Inhibition of adventitious rooting in Backhousia citriodora F. Muell cuttings correlate with the concentration of essential oil

Backhousia citriodora is typical of the many commercially valuable woody Australian Myrtaceae species that are recalcitrant in forming adventitious roots from cuttings after maturation. A series of experiments were conducted to identify an endogenous rooting inhibitor in line with established criteria. Endogenous levels of citral were correlated with the rooting capacities of juvenile versus mature, and easy- versus difficult-to-root genotypes of B. citriodora, in both winter and summer. The biological activity of citral was confirmed in bioassays on mung beans and easy-to-root B. citriodora seedlings. Evidence of a common mechanism of root inhibition with other species in the Myrtaceae and the role of action of citral are discussed.

Dry matter production and boron concentrations of vegetative and reproductive tissues of canola and sunflower plants grown in nutrient solution

Canola (Brassica napus L.) and sunflower (Helianthus annuus L.), two important oilseed crops, are sensitive to low boron (B) supply. Symptoms of B deficiency are often more severe during the reproductive stage, but it is not known if this is due to a decreased external B supply with time or an increased sensitivity to low B during this stage. Canola and sunflower were grown for 75 days after transplanting (DAT) in two solution culture experiments using Amberlite (IRA-743) B-specific resin to maintain constant B concentration in solution over the range 0.6 - 53 muM. Initially, the vegetative growth of both crops was good in all treatments. With the onset of the reproductive stage, however, severe B deficiency symptoms developed and growth of canola and sunflower was reduced with less than or equal to 0.9 and less than or equal to 0.7 muM B, respectively. At these concentrations, reproductive parts failed to develop. The critical B concentration (i.e. 90% of maximum shoot dry matter yield) in the youngest opened leaf was 18 mg kg(-1) in canola and 25 mg kg(-1) in sunflower at 75 DAT. The results of this study indicate that the reproductive stage of these two oilseed crops is more sensitive than the vegetative stage to low B supply.

Peanut resistance to Sclerotinia minor and S-sclerotiorum

The fungi Sclerotinia minor and S. sclerotiorum are the causal agents of two similar diseases of peanut (Arachis hypogaea L.). Both diseases cause significant losses in the Australian peanut industry. Development of cultivars with resistance to Sclerotinia will be an important component of integrated control. The aims of this project are to generate information that will assist in breeding for Sclerotinia resistance in peanut: to identify Sclerotinia-resistant peanut germplasm, to understand the inheritance and estimate heritability of resistance, and to test the effectiveness of identified sources of resistance against both S. minor and S. sclerotiorum. This study has clearly established that material that shows resistance to S. minor in the USA is resistant to S. minor and likely to be resistant to S. sclerotiorum in Australia. The high level of resistance to both S. minor and S. sclerotiorum in germplasm from Texas, particularly TxAG-4, was confirmed. VA 93B showed good resistance in the field, which is primarily due to the open bush type rather than physiological resistance. Physiological resistance to S. minor was also identified in a cultivar and a landrace from Indonesia and a rust-resistant line from Queensland. All germplasm found to have high physiological resistance to S. minor belonged to the Spanish type. Inheritance of physiological resistance to S. minor was studied using a Generation Means Analysis (GMA) of the cross TxAG-4/VA 93B and its reciprocal. The broad-sense heritability of physiological resistance on a single plant basis was estimated at 47%, much higher than earlier estimates obtained in field studies. The average gene action of Sclerotinia resistance genes from TxAG-4 was found to be additive. No dominance effects were detected in the GMA. A small but significant reciprocal effect between TxAG-4 and VA 93B indicated that VA 93B passed on some physiological resistance maternally. An experiment was conducted to confirm the value of resistance against both S. minor and S. sclerotiorum. TxAG-4 was found to have physiological resistance to both S. minor and S. sclerotiorum. This resistance was expressed against both Sclerotinia species by progeny that were selected for resistance to S. minor. On the basis of the information obtained, the comparative advantages of 3 strategies for Sclerotinia-resistant cultivar development are discussed: (1) introduction of germplasm; (2) recurrent backcrossing with screening and crossing in the BCnF1 generation; and (3) pedigree selection. At present, introduction and backcrossing are recommended as the preferred strategies.

Kinetics of baculovirus replication and release using real-time quantitative polymerase chain reaction

The study of viral-based processes is hampered by (a) their complex, transient nature, (b) the instability of products, and (c) the lack of accurate diagnostic assays. Here, we describe the use of real-time quantitative polymerase chain reaction to characterize baculoviral infection. Baculovirus DNA content doubles every 1.7 h from 6 h post-infection until replication is halted at the onset of budding. No dynamic equilibrium exists between replication and release, and the kinetics are independent of the cell density at the time of infection. No more than 16% of the intracellular virus copies bud from the cell. (C) 2002 John Wiley & Sons, Inc. Biotechnol Bioeng 77: 476-480, 2002; DOI 10.1002/bit.10126.

A pilot study of the manual force levels required to produce manipulation induced hypoalgesia

Objective. A pilot investigation of the influence of different force levels on a treatment technique's hypoalgesic effect. Design. Randomised single blind repeated measures. Background. Optimisation of such biomechanical treatment variables as the point of force application, direction of force application and the level of applied manual force is classically regarded as the basis of best practice manipulative therapy. Manipulative therapy is frequently used to alleviate pain, a treatment effect that is often studied directly in the neurophysiological, paradigm and seldom in biomechanical research. The relationship between the level of force applied by a technique (e.g. biomechanics) and its hypoalgesic effect was the focus of this study. Method. The experiment involved the application of a lateral glide mobilisation with movement treatment technique to the symptomatic elbow of six subjects with lateral epicondylalgia. Four different levels of force, which were measured with a flexible pressure-sensing mat, were randomly applied while the subject performed a pain free grip strength test. Results. Standardised manual force data varied from 0.76 to 4.54 N/cm, lower-upper limits 95 Cl, respectively. Pain free grip strength expressed as a percentage change from pre-treatment values was significantly greater with manual forces beyond 1.9 N/cm (P = 0.014). Conclusions. This study, albeit a pilot, provides preliminary evidence that in terms of the hypoalgesic effect of a mobilisation with movement treatment technique, there may be an optimal level of applied manual force.

On the overabundance of light rare earth elements in terrestrial zircons and its implication for Earth's earliest magmatic differentiation

We present whole-rock and zircon rare earth element (REE) data from two early Archaean gneisses (3.81 Ga and 3.64 Ga) from the Itsaq gneiss complex, south-west Greenland. Both gneisses represent extremely rare examples of unaltered, fresh and relatively undeformed igneous rocks of such antiquity. Cathodoluminescence imaging of their zircons indicates a single crystallisation episode with no evidence for either later metamorphic and/or anatectic reworking or inheritance of earlier grains. Uniform, single-population U/Pb age data confirm the structural simplicity of these zircons. One sample, a 3.64 Ga granodioritic gneiss from the Gothabsfjord, yields a chondrite-normalised REE pattern with a positive slope from La to Lu as well as substantial positive Ce and slight negative Eu anomalies, features generally considered to be typical of igneous zircon. In contrast, the second sample, a 3.81 Ga tonalite from south of the Isua Greenstone Belt, has variable but generally much higher light REE abundances, with similar middle to heavy REE. Calculation of zircon/melt distribution coefficients (D-REE(zircon/melt)) from each sample yields markedly different values for the trivalent REE (i.e. Ce and Eu omitted) and simple application of one set of D-REE(zircon/melt) to model the melt composition for the other sample yields concentrations that are in error by up to two orders of magnitude for the light REE (La-Nd). The observed light REE overabundance in the 3.81 Ga tonalite is a commonly observed feature in terrestrial zircons for which a number of explanations ranging from lattice strain to disequilibrium crystallisation have been proposed and are further investigated herein. Regardless of the cause of light REE overabundance, our study shows that simple application of zircon/melt distribution coefficients is not an unambiguous method for ascertaining original melt composition. In this context, recent studies that use REE data to claim that > 4.3 Ga Hadean detrital zircons originally crystallised from an evolved magma, in turn suggesting the operation of geological processes in the early Earth analogous to those of the present day (e.g. subduction and melting of hydrated oceanic crust), must be regarded with caution. Indeed, comparison of terrestrial Hadean and > 3.9 Ga lunar highland zircons shows remarkable similarities in the light REE, even though subduction processes that have been used to explain the terrestrial zircons have never operated on the Moon. (C) 2002 Elsevier Science B.V. All rights reserved.

Instabilities and drop formation in cylindrical liquid jets in reduced gravity

The effects of convective and absolute instabilities on the formation of drops formed from cylindrical liquid jets of glycerol/water issuing into still air were investigated. Medium-duration reduced gravity tests were conducted aboard NASA's KC-135 and compared to similar tests performed under normal gravity conditions to aid in understanding the drop formation process. In reduced gravity, the Rayleigh-Chandrasekhar Equation was found to accurately predict the transition between a region of absolute and convective instability as defined by a critical Weber number. Observations of the physics of the jet, its breakup, and subsequent drop dynamics under both gravity conditions and the effects of the two instabilities on these processes are presented. All the normal gravity liquid jets investigated, in regions of convective or absolute instability, were subject to significant stretching effects, which affected the subsequent drop and associated geometry and dynamics. These effects were not displayed in reduced gravity and, therefore, the liquid jets would form drops which took longer to form (reduction in drop frequency), larger in size, and more spherical (surface tension effects). Most observed changes, in regions of either absolute or convective instabilities, were due to a reduction in the buoyancy force and an increased importance of the surface tension force acting on the liquid contained in the jet or formed drop. Reduced gravity environments allow better investigations to be performed into the physics of liquid jets, subsequently formed drops, and the effects of instabilities on these systems. In reduced gravity, drops form up to three times more slowly and as a consequence are up to three times larger in volume in the theoretical absolute instability region than in the theoretical convective instability region. This difference was not seen in the corresponding normal gravity tests due to the masking effects of gravity. A drop is shown to be able to form and detach in a region of absolute instability, and spanning the critical Weber number (from a region of convective to absolute instability) resulted in a marked change in dynamics and geometry of the liquid jet and detaching drops. (C) 2002 American Institute of Physics.

Isolation and characterisation of Indian Ocean ciguatoxin

We report the isolation and initial characterisation of Indian Ocean ciguatoxin (I-CTX) present in toxic lipid soluble extracts isolated from ciguateric fishes collected off the Republic of Mauritius in the Indian Ocean. Following i.p. injection of this extract, mice displayed symptoms that were similar, though not identical, to those produced by Pacific and Caribbean ciguatoxins (P-CTXs and C-CTXs). Using a radiolabelled brevetoxin (PbTx) binding assay and mouse bioassay guided fractionation, I-CTX was purified by Florisil, Sephadex LH-20 and TSK HW-40S chromatography with good recovery. Isolation to purity was not possible by preparative reversed phase high-performance liquid chromatography (HPLC) due to significant losses of toxicity. However, analytical reversed phase HPLC coupled to an electrospray mass spectrometry detector identified a [M + H](+) ion at m/z 1141.58 which co-eluted with activity that displaced [3 H]-PbTx binding to rat brain. This mass corresponded to C-CTX-1, but the fragmentation pattern of I-CTX showed a different ratio of pseudo molecular and product ions. I-CTX was found to elute later than P-CTX-1 but was practically indistinguishable from C-CTX-1 on reversed phase HPLC, while the TSK HW-40S column chromatography differentiated I-CTX from the later eluting C-CTX-1. Taken together, these results indicate that I-CTX is a new ciguatoxin (CTX) responsible for ciguatera caused by reef fish in the Indian Ocean. (C) 2002 Elsevier Science Ltd. All rights reserved.

Effect of Metalaxyl Resistance and Cultivar Resistance on Control of Phytophthora nicotianae in Tobacco

Phytophthora nicotianae is a devastating root and stem pathogen of tobacco (Nicotiana tabacum) in South Africa. Growers strive to control the resulting disease, known as black shank, with metalaxyl treatments and resistant cultivars. The aim of this study was to consider whether development of metalaxyl resistance in P. nicotianae has contributed to poor disease control and if recently developed cultivars with high levels of resistance require metalaxyl for effective control. One hundred and thirty-two isolates of P. nicotianae were screened for sensitivity to metalaxyl. P. nicotianae isolates from most tobacco farms were metalaxyl sensitive. Growth of most isolates was inhibited completely at 1.0 μg a.i./ml. However, isolates from the MKTV tobacco producing area showed EC50 values ranging from 1.02 μg a.i./ml to 3.57 μg a.i./ml. Twenty-one tobacco cultivars were planted and treated with and without metalaxyl in two different growing seasons to evaluate their resistance to P. nicotianae and the value of using metalaxyl. Hicks was the most susceptible cultivar. Vuma/3/46, LK30/40/60-1, and LK33/60 exhibited the greatest resistance to P. nicotianae. Use of metalaxyl in combination with moderately resistant cultivars such as NC60 × TL33 and LK10/80/60 effectively reduced black shank in the field. Resistant cultivars were healthy and no significant difference between metalaxyl treated and untreated plants was observed.

Extensive multiple test centre evaluation of the VecTest malaria antigen panel assay

To determine which species and populations of Anopheles transmit malaria in any given situation, immunological assays for malaria sporozoite antigen can replace traditional microscopical examination of freshly dissected Anopheles. We developed a wicking assay for use with mosquitoes that identifies the presence or absence of specific peptide epitopes of circumsporozoite (CS) protein of Plasmodium falciparum and two strains of Plasmodium vivax (variants 210 and 247). The resulting assay (VecTest(TM) Malaria) is a rapid, one-step procedure using a 'dipstick' test strip capable of detecting and distinguishing between P. falciparum and P. vivax infections in mosquitoes. The objective of the present study was to test the efficacy, sensitivity, stability and field-user acceptability of this wicking dipstick assay. In collaboration with 16 test centres world-wide, we evaluated more than 40 000 units of this assay, comparing it to the standard CS ELISA. The 'VecTest(TM) Malaria' was found to show 92% sensitivity and 98.1% specificity, with 97.8% accuracy overall. In accelerated storage tests, the dipsticks remained stable for >15 weeks in dry conditions up to 45degreesC and in humid conditions up to 37degreesC. Evidently, this quick and easy dipstick test performs at an acceptable level of reliability and offers practical advantages for field workers needing to make rapid surveys of malaria vectors.

Randomized comparison of the quality of realtime fetal ultrasound images transmitted by ISDN and by IP videoconferencing

We compared the quality of realtime fetal ultrasound images transmitted using ISDN and IP networks. Four experienced obstetric ultrasound specialists viewed standard recordings in a randomized trial and rated the appearance of 30 fetal anatomical landmarks, each on a seven-point scale. A total of 12 evaluations were performed for various combinations of bandwidths (128, 384 or 768 kbit/s) and networks (ISDN or IF). The intraobserver coefficient of variation was 2.9%, 5.0%, 12.7% and 14.7% for the four observers. The mean overall ratings by each of the four observers were 4.6, 4.8, 5.0 and 5.3, respectively (a rating of 4 indicated satisfactory visualization and 7 indicated as good as the original recording). Analysis of variance showed that there were no significant interobserver variations nor significant differences in the mean scores for the different types of videoconferencing machines used. The most significant variable affecting the mean score was the bandwidth used. For ISDN, the mean score was 3.7 at 128 kbit/s, which was significantly worse than the mean score of 4.9 at 384 kbit/s, which was in turn significantly worse than the mean score of 5.9 at 768 kbit/s. The mean score for transmission using IP was about 0.5 points lower than that using ISDN across all the different bandwidths, but the differences were not significant. It appears that IP transmission in a private (non-shared) network is an acceptable alternative to ISDN for fetal tele-ultrasound and one deserving further study.

A quantitative state and transition model for the Mitchell grasslands of central western Queensland

Concerns of reduced productivity and land degradation in the Mitchell grasslands of central western Queensland were addressed through a range monitoring program to interpret condition and trend. Botanical and eclaphic parameters were recorded along piosphere and grazing gradients, and across fenceline impact areas, to maximise changes resulting from grazing. The Degradation Gradient Method was used in conjunction with State and Transition Models to develop models of rangeland dynamics and condition. States were found to be ordered along a degradation gradient, indicator species developed according to rainfall trends and transitions determined from field data and available literature. Astrebla spp. abundance declined with declining range condition and increasing grazing pressure, while annual grasses and forbs increased in dominance under poor range condition. Soil erosion increased and litter decreased with decreasing range condition. An approach to quantitatively define states within a variable rainfall environment based upon a time-series ordination analysis is described. The derived model could provide the interpretive framework necessary to integrate on-ground monitoring, remote sensing and geographic information systems to trace states and transitions at the paddock scale. However, further work is needed to determine the full catalogue of states and transitions and to refine the model for application at the paddock scale.

The first strand transfer reaction of HIV-1 reverse transcription is more efficient in infected cells than in cell-free natural endogenous reverse transcription reactions

Background: In the presence of dNTPs, intact HIV-1 virions are capable of reverse transcribing at least part of their genome, a process known as natural endogenous reverse transcription (NERT). PCR analysis of virion DNA produced by NERT revealed that the first strand transfer reaction (1stST) was inefficient in intact virions, with minus strand (-) strong stop DNA (ssDNA) copy numbers up to 200 times higher than post-1stST products measured using primers in U3 and U5. This was in marked contrast to the efficiency of 1stST observed in single-round cell infection assays, in which (-) ssDNA and U3-U5 copy numbers were indistinguishable. Objectives: To investigate the reasons for the discrepancy in first strand transfer efficiency between intact cell-free virus and the infection process. Study design: Alterations of both NERT reactions and the conditions of cell infection were used to test whether uncoating and/or entry play a role in the discrepancy in first strand transfer efficiency. Results and Conclusions: The difference in 1stST efficiency could not be attributed simply to viral uncoating, since addition of very low concentrations of detergent to NERT reactions removed the viral envelope without disrupting the reverse transcription complex, and these conditions resulted in no improvement in 1stST efficiency. Virus pseudotyped with surface glycoproteins from either vesicular stomatitis virus or amphotrophic murine leukaemia virus also showed low levels of 1stST in low detergent NERT assays and equivalent levels of (-) ssDNA and 1stST in single-round infections of cells, demonstrating that the gp120-mediated infection process did not select for virions capable of carrying out 1stST. These data indicate that a post-entry event or factor may be involved in efficient HIV-1 reverse transcription in vivo. (C) 2002 Elsevier Science B.V. All rights reserved.

Serum inhibin A and B concentrations during the menstrual cycle in mothers of spontaneous dizygotic twins

Dizygotic twinning in humans is influenced by genetic factors suggesting inherited variation affects follicle development and predisposes to double ovulations. In a previous study, we conducted a detailed examination of follicle development and variation in hormone concentrations during the menstrual cycle in mothers of DZ twins (MODZT) compared with an age-matched control group of mothers of singletons. We did not detect differences in FSH concentrations between mothers of twins and mothers of singletons. Serum inhibin concentrations were measured by a radioimmunoassay that did not distinguish between dimeric inhibin A and B forms and free inhibin alpha subunit. We therefore analyzed the samples from this study with specific assays to determine whether concentrations of inhibin A and B were different between MODZT and controls and therefore contribute to the twinning phenotype. There were no significant differences between MONT with single ovulations and control women in inhibin A and B concentrations during the cycle, including the critical period for the selection of the dominant follicle. These data suggest that the genetic cause of twinning is not associated with changes in FSH concentrations or recognised feedback mechanisms regulating FSH release.