Callus induction and plant regeneration from broccoli (Brassica oleracea var. italica) for transformation

We evaluated the efficiency of callus induction and plantlet regeneration from hypocotyl explants of broccoli (Brassica oleracea var. italica). The cultivars were ‘Marathon’, ‘Greenbelt’, and ‘Shogun’. Transformation success was not affected by the presence of tobacco feeder-cell layers on the culture media. The frequency of shoot regeneration was greater from 10-d-old hypocotyls than from 14-d-old hypocotyls. Both ‘Marathon’ and ‘Greenbelt’ had higher potentials for tissue regeneration than did ‘Shogun’. We found that for transformation selection, the optimum concentration was either 50 mg/L kanamycin or 100 mg/L genetkin.

Hydrolytic kinetic resolution of terminal mono- and bis-epoxides in the synthesis of insect pheromones: routes to (-)-(R)- and (+)-(S)-10-methyldodecyl acetate, (-)-(R)-10-methyl-2-tridecanone, (-)-(R)-(Z)-undec-6-en-2-ol (Nostrenol), (-)-(1R,7R)-1,7-dime

Hydrolytic kinetic resolution (HKR) of functionalised epoxides using (salen)Co(OAc) complexes provides enantiomerically enriched epoxides and diols, which have been transformed into important insect sex pheromones. In this general approach, (-)-(R)- and (+)-(S)-10-methyldodecyl acetates from the smaller tea tortrix moth were obtained, as was (-)-(R)-10-methyltridecan-2-one from the southern corn rootworm. The (S)-epoxide obtained from undec-1-en-6-yne was transformed to (-)-(R)-(Z)-undec-6-en-2-ol (Nostrenol) from ant-lions. HKR of appropriate bisepoxides was also investigated, and transformations of the resulting bisepoxides and epoxydiols provided (-)-(1R,7R)-1,7-dimethylnonylpropanoate from corn rootworms, (-)-(6R,12R)-6,12-dimethylpentadecan-2-one from the female banded cucumber beetle, and (-)-(2S,11S)-2,11-diacetoxytridecane and (+)-(2S,12S)-2,12-diacetoxytridecane from female pea-midges. (C) 2002 Elsevier Science Ltd. All rights reserved.

Nitrification in a Vertisol subsoil and its relationship to the accumulation of ammonium-nitrogen at depth

Unusually high concentrations of ammonium have been observed in a Vertisol below 1 m depth in southeast Queensland. This study investigated the possibility that an absence of nitrification is allowing this ammonium to accumulate and persist over time, and examined the soil environmental characteristics that may be responsible for limiting nitrifying organisms. The possibility that anaerobiosis, soil acidity, soil salinity, low organic carbon concentrations, and/or an absence of active nitrifying microorganisms were responsible for limiting nitrification was examined in laboratory and field studies. The presence/absence of anaerobic conditions was determined qualitatively using a field test to give an indication of electron lability. In addition, an incubation study was conducted and soil environmental conditions were improved for nitrifying organisms by adjusting the pH from 4.4 to 7, adjusting the electrical conductivity from 1.6 to 0.5 dS/m, amending with a soluble carbon substrate at a rate of 500 mg/kg, and using microorganisms from the surface horizon to inoculate to the subsoil. Over a 180-day period no nitrification was detected in the control samples from the incubation study, indicating that an extremely low rate of nitrification is likely to be responsible for allowing ammonium to accumulate in this soil. Analysis of the effect of soil environmental conditions on nitrification revealed that anaerobic conditions did not exist at depth and that pH, EC, organic carbon, and inoculation treatments added in isolation had no effect on nitrification. However, when inoculum was added to the soil in combination with pH, a significant increase in nitrification was observed, and the greatest amount of nitrification was observed when inoculum, pH, and EC treatments were added in combination. It was concluded that the reason for the low rate of nitrification in this soil is primarily the absence of a significant population of active nitrifying microorganisms, which may have been unable to colonise the subsoil environment due to its acidic, and to a lesser extent, its saline environment.

Molecular evidence for the common origin of snap-traps among carnivorous plants

The snap-trap leaves of the aquatic waterwheel plant (Aldrovanda) resemble those of Venus' flytrap (Dionaea), its distribution and habit are reminiscent of bladderworts (Utricularia), but it shares many reproductive characters with sundews (Drosera). Moreover, Aldrovanda has never been included in molecular phylogenetic studies, so it has been unclear whether snap-traps evolved only once or more than once among angiosperms. Using sequences from nuclear 18S and plastid rbcL, atpB, and matK genes, we show that Aldrovanda is sister to Dionaea, and this pair is sister to Drosera. Our results indicate that snap-traps are derived from flypaper-traps and have a common ancestry among flowering plants, despite the fact that this mechanism is used by both a terrestrial species and an aquatic one. Genetic and fossil evidence for the close relationship between these unique and threatened organisms indicate that carnivory evolved from a common ancestor within this caryophyllid clade at least 65 million years ago.

Testing the relationship between morphological and molecular rates of change along phylogenies

Molecular evolution has been considered to be essentially a stochastic process, little influenced by the pace of phenotypic change. This assumption was challenged by a study that demonstrated an association between rates of morphological and molecular change estimated for total-evidence phylogenies, a finding that led some researchers to challenge molecular date estimates of major evolutionary radiations. Here we show that Omland's (1997) result is probably due to methodological bias, particularly phylogenetic nonindependence, rather than being indicative of an underlying evolutionary phenomenon. We apply three new methods specifically designed to overcome phylogenetic bias to 13 published phylogenetic datasets for vertebrate taxa, each of which includes both morphological characters and DNA sequence data. We find no evidence of an association between rates of molecular and morphological rates of change.

New type of dual-channel PAM chlorophyll fluorometer for highly sensitive water toxicity biotests

A new type of dual-channel PAM chlorophyll fluorometer has been developed, which is specialised in the detection of extremely small differences in photosynthetic activity in algae or thylakoids suspensions. In conjunction with standardised algae cultures or isolated thylakoids, the new device provides an ultrasensitive biotest system for detection of toxic substances in water samples. In this report, major features of the new device are outlined and examples of its performance are presented using suspensions of Phaeodactylum tricornutum (diatoms) and of freeze-dried thylakoids of Lactuca sativa (salad). Investigated and reference samples are exposed to the same actinic intensity of pulse-modulated measuring light. The quantum yields are assessed by the saturation pulse method. Clock-triggered repetitive measurements of quantum yield typically display a standard deviation of 0.1%, corresponding to the inhibition induced by 0.02 mug diuron l(-1). Hence, for diuron or compounds with similar toxicity, the detection limit is well below the 0.1 mug l(-1) defined as the limit for the presence of a single toxic substance in water by the European Commission drinking water regulation. The amounts of water and biotest material required for analysis are very small, as a single assay involves two 1 ml samples, each containing ca. 0.5 mug chlorophyll. Both with Phaeodactylum and thylakoids the relationship between inhibition and diuron concentration is strictly linear up to 10% inhibition, with very similar slopes. Apparent inhibition depends on the actinic effect of the measuring light, showing optima at 6 and 4 mumol quanta m(-2) s(-1) with Phaeodactylum and thylakoids, respectively.

The relationship of natural killer cell counts, perforin mRNA and CD2 expression to post-exercise natural killer cell activity in humans

The aim of this study was to further investigate the mechanism of suppression of natural killer (NK) cell cytotoxic activity In peripheral blood following strenuous exercise. Blood was collected for analysis of NK cell concentration, cytotoxic activity, CD2 surface expression and perforin gene expression from runners (RUN, n = 6) and resting controls (CONTROL, n = 4) pre-exercise, 0, 1.5, 5, and 24 h following a 60-min treadmill run at 80% of VO2 peak. Natural killer cytotoxic activity, measured using a whole blood chromium release assay, fluctuated minimally in the CONTROL group and increased by 63% and decreased by 43% 0 and 1.5 h post-exercise, respectively, in the RUN group (group x time, P < 0.001). Lytic index (cytotoxic activity per cell) did not change. Perforin mRNA, measured using quantitative real-time polymerase chain reaction (ORT-PCR) decreased from pre- to post-exercise and remained decreased through 24 h, The decrease from pre- to 0 In post-exercise was seen predominately in the RUN group and was inversely correlated r = - 0.95) to pre-exercise perform mRNA. The NK cell surface expression of CD2 (lymphocyte function-associated antigen-2) was determined using fluorescent antibodies and flow cytometry, There was no change in the proportion of NK cells expressing CD2 or CD2 density, We conclude that (1) numerical redistribution accounted for most of the change in NK cytotoxic activity following a strenuous run, (2) decrease in perforin gene expression during the run was inversely related to pre-exercise levels but did not parallel changes in cytotoxic activity, and (3) CD2 surface expression was not affected by exercise.

Structure and function of plant toxins (with emphasis on cystine knot toxins)

Plant toxins are substances produced and secreted by plants to defend themselves against predators. In a broad sense, this includes all substances that have a toxic effect on targeted organisms, whether they are microbes, other plants, insects, or higher animals. Plant toxins have a diverse range of structures, from small organic molecules through to proteins. This review gives an overview of the various classes of plant toxins but focuses on an interesting class of protein-based plant toxins containing a cystine knot motif. This structural motif confers exceptional stability on proteins containing it and is associated with a wide range of biological activities. The biological activities and structural stability offer many potential applications in the pharmaceutical and agricultural fields. One particularly exciting prospect is in the use of protein-based plant toxins as molecular scaffolds for displaying pharmaceutically important bioactivities. Future applications of plant toxins are likely to involve genetic engineering techniques and molecular pharming approaches.

Accurate orientation relationship between ferrite and austenite in low carbon martensite and granular bainite

Convergent beam Kikuchi diffraction was used to accurately determine the orientation relationships (ORs) between austenite and martensite, and between austenite and granular bainite in two Fe-Ni-Mn-C alloys. Both martensite and granular bainite have the same crystallographic characteristics with the OR: (111)(A)parallel to(101)(F), [1 (1) over bar0](A) 2.5degrees +/- 2degrees from [1 (1) over bar(1) over bar](B).

Relationship between stress-induced martensitic transformation and impact toughness in low carbon austenitic steels

The effect of test temperature, which controls the stability of austenite, on the impact toughness of a low carbon Fe-Ni-Mn-C austenitic steel and 304 stainless steel, has been investigated. Under impact conditions, stress-induced martensitic transformation occurred, in a region near the fracture surface, at test temperatures below 80degreesC for the Fe-Ni-Mn-C steel and below -25degreesC for 304 stainless steel. The former shows significant transformation toughening and the highest impact toughness was obtained at 10degreesC, which corresponds to the maximum amount of martensite formed by stress-induced transformation above the Ms temperature. The stress-induced martensitic transformation contributes negatively to the impact toughness in the 304 stainless steel. Increasing the amount of stress-induced transformation to martensite, lowered the impact toughness. The experimental results can be well explained by the Antolovich theory through the analysis of metallography and fractography. The different effect of stress-induced transformation on the impact toughness in Fe-Ni-Mn-C steel and 304 stainless steel has been further understood by applying the crystallographic model for stress-induced martensitic transformation to these two steels. (C) 2002 Kluwer Academic Publishers.

Outcomes after trans-tibial amputation: The relationship between quiet stance ability, strength of the hip abductor muscles and gait

Background and Purpose Previous research regarding the symmetry of trans-tibial amputees has examined weight distribution and various gait parameters between prosthetic and sound limbs. However, to date, no known research has determined if asymmetry is present in the strength of the hip abductor muscles or if correlations exist between these categories of symmetry. The purpose of the present study was, therefore, to document asymmetry present in stance, strength and gait measures, and to determine the relationship between these variables. Method Twenty-three elderly, unilateral trans-tibial amputees stood on two adjacent forceplates whilst the weight distribution and standard deviation (SD) of the anterior-posterior and the medio-lateral centre of pressure excursion (COPE) under each limb was recorded during four 40 s trials: quiet stance (QS), with eyes open and eyes closed; and even stance (ES), with eyes open and eyes closed. Gait measures (velocity, cadence, step and stride lengths, stance:swing ratio and period of double support) over 10 m of fast, yet safe walking and measures of the strength of hip abductor muscles were also obtained by use of a stride analyser and a dynamometer, respectively. Results No significant differences were found between QS and ES measures. However, significantly more weight was taken on the sound limb than on the amputated limb. Notably, more anterior-posterior movement occurred under the sound limb than the amputated limb, with this becoming more apparent with the eyes closed. Movement in the medio-lateral direction was found to be the same between sides. No differences in muscle strength or gait measures between limbs were demonstrated. However. strong hip abductor muscles were correlated with increased weight-bearing on the amputated limb, improved gait parameters and reduced medio-lateral COPE under the amputated limb. Conclusions This research confirms the asymmetrical nature of amputee stance and demonstrates symmetry of strength and gait measures between limbs. The correlations between hip abductor muscle strength, weight distribution and gait measures illustrates the importance of pre- and postoperative training of these muscles. Copyright © 2002 Whurr Publishers Ltd.

Zeolite/rock phosphate - a novel slow release phosphorus fertiliser for potted plant production

A glasshouse study was undertaken to determine if the zeolite mineral clinoptilolite from an Australian deposit in combination with rock phosphate (RP) could significantly enhance the uptake of P by sunflowers. The zeolite/RP combination was intended to act as an exchange-fertiliser, with Ca2+ exchanging onto the zeolite in response to plant uptake of nutrient cations (NH4+ or K) enhancing the dissolution of the RP. A reactive RP (Sechura) and a relatively non-reactive RP (Duchess) were examined. Zeolite was used in Ca2+-, K+- and NH4+-saturated forms at ratios of 3.5:1 and 7:1 with RP; Ca2+-zeolite was considered the control, with exchange-induced dissolution possible from K+-and NH4+-zeolite, The zeolite/RP mixture was applied as a vertical band adjacent to the sunflower seedling. In addition, N was supplied as urea in an effort to determine if RP dissolution resulted from H+ release by nitrification. Phosphorus supply from the zeolite/RP system was compared with an available P source (KH2PO4). The experiment clearly demonstrated greatly enhanced plant uptake of P from RP when applied in combination with NH4-zeolite, though the P uptake was lower than that from the soluble P source. The zeolite/RP interaction was much more effective with the reactive R-P than the non-reactive material, Within the NH4+-zeolite/RP band, root proliferation was greatly increased, as would be expected in an exchange-fertiliser system. The K+-zeolite system did not produce a significantly greater yield than the Ca2+-zeolite control, probably because adequate K+ supply from the basal application reduced uptake within the zeolite/RP band, thus reducing the extent of exchange-induced dissolution. Nevertheless, increased root proliferation within the band was observed, implying that exchange-induced dissolution may also be possible from this system. The zeolite/RP system offers the considerable advantage of P release in response to plant demand and is unique in this regard. (C) 2002 Elsevier Science B.V. All rights reserved.

Attachment, marital interaction, and relationship satisfaction: A diary study

The relations among measures of attachment, spouse behavior, and marital satisfaction were assessed in a broad sample of 193 married couples, using both questionnaire and diary methods. Insecure attachment was associated with less favorable reports of spouse behavior, as assessed by diary checklists. Marital satisfaction was predicted by attachment treasures and reports of spouse behavior. The relation between attachment security and marital satisfaction was moderated, but not mediated, by reported spouse behavior. Specifically, insecure individuals' evaluations of their relationships were more reactive to recent spouse behavior, an effect that was especially marked for fearful participants and for those in longer-term marriages. Some gender differences in patterns of prediction were obtained. The results are discussed in terms of the working models associated with attachment styles, and the processes by which relationship satisfaction may be eroded over time.

Leaf water potential and osmotic adjustment as physiological traits to improve drought tolerance in rice

A sample of recombinant inbred lines (RILs) was derived from a bi-parental cross between Lemont and BK88-BR6, which contrasted in maintenance of leaf water potential (LWP) and expression of osmotic adjustment (OA). Genotypic variation for LWP and OA, and their associations with yield determination under water deficit, was studied in a series of five field experiments. Genotypic variation in the maintenance of high LWP was consistent across water deficit experiments. In the determination of genotypic variation in the maintenance of LWP, rate of water deficit was not an important factor influencing ranking, but degree of water deficit, and phenological development stage were important, particularly around heading. Genotypic variation in expression of OA was also observed under water deficits during both vegetative and flowering stages but ranking was inconsistent across experiments. This was in part because of large experimental errors associated with its measurement, but also because the expression of OA was associated with extent of decline of LWP. The relationship between OA and LWP was demonstrated when data were combined across experiments for vegetative and flowering stages. Under water-limited conditions around flowering, grain yield reduction was mainly due to a increased spikelet sterility. Variation in OA was not related to grain yield nor yield components. There were however, negative phenotypic and genetic correlations between LWP and percentage spikelet sterility measured at flowering stage on panicles at the same development stage during a water deficit treatment. This suggests that traits contributing to the maintenance of high LWP minimized the effects of water deficit on spikelet sterility and consequently grain yield. (C) 2002 Elsevier Science B.V. All rights reserved.