Acetylation proteomics and metabolomics analyses reveal the involvement of starch synthase undergoing acetylation modification during UV-B stress resistance in Rhododendron Chrysanthum Pall.

BACKGROUND: Rhododendron chrysanthum Pall. (R. chrysanthum) is a plant that lives in high mountain with strong UV-B radiation, so R. chrysanthum possess resistance to UV-B radiation. The process of stress resistance in plants is closely related to metabolism. Lysine acetylation is an important post-translational modification, and this modification process is involved in a variety of biological processes, and affected the expression of enzymes in metabolic processes. However, little is known about acetylation proteomics during UV-B stress resistance in R. chrysanthum. RESULTS: In this study, R. chrysanthum OJIP curves indicated that UV-B stress damaged the receptor side of the PSII reaction center, with a decrease in photosynthesis, a decrease in sucrose content and an increase in starch content. A total of 807 differentially expressed proteins, 685 differentially acetylated proteins and 945 acetylation sites were identified by quantitative proteomic and acetylation modification histological analysis. According to COG and subcellular location analyses, DEPs with post-translational modification of proteins and carbohydrate metabolism had important roles in resistance to UV-B stress and DEPs were concentrated in chloroplasts. KEGG analyses showed that DEPs were enriched in starch and sucrose metabolic pathways. Analysis of acetylation modification histology showed that the enzymes in the starch and sucrose metabolic pathways underwent acetylation modification and the modification levels were up-regulated. Further analysis showed that only GBSS and SSGBSS changed to DEPs after undergoing acetylation modification. Metabolomics analyses showed that the metabolite content of starch and sucrose metabolism in R. chrysanthum under UV-B stress. CONCLUSIONS: Decreased photosynthesis in R. chrysanthum under UV-B stress, which in turn affects starch and sucrose metabolism. In starch synthesis, GBSS undergoes acetylation modification and the level is upregulated, promotes starch synthesis, making R. chrysanthum resistant to UV-B stress.

Alpine Rhododendron population contractions lead to spatial distribution mismatch with their pollinators under climate change.

The effect of global climate change on plant-pollinator interaction is not limited to changes in phenology and richness within communities but also includes the spatial mismatch caused by the inconsistency of geographical distribution changes. Subsequently, the pollinator interaction network may be remodeled or even disrupted. In this study, we simulated the suitable habitat niche of 15 Rhododendron species and their eight pollinator species as well as their overlapping versus geographical mismatch under the current and three future climate change scenarios in 2090s, using MaxEnt. Results showed that the suitable habitat of all Rhododendron species would decrease in 2090s. In particular, 10, 8, and 13 Rhododendron-pollinator assemblages would have a reduced spatial match region under the climate change scenarios, mainly due to the contraction of the suitable habitat of Rhododendron species. The results provide novel insights into the response of plant-pollinator interactions to global warming, useful to prioritize conservation actions of alpine plant ecosystems.

Phenocam observed flowering anomaly of Rhododendron arboreum Sm. in Himalaya: a climate change impact perspective.

Flowering exhibits a significant relationship with environmental stimuli and changes. Effect of photoperiodism and vernalization have been well studied in flowering phenology; however, the effect of soil temperature on flowering is less explored which is one of the major factors of vegetation growth in alpine ecosystem. This study thus focuses on the effects of soil and air temperature on flowering response of Rhododendron arboreum Sm., a Himalayan tree species, which is also an indicator of spring initiation in high altitude regions. To monitor the flowering pattern, we employed automated phenocam, which was set up at 3356 masl in Tungnath (Indian Alpine region of Uttarakhand) for time-lapse photography of timberline ecotone. Soil and air temperature were recorded continuously at the timberline ecotone. Three years (2017 to 2020) of datasets were used for the present study. The phenocam observations displayed an interesting event in the year 2019-2020 with complete absence of flowering in R. arboreum population at Tungnath timberline ecotone. From the soil temperature data, an increase in winter (Dec-Jan, during which floral buds form) soil temperature, by > 1 °C, and no accumulation of freezing degree-days were found for the year 2019-2020. Air temperature however did not display any relationship with the failure of flowering, ruling out aerial chilling or frost injury of floral buds. From the results, a possible relationship between soil temperature and flowering can be suggested pointing towards necessary root apex vernalization stimulus in shallow rooted Rhododendrons. However, the dependency of flowering in Rhododendrons on winter soil temperature further requires continuous monitoring and more observations to make concrete inferences.

Factors affecting freezing tolerance: a comparative transcriptomics study between field and artificial cold acclimations in overwintering evergreens.

Cold acclimation (CA) is a well-known strategy employed by plants to enhance freezing tolerance (FT) in winter. Global warming could disturb CA and increase the potential for winter freeze-injury. Thus, developing robust FT through complete CA is essential. To explore the molecular mechanisms of CA in woody perennials, we compared field and artificial CAs. Transcriptomic data showed that photosynthesis/photoprotection and fatty acid metabolism pathways were specifically enriched in field CA; carbohydrate metabolism, secondary metabolism and circadian rhythm pathways were commonly enriched in both field and artificial CAs. When compared with plants in vegetative growth in the chamber, we found that the light signals with warm air temperatures in the fall might induce the accumulation of leaf abscisic acid (ABA) and jasmonic acid (JA) concentrations, and activate Ca2+ , ABA and JA signaling transductions in plants. With the gradual cooling occurrence in winter, more accumulation of anthocyanin, chlorophyll degradation, closure/degradation of photosystem II reaction centers, and substantial accumulation of glucose and fructose contributed to obtaining robust FT during field CA. Moreover, we observed that in Rhododendron 'Elsie Lee', ABA and JA decreased in winter, which may be due to the strong requirement of zeaxanthin for rapid thermal dissipation and unsaturated fatty acids for membrane fluidity. Taken together, our results indicate that artificial CA has limitations to understand the field CA and field light signals (like short photoperiod, light intensity and/or light quality) before the low temperature in fall might be essential for complete CA.

Photorespiration is the major alternative electron sink under high light in alpine evergreen sclerophyllous Rhododendron species.

Owing to the high leaf mass per area, alpine evergreen sclerophyllous Rhododendron have low values of mesophyll conductance (gm). The resulting low chloroplast CO2 concentration aggravates photorespiration, which requires a higher ATP/NADPH ratio. However, the significance of photorespiration and underlying mechanisms of energy balance in these species are little known. In this study, eight alpine evergreen sclerophyllous Rhododendron species grown in a common garden were tested for their gm, electron flow to photorespiration, and energy balancing. Under saturating light, gm was the most limiting factor for net photosynthesis (AN) in all species, and the species differences in AN were primarily driven by gm rather than stomatal conductance. The total electron flow through photosystem II (ETRII) nearly equaled the electron transport required for Rubisco carboxylation and oxygenation. Furthermore, blocking electron flow to photosystem I with appropriate inhibitors showed that electron flow to plastic terminal oxidase was not observed. As a result, these studied species showed little alternative electron flow mediated by water-water cycle or plastic terminal oxidase. By comparison, the ratio of electron transport consumed by photorespiration to ETRII (JPR/ETRII), ranging from 43%∼55%, was negatively correlated to AN and gm. Furthermore, the increased ATP production required by enhanced photorespiration was regulated by cyclic electron flow around photosystem I. These results indicate that photorespiration is the major electron sink for dissipation of excess excitation energy in the alpine evergreen sclerophyllous Rhododendron species. The coordination of gm, photorespiration and cyclic electron flow is important for sustaining leaf photosynthesis.

Climate and land use changes will degrade the distribution of Rhododendrons in China.

Biodiversity loss and variation in species responses to climate and land use change have been found across broad taxonomic groups. However, whether species from the same taxonomic group with distinct geographical ranges will respond differently is poorly understood. The aim of this study is to predict the potential impacts of future climate and land use change on the distribution of narrow- and wide-ranging Rhododendron species, and estimate their relative contribution in China. We applied the presence-only ecological niche model MaxEnt to predict the distribution of 10 narrow-ranging and 10 wide-ranging Rhododendron species for the year 2070, using three general circulation models and three scenarios of climate and land use change. We measured the predicted distribution change of each species using change ratio, distance and direction of core range shifts, and niche overlap using Schoener's D. We found that the distribution areas of six narrow-ranging species would decrease, of which one species would go extinct. The remaining four narrow-ranging species would experience range expansion. Distribution of all the wide-ranging Rhododendron species would decrease. All Rhododendrons will shift to the northwest. We conclude that Rhododendron species generally will be negatively affected by the climatic and land use change expected in 2070 from the three scenarios evaluated in this study, but some narrow-ranging species may be positively influenced. Narrow-ranging Rhododendron species are more vulnerable compared to wide-ranging Rhododendron species. This study demonstrated that the effects of climate and land use change on alpine and subalpine plant species is species-specific, thereby strengthening our understanding of the impacts of climate and land use change on plant distribution.

Pollen aggregation by viscin threads in Rhododendron varies with pollinator.

Pollen grains can be dispersed singly or variously aggregated in groups. Whether the evolution of pollen aggregation is driven by the pollinator remains unexplored. We hypothesize that an extensive pollen aggregation is favored under a scarcity of pollinators. Variation in pollen aggregation by viscin threads in 13 Rhododendron species was measured as it is related to pollen removal in a visit. Visitation rates of functional pollinator groups that vary in their grooming behavior were investigated in each species. Pollen deposited on stigmas in the field was also sampled. Seven Rhododendron species were infrequently pollinated by low-intensity grooming animals, including birds, butterflies and moths. The other six species were more frequently pollinated by bees with a high intensity of pollen grooming. Bird- and Lepidoptera-pollinated species produced longer pollen-connecting threads that connected more pollen grains. Phylogenetically independent contrast analysis of the 13 species showed that pollinator visitation frequency was negatively related to amounts of pollen removal per visit but not to stigmatic pollen loads. The finding of interspecific patterns in pollen removal related to pollinator visitation frequency suggests pollinator-mediated selection on pollen packaging strategies, supporting the hypothesis of floral evolution via pollen export.

Gene and metabolite profiling reveals flowering and survival strategies in Himalayan Rhododendron arboreum.

Rhododendron arboreum inhabits the Himalayan climate otherwise detrimental to many species, though the underlying survival mechanism remains unclear. Such temperate species have an inherent endurance towards freezing temperature which is prerequisite for an initiation and transition to flowering phase. Orchestrating the molecular architecture is vital towards managing distinct abiotic signals. To determine the molecular factors directing growth, development, and tolerance under environmental extremes in the species, the high-throughput transcriptome and metabolome data from vegetative as well as cold-acclimatized flowering season tissues were generated. Firstly, the de novo assembly pertaining to the foliar and floral tissues comprising of 157,427 unigenes was examined for a comparative analysis. 4149 of 12,577 transcripts observed with a significant fluctuating expression corresponded to seasonal retorts. Following the interactive network, 525 genes were distinguished as the epicenters of sense, response, and tolerance. Secondly, liquid chromatography coupled to mass spectrometry was adopted to profile the extent of metabolite richness across the tissues of two seasons. Taking into account the formula-based mappings offered by MetaboSearch tool, 421 unique ions obtained were annotated to 173 KEGG compounds, especially secondary metabolites. Moreover, by integrating the transcript and metabolite annotations, it was found that right from active metabolism, signaling, development, and their regulations, supplementary response to abiotic/biotic stimuli was induced. A multifaceted response displayed during flowering not only sponsored the climatic encounters but brought the shift from vegetative to reproductive growth. Overall, this comprehensive approach following transcriptome and non-targeted metabolome elucidated the contribution of genetic and metabolic factors in environmental responses.

Seasonal variations coupled with elevation gradient drives significant changes in eco-physiological and biogeochemical traits of a high altitude evergreen broadleaf shrub, Rhododendron anthopogon.

Higher elevations and, early as well as late phase of growing season are expected to be more stressful for plants in high altitudes. The present study was carried out on Rhododendron anthopogon D. Don, an evergreen shrub of Himalaya to understand variation in eco-physiological and biogeochemical traits due to combined effect of elevation gradient and growing season. We conducted our study at Rohtang, India (32°22'04″ N 77°15'17″ E) and undertook random sampling of leaves at four elevations (3200 m, 3600 m, 4000 m and 4250 m), and three time periods (late June, early August and late September) during growing season. We assessed 12 eco-physiological and biogeochemical variables and analysed results through ANOVA and multivariate analysis. It was found that leaf relative water content, nitrogen percentage (N%), carbon/nitrogen ratio (C/N ratio), total chlorophyll, malondialdehyde equivalents and proline content varied along two gradients (factors) with their interaction being statistically significant. Variance partitioning analysis of studied traits revealed that both factors contribute significantly, with 'season' component ranging between 55.75 % and 94.03 % for most of the parameters, whereas, 'elevation' component contributed more for leaf area, N% and C/N ratio (48.08 %-75.03 %). Our results suggest that eco-physiology of R. anthopogon is significantly influenced by interaction of seasonal variations coupled with elevation gradient. The study highlights the importance of examining both seasonal and elevational gradients in understanding plant adaptation strategies. Overall, our findings revealed that plasticity in eco-physiological and biogeochemical traits underline the wide distribution of R. anthopogon in the high altitudes.

Ozone disrupts adsorption of Rhododendron tomentosum volatiles to neighbouring plant surfaces, but does not disturb herbivore repellency.

The perennial evergreen woody shrub, Rhododendron tomentosum, confers associational resistance against herbivory and oviposition on neighbouring plants through passive adsorption of some of its constitutively emitted volatile organic compounds (VOCs). The adsorption process is dependent on transport of VOCs in the air. In polluted atmospheres, the VOCs may be degraded and adsorption impeded. We studied the effect of elevated ozone regimes on the adsorption of R. tomentosum volatiles to white cabbage, Brassica oleracea, and the oviposition of the specialist herbivore Plutella xylostella on the exposed plants. We found evidence for adsorption and re-emission of R. tomentosum volatiles by B. oleracea plants. Ozone changed the blend of R. tomentosum volatiles and reduced the amount of R. tomentosum volatiles recovered from B. oleracea plants. However, plants exposed to R. tomentosum volatiles received fewer P. xylostella eggs than control plants exposed to filtered air irrespective of whether R. tomentosum volatiles mixed with ozone. Ozone disrupts a volatile mediated passive plant-to-plant interaction by degrading some compounds and reducing the quantity available for adsorption by neighbouring plants. The change, however, did not affect the deterrence of oviposition by P. xylostella, suggesting that aromatic companion plants of Brassica crops may confer pest-deterring properties even in ozone-polluted environments.

In situ modeling of multimodal floral cues attracting wild pollinators across environments.

With more than 80% of flowering plant species specialized for animal pollination, understanding how wild pollinators utilize resources across environments can encourage efficient planting and maintenance strategies to maximize pollination and establish resilience in the face of environmental change. A fundamental question is how generalist pollinators recognize "flower objects" in vastly different ecologies and environments. On one hand, pollinators could employ a specific set of floral cues regardless of environment. Alternatively, wild pollinators could recognize an exclusive signature of cues unique to each environment or flower species. Hoverflies, which are found across the globe, are one of the most ecologically important alternative pollinators after bees and bumblebees. Here, we have exploited their cosmopolitan status to understand how wild pollinator preferences change across different continents. Without employing any a priori assumptions concerning the floral cues, we measured, predicted, and finally artificially recreated multimodal cues from individual flowers visited by hoverflies in three different environments (hemiboreal, alpine, and tropical) using a field-based methodology. We found that although "flower signatures" were unique for each environment, some multimodal lures were ubiquitously attractive, despite not carrying any reward, or resembling real flowers. While it was unexpected that cue combinations found in real flowers were not necessary, the robustness of our lures across insect species and ecologies could reflect a general strategy of resource identification for generalist pollinators. Our results provide insights into how cosmopolitan pollinators such as hoverflies identify flowers and offer specific ecologically based cues and strategies for attracting pollinators across diverse environments.

The role of idioblasts in leaf water relations of tropical Rhododendron.

PREMISE OF THE STUDY: Although much is known about the anatomy of idioblasts, the relationship of leaf idioblasts to leaf physiology is poorly known. Our goal was to understand the relationship between idioblast abundance and leaf water relations. Tropical epiphytic and shrub Rhododendron species were the model system. METHODS: Leaf succulence and idioblast metrics of 61 plants representing 17 species were compared with stomatal and water relation metrics. Correlation, ANOVA, and regression were used to understand which water relation traits were best aligned with the proportion of the leaf occupied by idioblasts for epiphytic and shrub species. KEY RESULTS: Idioblast volume per leaf area varied from 1.4-9.5 mm3·cm-2 among accessions, and an index of stomatal area per leaf area varied from 0.08-3.3. Succulence, stomatal, and water relation metrics varied significantly among species. Total idioblast volume was negatively correlated with leaf succulence metrics, and positively correlated with relative water content at the turgor loss point. Idioblasts were better related to capacitance in thin leaves. All idioblast metrics were significantly greater in epiphytic species. CONCLUSIONS: Leaf idioblasts can have a significant effect on leaf-lamina water relations. Idioblasts likely function as a water buffering system for thin leaves in tropical Rhododendron species. The association of idioblast prominence with leaf water buffering is greatest for epiphytic species with thin leaves. Thus, the proliferation of leaf idioblasts may have been an important innovation for Rhododendron species diversification into tropical epiphytic habitats.

Biochemical and proteomics analyses of antioxidant enzymes reveal the potential stress tolerance in Rhododendron chrysanthum Pall.

BACKGROUND: Rhododendron chrysanthum Pall., an endangered species with significant ornamental and medicinal value, is endemic to the Changbai Mountain of China and can also serve as a significant plant resource for investigating the stress tolerance in plants. Proteomics is an effective analytical tool that provides significant information about plant metabolism and gene expression. However, no proteomics data have been reported for R. chrysanthum previously. In alpine tundra, the abiotic stress will lead to a severe over-accumulation of reactive oxygen species (ROS). Many alpine plants overcome the severe stresses and protect themselves from the oxidative damage by increasing the ratio and activity of antioxidant enzymes. RESULTS: In our study, wild type and domesticated Rhododendron chrysanthum Pall. were used as experimental and control groups, respectively. Proteomics method combined with biochemical approach were applied for the stress tolerance investigation of R. chrysanthum at both protein and molecular level. A total of 1,395 proteins were identified, among which 137 proteins were up-regulate in the experimental group. The activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidases (APXs), and glutathione peroxidase (GPX) were significantly higher and the expression of APXs and GPX were also increased in the experimental group. Moreover, the interaction network analysis of these enzymes also reveals that the antioxidant enzymes play important roles in the stress resistance in plants. CONCLUSIONS: This is the first report of the proteome of Rhododendron chrysanthum Pall., and the data reinforce the notion that the antioxidant system plays a significant role in plant stress survival. Our results also verified that R. chrysanthum is highly resistant to abiotic stress and can serve as a significant resource for investigating stress tolerance in plants. REVIEWERS: This article was reviewed by George V. (Yura) Shpakovski and Ramanathan Sowdhamini.

Proteome dynamics of cold-acclimating Rhododendron species contrasting in their freezing tolerance and thermonasty behavior.

To gain a better understanding of cold acclimation in rhododendron and in woody perennials in general, we used the 2D-DIGE technique to analyze the rhododendron proteome during the seasonal development of freezing tolerance. We selected two species varying in their cold acclimation ability as well as their thermonasty response (folding of leaves in response to low temperature). Proteins were extracted from leaves of non-acclimated (NA) and cold acclimated (CA) plants of the hardier thermonastic species, R. catawbiense (Cata.), and from leaves of cold acclimated plants of the less hardy, non-thermonastic R. ponticum (Pont.). All three protein samples (Cata.NA, Cata.CA, and Pont.CA) were labeled with different CyDyes and separated together on a single gel. Triplicate gels were run and protein profiles were compared resulting in the identification of 72 protein spots that consistently had different abundances in at least one pair-wise comparison. From the 72 differential spots, we chose 56 spots to excise and characterize further by mass spectrometry (MS). Changes in the proteome associated with the seasonal development of cold acclimation were identified from the Cata.CA-Cata.NA comparisons. Differentially abundant proteins associated with the acquisition of superior freezing tolerance and with the thermonastic response were identified from the Cata.CA-Pont.CA comparisons. Our results indicate that cold acclimation in rhododendron involves increases in abundance of several proteins related to stress (freezing/desiccation tolerance), energy and carbohydrate metabolism, regulation/signaling, secondary metabolism (possibly involving cell wall remodeling), and permeability of the cell membrane. Cold acclimation also involves decreases in abundance of several proteins involved in photosynthesis. Differences in freezing tolerance between genotypes can probably be attributed to observed differences in levels of proteins involved in these functions. Also differences in freezing tolerance may be attributed to higher levels of some constitutive protective proteins in Cata. than in Pont. that may be required to overcome freeze damage, such as glutathione peroxidase, glutamine synthetase, and a plastid-lipid-associated protein.

Ecophysiological responses of Betula pendula, Pinus uncinata and Rhododendron ferrugineum in the Catalan Pyrenees to low summer rainfall.

Climate change is producing modifications in the intensity and frequency of rainfall in some regions of the planet. According to predictions, annual rainfall distribution in Western Europe will result in an increase in episodes of drought, thereby negatively affecting nutrient availability. Since high mountain systems will be particularly vulnerable, the physiological and nutritional responses to changes in summer rainfall were monitored over the course of two consecutive summers on three species, which are representative of subalpine forests: birch (Betula pendula Roth.), rhododendron (Rhododendron ferrugineum L.) and mountain pine (Pinus uncinata Mill.). Birch was especially susceptible to scarce precipitation showing alterations in leaf morphology and a decline in net photosynthesis (A) due to stomatal closure, which led to photoinhibition and to early leaf senescence as shown by the photosynthetic nitrogen-use efficiency (PNUE), carbon/nitrogen (C/N) ratio, foliar N and 13C isotope discrimination (Δ13C) results. The Δ13C of the soluble fraction is a good estimator of intrinsic water-use efficiency in this species. Rhododendron and mountain pine had sclerophyllous leaves, as indicated by leaf mass per area, Δ13C, PNUE and C/N results. Rhododendron was particularly affected by short periods of scarce rainfall, which negatively affected gas exchange and photochemistry, and reduced the remobilization of leaf N and P. Mountain pine was the most tolerant species since alterations of gas exchange, photochemistry and Δ13C were not observed. Its highest investment of N in RuBisCo and highest potassium, iron and magnesium leaf concentration contributed to the highest A rates observed.

Fast and Cheap in the Fall: Phylogenetic determinants of late flowering phenologies in Himalayan Rhododendron.

PREMISE OF THE STUDY: Biotic and abiotic pressures affect the beginning and end of phenologies differently, but little is known about how these temporal components may be determined by phylogeny. We tested for phylogenetic signal in the phenological components and related traits among Himalayan Rhododendron species. METHODS: We constructed a phylogeny with trnL-trnL-trnF, atpH-I, RPB2-I (3F-4R), and ITS 4-5, and examined it in combination with trait data recorded for a nine-species assemblage on Mt. Yulong, Yunnan, China. KEY RESULTS: Uniquely among phenological traits, 'last flowering day' had a significant phylogenetic signal. Last flowering day was latest in the clade with the smallest fruits. A similar association between the end of flowering and reproductive investment existed in data from Flora of China (Wu et al., 2005) for 160 Yunnan Rhododendron species, for which last flowering month was correlated with fruit size. CONCLUSIONS: Phylogenetic determinants of last flowering day and fruit size may be driven by limited time for fruit development before the onset of cold temperatures in autumn-a temporal niche that only small-fruited species can occupy. This strategy is analogous to 'fast and cheap' spring ephemerals. More may be gained from late-phenology studies, both within and among species and across seasons.

Photosynthetic limitation of several representative subalpine species in the Catalan Pyrenees in summer.

Information on the photosynthetic process and its limitations is essential in order to predict both the capacity of species to adapt to conditions associated with climate change and the likely changes in plant communities. Considering that high-mountain species are especially sensitive, three species representative of subalpine forests of the Central Catalan Pyrenees: mountain pine (Pinus uncinata Mill.), birch (Betula pendula Roth) and rhododendron (Rhododendron ferrugineum L.) were studied under conditions associated with climate change, such as low precipitation, elevated atmospheric [CO2 ] and high solar irradiation incident at Earth's surface, in order to detect any photosynthetic limitations. Short-term high [CO2 ] increased photosynthesis rates (A) and water use efficiency (WUE), especially in birch and mountain pine, whereas stomatal conductance (gs ) was not altered in either species. Birch showed photosynthesis limitation through stomatal closure related to low rainfall, which induced photoinhibition and early foliar senescence. Rhododendron was especially affected by high irradiance, showing early photosynthetic saturation in low light, highest chlorophyll content, lowest gas exchange rates and least photoprotection. Mountain pine had the highest A, photosynthetic capacity (Amax ) and light-saturated rates of net CO2 assimilation (Asat ), which were maintained under reduced precipitation. Furthermore, maximum quantum yield (Fv /Fm ), thermal energy dissipation, PRI and SIPI radiometric index, and ascorbate content indicated improved photoprotection with respect to the other two species. However, maximum velocity of carboxylation of RuBisco (Vcmax ) indicated that N availability would be the main photosynthetic limitation in this species.

Modelization of the Current and Future Habitat Suitability of Rhododendron ferrugineum Using Potential Snow Accumulation.

Mountain areas are particularly sensitive to climate change. Species distribution models predict important extinctions in these areas whose magnitude will depend on a number of different factors. Here we examine the possible impact of climate change on the Rhododendron ferrugineum (alpenrose) niche in Andorra (Pyrenees). This species currently occupies 14.6 km2 of this country and relies on the protection afforded by snow cover in winter. We used high-resolution climatic data, potential snow accumulation and a combined forecasting method to obtain the realized niche model of this species. Subsequently, we used data from the high-resolution Scampei project climate change projection for the A2, A1B and B1 scenarios to model its future realized niche model. The modelization performed well when predicting the species's distribution, which improved when we considered the potential snow accumulation, the most important variable influencing its distribution. We thus obtained a potential extent of about 70.7 km(2) or 15.1% of the country. We observed an elevation lag distribution between the current and potential distribution of the species, probably due to its slow colonization rate and the small-scale survey of seedlings. Under the three climatic scenarios, the realized niche model of the species will be reduced by 37.9-70.1 km(2) by the end of the century and it will become confined to what are today screes and rocky hillside habitats. The particular effects of climate change on seedling establishment, as well as on the species' plasticity and sensitivity in the event of a reduction of the snow cover, could worsen these predictions.

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In this study, 17 anatomical structure indexes of 15 Rhododendron cultivars were mea-sured by scanning electron microscope (SEM). Leaf anatomical structure indexes were screened via coefficient of variation, analysis of correlation and hierarchical cluster analysis, and comprehensive evaluation on heat resistance for each cultivar was conducted by the subordinate function. The results showed that the leaves of Rhododendron cultivars were typical bifacial leaf and the epidermal anticlinal walls showed slightly sinuate. The stomata only distributed in the lower epidermis and the shape was ruleless. The anatomical structure indexes all reached a significant level difference among 15 cultivars (P<0.01), except for lower epidermis thickness (P<0.05). Thickness of lamina corneum, stomatal density, stomatal width, the thickness palisade tissue and looseness of leaf spongy tissue were the main factors related to the hardness, while other indexes, such as stomatal length, stoma aperture, stomatal opening, length and thickness of upper epidermis, length and thickness of lower epidermis, thickness of spongy tissue, the ratio of the palisade tissue to spongy tissue, tightness of leaf palisade tissue, leaf thickness and media thickness didn't show much effect on heat resistance. There were some differences among 15 cultivars in heat resistance, and the order was Rhododendron 'Song Jiang Da Tao Hong' > Rhododendron 'Zhuang Yuan Hong' > Rhododendron 'Lv Se Guang Hui' > Rhododendron 'Fen Zhen Zhu' > Rhododendron 'Wai Guo Hong' > Rhododendron 'Lan Yin' > Rhododendron 'Bi Zhi' >Rhododendron 'Da He Zhi Chun' > Rhododendron 'Guo Qi Hong' > Rhododendron 'Yu Ling Long' > Rhododendron 'Hong Shan Hu' > Rhododendron 'Ning Bo Hong' > Rhododendron 'Tao Ban Zhu Sha' > Rhododendron 'Ai Ding Bao' > Rhododendron 'Liu Qiu Hong'. According to the heat hardiness, the cultivars could be divided into 4 groups: R. 'Song Jiang Da Tao Hong', R. 'Zhuang Yuan Hong' and R. 'Lv Se Guang Hui' with high heat resistance, R. 'Fen Zhen Zhu', R. 'Wai Guo Hong', R. 'Lan Yin', R. 'Bi Zhi', R. 'Da He Zhi Chun', R. 'Guo Qi Hong' and R. 'Yu Ling Long' with medium heat resistance, R. 'Hong Shan Hu', R. 'Ning Bo Hong', R. 'Tao Ban Zhu Sha' and R. 'Ai Ding Bao' with lower heat resistance, R. 'Liu Qiu Hong' without heat resistance. However, the accurate heat hardiness evaluation of Rhododendron still needs to consider other factors, including morphological structure, physiological and biochemical indicators and genetic factor of heat resistance, the harmfulness of Rhododendron, and the recovery state after being injured by high temperature.

Reproduction in Flame Azalea (Rhododendron calendulaceum, Ericaceae): A Rare Case of Insect Wing Pollination.

Although many angiosperms are serviced by flying pollinators, reports of wings as pollen vectors are rare. Flame azalea (Rhododendron calendulaceum) is visited by diverse insects, yet previous observations suggested that only butterfly wings may transfer pollen to stigmas. We used an experimental approach to determine whether butterfly wings are the primary vehicle of pollination in flame azalea. Over two seasons of observations, only butterflies (Papilio glaucus and Speyeria cybele) contacted both anthers and stigmas, yet because of differences in wing-flapping behavior, P. glaucus transferred pollen most efficiently. In contrast, bee species specialized either on pollen or nectar but did not contact both anthers and stigmas. A field experiment revealed that flowers excluding butterflies experienced almost complete fruit failure, whereas fruit set in open flowers did not differ from those that were hand pollinated. Additionally, butterflies had 56-fold more azalea pollen on their wings than bodies, while azalea stigmas bore both pollen and wing scales. These results suggest that plants with many visitors contacting reproductive organs may still specialize on a single guild of visitors for pollination and that wing-borne pollen transfer is a key mode of flame azalea pollination.