Exogenous application of IAA alleviates effects of supplemental ultraviolet‐B radiation in the medicinal plant Withania somnifera Dunal

• Supplemental (s)‐UV‐B radiation has adverse effects on the majority of plants. The present study was conducted to evaluate the effects of exogenous application of the growth hormone indole acetic acid (IAA) on various morphological, physiological and biochemical characteristics of Withania somnifera, an indigenous medicinal plant, subjected to s‐UV‐B.
• The s‐UV‐B‐treated plants received ambient + 3.6 kJm^?2·day^?1 biologically effective UV‐B, and IAA was applied at two doses (200 and 400 ppm) to s‐UV‐B‐exposed plants.
• The plant was forced to compromise its growth, development and photosynthetic patterns to survive under s‐UV‐B by increasing concentrations of secondary metabolites and antioxidants (thiol, proline, ascorbic acid, α‐tocopherol, ascorbate peroxidase, catalase, glutathione reductase, peroxidase, polyphenol oxidase, superoxide dismutase) to counteract oxidative stress. Increases in secondary metabolites were evidenced as increased activity of phenylpropanoid pathway enzymes: phenylalanine ammonia lyase, cinnamyl alcohol dehydrogenase, 4‐coumarate CoA ligase, chalcone isomerase and dihydroflavonol reductase. Application of different IAA doses reversed the detrimental effects of s‐UV‐B on W. somnifera by improving growth and photosynthesis and reducing concentrations of secondary metabolites and non‐enzymatic antioxidants. Antioxidant enzymes, however, had a synergistic effect on s‐UV‐B treatment and IAA application.
• The effects of s‐UV‐B on W. somnifera are ameliorated to varying degrees upon exogenous IAA application, and synergistic enhancement of antioxidant enzymes under s‐UV‐B+IAA treatment might be responsible for the partial recuperation of growth and plant protein content, as a UV‐B‐exposed plant is forced to allocate most of its photosynthate towards production of enzymes related to antioxidant defence.

     

Different physiological responses of cyanobacteria to ultraviolet‐B radiation under iron‐replete and iron‐deficient conditions: Implications for underestimating the negative effects of UV‐B radiation

Iron deficiency has been considered one of the main limiting factors of phytoplankton productivity in some aquatic systems including oceans and lakes. Concomitantly, solar ultraviolet‐B radiation has been shown to have both deleterious and positive impacts on phytoplankton productivity. However, how iron‐deficient cyanobacteria respond to UV‐B radiation has been largely overlooked in aquatic systems. In this study, physiological responses of four cyanobacterial strains (Microcystis and Synechococcus), which are widely distributed in freshwater or marine systems, were investigated under different UV‐B irradiances and iron conditions. The growth, photosynthetic pigment composition, photosynthetic activity, and nonphotochemical quenching of the different cyanobacterial strains were drastically altered by enhanced UV‐B radiation under iron‐deficient conditions, but were less affected under iron‐replete conditions. Intracellular reactive oxygen species (ROS) and iron content increased and decreased, respectively, with increased UV‐B radiation under iron‐deficient conditions for both Microcystis aeruginosa FACHB 912 and Synechococcus sp. WH8102. On the contrary, intracellular ROS and iron content of these two strains remained constant and increased, respectively, with increased UV‐B radiation under iron‐replete conditions. These results indicate that iron‐deficient cyanobacteria are more susceptible to enhanced UV‐B radiation. Therefore, UV‐B radiation probably plays an important role in influencing primary productivity in iron‐deficient aquatic systems, suggesting that its effects on the phytoplankton productivity may be underestimated in iron‐deficient regions around the world.     

Dimer/monomer status and in vivo function of salt‐bridge mutants of the plant UV‐B photoreceptor UVR8

UV RESISTANCE LOCUS8 (UVR8) is a photoreceptor for ultraviolet‐B (UV‐B) light that initiates photomorphogenic responses in plants. UV‐B photoreception causes rapid dissociation of dimeric UVR8 into monomers that interact with CONSTITUTIVELY PHOTOMORPHOGENIC1 (COP1) to initiate signal transduction. Experiments with purified UVR8 show that the dimer is maintained by salt‐bridge interactions between specific charged amino acids across the dimer interface. However, little is known about the importance of these charged amino acids in determining dimer/monomer status and UVR8 function in plants. Here we evaluate the use of different methods to examine dimer/monomer status of UVR8 and show that mutations of several salt‐bridge amino acids affect dimer/monomer status, interaction with COP1 and photoreceptor function of UVR8 in vivo. In particular, the salt‐bridges formed between arginine 286 and aspartates 96 and 107 are key to dimer formation. Mutation of arginine 286 to alanine impairs dimer formation, interaction with COP1 and function in vivo, whereas mutation to lysine gives a weakened dimer that is functional in vivo, indicating the importance of the positive charge of the arginine/lysine residue for dimer formation. Notably, a UVR8 mutant in which aspartates 96 and 107 are conservatively mutated to asparagine is strongly impaired in dimer formation but mediates UV‐B responses in vivo with a similar dose–response relationship to wild‐type. The UV‐B responsiveness of this mutant does not correlate with dimer formation and monomerisation, indicating that monomeric UVR8 has the potential for UV‐B photoreception, initiating signal transduction and responses in plants.     

Acute exposure to ultraviolet‐B radiation modulates sex steroid hormones and receptor expression in the skin and may contribute to the sex bias of melanoma in a fish model

Using the Xiphophorus fish melanoma model, we show a strong male bias for sunlight‐induced malignant melanoma, consistent with that seen in the human population. To examine underlying factors, we exposed adult X. couchianus fish to a single, sublethal dose of UVB and measured circulating sex steroid hormones and expression of associated hormone receptor genes over a 24‐h period. We found that a single exposure had profound effects on circulating levels of steroid hormones with significant decreases for all free sex steroids at 6 and 24 h and increases in conjugated 2‐estradiol and 11‐ketotestosterone at 6 and 24 h, respectively. Whereas ARα expression increased in male and female skin, neither ARβ nor either of the ERs showed significant responses to UVB in either sex. The rapid response of male androgens and their receptors in the skin after UVB irradiation implicates hormones in the male bias of skin cancer and suggests that the photoendocrine response immediately after UV exposure may be relevant to melanomagenesis.     

ATR and MKP1 play distinct roles in response to UV‐B stress in Arabidopsis

Ultraviolet‐B (UV‐B) stress activates MAP kinases (MAPKs) MPK3 and MPK6 in Arabidopsis. MAPK activity must be tightly controlled in order to ensure an appropriate cellular outcome. MAPK phosphatases (MKPs) effectively control MAPKs by dephosphorylation of phosphothreonine and phosphotyrosine in their activation loops. Arabidopsis MKP1 is an important regulator of MPK3 and MPK6, and mkp1 knockout mutants are hypersensitive to UV‐B stress, which is associated with reduced inactivation of MPK3 and MPK6. Here, we demonstrate that MPK3 and MPK6 are hyperactivated in response to UV‐B in plants that are deficient in photorepair, suggesting that UV‐damaged DNA is a trigger of MAPK signaling. This is not due to a block in replication, as, in contrast to atr, the mkp1 mutant is not hypersensitive to the replication‐inhibiting drug hydroxyurea, hydroxyurea does not activate MPK3 and MPK6, and atr is not impaired in MPK3 and MPK6 activation in response to UV‐B. We further show that mkp1 leaves and roots are UV‐B hypersensitive, whereas atr is mainly affected at the root level. Tolerance to UV‐B stress has been previously associated with stem cell removal and CYCB1;1 accumulation. Although UV‐B‐induced stem cell death and CYCB1;1 expression are not altered in mkp1 roots, CYCB1;1 expression is reduced in mkp1 leaves. We conclude that the MKP1 and ATR pathways operate in parallel, with primary roles for ATR in roots and MKP1 in leaves.     

Interactive direct and plant‐mediated effects of elevated atmospheric [CO2] and temperature on a eucalypt‐feeding insect herbivore

Understanding the direct and indirect effects of elevated [CO₂] and temperature on insect herbivores and how these factors interact are essential to predict ecosystem‐level responses to climate change scenarios. In three concurrent glasshouse experiments, we measured both the individual and interactive effects of elevated [CO₂] and temperature on foliar quality. We also assessed the interactions between their direct and plant‐mediated effects on the development of an insect herbivore of eucalypts. Eucalyptus tereticornis saplings were grown at ambient or elevated [CO₂] (400 and 650 μmol mol^?1 respectively) and ambient or elevated ( + 4 °C) temperature for 10 months. Doratifera quadriguttata (Lepidoptera: Limacodidae) larvae were feeding directly on these trees, on their excised leaves in a separate glasshouse, or on excised field‐grown leaves within the temperature and [CO₂] controlled glasshouse. To allow insect gender to be determined and to ensure that any sex‐specific developmental differences could be distinguished from treatment effects, insect development time and consumption were measured from egg hatch to pupation. No direct [CO₂] effects on insects were observed. Elevated temperature accelerated larval development, but did not affect leaf consumption. Elevated [CO₂] and temperature independently reduced foliar quality, slowing larval development and increasing consumption. Simultaneously increasing both [CO₂] and temperature reduced these shifts in foliar quality, and negative effects on larval performance were subsequently ameliorated. Negative nutritional effects of elevated [CO₂] and temperature were also independently outweighed by the direct positive effect of elevated temperature on larvae. Rising [CO₂] and temperature are thus predicted to have interactive effects on foliar quality that affect eucalypt‐feeding insects. However, the ecological consequences of these interactions will depend on the magnitude of concurrent temperature rise and its direct effects on insect physiology and feeding behaviour.     

Effects of the Epichloë fungal endophyte symbiosis with Schedonorus pratensis on host grass invasiveness

Initial studies of grass–endophyte mutualisms using Schedonorus arundinaceus cultivar Kentucky‐31 infected with the vertically transmitted endophyte Epichloë coenophiala found strong, positive endophyte effects on host‐grass invasion success. However, more recent work using different cultivars of S. arundinaceus has cast doubt on the ubiquity of this effect, at least as it pertains to S. arundinaceus–E. coenophiala. We investigated the generality of previous work on vertically transmitted Epichloë‐associated grass invasiveness by studying a pair of very closely related species: S. pratensis and E. uncinata. Seven cultivars of S. pratensis and two cultivars of S. arundinaceus that were developed with high‐ or low‐endophyte infection rate were broadcast seeded into 2 × 2‐m plots in a tilled, old‐field grassland community in a completely randomized block design. Schedonorus abundance, endophyte infection rate, and co‐occurring vegetation were sampled 3, 4, 5, and 6 years after establishment, and the aboveground invertebrate community was sampled in S. pratensis plots 3 and 4 years after establishment. Endophyte infection did not enable the host grass to achieve high abundance in the plant community. Contrary to expectations, high‐endophyte S. pratensis increased plant richness relative to low‐endophyte cultivars. However, as expected, high‐endophyte S. pratensis marginally decreased invertebrate taxon richness. Endophyte effects on vegetation and invertebrate community composition were inconsistent among cultivars and were weaker than temporal effects. The effect of the grass–Epichloë symbiosis on diversity is not generalizable, but rather specific to species, cultivar, infection, and potentially site. Examining grass–endophyte systems using multiple cultivars and species replicated among sites will be important to determine the range of conditions in which endophyte associations benefit host grass performance and have subsequent effects on co‐occurring biotic communities.     

Integration and scaling of UV‐B radiation effects on plants: from molecular interactions to whole plant responses

A process based model integrating the effects of UV‐B radiation to molecular level processes and their consequences to whole plant growth and development was developed from key parameters in the published literature. Model simulations showed that UV‐B radiation induced changes in plant metabolic and/or photosynthesis rates can result in plant growth inhibitions. The costs of effective epidermal UV‐B radiation absorptive compounds did not result in any significant changes in plant growth, but any associated metabolic costs effectively reduced the potential plant biomass. The model showed significant interactions between UV‐B radiation effects and temperature and any factor leading to inhibition of photosynthetic production or plant growth during the midday, but the effects were not cumulative for all factors. Vegetative growth were significantly delayed in species that do not exhibit reproductive cycles during a growing season, but vegetative growth and reproductive yield in species completing their life cycle in one growing season did not appear to be delayed more than 2–5 days, probably within the natural variability of the life cycles for many species. This is the first model to integrate the effects of increased UV‐B radiation through molecular level processes and their consequences to whole plant growth and development.     

Effects of prohydrojasmon‐treated corn plants on attractiveness to parasitoids and the performance of their hosts

We investigated the effect of prohydrojasmon [propyl (1RS,2RS)‐(3‐oxo‐ 2‐pentylcyclopentyl) acetate] (PDJ) treatment of intact corn plants, on their attractiveness to the specialist endoparasitoid, Cotesia kariyai Watanabe (Hymenoptera: Braconidae), and on the performance of the common armyworm, Mythimna separata (Walker) (Lepidoptera: Noctuidae) under laboratory conditions. Attractiveness of C. kariyai to PDJ‐treated plants was studied in a wind tunnel, whereas performance of M. separata larvae was tested in plastic cages. The attractiveness of the treated plants increased with concentrations of PDJ increasing to 2 mm , which was equivalent to the attractiveness of host‐infested plants. PDJ‐treated corn plants emitted 16 volatile compounds (α‐pinene, β‐myrcene, (Z)‐3‐hexenyl acetate, limonene, (E)‐β‐ocimene, linalool, (E)‐4,8‐dimethyl‐1,3,7‐nonatriene, (+)‐cyclosativene, ylangene, (E)‐β‐farnesene, (E, E)‐4,8,12‐trimethyl‐1,3,7,11‐tridecatetraene, α‐bergamotene, γ‐cadinene, δ‐cadinene, α‐muulolene and nerolidol), most of which were observed in the headspace of host‐infested corn plants with some quantitative and qualitative differences. We also tested the effects of PDJ treatment on the performance of M. separata larvae. The survival rates of the larval and pupal stages were significantly lower at 2 mm level of PDJ. A significant decrease in weight at 6th stadium larvae was observed only at 2 mm level of PDJ. In contrast, PDJ treatment at all PDJ concentration levels caused significant reduction in weight of pupal stage as compared to control. These data suggested that PDJ, originally developed as a plant growth regulator, especially to induce coloring of fruits, has the potential to induce direct and indirect defenses in corn plants against common armyworm, M. separata.     

Qualitative HPLC‐DAD/ESI‐TOF‐MS Analysis,Cytotoxic, and Apoptotic Effects of Croatian Endemic Centaurea ragusina L. Aqueous Extracts

Centaurea ragusina L., an endemic Croatian plant species, revealed a good cytotoxic activity of aqueous extracts (AE) on human bladder (T24) and human glioblastoma (A1235) cancer cell lines. The chemical constituents were tentatively identified using high performance liquid chromatography HPLC‐DAD/ESI‐TOF‐MS in negative ionization mode. The main compounds of herba extract were sesquiterpene lactones: solstitialin A 3,13‐diacetate and epoxyrepdiolide; organic acid: quinic acid. The main compounds of flower extract were organic acids: quinic acid, citric acid, and malic acid; sesquiterpene lactone: cynaropicrin; phenolic compounds: chlorogenic acid and phenylpropanoid: syringin. The AE of C. ragusina were investigated for correlation of their effects on human bladder (T24) and human glioblastoma (A1235) cancer cell lines using the MTT assay. Although both extracts showed significant dose‐ and time‐dependent cytotoxic activity against both cancer cell lines, the flower extract exhibited slightly higher activity. In order to determine type of cell death induced by treatment, cell lines were exposed subsequently to a treatment with both flower and herba AE. The majority of the cells died by induced apoptosis treatment. Flower AE (26.25%), compared to a leaf AE (22.15%) showed slightly higher percentage of an apoptosis in T24 cells, when compared to a non‐treated cells (0.04%).     

Host plant preference and offspring performance of a leaf‐mining moth,Caloptilia fraxinella,on two Fraxinus species

The preference‐performance or ‘mother‐knows‐best’ hypothesis states that female insects choose to oviposit on a host plant that increases the performance of their offspring. This positive link between host plant choice and larval performance is especially important for leaf miners with non‐motile larvae that are entirely dependent upon the oviposition choice of the female for host plant location. Preference and performance of the ash leaf coneroller, Caloptilia fraxinella (Ely) (Lepidoptera: Gracillariidae), a specialist on ash trees, Fraxinus spp. (Oleaceae), were tested in a series of laboratory and field experiments. Female C. fraxinella were exposed to two closely related hosts, black ash, Fraxinus nigra Marshall, and green ash, Fraxinus pennsylvanica Marshall var. subintegerrima (Vahl), in oviposition choice and wind tunnel flight experiments to determine which host is most attractive for oviposition. Caloptilia fraxinella females were inconsistent in host choice, yet performance of larvae was greater on green than black ash. In preference studies, C. fraxinella preferred to oviposit on black ash when leaflets were removed from the tree, but preferred intact green ash over black ash seedlings for oviposition and host location in a wind tunnel. In the field, however, more C. fraxinella visited black ash var. ‘Fallgold’ at leaf flush than green ash at the same sites. Age of the ash leaflet also influences oviposition in this leaf miner and females preferred new over old leaflets for oviposition. Performance of C. fraxinella larvae was evaluated in field and laboratory experiments and was greater on green ash than on black ash in both experiments based on larval survival and development time parameters. The stronger oviposition and host location preference in the field for black ash were not linked to enhanced performance of offspring, as green ash was the superior host, supporting higher larval survival and faster development. A stronger host location preference in the wind tunnel for green ash over black ash, however, suggests that under certain circumstances with this moth species, ‘mother (may) know best’.     

Regulation of polyamine metabolism in Pyropia cinnamomea (W.A. Nelson), an important mechanism for reducing UV‐B‐induced oxidative damage

It is generally accepted that ultraviolet (UV) radiation can have adverse affects on phototrophic organisms, independent of ozone depletion. The red intertidal seaweed Pyropia cinnamomea W.A. Nelson (previously Porphyra cinnamomea Sutherland et al. 2011), similar to many other intertidal macrophytes, is exposed to high levels of UV radiation on a daily basis due to emersion in the upper littoral zone. It has been shown that seaweeds, like higher plants, respond to an increased activity of antioxidative enzymes when exposed to stress. However, earlier investigations have shown that P. cinnamomea also compensates for stress due to UV radiation by increasing polyamine (PA) levels, especially bound‐soluble and bound‐insoluble PAs. The PA precursor putrescine (PUT) can be synthesized via two enzymatic pathways: arginine decarboxylase (ADC) and ornithine decarboxylase (ODC). Both of these enzymes showed increased activity in P. cinnamomea under UV stress. In higher plants, ADC is the enzyme responsible for increased PA levels during stress exposure, while ODC is correlated with cell division and reproduction. However, there are contrary findings in the literature. Using two irreversible inhibitors, we identified the enzyme most likely responsible for increased PUT synthesis and therefore increased stress tolerance in P. cinnamomea. Our results show that changes in the PA synthesis pathway in P. cinnamomea under UV stress are based on an increased activity of ADC. When either inhibitor was added, lipid hydroperoxide levels increased even under photosynthetically active radiation, suggesting that PAs are involved in protection mechanisms under normal light conditions as well. We also show that under optimum or low‐stress conditions, ODC activity is correlated with PUT synthesis.     

Time‐since‐fire and climate interact to affect the structural recovery of an Australian semi‐arid plant community

How does time‐since‐fire influence the structural recovery of semi‐arid, eucalypt‐dominated Murray‐Mallee shrublands after fire, and is recovery affected by spatial variation in climate? We assessed the structure and dynamics of a hummock grass, Triodia scariosa N.T. Burb, and mallee eucalypts – two key structural components of mallee shrublands – using a >100 year time‐since‐fire chronosequence. The relative influence of climatic variables, both individually and combined with time‐since‐fire, was modelled to account for spatial variation in the recovery of vegetation structural components. Time‐since‐fire was the primary determinant of the structural recovery of T. scariosa and eucalypts. However, climate, notably mean annual rainfall and rainfall variability, also influenced the recovery of the eucalypt overstorey, T. scariosa cover and mean hummock height. We observed that (i) the mean number of live eucalypt stems per individual decreased while mean individual basal area increased, (ii) cover of T. scariosa peaked at ~30 years post‐fire and gradually decreased thereafter, and (iii) the ‘hummock’ form of T. scariosa occurred throughout the chronosequence, whereas the ‘ring’ form tended not to occur until ~30 years post‐fire. Time‐since‐fire was the key determinant of the structural recovery of eucalypt‐dominated mallee shrublands, but there is geographical variation in recovery related to rainfall and its variability. Fire regimes are likely to have different effects across the geographic range of mallee shrublands.     

Isoalantolactone restores the sensitivity of gram‐negative Enterobacteriaceae carrying MCR‐1 to carbapenems

Polymyxin B has been re‐applied to the clinic as the final choice for the treatment of multidrug‐resistant gram‐negative pathogenic infections, but the use of polymyxin B has been re‐assessed because of the emergence and spread of the plasmid‐mediated mcr‐1 gene. The purpose of this study was to search for an MCR inhibitor synergistically acting with polymyxin to treat the infection caused by this pathogen. In this study, we used the broth microdilution checkerboard method to evaluate the synergistic effect of isoalantolactone (IAL) and polymyxin B on mcr‐1‐positive Enterobacteriaceae. Growth curve analysis, time‐killing assays and a combined disc test were used to further verify the efficacy of the combined drug. Colonization of the thigh muscle in mice, survival experiments and lung tissue section observations was used to determine the effect of synergy in vivo after Klebsiella pneumoniae and Escherichia coli infection. We screened a natural compound, IAL, which can enhance the sensitivity of polymyxin B to mcr‐1‐positive Enterobacteriaceae. The results showed that the combined use of polymyxin B and IAL has a synergistic effect on mcr‐1‐positive Enterobacteriaceae, such as K pneumoniae and E coli, not only in vitro but also in vivo. Our results indicate that IAL is a natural compound with broad application prospects that can prolong the service life of polymyxin B and make outstanding contributions to the treatment of gram‐negative Enterobacteriaceae infections resistant to polymyxin B.     

Effect of UV‐C Radiation on Resistance of Romaine Lettuce (Lactuca sativa L.) Against Botrytis cinerea and Sclerotinia minor

The aim of this research was to examine the effect of UV‐C on resistance of lettuce to Botrytis cinerea and Sclerotinia minor. Analysis of the lesion surfaces showed that plants exposed to UV‐C were less susceptible to the two pathogens, especially on the fourth day after inoculation. Chlorophyll, carotenoid contents and malondialdehyde and hydrogen peroxide were assayed after 1 day and 4 days. Lettuces treated with UV‐C and inoculated showed an increase in chlorophyll and carotenoid content, especially 24 h after inoculation, and low values of the two indicators of oxidative stress as compared with lettuces which were inoculated but did not receive UV‐C treatment.     

Effect of UV‐B radiation on the content of UV‐B absorbing compounds and photosynthetic parameters in Parmotrema austrosinense from two contrasting habitats

• We studied the resistance of Parmotrema austrosinense to UV‐B stress. We focused on the effects of a high dose UV‐B radiation on the content of chlorophylls, carotenoids and UV‐B screening compounds.
• Photosynthetic parameters were measured by chlorophyll fluorescence (potential and effective quantum yields, photochemical and non‐photochemical quenching) and evaluated in control and UV‐B‐treated lichens. Lichens from two different locations in Cordoba, Argentina, were selected: (i) high altitude and dry plots at (Los Gigantes) and (ii) lowland high salinity plots (Salinas Grandes).
• UV‐B treatment led to a decrease in the content of photosynthetic pigments and UV‐B screens (absorbance decrease in 220–350 nm) in the samples from Salinas Grandes, while in Los Gigantes samples, an increase in UV‐B screen content was observed. Chlorophyll fluorescence parameters showed a UV‐B‐induced decline in F_V/F_M, Φ_PSII and qP indicating limitation of primary photosynthetic processes in photosystem II (PSII) of symbiotic alga, more pronounced in Salinas Grandes samples. Protective mechanism of PSII were activated by the UV‐B treatment to a higher extent in samples from Salinas Grandes (NPQ 0.48) than in Los Gigantes samples (NPQ 0.26).
• We concluded that site‐related characteristics, and in particular different UV‐B radiation regimen, had a strong effect on resistance of the photosynthetic apparatus of P. austrosinense to UV‐B radiation.