RESUMO
BACKGROUND: Cellular components are controlled by genetic and physiological factors that define their shape and size. However, quantitively capturing the morphological characteristics and movement of cellular organelles from micrograph images is challenging, because the analysis deals with complexities of images that frequently lead to inaccuracy in the estimation of the features. Here we show a unique quantitative method to overcome biases and inaccuracy of biological samples from confocal micrographs. RESULTS: We generated 2D images of cell walls and spindle-shaped cellular organelles, namely ER bodies, with a maximum contrast projection of 3D confocal fluorescent microscope images. The projected images were further processed and segmented by adaptive thresholding of the fluorescent levels in the cell walls. Micrographs are composed of pixels, which have information on position and intensity. From the pixel information we calculated three types of features (spatial, intensity and Haralick) in ER bodies corresponding to segmented cells. The spatial features include basic information on shape, e.g., surface area and perimeter. The intensity features include information on mean, standard deviation and quantile of fluorescence intensities within an ER body. Haralick features describe the texture features, which can be calculated mathematically from the interrelationship between the pixel information. Together these parameters were subjected to multivariate analysis to estimate the morphological diversity. Additionally, we calculated the displacement of the ER bodies using the positional information in time-lapse images. We captured similar morphological diversity and movement within ER body phenotypes in several microscopy experiments performed in different settings and scanned under different objectives. We then described differences in morphology and movement of ER bodies between A. thaliana wild type and mutants deficient in ER body-related genes. CONCLUSIONS: The findings unexpectedly revealed multiple genetic factors that are involved in the shape and size of ER bodies in A. thaliana. This is the first report showing morphological characteristics in addition to the movement of cellular components and it quantitatively summarises plant phenotypic differences even in plants that show similar cellular components. The estimation of morphological diversity was independent of the cell staining method and the objective lens used in the microscopy. Hence, our study enables a robust estimation of plant phenotypes by recognizing small differences in complex cell organelle shapes and their movement, which is beneficial in a comprehensive analysis of the molecular mechanism for cell organelle formation that is independent of technical variations.
RESUMO
The anthropogenic pressure on the environment depends on the spatial scale. It is crucial to prioritise conservation actions at different spatial scales to be cost-efficient. Using horizon scanning with the Delphi technique, we asked what the most important conservation problems are in Poland at local and national scales. Twenty-six participants, PhD students, individually identified conservation issues important at the local and national scales. Each problem was then scored and classified into broader categories during the round discussions. Text mining, cross-sectional analyses, and frequency tests were used to compare the context, importance scores, and frequency of identified problems between the two scales, respectively. A total of 115 problems were identified at the local scale and 122 at the national scale. Among them, 30 problems were identical for both scales. Importance scores were higher for national than local problems; however, this resulted from different sets of problems identified at the two scales. Problems linked to urbanisation, education, and management were associated with the local scale. Problems related to policy, forestry, and consumerism were more frequent at the national scale. An efficient conservation policy should be built hierarchically (e.g. introducing adaptive governance), implementing solutions at a national scale with the flexibility to adjust for local differences and to address the most pressing issues.
