Somaclonal variation as a tool to develop pest resistant plants of Torenia fournieri ‘Compacta Blue’

Callus cultures of Torenia fournieri Compacta Blue were initiated on a modified Murashige and Skoog salt medium (MS) with 2.26 M 2,4-dichlorophenoxyacetic acid. Shoots were regenerated from these cultures using MS medium amended with 2.46 M indolebutyric acid and 8.88 M benzyladenine. These shoot cultures were subjected to two-spotted spidermite (Tetranychus urticae Koch.) and the greenhouse whitefly [Trialeurodes vaporariorum (Westwood)]. Pests were allowed to feed until such time that their populations started to decrease due to lack of food. The remaining live tissue of the Torenia was placed on MS medium amended with 2.28 M zeatin to induce new adventitious shoots and plantlets. Newly regenerated plantlets were acclimated to greenhouse conditions and evaluated for resistance to the pest to which they were subjected in vitro. Highly significant differences in pest numbers were found in somaclones for both the two-spotted spidermite and greenhouse whitefly when compared to control plants. A wide range of variability was observed among the somaclonal population. There were significantly fewer mite eggs laid on plants regenerated from in vitro cultures screened with two-spotted spidermites than on seed sown controls. Regenerants from cultures screened with whiteflies in vitro had fewer eggs, immatures and live adults than controls.Abbreviations BA benzyladenine - IBA indolebutyric acid - 2,4-d 2,4-dichlorophenoxyacetic acid - MS Murashige and Skoog salt medium Storrs Agricultural Research Station Scientific Publication 1641.     

Arachidonic acid-induced hypersensitive cell death as an assay of downy mildew resistance in pearl millet

Arachidonic acid (AA) induces hypersensitive response (HR) on coleoptile/root regions of two-day-old pearl millet seedlings. The response is comparable to the HR induced by the downy mildew pathogen, Sclerospora graminicola. A time gap in the appearance of cell necrosis among genotypes of pearl millet was related to the degree of resistance to downy mildew. Based on the time required for the development of necrotic spots induced by AA, the pearl millet genotypes were categorised as highly resistant/resistant (HR in 3–6 h), susceptible (HR in 7–12 h) and highly susceptible (HR in 13 h and above). The percentage disease incidence in each genotype was compared with the time required for the development of AA-induced HR. The appearance of hypersensitive cell necrosis was rapid in genotypes having high resistance to downy mildew and was slow in genotypes with high susceptibility. This simple method of screening various pearl millet genotypes in the absence of the pathogen aids in identifying the downy mildew resistant/susceptible host cultivars without the risk of introducing the virulent race of the pathogen.     

Determining the effectiveness of resistance to subterranean clover mottle sobemovirus in different genotypes of subterranean clover in the field using the grazing animal as virus vector

The effectiveness of resistance to subterranean clover mottle sobemovirus (SCMoV) previously identified in different genotypes of subterranean clover (Trifolium subterraneum) inoculated with infective sap in the glasshouse, was tested in two field experiments which used the grazing animal as virus vector. Replicated plots each consisting of paired test rows of 20 different genotypes were used. Clover plants infected with SCMoV were transplanted in between the paired test rows and these acted as sources of the virus for spread by grazing sheep. Although used in different years at different sites with different virus isolates, the field exposure methodology employed produced consistent results. The genotypes each behaved similarly in both experiments as regards the relative extents of SCMoV infection that developed, levels ranging from 0–98%. The previously identified resistance in six ‘highly resistant’ and three ‘partially resistant’ cultivars was effective under field conditions. However, the ‘partial resistance’ in three others was overcome, cvs Green Range and Mt Barker developing levels of infection approaching those in ‘susceptible’ cultivars, while an intermediate infection level developed in cv. Karridale. The three cultivars in which partial resistance was not effective all belonged to ssp. subterraneum. In subterranean clover breeding programmes, field screening using the grazing animal as a vector is advisable to determine whether SCMoV resistance found by sap inoculation is still effective under field conditions.     

A protocol for the efficient screening of putatively transformed plants forbar,the selectable marker gene,using the polymerase chain reaction

Amplification of thebar gene usingTaq DNA polymerase in PCR is often not successful, possibly due tobar's high GC content. We describe a PCR protocol in which reliable amplification at a sensitivity of one gene copy per genome (in this study, barley) present in the reaction was achieved using a novel pair of primers and Expand^tm High Fidelity DNA polymerase mix (Boehringer Mannheim). This method should allow for rapid screening of plants putatively transformed withbar.     

An impedimetric method for rapid screening of cosmetic preservatives

An efficient impedance method was developed for rapid evaluation of cosmetic preservatives. The method used decimal reduction time or D-value to assess preservative efficacies. The D-value, which was calculated from the plot of Log CFU ml^–1 versus time by linear regression analysis, could be obtained within 48 h. Thus, the time required for the challenge test was reduced from 4–8 weeks with the standard procedures (eg US Pharmacopeia), to 2 days with the current method. A calibration curve (r=-0.95) was established by plotting the Log CFU ml^–1 versus capacitance detection time (DT) of 108 samples. With the calibration, CFU can be estimated directly from the impedance test without plating. Two commercial biocides and several other chemicals were evaluated in a shampoo by the impedance procedure againstPseudomonas aeruginosa. The D-values obtained from the impedance test were not significantly different from those produced by the conventional plate count method. The technique was found to be particularly useful when screening a large number of compounds to find novel preservatives and synergistic preservative combinations.     

Determinants of peak muscle power: effects of age and physical conditioning

The relationships between absolute peak muscle power (W _peak), muscle cross sectional area (CSA_tot, i.e. the sum of both thigh and calf CSA) and muscle high energy phosphate concentration (adenosine 5-triphosphate [ATP] and phosphocreatine concentrations [PC]) were studied in 47 subjects classified into five groups: A, 10 sedentary (S) subjects aged 20–35 years; B, 9 S aged 35–50 years; C, 9 S aged more than 50 years; D, 13 children aged 8–13 years; and E, 6 athletes (top level volleyball players) aged 24 (SD 3) years. The W _peak was measured during a maximal vertical high jump off both feet on a force platform. The CSA_tot was measured anthropometrically. The [ATP] and [PC] were determined by ³¹Phosphorus nuclear magnetic resonance spectroscopy. The W _peak decreased with age, was 65% lower in D than in A, and 43% higher in E than in A. The CSA_tot did not vary with age, was 45% smaller in D than in A, and 15% greater in E than in A. The [ATP] and [PC] were essentially the same in all groups. The changes observed in W _peak were only partially accounted for by changes in CSA_tot. Therefore, in addition to the variables investigated, other factors appear to have been involved in the determination of W _peak with increasing age and training. An important role may be played by hormonal, particularly at puberty, and neural factors.     

The effects of tail autotomy on survivorship and body growth of Uta stansburiana under conditions of high mortality

We examined the effects of tail autotomy on survivorship and body growth of both adult and juvenile Uta stansburiana by directly manipulating tail condition. Tail loss decreased neither survivorship nor rate of body growth for individuals in two natural populations. Lack of an influence of tail loss on survivorship in these two populations may be the result of high mortality. Under high mortality any differential effects of tail loss will be lower than in populations facing lower mortality. Growth experiments in the laboratory demonstrated that, under conditions of minimal environmental variation and social interactions, there is no tradeoff between body growth and tail regeneration as has been suggested for other species of lizards. One possible reason for this difference is that U. stansburiana does not use the tail as a storage organ for lipids. The original and regenerated tails are composed mainly of protein. In general, any differential body growth between tailed and tailless individuals may be due to social interactions and not a diversion of limited energy into tail regeneration.     

Cell Lysis Due to Ultrasound Gel In Fine Needle Aspirates; an Important New Artefact In Cytology

An important new artefact in cytopathology is described. the initial observation of the artefact followed contamination of a breast fine needle aspiration (FNA) sample by ultrasound gel which was used to localize the lesion. the changes proved reproducible ex vivo. the changes varied depending on the conditions and degree of contamination, and ranged from cell swelling to leakage of nuclear chromatin and cell lysis. This artefact is discussed in the context of other major sources of cytology artefact. Pathologists and radiologists should beware of the detrimental effects of ultrasound gel on cytology specimens.     

High pressure conditions stimulate expression of chloramphenicol acetyltransferase regulated by the lac promoter in Escherichia coli

Abstract Recombinant plasmids with the chloramphenicol acetyltransferase (CAT) structural gene behind several kinds of promoters were tested for expression in Escherichia coli during growth at atmospheric pressure (0.1 MPa) and at high pressure (30 MPa). Expression of the CAT gene from the lac promoter was remarkably activated (approx. 78-fold) by high pressure in the absence of the inducer isopropyl-β-d-thiogalactopyranoside (IPTG). The stimulation of the CAT activity by the lac promoter at high pressure did not simply result from an increased plasmid copy number, because the CAT activities from the other promoters and β-lactamase activities were unaffected at high pressure.