PCR-based detection of Xanthomonas campestris pathovars in Brassica seed

Xanthomonas campestris is a seedborne bacterium that causes black rot of crucifers. Substantial crop losses may result from the rapid spread of the bacteria under favourable conditions, especially those occurring during seedling production. A PCR-based method has been developed for the rapid and sensitive detection of the pathovars of X. campestris that affect crucifers. Primers were designed to specifically amplify a 619 bp fragment of the hrpF gene from X. campestris . Amplification products were not detected from other Xanthomonas species, or from other pathogenic or epiphytic bacteria occurring on these plants. To avoid false-negative results arising from the presence of amplification inhibitors in plant extracts, primers targeting a 360 bp section of the internal transcribed spacer (ITS) region from Brassica spp. were included in a multiplex PCR. The assay readily detected X. campestris pv. campestris infections in diseased plants and from bacterial colonies isolated on growth media, and was more sensitive and specific than traditional plating methods and a commercially available ELISA. A seed-washing protocol was optimized to allow the detection of a single artificially infected seed among 10 000 healthy seeds using the multiplex PCR.     

Effects of delay in reapplication of nitrogen fertilizer following cutting silage from a ryegrass sward

Plots of a 2-year-old sward of Merlinda perennial ryegrass received a routine dressing of 100 kg N ha⁻¹ as compound fertilizer in March 1991, followed by further dressings of 100 kg N ha⁻¹ after cuts 1, 2 and 3 in a simulated four-cut silage system, either the same day as cutting or with a delay of 3, 7, 10 or 14 d. Partial irrigation ensured that fertilizer could be taken up immediately. Annual total dry matter yield, the dry matter yield for the experimental cuts 2–4 and their apparent response to N showed no effect of delays of 0–10 d in reapplication of fertilizer, but a significant ( P < 0·01) reduction for 14 d delay. Significantly ( P = 0·001) higher apparent recovery of N in the cut herbage was recorded where reapplication of fertilizer had been delayed by 7 or 10 d (93% and 91% respectively) in comparison with either smaller (0 delay, 78%; 3 d delay, 87%) or greater delay (14 d, 86%). It is suggested that lower recoveries at 0 and 3 d result from the sward's reduced NO₃ uptake following defoliation and that, while in practical situations the conditions may not be as rigorous as those imposed in this experiment, further experiments to determine the fate of N not recovered in the herbage are warranted, so defining management circumstances in which immediate reapplication of N might not be advisable.     

The effect of cooking methods on the lipid composition of potatoes

The effect of boiling and baking in either a convential or microwave oven on the lipid content of potatoes was investigated. The crude lipid content of potatoes was lowered significantly by all methods of cooking. The loss was highest in potatoes cooked in a microwave oven, least in boiled tubers, and intermediate in tubers baked in a conventional oven. Similar trends were observed for both cortex and pith tissues. Generally, the phospholipid content followed the same trend as the crude lipid, the exception being the cortex tissue of tubers baked in the conventional oven, in which the greatest loss of phospholipid occurred. Cooking did not significantly alter the fatty acid composition of tubers. A possible relationship between the degree of cell rupturing, known to be influenced by different mechanisms of heat transfer, and the amounts of tuber lipid associated with different cooking methods was discussed.     

Inheritance of genes coding for gliadin proteins and glume colour introgressed into Triticum aestivum from a synthetic wheat

A synthetic hexaploid, Triticum timofeevii×T. tauschii, was used to transfer disease resistance genes to the commercial cultivar Saratovskaya 29 (S29) by backcrossing. After five backcrosses the resulting derivatives still showed some traits of the synthetic, namely brown spike glumes and several gliadin components. Genetic analysis showed that the derived forms had inherited the Gli-D1 allele of the synthetic, which was found to be tightly linked to a gene for glume colour. Recombination percentages between these genes was estimated to be 2.5 ± 1.7%. The development of the derivatives was also accompanied by a rearrangement within the Gli-B1 locus, resulting in the formation of a new variant of the allele in S29.     

Measured Soil Water Concentrations of Cadmium and Zinc in Plant Pots and Estimated Leaching Outflows from Contaminated Soils

Soil water concentrations of cadmium and zinc were measured in plant pots with 15 contaminated soils which differed in origin, texture, pH (5.1 – 7.8) and concentrations of cadmium (0.2 – 17 mg Cd kg-¹) and zinc (36 – 1300 mg Zn kg-¹). The soil waters contained total concentrations of 0.5 to 17 µg Cd L-¹ and 9 to 3600 µg Zn L-¹, which were dominated by free metal ions as measured by an ion exchange-resin method. Annual leaching outflows were estimated from soil water concentrations to be 0.5 – 17 g Cd ha-¹ y-¹ and 9 – 3600 g Zn ha-¹ y-¹ per 100 mm of net percolation, corresponding to 0.1% per year of the total soil content of cadmium and zinc. The measured soil water concentrations of cadmium and zinc did not correlate linearly with the corresponding soil concentrations but correlated fairly well with concentrations measured in Ca(NO₃)₂ extracts of the soils and with soil water concentrations estimated from soil concentrations and pH. Such concentration estimates may be useful for estimating amounts of cadmium and zinc being leached from soils.     

Porcine leptin inhibits lipogenesis in porcine adipocytes

The present study examined whether recombinant porcine leptin alters lipid synthesis in porcine adipocytes. The stromal-vascular cell fraction of neonatal pig subcutaneous adipose tissue was isolated by collagenase digestion, filtration, and subsequent centrifugation. These cells were seeded on 25-cm2 tissue culture flasks and proliferated to confluency in 10% (vol/vol) fetal bovine serum in Dulbecco's modified Eagle medium/F12 (DMEM/F12, 50:50). Cultures were differentiated using 2.5% pig serum (vol/vol), 10 nM insulin, 100 nM hydrocortisone. After 7 d of lipid filling, cultures were washed free of this medium, incubated overnight in DMEM/F12 containing 2% pig serum (vol/vol), and then used for experiments. Acute experiments assessed U-(14)C-glucose or 1-(14)C-palmitate metabolism in cultures exposed to porcine leptin (0 to 1,000 ng/mL medium) for 4 h. Chronic experiments used cultures incubated with 0 to 1,000 ng porcine leptin/mL medium for 44 h before measurements of U-(14)C-glucose and 1-(14)C-palmitate oxidation and incorporation into lipid. Another experiment examined whether chronic leptin treatment alters insulin responsiveness by including insulin (10 nM) with incubations containing leptin. Leptin had no acute effects on glucose oxidation or conversion to lipid (P > 0.05). Acute leptin treatment decreased palmitate incorporation into lipids up to 45% (P < 0.05). Chronic leptin exposure decreased glucose oxidation (21%), total lipid synthesis (18%), and fatty acid synthesis (23%) at 100 ng/mL medium (P < 0.05). Insulin increased rates of glucose oxidation, total lipid, and fatty acid synthesis (P < 0.05); however, chronic exposure to 10 ng leptin/mL medium decreased the effectiveness of 10 nM insulin to affect these measures of glucose metabolism by approximately 18 to 46% (P < 0.05). Higher concentrations of leptin inhibited all effects of insulin on glucose metabolism (P < 0.05). Chronic exposure to leptin increased palmitate oxidation by 36% (P < 0.05). Chronic leptin exposure decreased palmitate incorporation into total lipids by 40% at 100 ng/mL medium (P < 0.05). Lipoprotein lipase activity was not affected (P > 0.05) by leptin. These data indicate that leptin functions to promote partitioning of energy away from lipid accretion within porcine adipose tissue by inhibiting glucose oxidation and lipogenesis indirectly, by decreasing insulin-mediated stimulation of lipogenesis, and by stimulating fatty acid oxidation while inhibiting fatty acid esterification.     

PET/CT FOLLOWING INTENSITY-MODULATED RADIATION THERAPY FOR PRIMARY LUNG TUMOR IN A DOG

A primary lung tumor in a dog treated with intensity-modulated radiation therapy was imaged approximately 6 weeks and 1-year posttreatment with combined positron emission tomography (PET) and computed tomography, utilizing the radiotracers 18F-fluorodeoxyglucose and 18F-fluorothymidine. These two tracers allowed discrimination of tumor from inflammation, and demonstrated spread of tumor along airways over time after treatment. Fusion of functional imaging with anatomic imaging is a useful tool, particularly in the field of oncology, with the potential for PET markers that delineate tumor from normal or reactive tissue, and potential or actual response to therapy.