R B Roemer’s research while affiliated with University of Utah and other places

Premise: Although maintaining the appropriate mid-day timing of the diel thermogenic events of cones of the dioecious cycads Macrozamia lucida and M. macleayi is central to the survival of both plant and pollinator in this obligate pollination mutualism, the nature of the underlying mechanism remains obscure. We investigated whether it is under circadian control. Circadian mechanisms control the timing of many ecologically important processes in angiosperms, yet only a few gymnosperms have been studied in this regard. Methods: We subjected cones to different ambient temperature and lighting regimens (constant temperature and darkness; stepwise cool/warm ambient temperatures in constant darkness; stepwise dark/light exposures at constant temperature) to determine whether the resulting timing of their thermogenic events was consistent with circadian control. Results: Cones exposed to constant ambient temperature and darkness generated multiple temperature peaks endogenously, with an average interpeak-temperature period of 20.7 (±0.20) h that is temperature-compensated (Q10 = 1.02). Exposure to 24-h ambient temperature cycles (12 h cool/12 h warm, constant darkness) yielded an interpeak-temperature period of 24.0 (±0.05) h, accurately and precisely replicating the ambient temperature period. Exposure to 24-h photo-cycles (12 h light/12 h dark, constant ambient temperature) yielded a shorter, more variable interpeak-temperature period of 23 (±0.23) h. Conclusions: Our results indicate that cycad cone thermogenesis is under circadian clock control and differentially affected by ambient temperature and light cycles. Our data from cycads (an ancient gymnosperm lineage) adds to what little is known about circadian timing in gymnosperms, which have rarely been studied from the circadian perspective.

Cycad cone thermogenesis and its associated volatiles are intimately involved in mediating the behavior of their obligate specialist pollinators. In eastern Australia, thrips in the Cycadothrips chadwicki species complex are the sole pollinators of many Macrozamia cycads. Further, they feed and reproduce entirely in the pollen cones. M. miquelii, found only in the northern range of this genus, is pollinated only by a C. chadwicki cryptic species that is the most distantly related to others in the complex. We examined the volatile profile from M. miquelii pollen and ovulate (receptive and non-receptive) cones to determine how this mediates pollination mechanistically, using GC-MS (gas chromatography-mass spectrometry) and behavioral tests. Monoterpenes comprise the bulk of M. miquelii volatile emissions, as in other Macrozamia species, but we also identified compounds not reported previously in any cycad, including three aliphatic esters (prenyl acetate and two of uncertain identity) and two aliphatic alcohols. The two unknown esters were confirmed as prenyl (3-methylbut-2-enyl) esters of butyric and crotonic ((E))-but-2-enoic) acids after chemical synthesis. Prenyl crotonate is a major component in emissions from pollen and receptive ovulate cones, is essentially absent from non-receptive cones, and has not been reported from any other natural source. In field bioassays, Cycadothrips were attracted only to those volatile treatments containing prenyl crotonate. We discuss M. miquelii cone odorants relative to those of other cycads, especially with respect to prenyl crotonate being a species-specific signal to this northern C. chadwicki cryptic species, and how this system may have diversified.

Macrozamia macleayi Miq. (family Zamiaceae) pollen cones generate high thermogenic temperatures that are crucial to pollination of these dioecious plants. However, cone thermal behaviour has not been characterised with respect to developmental stage, so any links with the progression and development of their pollination processes remain unclear. Here we show that after growing to full girth, cones progress through thermally active stages of slow/no growth, rapid lengthening, dehiscence and early post-dehiscence, each with a distinct thermal response. During slow/no growth cones exhibit a small late afternoon peak thermogenic temperature elevation above peak ambient, and remain elevated overnight. During rapid lengthening the late afternoon/night-time temperature elevations disappear, and mid-day thermogenesis commences. During dehiscence the midday cone temperature elevations become large, approaching 10°C near the day of maximum dehiscence rate, and then decrease daily. Pollen cones generate their large, dehiscence stage thermogenic temperature elevations synchronously with the diel ambient temperature peak, thus maximising the peak cone temperature. This likely enhances the expulsion of their pollen bearing obligate mutualist thrips pollinator, thus boosting pollination rates. Thermogenic events are fuelled by carbohydrates only, and significantly increase the pollen cone water loss – yet the percentage of water in sporophylls remains nearly constant (~63%) throughout all developmental stages. Similar coordinated pollen cone developmental stage and thermogenic responses are also present in Cycas micronesica K.D. Hill (family Cycadaceae), suggesting a conserved physiological response across cycad families.

An important outcome of plant thermogenesis is increased emissions of volatiles that mediate pollinator behavior. We investigated whether the large increase in emissions, mainly the monoterpene ß-myrcene (>90%), during daily thermogenic events of Macrozamia macleayi and lucida cycad cones are due solely to the influence of high cone temperatures or are, instead, a result of increased respiratory rates during thermogenesis. We concurrently measured temperature, oxygen consumption, and ß-myrcene emission profiles during thermogenesis of pollen cones under typical environmental temperatures and during experimental manipulations of cone temperatures and aerobic conditions, all in the dark. The exponential rise in ß-myrcene emissions never occurred without a prior, large increase in respiration, whereas an increase in cone temperature alone did not increase emissions. When respiration during thermogenesis was interrupted by anoxic conditions, ß-myrcene emissions decreased. The increased emission rates are not a result of increased cone temperature per se (through increased enzyme activity or volatilization of stored volatiles), but are dependent on biosynthetic pathways associated with increased respiration during thermogenesis that provide the carbon, energy (ATP) and reducing compounds (NADPH) required for ß-myrcene production through the methylerythritol phosphate (MEP) pathway. These findings establish the significant contribution of respiration to volatile production during thermogenesis. This article is protected by copyright. All rights reserved.

We have investigated the potential movement on air currents of pollen from Guam's native cycad, Cycas micronesica, proposed as ambophilic. We measured wind velocities and directions in different cycad habitats that vary in their exposure to trade winds, determined pollen settling velocities, and then modeled the potential horizontal pollen drift distance in each habitat. Similar measurements were performed on several entomophilous Zamia cycads and six zoophilous tropical trees used in horticulture or landscaping. All cycad species' pollen exhibited relatively slow mean settling velocities (0.73–1.29 cm · s L1) with C. micronesica pollen in the middle of this range. Our models predicted that wind in more open habitats with wind directions east northeast (ENE) to northeast (NE) and velocities >2 m · s L1 could transport C. micronesica pollen, either single grains or clumps, hundreds of meters downwind from the pollen source before falling 1 or 2 m. In forested habitats and at typical heights of cycad cones in the understory, the mean wind velocities ranged from <0.03 m · s L1 to ' '1 m · s L1. In habitats with mean winds ‡0.2 m · s L1 , models predicted pollen transport distances of tens of meters from the pollen source. In sheltered habitats with velocities near 0.03 m · s L1 , the potential wind transport of pollen was limited to less than a few meters, suggesting that wind would be an ineffective vector in such areas. Pollen grains of all angiosperm species were larger except one, and the species with larger grains had settling velocities 3–26 times faster than that of cycad pollens. Even so, winds in most Guam environments could transport pollen of most angiosperm species over 50 m before falling 1 m. In summary, the results suggested that pollen size, clumping tendencies, and drift of most of these species do not preclude a role for wind in moving pollen in habitats exposed to trade winds, and that other physical and plant characteristics affect their pollination mode. For C. micronesica, these pollen and plant attributes do not preclude entomophily, and insects are likely required in the deep understory where cycads are present.

This study presents a new approach for evaluating bioheat transfer equation (BHTE) models used in treatment planning, control and evaluation of all thermal therapies. First, 3D magnetic resonance temperature imaging (MRTI) data are used to quantify blood flow-related energy losses, including the effects of perfusion and convection. Second, this information is used to calculate parameters of a BHTE model: in this paper the widely used Pennes BHTE. As a self-consistency check, the BHTE parameters are utilized to predict the temperatures from which they were initially derived. The approach is evaluated with finite-difference simulations and implemented experimentally with focused ultrasound heating of an ex vivo porcine kidney perfused at 0, 20 and 40 ml/min (n = 4 each). The simulation results demonstrate accurate quantification of blood flow-related energy losses, except in regions of sharp blood flow discontinuities, where the transitions are spatially smoothed. The smoothed transitions propagate into estimates of the Pennes perfusion parameter but have limited effect on the accuracy of temperature predictions using these estimates. Longer acquisition time periods mitigate the effects of MRTI noise, but worsen the effect of flow discontinuities. For the no-flow kidney experiments the estimates of a uniform, constant Pennes perfusion parameter are approximately zero, and at 20 and 40 ml/min the average estimates increase with flow rate to 3.0 and 4.2 kg/m(3) /s, respectively. When Pennes perfusion parameter values are allowed to vary spatially, but remain temporally constant, BHTE temperature predictions are more accurate than when using spatially uniform, constant Pennes perfusion values, with reductions in RMSE values of up to 79%. Locations with large estimated perfusion values correspond to high flow regions of the kidney observed in T1 -weighted MR images. This novel, MRTI-based technique holds promise for improving understanding of thermal therapy biophysics and for evaluating biothermal models. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

Purpose: Minimising treatment time and protecting healthy tissues are conflicting goals that play major roles in making magnetic resonance image-guided focused ultrasound (MRgFUS) therapies clinically practical. We have developed and tested in vivo an adaptive model-predictive controller (AMPC) that reduces treatment time, ensures safety and efficacy, and provides flexibility in treatment set-up. Materials and methods: The controller realises time savings by modelling the heated treatment cell's future temperatures and thermal dose accumulation in order to anticipate the optimal time to switch to the next cell. Selected tissues are safeguarded by a configurable temperature constraint. Simulations quantified the time savings realised by each controller feature as well as the trade-offs between competing safety and treatment time parameters. In vivo experiments in rabbit thighs established the controller's effectiveness and reliability. Results: In all in vivo experiments the target thermal dose of at least 240 CEM43 was delivered everywhere in the treatment volume. The controller's temperature safety limit reliably activated and constrained all protected tissues to <9 CEM43. Simulations demonstrated the path independence of the controller, and that a path which successively proceeds to the hottest untreated neighbouring cell leads to significant time savings, e.g. when compared to a concentric spiral path. Use of the AMPC produced a compounding time-saving effect; reducing the treatment cells' heating times concurrently reduced heating of normal tissues, which eliminated cooling periods. Conclusions: Adaptive model-predictive control can automatically deliver safe, effective MRgFUS treatments while significantly reducing treatment times.

Purpose: The use of correct tissue thermal diffusivity values is necessary for making accurate thermal modelling predictions during magnetic resonance-guided focused ultrasound (MRgFUS) treatment planning. This study evaluates the accuracy and precision of two non-invasive thermal diffusivity estimation methods, a Gaussian temperature method and a Gaussian specific absorption rate (SAR) method. Materials and methods: Both methods utilise MRgFUS temperature data obtained during cooling following a short (<25 s) heating pulse. The Gaussian SAR method can also use temperatures obtained during heating. Experiments were performed at low heating levels (ΔT∼10 °C) in ex vivo pork muscle and in vivo rabbit back muscle. The non-invasive MRgFUS thermal diffusivity estimates were compared with measurements from two standard invasive methods. Results: Both non-invasive methods accurately estimated thermal diffusivity when using MR temperature cooling data (overall ex vivo error <6%, in vivo <12%). Including heating data in the Gaussian SAR method further reduced errors (ex vivo error <2%, in vivo <3%). The significantly lower standard deviation values (p < 0.03) of the Gaussian SAR method indicated that it had better precision than the Gaussian temperature method. Conclusions: With repeated sonications, either MR-based method could provide accurate thermal diffusivity values for MRgFUS therapies. Fitting to more data simultaneously likely made the Gaussian SAR method less susceptible to noise, and using heating data helped it converge more consistently to the FUS fitting parameters and thermal diffusivity. These effects led to the improved precision of the Gaussian SAR method.

Most functions postulated for thermogenesis in plant reproductive organs are associated with pollination and pollinator activity. During thermogenesis other chemical changes occur, and these also may affect pollinator activity. We address how cone thermogenesis and the associated chemical emissions influence pollinator behaviour in the obligate mutualist Macrozamia lucida and M. macleayi cycad – Cycadothrips chadwicki thrips pollination system. Cones of these dioecious gymnosperms have a diel, mid‐day metabolic burst that is associated with increased cone temperatures, volatile emissions (primarily β‐myrcene), and CO 2 and water vapour emissions. Concurrently, thrips leave cones en masse and then return to cones later in the day. We investigated the effects of these cues, individually and in combinations, on their potential to induce thrips to leave cones in the dark and light with a suite of Y‐tube behavioural experiments. The results suggest that ambient light, and high cone temperatures, humidity, and β‐myrcene levels, but not CO 2 , each induce Cycadothrips to leave cones. At typical overnight temperatures (14 °C) thrips were relatively inactive and negatively phototactic. At typical daytime ambient temperatures, 22–26 °C (lower than typical thermogenic temperatures), thrips were active and positively phototactic. Thrips moved away from or avoided thermogenic temperatures and high concentrations of β‐myrcene, and they preferred dry (<8% RH ) to humidified (~88% RH ) air. Whereas several variables individually induce thrips to leave a cone's darker interior towards the daytime light, these signals are presented simultaneously during thermogenesis. There thus seems to be enhanced or redundant signalling that enforces thrips departure from pollen cones thus increasing the chances of vectoring pollen to ovulate cones. While high temperature appears necessary to mediate thrips movement, its concurrence with other plant signals that produce similar responses, suggests a dynamic multimodal signalling system that operates as a functional unit.