Origins of non-linear and dissimilar relationships between epidermal UV absorbance and UV absorbance of extracted phenolics in leaves of grapevine and barley

A recent review of climate patterns in Southern Germany has suggested significant increases in ultraviolet (UV) radiation due to decreases in cloud coverage and in cloud frequency which compound the effects of stratospheric ozone depletion. Whether such UV radiation increases result in UV damage of higher plant leaves depends partly on the capacity of UV-absorbing hydroxycinnamic acids and flavonoids located in the plant epidermis to screen out UV radiation. Epidermal UV screening is most often assessed from UV absorbance of whole-leaf extracts but in the present work, this method is critically examined. In grapevine (Vitis vinifera L.), hydroxycinnamic acid as well as mono-hydroxylated and ortho-dihydroxylated flavonoid concentrations increased in parallel with fluorometrically detected adaxial epidermal UV absorbance but only the latter class of flavonoids was associated with epidermal UV absorbance in barley (Hordeum vulgare L). For both species, curvilinear relationships between epidermal and total phenolic UV absorbance were established: initial slopes of the curves differed markedly between species. Modelling suggested that curvilinearity arises from UV-transparent epidermal areas located between vacuoles which are particularly UV-absorbing due to high levels of phenolics. The species-dependent differences were related to allocation of high amounts of phenolics in the mesophyll and abaxial epidermis in barley but not in grapevine. Both factors, optical heterogeneity and variable distribution of phenolics, severely restrict the use of phenolic absorbance to estimate true epidermal screening.     

UV-absorbing compounds in <Emphasis Type="Italic">Porphyra haitanensis</Emphasis> (Rhodophyta) with special reference to effects of desiccation

The intertidal red alga Porphyra haitanensis Chang et Zheng is episodically desiccated and exposed to high levels of solar radiation at low tide during emersion. However, little has been documented on the relationship between the stresses during desiccation and related chemical compounds. We found that P. haitanensis thalli, when desiccated under indoor (artificial radiation) or outdoor (solar radiation) conditions, with or without UV radiation (UVR: 280–400 nm), contained significantly higher concentrations of UV-absorbing compounds (peak at 336 nm) than those maintained submerged (without desiccation). Solar UVR had no effect on the content of UV-absorbing compounds. Even though the concentration of these compounds decreased with time in all treatments, a slower decrease was observed in the desiccated samples. The samples with higher levels of UV-absorbing compounds showed higher photochemical efficiency of photosystem II (PS II) during the exposure or subsequent recovering process than samples with low concentration of UV-absorbing compounds, reflecting their protective role. The concentration of these compounds varied in different parts of the thallus, with the middle and marginal parts containing 60–80% more UV-absorbing compounds than the basal parts in both female and male plants. In addition, the marginal parts of male thalli contained more UV-absorbing compounds than the corresponding parts of female thalli. Our data suggest that desiccation plays a key role in this alga to maintain high concentration of UV-absorbing compounds, and that this might provide a beneficial advantage to compete in the intertidal zone where the organism is normally exposed to high levels of UVR.     

Flavonoids in translucent bracts of the Himalayan<Emphasis Type="Italic"> Rheum nobile</Emphasis> (Polygonaceae) as ultraviolet shields

UV-absorbing substances were isolated from the translucent bracts of Rheum nobile, which grows in the alpine zone of the eastern Himalayas. Nine kinds of the UV-absorbing substances were found by high performance liquid chromatography (HPLC) and paper chromatography (PC) surveys. All of the five major compounds are flavonoids, and were identified as quercetin 3-O-glucoside, quercetin 3-O-galactoside, quercetin 3-O-rutinoside, quercetin 3-O-arabinoside and quercetin 3-O-[6-(3-hydroxy-3-methylglutaroyl)-glucoside] by UV, ¹H and ¹³C NMR, mass spectra, and acid hydrolysis of the original glycosides, and direct PC and HPLC comparisons with authentic specimens. The four minor compounds were characterised as quercetin itself, quercetin 7-O-glycoside, kaempferol glycoside and feruloyl ester. Of those compounds, quercetin 3-O-[6-(3-hydroxy-3-methylglutaroyl)-glucoside] was found in nature for the first time. The translucent bracts of R. nobile accumulate a substantial quantity of flavonoids (3.3–5 mg per g dry material for the major compounds). Moreover, it was clarified by quantitative HPLC survey that much more of the UV-absorbing substances is present in the bracts than in rosulate leaves. Although the flavonoid compounds have been presumed to be the important UV shields in higher plants, there has been little characterisation of these compounds. In this paper, the UV-absorbing substances of the Himalayan R. nobile were characterised as flavonol glycosides based on quercetin.     

Photomorphogenic responses to UV radiation IV: a comparative study of UVB effects on growth and pigment accumulation in etiolated and de-etiolated hypocotyls of wild-type and aurea mutant of tomato Lycopersicon esculentum Mill)

ABSTRACT

The question of how de-etiolation of tomato seedling under continuous monochromatic yellow light exerts an influence on UV radiation-induced responses has been studied. Hypocotyl extension and the accumulation of anthocyanins and UV-absorbing compounds was compared in the aurea mutant of tomato and its isogenic wild type. The data of the present paper provide evidence that, during de-etiolation of tomato seedlings, yellow light exerts its effects over seedlings responsiveness to subsequent UV irradiation through several mechanisms: 1) a significant enhancement of shorter UVB wavelength efficiency in both the genotypes; 2) the abolition of UVA -blue light-induced accumulation of UV-absorbing compounds that does not involve pnyA; 3) the disappearance of UVA-blue light-induced hypocotyl growth inhibition that does not involve phyA; 4) higher anthocyanin accumulation rate in response to UV radiation mediated by phyA. Yellow light-induced growth inhibition and accumulation of UV-absorbing compound both mediated by phyA and present only in wild type tomato, appear to be completely separate from the action of UV radiation on the same responses.     

Habitat‐Dependent Variations in Berberine Content of Berberis asiatica Roxb. ex. DC. in Kumaon,Western Himalaya

The variation of the berberine content in roots and stem bark of Berberis asiatica with altitude and edaphic conditions in the western Himalaya was estimated by HPLC. The comparative assessment revealed a significantly higher berberine content in roots than in stem barks. Moreover, the berberine content varied significantly with altitude and edaphic conditions both in root and stem bark samples. The populations growing at low altitude contained significantly more berberine than the ones growing at high altitude. Also the moisture and potassium (K) percentage of the soil significantly influenced the berberine content.     

Accumulation of UV-absorbing flavonoids induced by UV-B radiation inAmbidopsis thaliana L.

Irradiation ofArabidopsis with ultraviolet (UV) light resulted in intensity- and wavelength-dependent increases in the levels of a small family of UV-absorbing flavonoids, which accumulate in the aerial parts of the plants. A gradient of sensitivity to UV-B radiation is described in the different leaves of developingArabidopsis plants whereby the earliest formed leaves become damaged by UV-B faster and more extensively than later formed leaves. This UV-sensitivity gradient tightly parallels differences in constitutive as well as UV-induced levels of flavonoid accumulation among the various leaves, suggesting a direct role of flavonoids in protection against damage by UV radiation. The level of accumulated flavonoids, both constitutive and UV-induced, in each leaf appear to be dependent on the specific developmental state of each leaf as well as the overall developmental state of the plant. The UV-mediated flavonoid response, along with the observed UV-induced damage, appear not to be systemic inArabidopsis but restricted very closely to the irradiated areas of leaves.I am deeply indebted to Robert Fischer and Bob Buchanan for providing access to their laboratories and for their invaluable help without which this work could not have been realized. I especially wish to thank Lola Peñarrubia, Elena del Campillo, Patrick Neil and Julie Montgomery for innumerable and fruitful discussions. This work was supported by Cooperative State Research Service, U.S. Department of Agriculture, under Agreements Nos. 90-37280-5664 and 90-372780-5808.     

Altitudinal variation of secondary metabolites in flowering heads of the Asteraceae: trends and causes

Plants in alpine habitats are exposed to severe environmental stresses including temperature and radiation extremes. The observation that flowering heads from high altitude populations of Scorzoneroides helvetica (Mérat) J.Holub (synonym: Leontodon helveticus Mérat) contained higher amounts of luteolin derivatives than conspecific populations in lower altitudes prompted further investigations. An elevational increase of phenolics was confirmed in flowering heads of neophytic populations of three additional taxa of the Cichorieae tribe in New Zealand. A solely genetic basis of the altitudinal trends of flavonoids and phenolic acids was eliminated by field experiments with cultivars of the medicinal herbs Arnica montana L. and Matricaria chamomilla L. planted at nine different altitudes ranging from 600 to 2,200 m in the Tyrolean Alps. Parallel experiments on potted plants excluded soil characteristics as the factor causing the observed variation. The initial hypotheses that enhanced UV-B radiation in higher altitudes was triggering an increase in the ratio of B-ring-ortho-diphenolic versus B-ring-monophenolic flavonols in flowering heads of Arnica was disproved by climate chamber experiments resulting in no significant difference between plants grown in ambient and threefold ambient UV-B radiation regimes. In contrast, an increase of this ratio similar to the changes observed in higher altitudes resulted from a decrease in temperature by 5°C in a second climate chamber experiment. Conclusively, enhanced UV-B radiation is probably not the key factor inducing shifts in the phenolic composition in Asteraceae growing at higher altitudes but it is rather the temperature which decreases with altitude.     

Short communication. Do anthocyanins play a role in UV protection of the red juvenile leaves of Syzygium?

The biological function of juvenile leaves pigmented with anthocyanin is poorly understood. The role anthocyanins play in UV protection was assessed in juvenile leaves of two Syzygium species (S. luehmannii and S. wilsonii) which contain high anthocyanin concentrations. HPLC was used to separate UV-absorbing anthocyanins from other soluble UV-absorbing phenolic compounds. The isolated anthocyanins (predominantly malvidin-3,5-diglucoside) contributed little to the total absorbance of UV-A and UV-8 radiation. This was because the non-acylated anthocyanins only effectively absorbed shortwave UV-B radiation and the strong absorbance by other compounds. These results suggest that the UV protection hypothesis is not valid for anthocyanins in juvenile Syzygium leaves.     

Genomic and phenotypic differentiation of Arabidopsis thaliana along altitudinal gradients in the North Italian Alps

Altitudinal gradients in mountain regions are short‐range clines of different environmental parameters such as temperature or radiation. We investigated genomic and phenotypic signatures of adaptation to such gradients in five Arabidopsis thaliana populations from the North Italian Alps that originated from 580 to 2350 m altitude by resequencing pools of 19–29 individuals from each population. The sample includes two pairs of low‐ and high‐altitude populations from two different valleys. High‐altitude populations showed a lower nucleotide diversity and negative Tajima's D values and were more closely related to each other than to low‐altitude populations from the same valley. Despite their close geographic proximity, demographic analysis revealed that low‐ and high‐altitude populations split between 260 000 and 15 000 years before present. Single nucleotide polymorphisms whose allele frequencies were highly differentiated between low‐ and high‐altitude populations identified genomic regions of up to 50 kb length where patterns of genetic diversity are consistent with signatures of local selective sweeps. These regions harbour multiple genes involved in stress response. Variation among populations in two putative adaptive phenotypic traits, frost tolerance and response to light/UV stress was not correlated with altitude. Taken together, the spatial distribution of genetic diversity reflects a potentially adaptive differentiation between low‐ and high‐altitude populations, whereas the phenotypic differentiation in the two traits investigated does not. It may resemble an interaction between adaptation to the local microhabitat and demographic history influenced by historical glaciation cycles, recent seed dispersal and genetic drift in local populations.     

Differences in growth and physiological traits of two poplars originating from different altitudes as affected by UV‐B radiation and nutrient availability

Cuttings of Populus kangdingensis and Populus cathayana originating from altitudes of 3500 and 1500 m in southwestern China, respectively, were grown for one growing season in the field under ambient or ambient plus supplemental ultraviolet‐B (UV‐B) radiation with two levels of nutrients. In both species, enhanced UV‐B radiation significantly increased UV‐B absorbing compounds and guaiacol peroxidase (GPX) activity, while no significant effects were observed in photosynthetic pigments and proline content. On the other hand, cuttings grown with high‐nutrient availability had larger leaf area, higher total biomass and GPX activity as well as higher water use efficiency (WUE) (as measured by stable carbon isotope composition, δ¹³C) when compared with low‐nutrient conditions, while UV‐B absorbing compounds and ascorbic acid (AsA) content significantly decreased. Differences in responses to enhanced UV‐B radiation and nutrient availability were observed between the two species. Nutrient‐induced increases in chlorophyll a, chlorophyll b and total chlorophyll as well as in carotenoids were greater in P. kangdingensis than in P. cathayana. In P. cathayana, enhanced UV‐B radiation significantly decreased leaf area and total biomass, while it significantly increased WUE and ascorbate peroxidase (APX). In contrast, such changes were not observed in P. kangdingensis. In addition, the effects of enhanced UV‐B radiation on leaf area, total biomass and UV‐B absorbing compounds were closely related to the nutrient status. Our results indicated that P. kangdingensis, which originates from the altitude of 3500 m and is apparently adapted to low‐nutrient and high‐UV‐B habitats, exhibits better tolerance to enhanced UV‐B radiation and greater growth under low‐nutrient availability than does P. cathayana originating from the altitude of 1500 m.     

转CiCHS基因拟南芥的黄酮代谢及抗氧化能力分析

该研究克隆了中间锦鸡儿的查尔酮合成酶基因(CiCHS)并转入野生型拟南芥和tt4突变体,用qRT-PCR检测了转基因拟南芥中内源AtCHS基因的表达量,用分光光度法分析了转基因拟南芥的总黄酮、丙二醛含量及DPPH自由基清除能力,用HPLC法检测了转基因拟南芥的柚皮苷含量。结果显示:(1)转基因拟南芥中,内源AtCHS基因的表达量约为野生型的十分之一,总黄酮含量明显高于野生型;HPLC测得转基因株系中柚皮苷含量高于野生型;紫外照射处理前后转基因拟南芥中丙二醛积累量明显少于野生型。(2)转基因株系提取物对DPPH自由基清除能力显著高于野生型。(3)CiCHS基因互补拟南芥tt4突变体,转基因株系的种皮呈现浅棕色。研究表明,中间锦鸡儿CiCHS基因异源表达后生成了柚皮苷,使转基因植物的抗氧化性增强,部分恢复了tt4突变体的种皮颜色。     

Protection from uv radiation in the economic crop, <Emphasis Type="Italic">Opuntia</Emphasis> SPP.

Cacti of the genus puntia are an economically important crop. Understanding the mechanisms they possess to protect against UV radiation is important for assessing their possible response to climatic change. Measurements of the concentrations of UV-screening compounds and epidermal transmittance for two species of platyopuntia, Opuntia engelmannii Salm-Dyck. and O. phaeacantha Engelm. during 1998 and 1999 were used to investigate the UV-protection afforded by the cactus epidermis. A UV-radiative transfer model was used to investigate the interception of UV radiation on differently oriented surfaces. We show that vertical morphology itself confers significant protection against UV radiation compared to a horizontal surface. Concentrations of UV-screening flavonoids were found to vary depending on the UV exposure of different surfaces. West-facing surfaces had lower concentrations than east-facing surfaces, although theoretically they should be identical. This might be explained by the higher mean temperatures on west-facing surfaces. Although UV-absorbing soluble flavonoids in the epidermis block both UV-B and UV-A, the structure of the epidermis alone may be sufficient to remove up to 94% of the UV-B portion of the spectrum. These data yield insights into possible mechanisms of recent declines in cacti populations.     

The effects of salicylic acid on pigment contents in ultraviolet radiation stressed pepper plants

Pepper (Capsicum annuum L.) plants were sprayed with salicylic acid (SA) and treated with ultraviolet radiation UV-A (320–390 nm), UV-B (312 nm), and UV-C (254 nm) of 6.1, 5.8, and 5.7 W m⁻², respectively. UV significantly reduced contents of chlorophyll (Chl) a and b, and carotenoids (Car). SA treatment moderated Chl and Car reduction in plants treated with UV-B and UV-C. The quantity of antocyanins, flavonoids, rutin, and UV-absorbing compounds in plants that were treated with UV-B, UV-C, and SA were significantly increased. Foliar spray of SA counteracted the UV effects on pepper.     

Developmental responses to UV-B radiation in common frog <Emphasis Type="Italic">Rana temporaria</Emphasis> embryos from along an altitudinal gradient

Solar ultraviolet radiation (UV) is harmful for developing amphibians. As UV increases with altitude and latitude, it is suggested that high altitude and latitude populations have evolved tolerance to high levels of UV. Using laboratory experiments, we tested the hypothesis that Rana temporaria populations from several altitudes (438–2,450 m above sea level) were adapted to UV by assessing the effects of artificial UV on embryos mortality rate, malformations, and body length at hatching. We also tested the protective role of the jelly surrounding the embryos. Without artificial UV exposure, hatching success decreased with altitude of the population. Malformation rates were low for all populations (mean 1.36%), and hatching size increased with altitude. The artificial UV (UV-B, UV-A, and visible) used was similar to the solar spectrum received at high altitude. Exposed embryos had performance similar to that of embryos without exposure: a decreased hatching success with altitude and a low malformation rate (mean 0.85%). However, hatching size did not vary with altitude, and UV-exposed embryos tended to be smaller at hatching than non-exposed embryos. Removal of the protective jelly envelope greatly decreased the performance of UV-exposed embryos: hatching success strongly decreased with altitude and embryos of the highest population (2,450 m asl) did not develop. Malformation reached 4.98%, without population differences, and hatching size of embryos without jelly was smaller than hatching size of non-manipulated embryos with no population effect. This study demonstrates that lowland jellyless embryos were less sensitive (i.e., exhibited a higher survival rate) than highland embryos and, on the other hand, that the jelly envelope was a more efficient protection for embryos in highland than in lowland populations. A trade-off hypothesis is presented to explain this difference in UV tolerance and protection among populations: in a harsh, highland environment (i.e., with a curtailed activity period), embryos have to invest in development and growth and, thus, embryo protection should be a female investment (via the jelly). In a lowland environment, embryonic growth and development are less constrained and embryos are able to invest and use protection or repair mechanisms inherited from their parents.     

Effect of irradiation on the protein profile,protein content,and quality of agar from <Emphasis Type="Italic">Gracilaria asiatica</Emphasis> Zhang et Xia (Rhodophyta)

There are about 15 species of Gracilaria reported in Vietnam. Of these, Gracilaria asiatica Zhang et Xia is being cultivated on a large scale in Northern Vietnam, which has a subtropical climate. During the rainy season, from May to October, the growth of G. asiatica is drastically reduced or even ceases due to very low salinity and high temperature. Therefore, it is important to improve the tolerance of G. asiatica to a wide range of salinity and temperatures. This paper presents the results of research on strain improvement of G. asiatica using irradiation and selection media. Three irradiation doses of 20, 60, and 100 krad were tested against the control (with no irradiation). Afterward, the seaweed biomass was cultivated on a selected medium, ESS-1, containing NaCl in concentrations of 23‰ (C1) and 0‰ (C2). The results showed that a higher survival rate of G. asiatica was observed with the 20- and 60-krad doses. The protein content and composition of selected seaweeds were analyzed and compared with the control. SDS-PAGE showed no remarkable difference in the protein composition between the control and irradiated samples. However, the 67-kDa protein band of seaweed treated with 20 and 60 krad, then grown on ESS-1 medium with 23% NaCl, had a higher density than other samples. This protein was reported to play an important role in G. asiatica, by enhancing its tolerance to variable salinity and temperature. Although the organic and inorganic content of all samples remained almost the same, the content and quality of agar extracted from irradiated seaweeds were higher than those of the controls. Due to the high amount of 3.6 anhydro-α-L-galactose combined with low amounts of sulfate found in irradiated seaweeds, the freezing and melting points of extracted agar were lower. Eventually, this resulted in higher condensation and better quality of agar, such as in its gel-forming ability. The quality of fluid agar extracted from selected seaweeds improved as shown in the remarkable decrease in Ca²⁺, Mg²⁺, and total Fe ion content, thus lowering its melting point compared with the control. Presented at the 6th Meeting of the Asian Pacific Society of Applied Phycology, Manila, Philippines.     

Acclimation to UV irradiance in Gracilaria chilensis Bird,McLachlan & Oliveira (Gigartinales,Rhodophyta)

Photoautotrophs can cope with an increase in ultraviolet (UV) irradiance in the aquatic environment, through protection and acclimation mechanisms (i.e. synthesis of UV-absorbing compounds). This capacity has been proven to vary according to the organism's sensitivity. To quantify variations of this capacity between the different parts of macroalgae, an in vitro study was performed with the tips, cystocarps and thalli segments of Gracilaria chilensis. Whole algae incubated during 3 days at high and low PAR, supplying UV B (4.6 µW cm^–2) during 2 hours showed, as predicted, an increase in absorption (OD) at 320 nm of the different parts, after the first day of exposure to UV B. The tips presented the highest increase in the standardized OD at 320 nm relative to cystocarps and thalli segments; their mean percentage of increase was 38% and 29% at low and high PAR, respectively. The lowest sensitivity was consistently found in the thalli segments, while the highest was in the tips. The tips are important for growth and therefore they play a major role in the maintenance of the Gracilaria populations. Acclimation mechanisms that occurred in a short time scale, and mainly in the tips, may allow Gracilaria to have an almost immediate protection to increases in UV B fluxes.     

Growth,pigments, UV-absorbing compounds and agar yield of the economic red seaweed <Emphasis Type="Italic">Gracilaria lemaneiformis</Emphasis> (Rhodophyta) grown at different depths in the coastal waters of the South China Sea

The economic red alga, Gracilaria lemaneiformis Bory, was grown at different depths in the coastal waters of the South China Sea, and its growth, pigments, ultra-violet (UV)-absorbing compounds and agar yield were investigated in order to see the impacts of depth change. Gracilaria lemaneiformis grew slower at greater depths in March, while the highest relative growth rate (RGR) was found at about 1.0 m depth in April, about 9% higher than that at surface water (0.5 m below the surface). The RGR increased with the increasing daily photosynthetically active radiation (PAR) dose received by the thalli at different depths. The contents of phycoerythrin and chlorophyll a increased, while that of UV-absorbing compounds (UVAC, absorption peak at 325 nm) decreased with increased depth. The highest levels of the UVAC in the thalli grown in surface seawater played a protective role against solar UV radiation (280–400 nm). The content of UVAC declined at deeper depths and under indoor low PAR. The agar yield of the thalli increased with the increasing depths, with the highest content found at 3.5 m depth.     

The lizard Psammodromus algirus (Squamata: Lacertidae) is darker at high altitudes

Altitudinal gradients offer a good opportunity to study organisms' adaptations to clinal environmental variables. Regarding altitude, the most influential variables on organisms are temperature and ultraviolet (UV) solar radiation, the first decreasing and the second increasing with altitude. Both variables affect ectotherms' biology, as ectotherms depend on environmental temperature for thermoregulation, frequently being heliotherms. Here, we studied dorsal coloration in the lizard Psammodromus algirus (Linnaeus, 1758) along a wide altitudinal gradient (2200 m) in Sierra Nevada (south‐east Spain). We hypothesize that the skin will be darker with altitude, i.e. in environments with lower temperatures and higher UV radiation intensity. Results show that individual dorsal colorations became darker at high altitude. We propose two non‐mutually exclusive explanations for this result: (1) darker dorsal surface would favour faster warming at high altitudes, where temperature is lower, and (2) darker dorsal surface would protect against UV radiation, stronger at high altitudes. We found significant relationships between both temperature and UV radiation and population dorsal darkness, giving mixed support for the two explanations. Moreover, dorsal hue was positively correlated with substrate hue, suggesting that hue evolved to maximize crypsis. Our study therefore suggests that geographical variation in dorsal coloration in this lizard is adaptive, and darkness coloration might have evolved in response to adverse conditions (low temperature and high UV radiation) at high altitudes. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 112 , 132–141.