Combined Effect of Microplastics and Cd Alters the Enzymatic Activity of Soil and the Productivity of Strawberry Plants.

The synergistic effect between heavy metals and microplastics can affect soil properties as well as plant performance and yield. The objective of this study was to evaluate the combined effect of microplastics and cadmium on a soil-plant system. Specifically, we proposed to explore changes in soil microbiological activity, the growth and yield parameters of strawberry plants, and to evaluate the accumulation of these pollutants in the soil and root system. Plants were planted in clay pots under greenhouse conditions. The experiment was set up as a completely randomized design, with four treatments (Control; MPs; Cd; and Cd + MPs) and five replicates. The results showed that MPs and/or Cd affected plant growth, plant biomass, the number of fruits, root characteristics, dehydrogenase activity, acid phosphatase, and microbial biomass, and increased the accumulation of Cd in the roots and soil. The increased bioavailability of Cd, due to the presence of microplastics, could explain the observed negative effects on soil properties and the performance of strawberry plants.

Relieving your stress: PGPB associated with Andean xerophytic plants are most abundant and active on the most extreme slopes.

Introduction: Plants interact with plant growth-promoting bacteria (PGPB), especially under stress condition in natural and agricultural systems. Although a potentially beneficial microbiome has been found associated to plants from alpine systems, this plant- PGPB interaction has been scarcely studied. Nevados de Chillán Complex hold one of the southernmost xerophytic formations in Chile. Plant species living there have to cope with drought and extreme temperatures during the growing season period, microclimatic conditions that become harsher on equatorial than polar slopes, and where the interaction with PGPB could be key for plant survival. Our goal was to study the abundance and activity of different PGPB associated to two abundant plant species of Andean xerophytic formations on contrasting slopes. Methods: Twenty individuals of Berberis empetrifolia and Azorella prolifera shrubs were selected growing on a north and south slope nearby Las Fumarolas, at 2,050 m elevation. On each slope, microclimate based on temperature and moisture conditions were monitored throughout the growing period (oct. - apr.). Chemical properties of the soil under plant species canopies were also characterized. Bacterial abundance was measured as Log CFU g-1 from soil samples collected from each individual and slope. Then, the most abundant bacterial colonies were selected, and different hormonal (indoleacetic acid) and enzymatic (nitrogenase, phosphatase, ACC-deaminase) mechanisms that promote plant growth were assessed and measured. Results and Discussion: Extreme temperatures were observed in the north facing slope, recording the hottest days (41 vs. 36°C) and coldest nights (-9.9 vs. 6.6°C). Moreover, air and soil moisture were lower on north than on south slope, especially late in the growing season. We found that bacterial abundance was higher in soils on north than on south slope but only under B. empetrifolia canopy. Moreover, the activity of plant growth-promoting mechanisms varied between slopes, being on average higher on north than on south slope, but with plant species-dependent trends. Our work showed how the environmental heterogeneity at microscale in alpine systems (slope and plant species identity) underlies variations in the abundance and plant growth promoting activity of the microorganisms present under the plant canopy of the Andean xerophytic formations and highlight the importance of PGPB from harsh systems as biotechnological tools for restoration.

Warmer Temperatures Affect the in situ Freezing Resistance of the Antarctic Vascular Plants.

Although positive effects on growth and reproduction of Antarctic vascular plants have been reported under warmer temperatures, it could also increase the vulnerability of these plants to freezing. Thus, we assessed in situ whether warming decreases the freezing resistance of Colobanthus quitensis and Deschampsia antarctica, and we compared the level and mechanism of freezing resistance of these species in the field with previous reports conducted in lab conditions. We assessed the freezing resistance of C. quitensis and D. antarctica by determining their low temperature damage (LT50), ice nucleation temperature (NT) and freezing point (FP) in three sites of the King George Island. Plants were exposed during two growing seasons to a passive increase in the air temperature (+W). +W increased by 1K the mean air temperatures, but had smaller effects on freezing temperatures. Leaf temperature of both species was on average 1.7K warmer inside +W. Overall, warming decreased the freezing resistance of Antarctic species. The LT50 increased on average 2K for C. quitensis and 2.8K for D. antarctica. In contrast, NT and FP decreased on average c. 1K in leaves of warmed plants of both species. Our results showed an averaged LT50 of -15.3°C for C. quitensis, and of -22.8°C for D. antarctica, with freezing tolerance being the freezing resistance mechanism for both species. These results were partially consistent with previous reports, and likely explanations for such discrepancies were related with methodological differences among studies. Our work is the first study reporting the level and mechanisms of freezing resistance of Antarctic vascular plants measured in situ, and we demonstrated that although both plant species exhibited a great ability to cope with freezing temperatures during the growing season, their vulnerability to suffer freezing damage under a warming scenario increase although the magnitude of this response varied across sites and species. Hence, freezing damage should be considered when predicting changes in plant responses of C. quitensis and D. antarctica under future climate conditions of the Antarctic Peninsula.

Winter is coming: plant freezing resistance as a key functional trait for the assembly of annual Mediterranean communities.

Background and Aims: In Mediterranean annual plants, germination mainly occurs during the autumn and only those seedlings that survive winter freezing can flower and produce seedlings in spring. Surprisingly, the effect of freezing events as an abiotic determinant of these communities remains unexplored. The present study aimed to investigate how freezing events affect annual Mediterranean communities and whether their functional structure as related to freezing resistance is linked to the main biotic and abiotic determinants of these communities. Methods: In 120 plots located on a semi-arid Mediterranean steppe (Spain), the community functional structure related to the lethal temperature causing 50 % frost damage (LT50 trait) in seedlings was estimated and summarized as the community-weighted mean (CWM-LT50) and its functional diversity (FD-LT50). Plots were stratified according to distance to rabbit shelters and latrines as a proxy for rabbit density, proximity to Stipa tenacissima and spring water availability, where annual species abundance was recorded in all plots over three consecutive years. Key Results: Annual species were able to resist a threshold temperature of -4 °C and most had LT50 values around the absolute minimum temperature (-9.5 °C) in the three years. Higher rabbit densities led to lower CWM-LT50 and higher FD-LT50 values. Plots close to Stipa tussocks had higher CWM-LT50 values whereas water availability had no effects. Conclusions: High freezing resistance was extended among winter annual species, suggesting the presence of an association between historical environmental filtering and low winter temperatures. However, the community functional structure related to freezing resistance remained variable among scenarios with differences in herbivory pressure and distance to perennial vegetation. The trends observed indicate that traits that allow plants to deal with herbivory may also promote freezing resistance, and that tussocks can act as nurses via microclimatic amelioration of harsher winter conditions.

Drought increases the freezing resistance of high-elevation plants of the Central Chilean Andes.

Freezing temperatures and summer droughts shape plant life in Mediterranean high-elevation habitats. Thus, the impacts of climate change on plant survival for these species could be quite different to those from mesic mountains. We exposed 12 alpine species to experimental irrigation and warming in the Central Chilean Andes to assess whether irrigation decreases freezing resistance, irrigation influences freezing resistance when plants are exposed to warming, and to assess the relative importance of irrigation and temperature in controlling plant freezing resistance. Freezing resistance was determined as the freezing temperature that produced 50 % photoinactivation [lethal temperature (LT50)] and the freezing point (FP). In seven out of 12 high-Andean species, LT50 of drought-exposed plants was on average 3.5 K lower than that of irrigated plants. In contrast, most species did not show differences in FP. Warming changed the effect of irrigation on LT50. Depending on species, warming was found to have (1) no effect, (2) to increase, or (3) to decrease the irrigation effect on LT50. However, the effect size of irrigation on LT50 was greater than that of warming for almost all species. The effect of irrigation on FP was slightly changed by warming and was sometimes in disagreement with LT50 responses. Our data show that drought increases the freezing resistance of high-Andean plant species as a general plant response. Although freezing resistance increases depended on species-specific traits, our results show that warmer and moister growing seasons due to climate change will seriously threaten plant survival and persistence of these and other alpine species in dry mountains.

Summer Freezing Resistance: A Critical Filter for Plant Community Assemblies in Mediterranean High Mountains.

Assessing freezing community response and whether freezing resistance is related to other functional traits is essential for understanding alpine community assemblages, particularly in Mediterranean environments where plants are exposed to freezing temperatures and summer droughts. Thus, we characterized the leaf freezing resistance of 42 plant species in 38 plots at Sierra de Guadarrama (Spain) by measuring their ice nucleation temperature, freezing point (FP), and low-temperature damage (LT50), as well as determining their freezing resistance mechanisms (i.e., tolerance or avoidance). The community response to freezing was estimated for each plot as community weighted means (CWMs) and functional diversity (FD), and we assessed their relative importance with altitude. We established the relationships between freezing resistance, growth forms, and four key plant functional traits (i.e., plant height, specific leaf area, leaf dry matter content (LDMC), and seed mass). There was a wide range of freezing resistance responses and more than in other alpine habitats. At the community level, the CWMs of FP and LT50 responded negatively to altitude, whereas the FD of both traits increased with altitude. The proportion of freezing-tolerant species also increased with altitude. The ranges of FP and LT50 varied among growth forms, and only leaf dry matter content was negatively correlated with freezing-resistance traits. Summer freezing events represent important abiotic filters for assemblies of Mediterranean high mountain communities, as suggested by the CWMs. However, a concomitant summer drought constraint may also explain the high freezing resistance of species that thrive in these areas and the lower FD of freezing resistance traits at lower altitudes. Leaves with high dry matter contents may maintain turgor at lower water potential and enhance drought tolerance in parallel to freezing resistance. This adaptation to drought seems to be a general prerequisite for plants found in xeric mountains.

Photosynthetic Light Responses May Explain Vertical Distribution of Hymenophyllaceae Species in a Temperate Rainforest of Southern Chile.

Some epiphytic Hymenophyllaceae are restricted to lower parts of the host (< 60 cm; 10-100 µmol photons m(-2) s(-1)) in a secondary forest of Southern Chile; other species occupy the whole host height (≥ 10 m; max PPFD > 1000 µmol photons m(-2) s(-1)). Our aim was to study the photosynthetic light responses of two Hymenophyllaceae species in relation to their contrasting distribution. We determined light tolerance of Hymenoglossum cruentum and Hymenophyllum dentatum by measuring gas exchange, PSI and PSII light energy partitioning, NPQ components, and pigment contents. H. dentatum showed lower maximum photosynthesis rates (A max) than H. cruentum, but the former species kept its net rates (An) near Amax across a wide light range. In contrast, in the latter one, An declined at PPFDs > 60 µmol photons m(-2) s(-1). H. cruentum, the shadiest plant, showed higher chlorophyll contents than H. dentatum. Differences in energy partitioning at PSI and PSII were consistent with gas exchange results. H. dentatum exhibited a higher light compensation point of the partitioning of absorbed energy between photochemical Y(PSII) and non-photochemical Y(NPQ) processes. Hence, both species allocated energy mainly toward photochemistry instead of heat dissipation at their light saturation points. Above saturation, H. cruentum had higher heat dissipation than H. dentatum. PSI yield (YPSI) remained higher in H. dentatum than H. cruentum in a wider light range. In both species, the main cause of heat dissipation at PSI was a donor side limitation. An early dynamic photo-inhibition of PSII may have caused an over reduction of the Qa+ pool decreasing the efficiency of electron donation to PSI. In H. dentatum, a slight increase in heat dissipation due to acceptor side limitation of PSI was observed above 300 µmol photons m(-2)s(-1). Differences in photosynthetic responses to light suggest that light tolerance and species plasticity could explain their contrasting vertical distribution.

Facilitative interactions do not wane with warming at high elevations in the Andes.

Positive interactions between species are known to play an important role in the structure and dynamics of alpine plant communities. The balance between negative and positive interactions is known to shift along spatial and temporal gradients, with positive effects prevailing over negative ones as the environmental stress increases. Thus, this balance is likely to be affected by climate change. We hypothesized that increases in temperature (a global warming scenario) should decrease the importance of positive interactions for the survival and growth of alpine plant species. To test this hypothesis, we selected individuals of the native grass species Hordeum comosum growing within the nurse cushion species Azorella madreporica at 3,600 m.a.s.l. in Los Andes (Chile), and performed nurse removal and seedling survival experiments under natural and warmer conditions. For warmer conditions, we used open-top chambers, which increased the temperature by 4 °C. After two growing seasons, we compared the effect of nurse removal on the survival, biomass, and photochemical efficiency of H. comosum individuals under warmer and natural conditions. Nurse removal significantly decreased the survival, biomass, and photochemical efficiency of H. comosum, demonstrating the facilitative effects of nurse cushions. Seedling survival was also enhanced by cushions, even under warmer conditions. However, warmer conditions only partially mitigated the negative effects of nurse removal, suggesting that facilitative effects of cushions do not wane under warmer conditions. Thus, facilitative interactions are vital to the performance and survival of alpine species, and these positive interactions will continue to be important in the warmer conditions of the future in high-alpine habitats.

Summer freezing resistance decreased in high-elevation plants exposed to experimental warming in the central Chilean Andes.

Alpine habitats have been proposed as particularly sensitive to climate change. Shorter snow cover could expose high-elevation plants to very low temperatures, increasing their risk of suffering damage by freezing, hence decreasing their population viability. In addition, a longer and warmer growing season could affect the hardening process on these species. Thus, understanding the ability of these species to withstand freezing events under warmer conditions is essential for predicting how alpine species may respond to future climate changes. Here we assessed the freezing resistance of 11 species from the central Chilean Andes by determining their low temperature damage (LT(50)) and freezing point (FP) after experimental warming in the field. Plants were exposed during two growing seasons to a passive increase in the air temperature using open top chambers (OTCs). OTCs increased by ca. 3 K the mean air and soil daytime temperatures, but had smaller effects on freezing temperatures. Leaf temperature of the different species was on average 5.5 K warmer inside OTCs at midday. While LT(50) of control plants ranged from -9.9 to -22.4, that of warmed plants ranged from -7.4 to -17.3 degrees C. Overall, high-Andean species growing inside OTCs increased their LT(50) ca. 4 K, indicating that warming decreased their ability to survive severe freezing events. Moreover, plants inside OTCs increased the FP ca. 2 K in some studied species, indicating that warming altered processes of ice crystal formation. Resistance of very low temperatures is a key feature of high-elevation species; our results suggest that current climate warming trends will seriously threaten the survival of high-elevation plants by decreasing their ability to withstand severe freezing events.

Freezing resistance varies within the growing season and with elevation in high-Andean species of central Chile.

Predicted increases in the length of the growing season as a result of climate change may more frequently expose high-elevation plants to severe frosts. Understanding the ability of these species to resist frosts during the growing season is essential for predicting how species may respond to changes in temperature regimes. Here, we assessed the freezing resistance of 24 species from the central Chilean Andes by determining their low temperature damage (LT(50)), ice nucleation temperature (NT), freezing point (FP) and freezing resistance mechanism (i.e. avoidance or tolerance). The Andean species were found to resist frosts from -8.2 to -19.5 degrees C during the growing season, and freezing tolerance was the most common resistance mechanism. Freezing resistance (LT(50)) varied within the growing season, decreasing towards the end of this period in most of the studied species. However, the FP showed the opposite trend. LT(50) increased with elevation, whilst FP was lower in plants from lower elevations, especially late in the growing season. Andean species have the potential to withstand severe freezing conditions during the growing season, and the aridity of this high-elevation environment seems to play an important role in determining this high freezing resistance.

Positive interactions between alpine plant species and the nurse cushion plant Laretia acaulis do not increase with elevation in the Andes of central Chile.

In alpine habitats, positive interactions among plants tend to increase with elevation as a result of altitudinal increase in environmental harshness. However, in mountains located in arid zones, lower elevations are also stressful because of scarce availability of water, suggesting that positive interactions may not necessarily increase with elevation. Here we analysed the spatial association of plant species with the nurse cushion plant Laretia acaulis at two contrasting elevations, and monitored the survival of seedlings of two species experimentally planted within and outside cushions in the semiarid Andes of central Chile. Positive spatial associations with cushions were more frequent at lower elevations. Species growing at the two elevations changed the nature of their association with cushions from neutral or negative at higher elevations to positive at lower elevations. Survival of seedlings was higher within cushions, particularly at lower elevations. The increased facilitation by cushions at lower elevations seems to be related to provision of moisture. This result suggests that cushion plants play a critical role in structuring alpine plant communities at lower elevations, and that climatic changes in rainfall could be very relevant for persistence of plant communities.