The stomata of plants mainly regulate gas exchange and water dispersion between the interior and external environments of plants and play a major role in the plants’ health. The existing methods of stomata segmentation and measurement are mostly for specialized plants. The purpose of this research is to develop a generic method for the fully automated segmentation and measurement of the living stomata of different plants. The proposed method utilizes level set theory and image processing technology and can outperform the existing stomata segmentation and measurement methods based on threshold and skeleton in terms of its versatility. The single stomata images of different plants were the input of the method and a level set based on the Chan-Vese model was used for stomatal segmentation. This allowed the morphological features of the stomata to be measured. Contrary to existing methods, the proposed segmentation method does not need any prior information about the stomata and is independent of the plant types. The segmentation results of 692 living stomata of black poplars show that the average measurement accuracies of the major and minor axes, area, eccentricity and opening degree are 95.68%, 95.53%, 93.04%, 99.46% and 94.32%, respectively. A segmentation test on dayflower (Commelina benghalensis) stomata data available in the literature was completed. The results show that the proposed method can effectively segment the stomata images (181 stomata) of dayflowers using bright-field microscopy. The fitted slope of the manually and automatically measured aperture is 0.993, and the R2 value is 0.9828, which slightly outperforms the segmentation results that are given in the literature. The proposed automated segmentation and measurement method for living stomata is superior to the existing methods based on the threshold and skeletonization in terms of versatility. The method does not need any prior information about the stomata. It is an unconstrained segmentation method, which can accurately segment and measure the stomata for different types of plants (woody or herbs). The method can automatically discriminate whether the pore region is independent or not and perform pore region extraction. In addition, the segmentation accuracy of the method is positively correlated with the stomata’s opening degree.

Research Highlights: Understanding of the spatial variation of root exudation on a regional scale can help understand the response of plant physiological activities to environmental changes. Background and Objectives: Although root exudation has become an important topic in belowground ecology, its relationship with root traits and environmental factors is poorly understood. Our objective was to explore how root traits and environmental factors influence root exudation. Materials and Methods: We used a multi-factorial design consisting of three tree species spanning across sites located at three latitudes to assess root exudation dynamics, which was measured using a syringe-basis incubation system. Results: The strongest and clearest effect observed in our study was a decrease in root exudation rates of Korean pine (Pinus koraiensis Sieb. et Zucc.) and larch (Larix gmelinii (Rupr.) Kuze.) at sites located in higher latitudes. Root exudation rates were positively related to mean annual temperature, mean annual precipitation, and negatively related to soil total organic carbon. Conclusions: Root exudation in coniferous species decreased at sites located in higher latitudes. Despite differences in root exudation rate among sites located at different latitudes and species with suitable variation in root morphological traits and environmental factors, we could not identify consistent influencing factors on root exudation rates.

The soil microbial community structure is critical to the cycling of carbon and nitrogen in forest soils. As afforestation practices increasingly promote different functional traits of tree species, it has become critical to understand how they influence soil microbial community structures, which directly influence soil biogeochemical processes. We used fungi ITS and bacteria 16S rDNA to investigate soil microbial community structures in three monoculture plantations consisting of a non-native evergreen conifer (Pinus sibirica), a native deciduous conifer (Larix gmelinii), and a native deciduous angiosperm (Betula platyphylla) and compared them with two 1:1 mixed-species plantations (P. sibirica and L. gmelinii, P. sibirica and B. platyphylla). The fungal community structure of the conifer–angiosperm mixed plantation was similar to that of the non-native evergreen conifer, and the bacterial community structure was similar to that of the angiosperm monoculture plantation. Fungal communities were strongly related to tree species, but bacterial communities were strongly related to soil nitrogen. The co-occurrence networks were more robust in the mixed plantations, and the microbial structures associated with soil carbon and nitrogen were significantly increased. Our results provide a comparative study of the soil microbial ecology in response to afforestation of species with different functional traits and enhance the understanding of factors controlling the soil microbial community structure.