Ultraviolet-B radiation effects on plants: induction of morphogenic responses

Plants raised under field conditions are acclimated to ambient levels of solar UV-B radiation. Morphogenic responses are part of the UV-B acclimation process and have been hypothesized to contribute to UV avoidance. UV-B induced morphogenic responses include inhibition of hypocotyl and stem elongation, leaf curling, leaf thickening and increased axillary branching. So far, neither the photosensory nor the signal transduction mechanism involved in UV-B mediated morphogenesis has been identified. The combination of classical photobiological techniques and Arabidopsis genetic resources comprises a powerful tool for the analysis of morphogenic responses. However, no morphogenic mutants, specifically altered in their response to UV-B, have yet been identified. In this paper we discuss the possibility that some UV-B driven morphogenic responses do not involve a dedicated photosensory system, but rather are a consequence of UV-B induced changes in secondary metabolism. UV-B induced flavonoid aglycones and phenol-oxidizing peroxidases can affect, respectively, polar auxin transport and auxin catabolism, and hence plant architecture. Integration of genetic, photobiological, biochemical and physiological approaches is necessary to fully appraise the ecophysiological role of UV-B radiation in controlling plant architecture.     

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.     

Contribution of phenolic compounds to the UV-B screening capacity of developing barley primary leaves in relation to DNA damage and repair under elevated UV-B levels

Epidermally located UV-absorbing hydroxycinnamic acid conjugates and flavonoid glycosides are known to be efficient UV-B protectants in higher plants, although important biological molecules are not always fully protected. However, repair mechanisms also exist, such as repair of damaged DNA by photolyases. To distinguish between the relative importance of the phenolic compounds and of DNA repair, developing primary leaves of two barley lines, mutant ant 30-310, deficient in flavonoids, and its parent line Ca 33787, were grown under relatively high visible light (650-700 micromol m(-2) s(-1) max for 6 h in a 13 h photoperiod) and supplemented with (+ UV-B) or without (-UV-B) 12 kJ m(-2) UV-B(BE) for 6 h daily. UV-B screening capacity of the leaf phenolics was determined at 315 nm during leaf development and compared with thymine dimers (TD) accumulation, as an indicator of UV-B-induced DNA damage and potential subsequent repair. The degree of damage was related to the phenolic contents of the leaves. UV-B screening capacity was increased ca. 4-fold in the parent line (+ UV-B), mainly due to UV-induced flavonoid (saponarin, lutonarin) accumulation in epidermal and subepidermal mesophyll tissue, relative to the flavonoid-deficient mutant. Nevertheless, in the parent line an 8-fold increase in TD levels occurred over the growth period of 18 days, whereas the mutant accumulated additional DNA damage, with 6- to 9-fold higher TD amounts. Surprisingly, under the high UV-B irradiation, growth and development of the primary leaves in both lines were only slightly reduced.     

Metabolomic and physiological responses reveal multi-phasic acclimation of Arabidopsis thaliana to chronic UV radiation

Biochemical changes in vivo and pathway interactions were investigated using integrated physiological and metabolic responses of Arabidopsis thaliana L. to ultraviolet (UV) radiation (280–400 nm) at 9.96 kJ m⁻² d⁻¹ over the entire life cycle from seed to seed (8 weeks). Columbia-0 (Col-0) and a UV-B sensitive accession ( fah-1 ) showed significant ( P  < 0.001) reductions in leaf growth after 6 weeks. Col-0 recovered growth after 8 weeks, with recovery corresponding to a switch from production of phenylpropanoids to flavonoids. fah-1 failed to recover, indicating that sinapate production is an essential component of recovery. Epidermal features show that UV radiation caused significant ( P  < 0.001) increases in trichome density, which may act as a structural defence response. Stomatal indices showed a significant ( P  < 0.0001) reduction in Col-0 and a significant ( P  < 0.001) increase in fah-1 . Epidermal cell density was significantly increased under UV radiation on the abaxial leaf surface, suggesting that that a fully functioning phenylpropanoid pathway is a requirement for cell expansion and leaf development. Despite wild-type acclimation, the costs of adaptation lead to reduced plant fitness by decreasing flower numbers and total seed biomass. A multi-phasic acclimation to UV radiation and the induction of specific metabolites link stress-induced biochemical responses to enhanced acclimation.     

Downsizing in plants—UV light induces pronounced morphological changes in the absence of stress

Ultraviolet (UV) light induces a stocky phenotype in many plant species. In this study, we investigate this effect with regard to specific UV wavebands (UV-A or UV-B) and the cause for this dwarfing. UV-A- or UV-B-enrichment of growth light both resulted in a smaller cucumber (Cucumis sativus L.) phenotype, exhibiting decreased stem and petiole lengths and leaf area (LA). Effects were larger in plants grown in UV-B- than in UV-A-enriched light. In plants grown in UV-A-enriched light, decreases in stem and petiole lengths were similar independent of tissue age. In the presence of UV-B radiation, stems and petioles were progressively shorter the younger the tissue. Also, plants grown under UV-A-enriched light significantly reallocated photosynthates from shoot to root and also had thicker leaves with decreased specific LA. Our data therefore imply different morphological plant regulatory mechanisms under UV-A and UV-B radiation. There was no evidence of stress in the UV-exposed plants, neither in photosynthetic parameters, total chlorophyll content, or in accumulation of damaged DNA (cyclobutane pyrimidine dimers). The abscisic acid content of the plants also was consistent with non-stress conditions. Parameters such as total leaf antioxidant activity, leaf adaxial epidermal flavonol content and foliar total UV-absorbing pigment levels revealed successful UV acclimation of the plants. Thus, the UV-induced dwarfing, which displayed different phenotypes depending on UV wavelengths, occurred in healthy cucumber plants, implying a regulatory adjustment as part of the UV acclimation processes involving UV-A and/or UV-B photoreceptors.

A stocky phenotype develops in healthy cucumber plants as a regulatory adjustment toward UV-A and UV-B-enriched light, revealing a strong interaction between UV acclimation and developmental processes.     

Ultraviolet-B radiation impacts light-mediated turnover of the photosystem II reaction center heterodimer in Arabidopsis mutants altered in phenolic metabolism

Ultraviolet-B (UV-B) radiation can have a negative impact on the growth and development of plants. Plants tolerant to UV-B alleviate these effects using UV-screening pigments that reduce the penetration of UV-B into mesophyll tissue. Little is known about the relative contribution of specific phenolic compounds to the screening capacity of leaves. The D1 and D2 proteins constituting the photosystem (PS) II reaction center heterodimer are targets of UV-B radiation and can be used as an in situ sensor for UV penetration into photosynthetic tissue. Degradation of these proteins occurs under very low fluences of UV-B, and is strongly accelerated in the presence of visible light. Using the D1-D2 degradation assay, we characterized UV-B sensitivity of Arabidopsis mutants (tt4, tt5, and fah1) that are genetically altered in their composition of phenolic compounds. We found that changes in phenol metabolism result in altered rates of PSII reaction center heterodimer degradation under mixtures of photosynthetically active radiation and UV-B. A comparison of D2 degradation kinetics showed increased UV sensitivity of the Landsberg (Landsberg erecta) tt5 mutant relative to the Landsberg tt4 mutant and the Landsberg wild type. Despite a lack of flavonoid accumulation, the tt4 mutant is not particularly UV sensitive. However, the tolerance of this mutant to UV-B may reflect the increased accumulation of sinapate esters that strongly absorb in the UV range, and may thus protect the plant against environmentally relevant UV-B radiation. This sinapate-mediated protection is less obvious for the tt4 mutant of Columbia ecotype, indicating that the relative contribution of particular phenolics to the total screening capacity varies with the genetic background. The role of sinapate esters in UV screening is further substantiated by the results with the fah1 mutant where absence of most of the sinapate esters results in a significantly accelerated degradation of D2 under mixed light conditions. Because the latter mutant is not expected to be deficient in flavonoids, the relative contribution of flavonoids as protectants of PSII reaction center heterodimer against UV-B damage in Arabidopsis needs to be re-evaluated vis-a-vis screening by simple phenolics like sinapate esters.     

How plants protect themselves from ultraviolet-B radiation stress

Ultraviolet-B (UV-B) radiation has a wavelength range of 280–315 nm. Plants perceive UV-B as an environmental signal and a potential abiotic stress factor that affects development and acclimation. UV-B regulates photomorphogenesis including hypocotyl elongation inhibition, cotyledon expansion, and flavonoid accumulation, but high intensity UV-B can also harm plants by damaging DNA, triggering accumulation of reactive oxygen species, and impairing photosynthesis. Plants have evolved “sunscreen” flavonoids that accumulate under UV-B stress to prevent or limit damage. The UV-B receptor UV RESISTANCE LOCUS 8 (UVR8) plays a critical role in promoting flavonoid biosynthesis to enhance UV-B stress tolerance. Recent studies have clarified several UVR8-mediated and UVR8-independent pathways that regulate UV-B stress tolerance. Here, we review these additions to our understanding of the molecular pathways involved in UV-B stress tolerance, highlighting the important roles of ELONGATED HYPOCOTYL 5, BRI1-EMS-SUPPRESSOR1, MYB DOMAIN PROTEIN 13, MAP KINASE PHOSPHATASE 1, and ATM- and RAD3-RELATED. We also summarize the known interactions with visible light receptors and the contribution of melatonin to UV-B stress responses. Finally, we update a working model of the UV-B stress tolerance pathway.

Recent findings that update our understanding of the molecular pathway for ultraviolet-B radiation stress responses in plants are summarized.     

Arabidopsis thaliana plants acclimated to low dose rates of ultraviolet B radiation show specific changes in morphology and gene expression in the absence of stress symptoms

Ultraviolet B (UV-B) acclimation comprises complex and poorly understood changes in plant metabolism. The effects of chronic and ecologically relevant UV-B dose rates on Arabidopsis thaliana were determined. The UV-B acclimation process was studied by measuring radiation effects on morphology, physiology, biochemistry and gene expression. Chronic UV-B radiation did not affect photosynthesis or the expression of stress responsive genes, which indicated that the UV-acclimated plants were not stressed. UV-induced morphological changes in acclimated plants included decreased rosette diameter, decreased inflorescence height and increased numbers of flowering stems, indicating that chronic UV-B treatment caused a redistribution rather than a cessation of growth. Gene expression profiling indicated that UV-induced morphogenesis was associated with subtle changes in phytohormone (auxins, brassinosteroids and gibberellins) homeostasis and the cell wall. Based on the comparison of gene expression profiles, it is concluded that acclimation to low, chronic dose rates of UV-B is distinct from that to acute, stress-inducing UV-B dose rates. Hence, UV-B-induced morphogenesis is functionally uncoupled from stress responses.     

Flavonoids and auxin transport: modulators or regulators?

Flavonoids are polyphenolic compounds found in all vascular and non-vascular plants. Although nonessential for plant growth and development, flavonoids have species-specific roles in nodulation, fertility, defense and UV protection. Flavonoids have been shown to modulate transport of the phytohormone auxin in addition to auxin-dependent tropic responses. However, flavonoids are not essential regulators of these processes because transport and tropic responses occur in their absence. Flavonoids modulate the activity of auxin-transporting P-glycoproteins and seem to modulate the activity of regulatory proteins such as phosphatases and kinases. Phylogenetic analysis suggests that auxin transport mechanisms evolved in the presence of flavonoid compounds produced for the scavenging of reactive oxygen species and defense from herbivores and pathogens.     

Arabidopsis Flavonoid Mutants Are Hypersensitive to UV-B Irradiation

Increases in the terrestrial levels of ultraviolet-B (UV-B) radiation (280 to 320 nm) due to diminished stratospheric ozone have prompted an investigation of the protective mechanisms that contribute to UV-B tolerance in plants. In response to UV-B stress, flowering plants produce a variety of UV-absorptive secondary products derived from phenylalanine. Arabidopsis mutants with defects in the synthesis of these compounds were tested for UV-B sensitivity. The transparent testa-4 (tt4) mutant, which has reduced flavonoids and normal levels of sinapate esters, is more sensitive to UV-B than the wild type when grown under high UV-B irradiance. The tt5 and tt6 mutants, which have reduced levels of UV-absorptive leaf flavonoids and the monocyclic sinapic acid ester phenolic compounds, are highly sensitive to the damaging effects of UV-B radiation. These results demonstrate that both flavonoids and other phenolic compounds play important roles in vivo in plant UV-B protection.     

Arabidopsis UVR8 regulates ultraviolet-B signal transduction and tolerance and contains sequence similarity to human regulator of chromatin condensation 1

To further our understanding of how plants defend against the harmful effects of ultraviolet (UV) light, we characterized an Arabidopsis mutant hypersensitive to UV-B. This mutant, UV resistance locus 8-1 (uvr8-1), contains a single recessive mutation at the bottom of chromosome 5. Fine-scale mapping localized uvr8-1 to a 21-kb locus containing five predicted open reading frames. Sequencing of this entire region revealed that the uvr8-1 allele contains a 15-nucleotide deletion in a gene similar to the human guanine nucleotide exchange factor regulator of chromatin condensation 1. This mutation reduces the UV-B-mediated induction of flavonoids and blocks chalcone synthase mRNA and protein induction. In contrast, uvr8-1 has enhanced induction of PR1 and PR5 proteins in response to UV-B, an indication of increased UV-B injury. These results suggest that UVR8 acts in a UV-B signal transduction pathway leading to induction of flavonoid biosynthesis.     

Effect of reduced arginine decarboxylase activity on salt tolerance and on polyamine formation during salt stress in Arabidopsis thaliana

Polyamines have been suggested to play an important role in stress protection. However, attempts to determine the function of polyamines have been complicated by the fact that, dependent on the conditions, polyamine contents increase or decrease during stress. To determine the importance of polyamine formation during salt stress, we analysed polyamine contents and salt tolerance in two Arabidopsis thaliana mutants, spe1-1 and spe2-1 (Watson et al. Plant J 13: 231–239, 1998), with reduced activity of arginine decarboxylase (EC 4.1.1.19), an important enzyme in polyamine synthesis. Polyamines accumulated in wild-type plants (Col-0 and Ler-0) that were pre-treated with 100 m M NaCl before transfer to 125 m M NaCl, but not in plants that were directly transferred to 125 m M NaCl without prior treatment with 100 m M NaCl. This shows that polyamine accumulation depends on acclimation to salinity. The salt treatment that induced polyamine accumulation in wild-type plants did not lead to polyamine accumulation in the spe1-1 and spe2-1 mutants. Decreased fresh weight, chlorophyll content and photosynthetic efficiency indicated that the spe1-1 mutant was more severely affected by salt stress than its wild type, Col-0. In the spe2-1 mutant decreased salt tolerance compared to its wild type, Ler-0, became apparent as bleaching under severe salt stress. The present results demonstrate that decreased polyamine formation due to lower arginine decarboxylase activity leads to reduced salt tolerance.     

Content of Osmolytes and Flavonoids under Salt Stress in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> Plants Defective in Jasmonate Signaling

The effects of the salt stress (200 mM NaCl) and exogenous jasmonic acid (JA) on levels of osmolytes and flavonoids in leaves of four-week-old Arabidopsis thaliana L. plants of the wild-type (WT) Columbia-0 (Col-0) and the mutant jin1 (jasmonate insensitive 1) with impaired jasmonate signaling were studied. The increase in proline content caused by the salt stress was higher in the Col-0 plants than in the mutant jin1. This difference was especially marked if the plants had been pretreated with exogenous 0.1 μM JA. The sugar content increased in response to the salt stress in the JA-treated WT plants but decreased in the jin1 mutant. Treatment with JA of the WT plants but not mutant defective in jasmonate signaling also enhanced the levels of anthocyanins and flavonoids absorbed in UV-B range in leaves. The presence of JA increased salinity resistance of the Col-0 plants, since the accumulation of lipid peroxidation products and growth inhibition caused by NaCl were less pronounced. Under salt stress, JA almost did not render a positive effect on the jin1 plants. It is concluded that the protein JIN1/MYC2 is involved in control of protective systems under salt stress.     

GmPIN1-mediated auxin asymmetry regulates leaf petiole angle and plant architecture in soybean

Crop breeding during the Green Revolution resulted in high yields largely due to the creation of plants with semi-dwarf architectures that could tolerate high-density planting. Although semi-dwarf varieties have been developed in rice, wheat and maize, none was reported in soybean(Glycine max), and few genes controlling plant architecture have been characterized in soybean. Here, we demonstrate that the auxin efflux transporter PINFORMED1(GmPIN1), which determines polar auxin transport, regulate...     

A flavonoid mutant of barley (Hordeum vulgare L.) exhibits increased sensitivity to UV-B radiation in the primary leaf

The aim of the present investigation was to define the role of soluble flavonoids as UV-B protectants in the primary leaf of barley (Hordeum vulgare L.). For this purpose we used a mutant line (Ant 287) from the Carlsberg collection of proanthocyanidin-free barley containing only 7% of total extractable flavonoids in the primary leaf as compared to the mother variety (Hiege 550/75). Seven-day-old leaves from plants grown under high visible light with or without supplementary UV-B radiation were used for the determination of UV-B sensitivity. UV-B-induced changes were assessed from parameters of chlorophyll fluorescence of photosystem II, including initial and maximum fluorescence, apparent quantum yield, and photochemical and non-photochemical quenching. A quartz fibre-optic microprobe was used to evaluate the amount of potentially harmful UV-B (310 nm radiation) penetrating into the leaf as a direct consequence of flavonoid deficiency. Our data indicate an essential role of flavonoids in UV-B protection of barley primary leaves. In leaves of the mutant line grown under supplementary UV-B, an increase in 310nm radiation in the mesophyll and a strong decrease in the quantum yield of photosynthesis were observed as compared to the corresponding mother variety. Primary leaves of liege responded to supplementary UV-B radiation with a 30% increase in the major flavonoid saponarin and a 500% increase in the minor compound lutonarin. This is assumed to be an efficient protective response since no changes in variable chlorophyll fluorescence were apparent. In addition, a further reduction in UV-B penetration into the mesophyll was recorded in these leaves.     

IBR5, a dual-specificity phosphatase-like protein modulating auxin and abscisic acid responsiveness in Arabidopsis

Auxin is an important plant hormone that plays significant roles in plant growth and development. Although numerous auxin-response mutants have been identified, auxin signal transduction pathways remain to be fully elucidated. We isolated ibr5 as an Arabidopsis indole-3-butyric acid-response mutant, but it also is less responsive to indole-3-acetic acid, synthetic auxins, auxin transport inhibitors, and the phytohormone abscisic acid. Like certain other auxin-response mutants, ibr5 has a long root and a short hypocotyl when grown in the light. In addition, ibr5 displays aberrant vascular patterning, increased leaf serration, and reduced accumulation of an auxin-inducible reporter. We used positional information to determine that the gene defective in ibr5 encodes an apparent dual-specificity phosphatase. Using immunoblot and promoter-reporter gene analyses, we found that IBR5 is expressed throughout the plant. The identification of IBR5 relatives in other flowering plants suggests that IBR5 function is conserved throughout angiosperms. Our results suggest that IBR5 is a phosphatase that modulates phytohormone signal transduction and support a link between auxin and abscisic acid signaling pathways.     

Flavonoids and Auxin Transport Inhibitors Rescue Symbiotic Nodulation in the Medicago truncatula Cytokinin Perception Mutant cre1

Initiation of symbiotic nodules in legumes requires cytokinin signaling, but its mechanism of action is largely unknown. Here, we tested whether the failure to initiate nodules in the Medicago truncatula cytokinin perception mutant cre1 (cytokinin response1) is due to its altered ability to regulate auxin transport, auxin accumulation, and induction of flavonoids. We found that in the cre1 mutant, symbiotic rhizobia cannot locally alter acro- and basipetal auxin transport during nodule initiation and that these mutants show reduced auxin (indole-3-acetic acid) accumulation and auxin responses compared with the wild type. Quantification of flavonoids, which can act as endogenous auxin transport inhibitors, showed a deficiency in the induction of free naringenin, isoliquiritigenin, quercetin, and hesperetin in cre1 roots compared with wild-type roots 24 h after inoculation with rhizobia. Coinoculation of roots with rhizobia and the flavonoids naringenin, isoliquiritigenin, and kaempferol, or with the synthetic auxin transport inhibitor 2,3,5,-triiodobenzoic acid, rescued nodulation efficiency in cre1 mutants and allowed auxin transport control in response to rhizobia. Our results suggest that CRE1-dependent cytokinin signaling leads to nodule initiation through the regulation of flavonoid accumulation required for local alteration of polar auxin transport and subsequent auxin accumulation in cortical cells during the early stages of nodulation.     

Salicylate accumulation inhibits growth at chilling temperature in Arabidopsis

The growth of Arabidopsis plants in chilling conditions could be related to their levels of salicylic acid (SA). Plants with the SA hydroxylase NahG transgene grew at similar rates to Col-0 wild types at 23 degrees C, and growth of both genotypes was slowed by transfer to 5 degrees C. However, at 5 degrees C, NahG plants displayed relative growth rates about one-third greater than Col-0, so that by 2 months NahG plants were typically 2.7-fold larger. This resulted primarily from greater cell expansion in NahG rosette leaves. Specific leaf areas and leaf area ratios remained similar in both genotypes. Net assimilation rates were similar in both genotypes at 23 degrees C, but higher in NahG at 5 degrees C. Chlorophyll fluorescence measurements revealed no PSII photodamage in chilled leaves of either genotype. Col-0 shoots at 5 degrees C accumulated SA, particularly in glucosylated form. SA in NahG shoots showed similar tendencies at 5 degrees C, but at greatly depleted levels. Catechol was not detected as a metabolite of the NahG transgene product. We also examined growth and SA levels in SA signaling and metabolism mutants at 5 degrees C. The partially SA-insensitive npr1 mutant displayed growth intermediate between NahG and Col-0, while the SA-deficient eds5 mutant behaved like NahG. In contrast, the cpr1 mutant at 5 degrees C accumulated very high levels of SA and its growth was much more inhibited than wild type. At both temperatures, cpr1 was the only SA-responsive genotype in which oxidative damage (measured as thiobarbituric acid-reactive substances) was significantly different from wild type.