Mapping of five resistance genes to sugar-beet powdery mildew using AFLP and anchored SNP markers.

Sugar-beet powdery mildew, caused by the fungus Erysiphe betae, now occurs in all sugar-beet growing areas and can reduce sugar yield by up to 30%. Powdery mildew resistant plants from three novel sources were crossed with sugar beet to generate segregating populations. Evaluation of resistance was carried out in artificially inoculated field and controlled environment tests. The resistance level in two of the sources was found to be significantly higher than that in currently available sugar-beet cultivars. AFLP analysis was used in combination with bulked segregant analysis to develop markers linked to the resistant phenotype in each population. Five dominant major resistance genes were identified and assigned the proposed symbols Pm2 to Pm6. Pm3 conferred complete resistance to powdery mildew; the other genes conferred high levels of partial resistance. From the use of anchoring SNP markers, two genes were located to chromosome II and three to chromosome IV. Two of the genes on chromosome IV mapped to the same location and one of the genes on chromosome II mapped to the same region as the previously identified Pm1 gene. With the availability of these genes there is now excellent potential for achieving durable resistance to sugar-beet powdery mildew, thus reducing or obviating the need for chemical control.

An anchored linkage map for sugar beet based on AFLP, SNP and RAPD markers and QTL mapping of a new source of resistance to Beet necrotic yellow vein virus.

Rhizomania, caused by Beet necrotic yellow vein virus (BNYVV), is an important sugar-beet disease worldwide and can result in severe losses of root yield and sugar content. We have identified a major QTL for BNYVV resistance from a new source in a segregating population of 158 individuals. The QTL explained an estimated 78% of the observed phenotypic variation and the gene conferring the partial resistance is referred to as Rz4. AFLP was used in combination with bulked segregant analysis (BSA) to develop markers linked to the resistance phenotype. AFLP marker analysis was extended to produce a linkage map that was resolved into nine linkage groups. These were anchored to the nine sugar-beet chromosomes using previously published SNP markers. This represents the first anchored sugar-beet linkage map to be published with non-anonymous markers. The final linkage map comprised 233 markers covering 497.2 cM, with an average interval between markers of 2.1 cM. The Rz4 QTL and an Rz1 RAPD marker were mapped to chromosome III, the known location of the previously identified BNYVV resistance genes Rz1, Rz2 and Rz3. The availability to breeders of new resistance sources such as Rz4 increases the potential for breeding durable disease resistance.

Comparison of application methods to prolong the survival of potential biocontrol bacteria on stored sugar-beet seed.

AIMS: To develop bacterial inoculation treatments on sugar-beet seed that will maintain a commercially acceptable degree of viability for a minimum of 4 months storage at ambient temperature. METHODS AND RESULTS: Single rifampicin-resistant (Rif(+)) strains of both Gram-positive and negative bacterial isolates (mostly pseudomonads) were applied in turn to sugar-beet seed in a comparative study by seed soaking, encapsulation in alginate, pelleting using an inoculated peat carrier or seed priming. The treated seed was assessed for bacterial survival over a time course by plating out homogenized samples onto a selective medium. Priming inoculation offered a significant improvement over all the other application strategies tested. After pelleting with fungicides and drying at 40 degrees C, Pseudomonas marginalis/putida P1W1 maintained populations of >6.6 log(10) CFU g(-1) seed during 4 months storage at 15 degrees C. Subsequent experiments verified a stabilized population under these storage conditions with commercial pellets at <7% moisture content. CONCLUSION: An inoculation method was established which allowed the survival on seed of a Gram-negative bacterium at ambient temperature with little loss in viability. SIGNIFICANCE AND IMPACT OF THE STUDY: This has promising implications for the delivery of beneficial bacteria, especially Gram-negative strains, on sugar beet.

Colonization of the developing rhizosphere of sugar beet seedlings by potential biocontrol agents applied as seed treatments.

AIMS: Poor colonization of the rhizosphere is a major constraint of seed treatment biological control. The objectives of this study were to; examine the colonization of the rhizosphere of sugar beet seedlings by selected rhizobacteria; determine the influence of the host rhizosphere and percolating water on the distribution of the bacteria; and deliver two biological control agents (BCAs) by co-inoculation. METHODS AND RESULTS: Rifampicin-resistant bacterial strains (Rif +) applied as single treatments to seed sown in columns of field soil produced persistent populations of 5-9 log10 cfu g-1 in the infection court of the damping-off pathogen Aphanomyces cochlioides in a controlled environment. However, isolates varied in their ability to colonize the lower rhizosphere. Percolating water significantly increased the colonization of the upper rhizosphere. Bacterial populations in the soil profiles of "non-rhizosphere" controls declined markedly with time. There was no interaction between the two selected BCAs applied as a seed treatment mixture. CONCLUSIONS: The distribution of the bacteria resulted primarily from root colonization although percolating water may modify the colonization profiles. Co-inoculation of the sugar-beet rhizosphere is a viable proposition. SIGNIFICANCE AND IMPACT OF THE STUDY: Potential BCAs were successfully delivered to the known infection court of A. cochloides and persisted for the infection period. This bioassay can be used as a tool for the selection of BCAs for field trials.

Nimodipine premedication and induction dose of propofol.

UNLABELLED: Antagonists at the L-type voltage sensitive calcium channel (L-VSCC) potentiate anesthetic potency in experimental models, suggesting that it may be a target site for IV anesthetics. Nimodipine is a 1, 4-dihydro- pyridine antagonist of L-VSCC which crosses the blood-brain barrier. We tested the hypothesis that premedication with oral nimodipine in healthy patients would reduce the induction dose of propofol, independently of its effects on the cerebral circulation. Sixty ASA physical status I or II patients (18-60 yr), undergoing knee arthroscopy or minor urological surgery, were randomized to receive either nimodipine 60 mg or placebo, orally 1-2 h before induction. Noninvasive mean blood pressure, heart rate, and time-averaged mean velocity in the middle cerebral artery by using transcranial Doppler ultrasonography were obtained before and 5 min after the induction of anesthesia. Propofol 1% was administered by an infusion pump at a rate of 10 mL/min. Both groups of patients had a reduction in mean blood pressure after the induction (P < 0.01), but there were no significant differences between the groups. The induction dose of propofol was 2.19 mg/kg (95% confidence interval [CI]: 1.97-2.42) in the nimodipine group, compared with 2.16 mg/kg (95% CI 1.98-2.34) in the control group, P = 0.8. Time-averaged mean velocity remained unchanged after the induction of anesthesia in both patients receiving nimodipine premedication (51% CI 43-59 cm/s to 52% CI 46-58 cm/s, P = 0.6) and those receiving placebo (50% CI 43-58 cm/s to 53% CI 45-59 cm/s, P = 0.3). Premedication with oral nimodipine 60 mg does not reduce the induction dose of propofol compared with placebo, casting doubt on the hypothesis that propofol has an anesthetic action at L-VSCC. IMPLICATIONS: Premedication with oral nimodipine 60 mg does not reduce the induction dose of propofol compared with placebo, casting doubt on the hypothesis that propofol has an anesthetic action at L-type voltage sensitive calcium channels.

Some failings of pulsatility index and damping factor.

Pulsatility index (PI) is a commonly used method of objective assessment of the Doppler waveform. PI falls with increasing proximal stenosis and is raised by increasing peripheral resistance. Damping factor (DF) for an arterial segment is calculated by dividing the proximal by the distal PI. DF rises with increasing severity of disease of the arterial segment. DF is not, however, sufficiently accurate to be used alone but is usually combined with transit time measurements to provide information of diagnostic use. Both PI and DF have been examined in a canine model of combined segment disease. With increasing stenosis, distal PI falls as expected but so also does proximal PI. Such a stenosis is, in effect, a flow-throttling resistance so that although the characteristics of blood flow are altered by its presence, similar changes are observed both above and below the stenosis. The reduction of PI by a stenosis distal to the insonation site may result in the false interpretation of a low PI as indicating disease proximal to the insonation site. The observed similarity between PI proximal and distal to a stenosis reduces the usefulness of pulsatility index damping factor, particularly in the assessment of the femoro-popliteal segment in combined segment disease.

Genetical analysis of microspore derived plants of barley (Hordeum vulgare).

From an F1 hybrid between the two barley (Hordeum vulgare L.) cultivars 'Golden Promise' and 'Mazurka' a series of doubled haploid (DH) lines were generated both from microspores by anther culture and from immature zygotic embryos after hybridization withH. bulbosum. The DH lines from both sources were used to monitor the segregation of the five major genes, rachilla hair length, DDT susceptibility, height, C hordein polymorphism and mildew resistance. Whereas the microspore-derived samples showed significant departures from the expected 1∶1 ratio for three of the five genes, theH. bulbosum lines showed deviation for only one gene. Analysis of linkage data also showed differences between the two series of DH lines. Cytogenetic analysis revealed a mean chiasma frequency in theH. bulbosum lines which was very similar to the F1 hybrid. In contrast, four of the ten microspore derived lines examined showed a reduced chiasma frequency. One showed evidence of translocation heterozygosity.

Factors influencing outcome in elderly patients with urinary incontinence and detrusor instability.

The majority of elderly incontinent patients with detrusor instability in whom various medical treatments had been tried and failed, became dry or very significantly improved following attendance at a Continence Clinic. The reason for this was not only because this clinic had access to specialized investigational equipment, but also because patients received individual psychological and practical management of their incontinence. All were given time to discuss their problems, had imipramine titrated against effect, and practised habit-retraining programmes. Treatment was started in hospital if there was any doubt of the patients misunderstanding or not complying with such regimens. Patients did best if they were sensible and mobile. Urodynamic measurements seemed less important individually, although overall final outcome correlated with the severity of the instability.

Haemodynamics of the adjunctive arteriovenous fistula in femorodistal bypass grafting: an experimental study.

The results of femorotibial bypass for limb salvage vary a great deal. One of the reasons for this might be the discrepancy between potential inflow and run-off into the foot. An arteriovenous fistula at the distal graft anastomosis may improve results but the best anatomical arrangement for the fistula is unknown. Ileofemoral grafts were performed in dogs after the hind limb was rendered ischaemic. The distal end of the graft was anastomosed proximal to, superimposed upon, or distal to an arteriovenous fistula between the femoral artery and its accompanying femoral vein. The effect of the fistula on graft/run-off haemodynamics was then measured. The adjunctive arteriovenous fistula increased inflow by a mean of 900 per cent and reduced systemic pressure by 10 per cent. Peripheral resistance was reduced by 85 per cent. Distal arterial run-off was maximized with respect to total graft flow when the graft was placed distal to the fistula (P less than 0.05). The venous steal of flow and perfusion pressure produced by the fistula was minimized with the same configuration compared to the two other arrangements (P less than 0.01 and P less than 0.05). Placement of the graft distal to the adjunctive arteriovenous fistula maximized distal arterial flow and pressure, and significantly increased graft flow.

Changes in Doppler ultrasound sonagrams at varying distances from stenoses.

The effect of different degrees of arterial stenosis on Doppler ultrasound sonagrams recorded from various distances downstream has been investigated in a canine model. It has been found that even mild stenoses give rise to considerable disturbances close to the stenosis, and that more severe stenoses give rise to greater disturbances which propagate further downstream. The distance the disturbances travelled was not related to stroke volume, and it was only for stenoses of 88% area reduction and greater that such disturbances ever propagated beyond one stroke-length. Quantitative assessment of moderate degrees of proximal stenosis using continuous wave Doppler ultrasound will not be reliable unless the distance between the major stenosis and the recording site is known.

Clinical and urodynamic studies in 100 elderly incontinent patients.

Clinical details were noted and urodynamic studies carried out on 100 elderly patients referred to an incontinence clinic, of whom 48 attended as day patients. Thirty patients had no problem apart from their incontinence, and only 38 had a clinically detectable neurological lesion. The average mental orientation score in 48 of the patients was 7.6, 23 patients scoring the top score of 10. Most patients were mobile without assistance from another person. Patients could be placed into one of four diagnostic groups according to the appearance of the cystometric tracings, but no bladder capacity or pressure was characteristic of any group. The maximum urethral closure pressure and functional profile length were similar for each group within each sex. There was no correlation between clinical and urodynamic findings, yet each of the four diagnostic categories have different therapeutic implications. It is concluded that urodynamic investigation is necessary in elderly incontinent patients before treatment.

Imipramine--a possible alternative to current therapy for urinary incontinence in the elderly.

Imipramine was given orally at night to 10 elderly patients with urinary incontinence associated with spontaneous unstable detrusor contractions. The dose was increased for each patient up to a maximum of 150 mg. at night, or until continence was achieved or side effects occurred. Of the 10 patients 6 became continent. In 3 of the 6 patients who underwent repeat cystometry bladder capacity had increased (mean 105 cc), bladder pressure at capacity decreased (mean 18 cm. water) and urethral pressure increased (mean 30 cm. water). There was no correlation between plasma desmethylimipramine and dose, or clinical or urodynamic effect.

The effect of proximal stenosis on Doppler waveforms: a comparison of three methods of waveform analysis in an animal model.

Methods of quantifying changes in spectrum analysed Doppler waveforms have been assessed in an experimental canine model. Artificial stenoses of varying (0-95%) area reduction have been inserted in the iliac artery of five anaesthetised greyhounds. 133 sets of Doppler waveforms recorded distally have been subjected to principal component analysis (PCA), transfer function analysis (TFA) and pulsatility index calculation (PI). Transfer function damping factor delta and PI were only able to distinguish those stenoses of greater than 85% area reduction. PCA, however, was more reliable in this range, and was usually able to distinguish stenoses of 65% and 77% area reduction. We conclude that PCA deserves further assessment in patients with aorto-iliac disease.

The effect of flavoxate on uninhibited detrusor contractions and urinary incontinence in the elderly.

The effect of flavoxate, a smooth muscle relaxant, was investigated in 6 elderly patients with uninhibited detrusor contractions associated with urinary incontinence. Cystometry was done initially to confirm the diagnosis, immediately after an intravenous injection of 100 mg. flavoxate and after 200 mg. flavoxate orally 4 times daily for 7 days. No consistent drug effect could be detected cystometrically and incontinence was unchanged clinically.

The relationship between ultrasonic pulsatility index and proximal arterial stenosis in a canine model.

Although the ultrasonic pulsatility index (PI) is used as a test of arterial inflow, the relationship between proximal arterial stenosis and PI has not been fully evaluated. In a series of experiments on dogs, over 200 measurements of pressure, flow (using an indwelling electromagnetic flowmeter), and PI were made distal to implanted arterial stenoses of varying length (0.5--9 mm) and cross-sectional area (36--94% reduction in area). It was shown that, whereas the reduction of PI correlates broadly with stenosis severity, the scatter of results is wide for all but the tightest stenoses. This scatter is due at least in part to the variability of the vascular bed distal to the site of measurement.