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1.  Diurnal pattern of stomatal conductance in the large-leaved temperate liana Aristolochia macrophylla depends on spatial position within the leaf lamina 
Annals of Botany  2013;111(5):905-915.
Background and Aims
The large distance between peripheral leaf regions and the petiole in large leaves is expected to cause stronger negative water potentials at the leaf apex and marginal zones compared with more central or basal leaf regions. Leaf zone-specific differences in water supply and/or gas exchange may therefore be anticipated. In this study, an investigation was made to see whether zonal differences in gas exchange regulation can be detected in large leaves.
Methods
The diurnal course of stomatal conductance, gs, was monitored at defined lamina zones during two consecutive vegetation periods in the liana Aristolochia macrophylla that has large leaves. Local climate and stem water potential were also monitored to include parameters involved in stomatal response. Additionally, leaf zonal vein densities were measured to assess possible trends in local hydraulic supply.
Key Results
It was found that the diurnal pattern of gs depends on the position within a leaf in A. macrophylla. The highest values during the early morning were shown by the apical region, with subsequent decline later in the morning and a further gradual decline towards the evening. The diurnal pattern of gs at the marginal regions was similar to that of the leaf tip but showed a time lag of about 1 h. At the leaf base, the diurnal pattern of gs was similar to that of the margins but with lower maximum gs. At the the leaf centre regions, gs tended to show quite constant moderate values during most of the day. Densities of minor veins were lower at the margin and tip compared with the centre and base.
Conclusions
Gas exchange regulation appears to be zone specific in A. macrophylla leaves. It is suggested that the spatial–diurnal pattern of gs expressed by A. macrophylla leaves represents a strategy to prevent leaf zonal water stress and subsequent vein embolism.
doi:10.1093/aob/mct061
PMCID: PMC3631344  PMID: 23606681
Stomatal conductance; heterogeneous stomatal conductance; macrophyll; leaf size; leaf venation; diurnal pattern; stem water potential; Aristolochia macrophylla
2.  Visualization of embolism formation in the xylem of liana stems using neutron radiography 
Annals of Botany  2013;111(4):723-730.
Background and Aims
Cold neutron radiography was applied to directly observe embolism in conduits of liana stems with the aim to evaluate the suitability of this method for studying embolism formation and repair. Potential advantages of this method are a principally non-invasive imaging approach with low energy dose compared with synchrotron X-ray radiation, a good spatial and temporal resolution, and the possibility to observe the entire volume of stem portions with a length of several centimetres at one time.
Methods
Complete and cut stems of Adenia lobata, Aristolochia macrophylla and Parthenocissus tricuspidata were radiographed at the neutron imaging facility CONRAD at the Helmholtz-Zentrum Berlin für Materialien und Energie, with each measurement cycle lasting several hours. Low attenuation gas spaces were separated from the high attenuation (water-containing) plant tissue using image processing.
Key results
Severe cuts into the stem were necessary to induce embolism. The formation and temporal course of an embolism event could then be successfully observed in individual conduits. It was found that complete emptying of a vessel with a diameter of 100 µm required a time interval of 4 min. Furthermore, dehydration of the whole stem section could be monitored via decreasing attenuation of the neutrons.
Conclusions
The results suggest that cold neutron radiography represents a useful tool for studying water relations in plant stems that has the potential to complement other non-invasive methods.
doi:10.1093/aob/mct014
PMCID: PMC3605950  PMID: 23393096
Xylem; plant water transport; embolism; neutron radiography; Adenia lobata; Aristolochia macrophylla; Parthenocissus tricuspidata.
3.  Sorting of droplets by migration on structured surfaces 
Summary
Background: Controlled transport of microdroplets is a topic of interest for various applications. It is well known that liquid droplets move towards areas of minimum contact angle if placed on a flat solid surface exhibiting a gradient of contact angle. This effect can be utilised for droplet manipulation. In this contribution we describe how controlled droplet movement can be achieved by a surface pattern consisting of cones and funnels whose length scales are comparable to the droplet diameter.
Results: The surface energy of a droplet attached to a cone in a symmetry-preserving way can be smaller than the surface energy of a freely floating droplet. If the value of the contact angle is fixed and lies within a certain interval, then droplets sitting initially on a cone can gain energy by moving to adjacent cones.
Conclusion: Surfaces covered with cone-shaped protrusions or cavities may be devised for constructing “band-conveyors” for droplets. In our approach, it is essentially the surface structure which is varied, not the contact angle. It may be speculated that suitably patterned surfaces are also utilised in biological surfaces where a large variety of ornamentations and surface structuring are often observed.
doi:10.3762/bjnano.2.25
PMCID: PMC3148036  PMID: 21977433
microdroplets; microfluidics; surface; surface energy; surface structures
4.  Computer-based Studies of Diffusion through Stomata of Different Architecture 
Annals of Botany  2007;100(1):23-32.
Background and Aims
The influence of stomatal architecture on stomatal conductance and on the developing concentration gradient was explored quantitatively by comparing diffusion rates of water vapour and CO2 occurring in a set of three-dimensional stoma models. The influence on diffusion of an internal cuticle, a sunken stoma, a partially closed stoma and of substomatal chambers of two different sizes was considered.
Methods
The study was performed by using a commercial computer program based on the Finite Element Method which allows for the simulation of diffusion in three dimensions. By using this method, diffusion was generated by prescribed gas concentrations at the boundaries of the substomatal chamber and outside of the leaf. The program calculates the distribution of gas concentrations over the entire model space.
Key Results
Locating the stomatal pore at the bottom of a stomatal antechamber with a depth of 20 µm decreased the conductance significantly (at roughly about 30 %). The humidity directly above the stomatal pore is significantly higher with the stomatal antechamber present. Lining the walls of the substomatal chamber with an internal cuticle which suppresses evaporation had an even stronger effect by reducing the conductance to 60 % of the original value. The study corroborates therefore the results of former studies that water will evaporate preferentially at sites in the immediate vicinity to the stomatal pore if no internal cuticle is present. The conductance decrease affects only water vapour and not CO2. Increasing the substomatal chamber increases CO2 uptake, whereas transpiration increases if an internal cuticle is present.
Conclusions
Variation of stomatal structure may, with unchanged pore size and depth, profoundly affect gas exchange and the pathways of liquid water inside the leaf. Equations for calculation of stomatal conductance which are solely based on stomatal density and pore depth and size can significantly overestimate stomatal conductance.
doi:10.1093/aob/mcm075
PMCID: PMC2735294  PMID: 17483152
Gas exchange; diffusion; stomata; stomatal conductance; internal cuticle; sunken stomata; stomatal antechamber

Results 1-4 (4)