogyne incognita on tomato. In vitro studies demonstrated that all tested isolates were effective in causing nematode mortality compared with the control. Trichoderma slightly reduced nematode damage to tomato in vivo. Treatment of the soil with the biocontrol agents before transplanting, improved control over treatment directly at transplanting. The Trichoderma isolates could not be re-isolated from the endorhiza, but were successfully re-isolated from the rhizosphere 45 days after fungal inoculation. Only slight increases in plant growth could be measured. The mutualistic endophyte F. oxysporum 162, used as positive control, was more effective in root-knot nematode biocontrol than the Trichoderma isolates.

Criteria for examiner-independent nocturnal penile tumescence and rigidity monitoring (NPTR): correlations to invasive diagnostic methods.

78 patients (pts.) with erectile dysfunction (ED) underwent conventional diagnostic evaluation including NPTR monitoring with the Rigiscan, intracavernous drug testing, dynamic cavernosometry, doppler evaluation, pharmaco-angiography and neurologic tests. Only erections of the best rigidity recorded over three nights served for classification into three classes of rigidity. Patients with neurogenic impotence showed significantly poorer and fewer erectile events per hour than vasculogenic impotent men. Patients with arteriogenic ED had better erections than patients with venogenic or mixed arterio-venogenic ED. Severity of organic ED during conventional diagnostic work-up correlated to loss of rigidity during NPTR recordings, but no differential diagnosis between arteriogenic, venogenic or neurogenic impotence could be made with NPTR data alone. The presented criteria for NPTR evaluation permit a time-saving, examiner-independent analysis.

Intracavernosal kinetics of eicosanoids and endothelin during erection. Data from human and animal studies on intrapenile and systemic prostaglandins.

The release of mediator substances of the arachidonic acid cascade is closely related to the functional state of the endothelium. A significant lower prostacyclin/thromboxane A2 ratio in penile plasma of organogenic impotent patients in comparison to patients with psychogenic erectile dysfunction has been described in the literature. We observed the time-related liberation of prostacyclin, thromboxane A2 and the vasoactive peptide endothelin for 16 minutes of a drug-induced erection. We compared kinetics of patients with penile deviation and transsexualism, to patients suffering from severe organogenic impotence. We assessed the usefulness of the prostacyclin-to-thromboxane A2 ratio as a possible indicator of corporal degeneration. An animal model has been created to observe differences between rabbits under 100 days of standard diet alimentation, rabbits under cholesterol enriched diet and rabbits with hereditary hyperlipidemia type II A. Hyperlipidemia is suspected to be one possible factor causing organogenic impotence. Enzyme-immuno-assays were used for the determination of all substances. The systemic prostacyclin-to-thromboxane A2 ratio differed significantly between control rabbits and rabbits with hyperlipidemia. Prostacyclin, thromboxane A2 and endothelin in corpus cavernosum plasma showed a typical profile during spontaneous and drug-induced erection. A significant difference between groups of patients suffering from organogenic or psychogenic impotence could not be found. The value of the determination of the studied substances in differential diagnosis seems to be dubious.

Endophytic Colonization of Plants by the Biocontrol Agent Rhizobium etli G12 in Relation to Meloidogyne incognita Infection.

ABSTRACT The external and internal colonization of potato and Arabidopsis roots by the biocontrol strain Rhizobium etli G12 containing a plasmidborne trp promoter green fluorescent protein transcriptional fusion, pGT-trp, was studied in the presence and absence of the root-knot nematode Meloidogyne incognita. Plant colonization behavior and biocontrol potential of the marked strain G12(pGT-trp) was not altered compared with the parental strain. Plasmid pGT-trp was stable for more than 80 generations without selection and conferred sufficient fluorescence to detect single bacterial cells in planta. Although bacteria were found over the entire rhizoplane, they preferentially colonized root tips, the emerging lateral roots, and galled tissue caused by Meloidogyne infestation. Internal colonization of potato roots was mainly observed in epidermal cells, especially root hairs. G12(pGT-trp) colonization was also observed in inner Arabidopsis root tissues in areas of vascularization. In the presence of M. incognita, G12(pGT-trp) colonized the interior of nematode galls in high numbers. In some cases, bacterial colonization even extended from the galled tissue into adjacent root tissue. The internally colonized sites in roots were often discontinuous. Fluorescence microscopy of gfp-tagged rhizobacteria was a sensitive and a rapid technique to study external and internal colonization of plant roots by bacteria interacting with nematodes.

Immunoglobulin E antibody against environmental allergens in subjects with trimellitic anhydride-induced asthma.

The purpose of this study was to assess the relationship between atopy and the development of occupational asthma as a consequence of exposure to trimellitic anhydride (TMA). A case-control study was performed, which comprised 16 employees identified as having TMA-induced asthma and 44 similarly exposed controls. Specific immunoglobulin E measurements in response to cat, dust mite, ryegrass, and ragweed antigens were performed. Fifty-six percent of cases and 29% of controls were found to be atopic (P = 0.098). We demonstrated that there was a trend toward employees with TMA asthma being more atopic than those without TMA asthma. Atopy as an assessment of risk for the development of TMA asthma is unlikely to be useful, although further investigation may be warranted.

[Angiography of the a. pudenda and its branches in erectile dysfunction. A modified angiographic technic and optimized use of vasodilator substances]. / Angiographie der A. pudenda und ihrer Aste bei erektiler Dysfunktion. Modifizierte angiographische Technik und optimierter Einsatz vasodilatierender Substanzen.

A new simplified technique for evaluating the internal pudendal artery and the penile vessels is described using a new catheter configuration with a very short 90 degrees-angled tip. In 30 consecutive patients, a superselective catheterisation of the internal pudendal artery was achieved in 93%. To improve visualisation of the penile arteries, the selective intraarterial application of 0.2 mg nitroglycerin in addition to the mandatory intracavernous injection of papaverin or prostaglandin E1 has proved to be of value. Examination of the blood blow velocity in the dorsal penile artery with Doppler ultrasound 2 minutes after intra-arterial application of 0.2 mg nitroclyerin showed a mean elevation of the Doppler amplitude by the factor 2.4 (range 0.4-9). With regard to the maximal increase of the blood velocity (range 1-6 minutes), a high interindividual difference was observed.

A genetic algorithm-based approach to flexible flow-line scheduling with variable lot sizes.

Genetic algorithms (GAs) have been used widely for such combinatorial optimization problems as the traveling salesman problem (TSP), the quadratic assignment problem (QAP), and job shop scheduling. In all of these problems there is usually a well defined representation which GA's use to solve the problem. We present a novel approach for solving two related problems-lot sizing and sequencing-concurrently using GAs. The essence of our approach lies in the concept of using a unified representation for the information about both the lot sizes and the sequence and enabling GAs to evolve the chromosome by replacing primitive genes with good building blocks. In addition, a simulated annealing procedure is incorporated to further improve the performance. We evaluate the performance of applying the above approach to flexible flow line scheduling with variable lot sizes for an actual manufacturing facility, comparing it to such alternative approaches as pair wise exchange improvement, tabu search, and simulated annealing procedures. The results show the efficacy of this approach for flexible flow line scheduling.

[Selective microsurgical therapy in vascular-induced erectile impotence]. / Differenzierte mikrochirurgische Therapie bei vaskulär bedingter erektiler Impotenz.

Patients with erectile dysfunction should be subjected to a complex and complete diagnostic procedure, including selective pharmaco-phalloangiography and dynamic pharmacocavernosonography only when SKAT has failed and after the exclusion of neurological disorders. Patients being offered these invasive procedures should be highly motivated and willing to undergo surgical correction of the vascular origin of their erectile impotence. If peripheral vascular occlusive disease is found affecting the pelvic arteries, the best operative technique must be selected. In our opinion, this decision should be made intraoperatively after visualization of the dissected dorsal penile vessels. Use of the Doppler probe can be extremely helpful during the operation. Intraoperative findings on the degree of arteriosclerosis and arterial flow can differ from those allowed by preoperative diagnostic procedures. Penile erectile function was restored in seven of nine patients after penile revascularization performed under microsurgical conditions. We used the operative methods described by Virag and Hauri, applying our own modification of the Hauri procedure in two patients. With careful selection of patients and methods it should be possible to resolve individual patients' problems with peripheral occlusion, even though the exact hemodynamic pathology remains obscure.

[Reflectory stimulation of the pudendal nerve in dynamic pharmaco-cavernosometry]. / Die reflektorische Stimulation des N. pudendus bei der dynamischen Pharmakokavernosometrie.

A prospective study was performed in 20 patients to assess the impact of vibratory glans stimulation on the course and results of dynamic pharmaco-cavernosography and cavernosometry. An intracorporeal pressure rise was achieved with lower amounts of saline when stimulation was applied and in some cases could not be provoked by saline infusion without glans stimulation. False-positive diagnoses of diffuse venous leakage could thus be eliminated. The data presented in this paper confirm the need for stimulation of the pudendal reflex arch to achieve physiological conditions. The simplicity of the procedure and the minimal time requirement suggest its clinical routine application of vibratory glans stimulation during cavernosography and cavernosometry.

Effects of Meloidogyne incognita on Growth and Storage-Root Formation of Cassava (Manihot esculenta).

Two-node cuttings of cassava cultivar SS4 were inoculated with 1,000 infective juveniles of Meloidogyne incognita at 1, 14, 40, 70, 88, and 127 days after planting (DAP). Plant growth and root damage were assessed at 150 DAP. Meloidogyne incognita significantly reduced the number of storageroots formed in plants inoculated at 14, 40, 70, and 88 DAP and the total weight of storage-roots in plants inoculated at 1, 14, 40, 70, and 88 DAP, compared to uninoculated plants. Individual storage-root weight and plant height were not affected by M. incognita. Storage-root formation in cassava is initiated when plants are 1 to 2 months old. The results of this experiment indicate that, at this time, young cassava plants are most prone to root-knot nematode damage in terms of storage-root formation. The production loss caused by M. incognita to young SS4 plants was due to a reduction of storage-root number rather than a reduction in individual storage-root weight.

Interaction of Meloidogyne naasi, Pratylenchus penetrans, and Tylenchorhynchus agri on Creeping Beentgrass.

The pathogenicity and interactions of Meloidogyne naasi, Pratylenchus penetrans, and Tylenchorhynchus agri on 'Toronto C-15' creeping bentgrass, Agrostis palustris, was studied in a long-term greenhouse experiment. Based on dry weights of roots and clippings, M. naasi alone and in all combinations with P. penetrans and T. agri was highly pathogenic to creeping bentgrass. P. penetrans and T. agri alone and in combination inhibited root growth but adversely affected top growth only when the two were co-inoculated. In combination, the effects of each species on top growth were additive, with M. naasi the dominant pathogen. Creeping bentgrass was an excellent host for M. naasi and T. agri, but a poor host for P. penetrans. T. agri inhibited population increase of M. naasi, indicating nematode-nematode competition, but neither T. agr/ nor P. penetrans was affected by any of the combinations.

Optimizing the efficacy of Paecilomyces lilacinus (strain 251) for the control of root-knot nematodes.

The egg pathogenic fungus Paecilomyces lilacinus (strain 251) is a biocontrol fungus with a potential range of activity to control the worldwide most important plant parasitic nematodes. This biological nematicide may be an useful tool in an integrated approach to control mainly sedentary nematodes. Greenhouse experiments were conducted with the root-knot nematodes Meloidogyne incognita and M. hapla on tomato. P. lilacinus, formulated as WG (BIOACT WG), was incorporated into soil inoculated with root-knot nematode eggs prior to transplanting the susceptible tomato cultivar "Hellfrucht". Furthermore, soil treatments were combined with seedling treatments 24 hours before transplanting and a soil drench 2 weeks after planting, respectively. Seedling and post planting treatment was also combined with a soil treatment at planting. All single or combination treatments tested decreased the gall index and the number of egg masses compared to the untreated control 12 weeks after planting. However, the combination of the seedling treatment with a pre- or at-planting application of P. lilacinus was necessary to achieve higher levels of control. Additional post plant drenching resulted in only a slight increase In efficacy. To the feasibility of this modified application system for the control of root-knot nematodes, a yield experiment was conducted with M. hapla and the susceptible cultivar "Gnom F1 Hybrid". It could be demonstrated that the above mentioned combination of pre-planting application plus the seedling and one post plant drench gave the best control and resulted in a significant fruit yield increase in concurrence with a decrease in number of galls per root.

Efficacy of Paecilomyces lilacinus (strain 251) for the control of root-knot nematodes.

The egg pathogenic fungus Paecilomyces lilacinus (strain 251), is a unique strain with a wide range of activity against the most important plant parasitic nematodes. Due to increased production capacity by solid state fermentation and a new water dispersible granule (WDG) formulation, this biological nematicide may be used in an integrated approach to control plant parasitic nematodes. Dose response experiments were conducted with the root-knot nematode Meloidogyne incognita on tomatoes using the new WDG formulation. The results revealed a clear correlation between rate applied and the degree of control concerning the reduction in damage to the root and multiplication of the nematode. Best control was achieved by applying the biological nematicide at rates of 2 to 4 times 10(9) conidia per plant as a soil treatment one week before planting. Monitoring the P. lilacinus population in the rhizosphere showed a decline after 2 to 3 month which can lead to insufficient control over a full growing season. Repeated application to maintain the antagonist population at a sufficient level could be used to secure long term control of root-knot nematodes.