Stress caused by environmental factors evokes dynamic changes in plant phenotypes. In this study, we deciphered simultaneously the reaction of plant growth and chlorophyll fluorescence related parameters using a novel approach which combines existing imaging technologies (GROWSCREEN FLUORO). Three different abiotic stress situations were investigated demonstrating the benefit of this approach to distinguish between effects related to (1) growth, (2) chlorophyll-fluorescence, or (3) both of these aspects of the phenotype. In a drought stress experiment with more than 500 plants, poly(ADP-ribose) polymerase (PARP) deficient lines of Arabidopsis thaliana (L.) Heynh showed increased relative growth rates (RGR) compared with C24 wild-type plants. In chilling stress, growth of PARP and C24 lines decreased rapidly, followed by a decrease in Fv/Fm. Here, PARP-plants showed a more pronounced decrease of Fv/Fm than C24, which can be interpreted as a more efficient strategy for survival in mild chilling stress. Finally, the reaction of Nicotiana tabacum L. to altered spectral composition of the intercepted light was monitored as an example of a moderate stress situation that affects chlorophyll-fluorescence related, but not growth-related parameters. The examples investigated in this study show the capacity for improved plant phenotyping based on an automated and simultaneous evaluation of growth and photosynthesis at high throughput.
Plants constantly adapt their leaf orientation in response to fluctuations in the environment, to maintain radiation use efficiency in the face of varying intensity and incidence direction of sunlight. Various methods exist for measuring structural canopy parameters such as leaf angle distribution. However, direct methods tend to be labour-intensive, while indirect methods usually give statistical information on stand level rather than on individual leaves. We present an area-based, binocular stereo system composed of commercially available components that allows three-dimensional reconstruction of small-to medium-sized canopies on the level of single leaves under field conditions. Spatial orientation of single leaves is computed with automated processes using modern, well-established stereo matching and segmentation techniques, which were adapted for the properties of plant canopies, providing high spatial and temporal resolution (angle measurements with an accuracy of approx. Ϯ5°and a maximum sampling rate of three frames per second). The applicability of our approach is demonstrated in three case studies: (1) the dihedral leaflet angle of an individual soybean was tracked to monitor nocturnal and daytime leaf movement showing different frequencies and amplitudes; (2) drought stress was diagnosed in soybean by quantifying changes in the zenith leaflet angle distribution; and (3) the diurnal course of the zenith leaf angle distribution of a closed soybean canopy was measured.
Neomycin and U73122 are known to suppress inositol 1,4,5-trisphosphate (InsP3) production by inhibition of phospholipase C. We studied the effects of these inhibitors on the excitatory currents, Iex, in Chara corallina under voltage-clamp conditions. Computer simulations of the experimental effects by a minimum model for the excitatory reaction pathway allow the assignment of the inhibitory effects to one specific reaction step, i.e. the release of Ca2+ from InsP3-sensitive internal stores. In contrast, the inhibitory effect of La3+ on Iex suggests inactivation of Cl- channels. Furthermore, ryanodine-sensitive Ca2+ stores seem to be irrelevant for electrical excitation in Chara.
Dicot leaves grow with pronounced diel (24-h) cycles that are controlled by a complex network of factors. It is an open question to what extent leaf growth dynamics are controlled by long-range or by local signals. To address this question, we established a stereoscopic imaging system, GROWSCREEN 3D, which quantifies surface growth of isolated leaf discs floating on nutrient solution in wells of microtiter plates. A total of 458 leaf discs of tobacco (Nicotiana tabacum) were cut at different developmental stages, incubated, and analyzed for their relative growth rates. The camera system was automatically displaced across the array of leaf discs; visualization and camera displacement took about 12 s for each leaf disc, resulting in a time interval of 1.5 h for consecutive size analyses. Leaf discs showed a comparable diel leaf growth cycle as intact leaves but weaker peak growth activity. Hence, it can be concluded that the timing of leaf growth is regulated by local rather than by systemic control processes. This conclusion was supported by results from leaf discs of Arabidopsis (Arabidopsis thaliana) Landsberg erecta wild-type plants and starch-free1 mutants. At night, utilization of transitory starch leads to increased growth of Landsberg erecta wild-type discs compared with starch-free1 discs. Moreover, the decrease of leaf disc growth when exposed to different concentrations of glyphosate showed an immediate dose-dependent response. Our results demonstrate that a dynamic leaf disc growth analysis as we present it here is a promising approach to uncover the effects of internal and external cues on dicot leaf development.
ive and are unsuitable for impaling electrodes. Methods for modelling the kinetics and permeation of Much of our knowledge of ion-transport mechanisms ion channels using single-channel recordings are prein plant cell membranes comes from experiments sented. How the ionic currents underlying an action using voltage-clamp. This technique allows the measpotential (AP) can be identified using the AP-clamp urement of ionic currents across the membrane, whilst method is discussed.
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