The freeze-thaw cycle exacerbates the ecotoxicity of polystyrene nanoplastics to Secale cereale L. seedlings.

In the context of global climate change, recurrent freeze-thaw cycles (FTC) and concurrent exposure to polystyrene nanoplastics (PSNPs) directly impact crop growth and indirectly affect resilience to abiotic stress. In January 2023, experiments at the Environmental Biology Laboratory, Jilin University, Changchun, China, exposed rye seedlings to 100 nm PSNPs at concentrations of 0, 10, 50, and 100 mg/L for seven days, followed by three FTC. Scanning electron microscopy (SEM) demonstrated that PSNPs migrated from the roots to the leaves, with FTC significantly exacerbating their accumulation within plant tissues. Transmission electron microscopy (TEM) observations showed that FTC disrupted normal cell division, and combined stress from NPs damaged plant organs, particularly chloroplasts, thereby substantially inhibiting photosynthesis. FTC delayed plant phenological stages. Under combined stress, malondialdehyde (MDA) accumulation in plant tissues increased by 15.6%, while hydrogen peroxide (H2O2) content decreased. Simultaneously, the activities of peroxidase (POD) and catalase (CAT) increased by 34.2% and 38.6%, respectively. Molecular docking unveiled that PSNPs could bind to the active center of POD/CAT through hydrogen bonding or hydrophobic interactions. The Integrated Biomarker Response (IBR) index highlighted FTC as a crucial determinant for pronounced effects. Moreover, an apparent dose-dependent effect was observed, with antioxidant enzyme activities in rye seedlings induced by low pollutant concentrations and inhibited by high concentrations. These results indicate that FTC and PSNPs can disrupt plant membrane systems and cause severe oxidative damage. Overall, this study provides compelling scientific evidence of the risks associated with NPs exposure in plants subjected to abiotic stress.

Artemisinin and Ambrosia trifida extract aggravate the effects of short freeze-thaw stress in winter rye (Secale cereale) seedlings.

The freeze-thaw and allelopathy from alien giant ragweed (Ambrosia trifida L.) and artemisinin have led to a serious stress to plants, influencing the agricultural quality and crop yield in north-east China. Yet, little is known how allelopathy affect plants under the freeze-thaw process. In this study, the characteristics in winter rye (Secale cereale L.) seedlings were investigated by laboratory simulation. The results showed that during the freezing process, application of artemisinin and A. trifida extract significantly increased the soluble protein content and accelerated lipid peroxidation, while they significantly inhibited antioxidant enzymes, photosynthesis and respiration (P <0.05). During the thawing process, the freezing pressure decreased, and activities of antioxidant enzymes were significantly improved to mitigate artemisinin and A. trifida extract induced stress (P <0.05). In addition, the sensitivity of the investigated metabolic processes in winter rye seedlings were highest to artemisinin and A. trifida extract in the freezing process. This study suggested that the stress response induced by artemisinin and A. trifida extract on winter rye seedlings in the freezing process was greater than that in the thawing process.

Impact of repeated freeze-thaw cycles environment on the allelopathic effect to Secale cereale L. seedlings.

Allelopathy, as environmental stress, plays a prominent role in stress ecotoxicity, and global warming directly increases freeze-thaw cycles (FTCs) frequency in the winter. Yet, the effect between FTCs environment and allelopathy stress is rarely known, and the interaction of allelopathy stresses lacks consideration. Here, we addressed interactions between artemisinin stress (AS) and A. trifida extract stress (AES) under Non-FTCs and FTCs environments. The results found that AS and AES had an antagonistic relation under Non-FTCs environment, while a strong synergism and cooperation under FTCs environment affect the growth and physiology in S. cereale seedlings. Besides, AS and AES under FTCs environment had more inhibition on the growth of roots and shoots, chlorophylls, photosynthetic parameters, and relative water content; while more promotion on malondialdehyde, soluble sugar, and soluble protein. Moreover, the antioxidant enzyme activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX) were increased by AS and AES, showing a good resistance of S. cereale seedlings to allelopathy stress, but FTCs environment significantly weakened this resistance. Thus, the allelopathic effect of AS and AES on S. cereale seedlings was significantly emphasized by FTCs environment.

Short-term effects of combined freeze­thaw and saline­alkali stresses on the physiological response in highland barley (Hordeum vulgare).

Highland barley (Hordeum vulgare L.), as the dominant crop on the Qinghai-Tibetan Plateau, is a typical representative of plants adapted to extreme environmental conditions. However, the harsh environment, severe salinisation and frequent freezing and thawing in the Qinghai-Tibetan Plateau are main limiting factor for crop growth in this region. The physiological response of highland barley to salinisation and freeze-thaw stresses was studied in this paper. Under the combined stresses of 60mmol/LNaCl·60mmol/LNaHCO3 and freeze-thaw cycles (10, -5, and 10°C), the changes in the relative moisture content, relative electrical conductivity, soluble protein, malondialdehyde (MDA) and photosynthetic indices Pn and E in seedling leaves of eight groups of treatments (CK, S, A, S-A, CK (FT), S (FT), A (FT), and S-A (FT)) were analysed. Results showed that a single stress did not cause a change in the MDA content. All of the combined stresses in S-A, CK (FT), S (FT), A (FT), and S-A (FT) treatments increased the MDA content of barley seedlings, and the MDA content of S-A (FT) reached 28.438 at T2 (-5°C) µmol/g. During the freeze-thaw cycle, the cell membrane of seedlings was damaged more seriously by alkali stress, which showed a significant increase in relative conductivity. The relative moisture content value of seedlings was more than 100% because the seedlings could absorb more moisture due to mechanical injury. The protein content of osmoregulatory substances in highland barley seedlings increased with increasing stress, indicating resistance to stress. Moreover, the effect of freeze-thaw stress on photosynthesis was more significant. The changes in indices proved that an appropriate amount of salt stress could improve the resistance of the plant cell membrane. Alkali stress had a significant effect on the growth of highland barley seedlings. Freezing and thawing can aggravate the damage of saline-alkali stress to highland barley seedlings, resulting in changes in the biological membrane permeability and photosynthesis of seedlings. The fluctuation of osmoregulation substance content confirmed that highland barley seedlings had a certain degree of stress resistance. Freeze-thaw cycles will aggravate the damage of land salinisation to highland barley seedlings. To better reduce the impact and loss of land salinisation and freeze-thaw disasters on agriculture in the Qinghai-Tibetan Plateau, priority should be given to solving freeze-thaw stress in the process of grain production.

Response characteristics of highland barley (Hordeum vulgare) seedlings to the stress of salinity and artemisinin under freeze­thaw environment.

In Qinghai-Tibet Plateau, crops are commonly subjected to freeze-thaw and salt stress factors simultaneously, and allelopathy is common, which affects the growth of highland barley (Hordeum vulgare L.), the largest food crop in Tibet. In order to explore the effects of artemisinin, salt and freeze-thaw (FAS) stress on physiological characteristics of highland barley seedlings, hydroponic experiment was carried out with the addition of 20mg/L artemisinin and 150mMNaCl as well as the simulation of freeze-thaw environment. The results suggested that under combined stress, the soluble protein content in combined stresses of artemisinin, FAS increased by 97.8%, the variation of relative conductivity in FAS group was lower than that in combined salt and freeze-thaw stress (FS), the relative water content decreased significantly (P <0.05), the malondialdehyde (MDA), H2 O2 and soluble sugar content in FAS group accumulated but less than those in FS group, and the superoxide dismutase (SOD) activity in combined artemisinin and freeze-thaw stress (FA) and FAS groups decreased. In addition, after freeze-thaw treatment, photosynthesis was weakened, and internal CO2 conentration (C i ) in FAS group significantly decreased (P <0.05). This study proved that appropriate amount of artemisinin can alleviate the damage of salt and freeze-thaw stress on barley seedlings.

Physiological response of barley seedlings to salinity and artemisinin combined stresses under freeze-thaw environment.

In the Qinghai-Tibet Plateau, both the large daily temperature difference and soil salinization make plants susceptible to abiotic stresses such as freeze-thaw and salinity. Meanwhile, crops in this area can be affected by artemisinin, an antimalarial secondary metabolite produced in Artemisia. Under freeze-thaw and salinity stresses, artemisinin was induced as an allelopathy stress factor to explore the physiological response of highland barley, including the relative electrical conductivity (RC), soluble protein (SP) content, malondialdehyde (MDA) content, antioxidant enzyme activity, and water use efficiency (WUE). Compared with the control group, the contents of RC and MDA in seedling leaves under stress were significantly increased by 24.74-402.37% and 20.18-77.95%, indicating that cell membrane permeability was greatly damaged, and WUE was significantly decreased by 15.77-238.59%. The activity of enzymes increased under single stress and decreased under combined stress. Salinity, artemisinin, and freeze-thaw stress show a synergistic relationship; that is, compound stresses were more serious than single stress. In summary, the results of this study revealed the physiological and ecological responses of barley seedlings under different habitat stresses and the interactions among different stress factors.

Response characteristics of highland barley under freeze-thaw, drought and artemisinin stresses.

The freeze-thaw of early spring in China's Qinghai-Tibet Plateau is often accompanied by severe droughts. Artemisia annua, widely distributed in China, releases allelopathic substances, mainly artemisinin, to the environment and exerts a wide range of effects on crops. This paper studied the physiological effects of highland barley under freeze-thaw, drought, and artemisinin stress through indoor simulation experiments. The physiological response characteristics of superoxide dismutase (SOD) activity, catalase (POD) activity, net photosynthetic rate, relative water content (RWC), relative electrical conductivity, malondialdehyde (MDA) content, and soluble protein content in highland barley were analyzed. The results showed that artemisinin and drought contributed to the increase of SOD activity and the decrease of POD activity. Under the freeze-thaw stress, the SOD and POD activities both decreased firstly and then increased, but the effect of compound stress on POD was more complicated. Either artemisinin, drought, or low temperature could reduce the net photosynthetic rate of highland barley. Low temperature had more significant impacts on photosynthesis, and compound stress would show a single stress superimposed effect. Artemisinin, drought, and low temperature could reduce the RWC of highland barley, and increase the relative electrical conductivity and the concentration of soluble protein (except for low temperature stress above zero, which reduces the concentration of soluble protein). However, the effect of compound stress on soluble protein is more complex. The single stress of artemisinin and drought had no obvious effect on MDA content, while the MDA content was increased significantly under the freeze-thaw stress and the compound stress of artemisinin and drought, and the MDA content reached its peak at T1. The results are helpful to explore the effects of freeze-thaw, drought and artemisinin stress on the growth of highland barley under the background of the aridification of the Qinghai-Tibet Plateau, and provide ideas for rational agricultural management.

Physiological effects of different stubble height and freeze-thaw stress on Secale cereale L. seedlings.

BACKGROUND: As a biennial plant, Secale cereale L is usually harvested in the autumn in the northern part of China where the temperature difference between day and night is of great disparity Through the pot experiment, the seedlings were cut to 2, 6 and 10 cm stubble height, and the simulated freeze-thaw (FT) stress (10/- 5 °C) was carried out after 6 days regrowth. The physiological effects of FT with different stubble height were revealed by analyzing the relative water content (RWC), osmotic adjustment substance concentration (soluble sugar and protein), membrane peroxidation (MDA) and catalase (CAT) activity. RESULTS: The results demonstrated that under freeze stress (- 5 °C), the content of soluble protein and MDA decreased and the seedlings of 2 cm treatment kept higher level of soluble protein and MDA, while the seedlings of 6 and 10 cm treatments kept higher level of the RWC, soluble sugar content, and CAT activity. After FT stress, the content of soluble sugar and protein, RWC in the 6 cm treatment were higher than those in 2 cm and 10 cm treatments, and the CAT activity in 10 cm treatment was the highest while the MDA content is lower. CONCLUSION: These data suggest that keeping high stubble height is more adaptive for short-term FT stress.

Physiological effects of the combined stresses of freezing-thawing, acid precipitation and deicing salt on alfalfa seedlings.

BACKGROUND: Frequent freeze-thaw phenomena, together with widely used deicing salt and intense acid precipitation, often occur in northeastern China, causing damage to various aspects of plants, such as the permeability of biological membranes, osmotic adjustment, and photosystems. Aiming to explore the resistance of alfalfa to freezing-thawing (F), acid precipitation (A) and deicing salt (D), this study used Medicago sativa cv. Dongmu-70 as the experimental material, and the contents of malondialdehyde (MDA), soluble protein, soluble sugars, proline and chlorophyll were evaluated. RESULTS: As the temperature decreased, the MDA content in the seedlings of the group under combined stress (A-D-F) increased and was significantly higher than that of group F (by 69.48 ~ 136.40%). Compared with those in the control (CK) group, osmotic substances such as soluble sugars and proline in the treatment groups were higher, while the soluble protein content was lower. The chlorophyll contents in the seedlings of the treatment groups were lower than those of the CK group; however, the chlorophyll content displayed a non-significant change during the free-thaw cycle. CONCLUSION: Injury to the permeability of the biological membranes and photosystems of alfalfa results from stress. Moreover, alfalfa maintains osmotic balance by adaptively increasing the potential of osmotic substances such as soluble sugars and proline. Furthermore, the influence of stress from freezing-thawing and deicing salt is highly substantial, but the combined stresses of acid precipitation with the two factors mentioned above had little effect on the plants.

Physiological response of Secale cereale L. seedlings under freezing-thawing and alkaline salt stress.

Freezing-thawing and saline-alkaline are the major abiotic stress for the pasture in most high-latitude areas, which are serious threats to the yield of pasture. In this study, the osmotic adjustment substances, membrane lipid peroxidation, and antioxidant enzymes activities of rye (Secale cereale L., cv. Dongmu-70) seedlings under different treatments: CK (no treatment), SC (Na2CO3 treatment), FT (freezing-thawing treatment), and FT+SC (combined Na2CO3 and freezing-thawing treatments), were investigated. At the freezing stage, the content of MDA and proline, the activity of APX, SOD, and POD increased with the decrease of the temperature in the leaves of rye seedlings in FT and FT+SC treatments and reached the maximum value at - 5 °C. In addition, the content of protein and H2O2, CAT activity reached the maximum value at 0 °C; the damage is larger under low temperature stress at 0 °C and - 5 °C in rye seedling. At the thawing stage, the content of MDA and H2O2 in seedling leaves decreased in FT and FT + SC treatments. These results demonstrated that proline content and antioxidant enzymes activities could play an important role in protecting cytomembrane and scavenging ROS respectively in rye under alkaline salt stress and freezing-thawing stress. The result also indicated rye seedlings were subjected to a freezing-thawing stress which resulted in a reversible (recoverable) injury.

Physiological response in the leaf and stolon of white clover under acid precipitation and freeze-thaw stress.

Freeze-thaw (FT) in northern China is a common event in spring and autumn, and the release of sulfur dioxide from coal-burning in winter is apt to trigger acid precipitation. Both these stresses can aggravate the wintering ability of white clover (Trifolium repens L.). Acid precipitation and FT simulation experiments were carried out in the field and an indoor alternation refrigerator, respectively. The contents of soluble protein, soluble sugar, malondialdehyde (MDA), proline and antioxidant activity were tested under acid precipitation and FT stress. The results showed that under acid precipitation stress, the content of MDA, peroxidase and superoxide dismutase increased in both leaves and stolons, whereas soluble protein and soluble sugar content declined compared with the control groups. During the freezing period, the content of antioxidant enzyme activity, soluble protein and proline increased at first and then dropped, whereas MDA and soluble sugar content both increased. As a conclusion, the stolon of white clover is more sensitive than the leaf to short-term stress, either as the single FT stress or the combined stress of FT and acid precipitation, suggesting that maintaining more leaves can contribute to the resistance of white clover to these stresses.

[Effects of grazing on architecture and small-scale pattern of grasses on artificial grassland in subtropical zone].

This study was conducted on a 5-year artificial grassland in subtropical zone of South China. The main types of established artificial grassland there were Dactylis glomerata-Lolium prenne-Trifolium repens and D. glomerata-T. repens pastures. Four grazing intensities were designed, i.e., CK (no grazing), G1 (6 adult sheep x hm(-2)), G2 (7.5 adult sheep x hm(-2)) and G3 (10 adult sheep x hm(-2)), and all the grazing plots were rotationally grazed. The architecture and small-scale pattern of grasses on the grazed and ungrazed grassland were discussed. After a period of 5-year grazing, the plant basal width and sward height of D. glomerata and T. pratense decreased gradually. In treatments CK, G1, G2 and G3, the basal width of D. glomerata was 6-8, 2-4, 0-2 and 0-2 cm, and that of T. pratense was 1-1.2, 6-8, 4-6 and 2-4 cm, respectively. The tuft density of D. glomerata in treatments CK, G1, G2 and G3 was 60, 95.1, 210.2 and 160 tufts x m(-2), respectively. The tiller number per plant of D. glomerata decreased, while its tuft density increased significantly due to the increased grazing intensity. With the increase of grazing intensity, the internode length of T. repens decreased, while its branching angle increased. The average internode length in treatments CK, G1, G2 and G3 was 2.04, 1.69, 1.64 and 1.51 cm, while the branching angle was 46.5, 65, 73 and 77.3 degrees, respectively. The average leaf density of T. repens in treatments CK, G1, G2 and G3 was 2.9, 13.0, 4.7 and 1.0 x m(-2), while the relevant stolon density was 19.9, 101, 142 and 82.6 m x m(-2), respectively. Under moderate grazing intensity, both the leaf and stolon densities of T. repens increased. The main scale on small pattern of D. glomerata, T. repens and T. pratense was 2 cm x 2 cm, which was further decreased under higher grazing intensity in the treatments of D. glomerata and T. pratense. Considering the heterogeneity of environmental resources, the change of the architecture and small-scale pattern could be regarded as an adaptation of grasses under grazing disturbance.

[Effects of grazing, cutting and decapitating on grass populations on the artificial grassland in subtropical zone of China].

Studies on the effects of grazing, cutting and decapitating on grass populations on the artificial grassland in the subtropical zone of China showed that moderate grazing and cutting could increase the adaptability of Dactylis glomerata and Lolium prenn due to decapitating. Decapitating could rescind the apical dominance and reproduction growth of grass, and hence, the ecological effects of grazing and cutting on grass could be realized by decapitating, and the population density, caloric value and energy accumulation of Dactylis glomerata and Lolium prenn increased obviously. Apical dominance of grasses could lower the population density, energy accumulation and intra-population competitive ability.

[Effect of cutting periodicity on energy allocation and architecture of Trifolium repens].

On the subtropical mountain swards, the effect of cutting periodicity on energy allocation, module density, number of branches and branching angle of T. repens were significant, With cutting periodicity raised, the energy allocation of stolon increased steadly, the caloric content of unit stolon dropped gradually, and the changing pattern of leaf density, stem density, branch number and internode's length was from low to high, and then to low. The branching intensity of T. repens ranged from 15 to 23.7 branches.m-2, and branching angle raried from 49.5 degrees to 60.2 degrees while the cutting periodicity differed.