Although Liliaceae are valuable resource plants with medicinal and edible uses, techniques for evaluating their tolerance to various abiotic stresses are very limited. This study evaluated the cold tolerance using visual inspection, electrolyte leakage, and Evan’s blue assay. Visual inspection of the responses to different temperatures, using a temperature range of 4 to −12 °C, showed that Scilla scilloides was receive the least damage. However, electrolyte leakage tests showed slightly different results from visual inspection. The median lethal temperature (LT₅₀) was expected to be between −4 and −8 °C. The LT₅₀ was considered a measure of damage due to electrolyte leakage in plant cold tolerance evaluation. As a result of predicting the lethal temperature using the logistic regression equation, the average LT₅₀ of the four plants was −9.0 °C. The species with the lowest LT₅₀ was measured for Hosta plantagines (−11.14°C), whereas the highest LT₅₀ was measured for Hemerocallis fulva (−7.14°C). As a result of the Evan’s blue assay, it was found that cell necrosis occurred when the plants were exposed to low temperatures. Visual observation showed that more than 50% of the three plants’ cells, except for H. plantaginea, were stained blue even at 8 °C. From this result, H. plantaginea was judged to have strong low-temperature tolerance. At −12 °C, more than 50% of the four Liliaceae plants were colored blue, and the LT₅₀ value was expected to be below −12 °C. The reducing sugar content, an indicator of plant cold tolerance, was the highest in H. plantaginea, followed by S. scilloide and H. longipes. Combining the three methods, H. plantaginea had the highest cold tolerance, followed by H. longipes, S. scilloides, and H. fulva. The results of this study will be widely used in selecting cold-tolerant useful resource plants.

Sequential drought and heat stress pose a growing threat to forest ecosystems in the context of climate change, yet systematic evaluation methods for woody plants remain limited. This study aimed to develop a comprehensive screening platform for identifying woody plant species tolerant to sequential drought and heat stress among 27 native species growing in Korea. A sequential stress protocol was applied: drought stress for 2 weeks, followed by high-temperature exposure at 45 °C. Physiological indicators, including relative water content (RWC) and electrolyte leakage index (ELI), were used for preliminary screening, supported by phenotypic observations, Evans blue staining for cell death, and DAB staining to assess oxidative stress and recovery ability. The results revealed clear differences among species. Chamaecyparis obtusa, Quercus glauca, and Q. myrsinaefolia exhibited strong tolerance, maintaining high RWC and low ELI values, while Albizia julibrissin was highly susceptible, showing severe membrane damage and low survival. DAB staining successfully distinguished tolerance levels based on oxidative recovery. Additional species such as Camellia sinensis, Q. acuta, Q. phillyraeoides, Q. salicina, and Ternstroemia japonica showed varied responses: Q. phillyraeoides demonstrated high tolerance, T. japonica showed moderate tolerance, and Q. salicina was relatively sensitive. The integrated screening system effectively differentiated tolerant species through multiscale analysis—physiological, cellular, and morphological—demonstrating its robustness and applicability. This study provides a practical and reproducible framework for evaluating sequential drought and heat stress in trees and offers valuable resources for urban forestry, reforestation, and climate-resilient species selection.

Cold stress is a critical factor restricting the cultivation of subtropical evergreen species such as Camellia sinensis and C. japonica in temperate climates. This study aimed to develop an integrated framework for evaluating cold tolerance by combining visual assessment, electrolyte leakage (EL), Evans blue staining, and nonlinear regression modeling. All experiments were conducted with n = 3 samples per treatment, and statistical analyses were performed at a significance level of α = 0.05. Under freezing treatment at −6 °C, C. japonica exhibited faster and more severe damage, including leaf curling and vein darkening, compared to C. sinensis. Electrolyte leakage and cell death increased rapidly in C. japonica, and a sharp rise in cell death occurred in both species when EL exceeded 55%. Logistic regression of EL data estimated LT₅₀ values of −10.96 °C for C. sinensis and −9.38 °C for C. japonica, while EL Temp₅₀ values were −9.59 °C and −8.97 °C, respectively, indicating higher membrane stability in C. sinensis. The difference in LT₅₀ between the two species was statistically significant (p < 0.05). Biochemical and heatmap analyses from 25 °C to −12 °C showed that C. sinensis maintained higher chlorophyll, antioxidant activity, and sugar levels, reflecting stronger cold tolerance. In contrast, C. japonica accumulated more proline and MDA, indicating higher stress sensitivity and membrane damage. This study presents a reproducible, quantitative framework for evaluating cold tolerance in Camellia species, offering valuable insights for breeding and expanding cultivation under climate change.

Moss in container seedling nurseries competes with seedlings for water and nutrients while blocking light, thereby inhibiting growth. This study aimed to address this issue by evaluating the moss control efficacy of 11 chemical compounds, including terpinyl acetate (TA), limonene, and Hinoki essential oil (HEO). The plate experiment results led to the selection of 6 substances (TA, limonene, HEO, pine leaf extract, baking soda, pelargonic acid) that stably controlled both Polytrichum commune Hedw. and Marchantia. Polymorpha L. When TA, limonene, and HEO were combined with surfactants, moss control rates increased and showed stable performance. In the container seedling experiment, TA, limonene, and HEO demonstrated high moss control effects while exhibiting low growth inhibition. When these three substances were combined with surfactants, the electrolyte leakage index (ELI) decreased, indicating minimal cell membrane damage. Additionally, TA treatment maintained stable soil physicochemical properties with no significant changes in pH or nutrient levels. Microscopic analysis of moss cells showed cell wall deformation and expansion of intercellular spaces in the three substance treatment groups. Future verification of long-term effectiveness, expansion of application targets, and assessment of economic feasibility could lead to the development of eco-friendly moss removal agents for improving container seedling quality.

This study evaluated cold tolerance and selected superior individuals from 100 Zoysia japonica ‘Zenith’ plants. Following pre-cold treatment at 4 °C for 24 h, candidates were screened using electrolyte leakage (EL) with a 10% selection intensity. Selected candidates and cold-sensitive controls were exposed to 0, −5, and −10 °C for 24 h and assessed using visual injury, EL, Evans blue staining, and 3,3′-diaminobenzidine (DAB) staining. Visual injury increased with decreasing temperature but did not clearly distinguish cold tolerance. Individual 11 exhibited the lowest lethal temperature causing 50% injury (LT₅₀) based on both EL (−10.4 °C) and Evans blue staining (−11.0 °C). The two LT₅₀ estimates were strongly correlated (r = 0.996) and concordant (Lin’s concordance correlation coefficient = 0.909). DAB-based recovery analysis showed effective hydrogen peroxide (H₂O₂) clearance in individuals 5, 11, 21, 23, and 40, with individual 23 exhibiting the lowest reactive oxygen species area under the curve (121.5). A modified Generalized Value Index integrating visual injury, mean LT₅₀, oxidative stress, and recovery slope identified individuals 5, 11, 18, 23, and 30 as superior cold-tolerant selections. Therefore, combining membrane-damage and oxidative-recovery indicators can provide a robust framework for selecting cold-tolerant Z. japonica ‘Zenith’ for field validation and breeding.