Impacts of enhanced UVB radiation on photosynthetic characteristics of the marine diatom <Emphasis Type="Italic">Phaeodactylum tricornutum</Emphasis> (Bacillariophyceae,Heterokontophyta)

Solar ultraviolet B (UVB) irradiance at the Earth’s surface is increasing due to anthropogenic influences. To evaluate the effects of enhanced UVB radiation on photosynthetic characteristics of the marine diatom Phaeodactylum tricornutum, the species was exposed to four levels of UVB radiation, 0, 0.25, 0.75, and 1.50 KJ m^?2 day^?1 for 7 days. Effects of UVB stress on net photosynthetic rate, net respiration rate, variable chlorophyll (Chl) fluorescence parameters, Chl a and carotenoid contents, and UV-absorbing compounds (UVACs) were investigated. Results showed that there were no significant differences in terms of net respiration rate or maximal photochemical efficiency of photosystem II (F_v/F_m) between the treatments in the short or long term. However, enhanced UVB radiation at an intensity of 0.16 W m^?2 had a negative effect on the net photosynthetic rate, electron transport rate, and on the pathway of excess energy dissipation over the short term (1 to 5 days). Carotenoid and UVACs content increased under UVB radiation. Photosynthetic parameters were unaffected by UVB radiation on the seventh day indicating that P. tricornutum can adapt to UVB radiation in the long term. Results of the present study indicate that there is a dynamic balance between damage and adaptation in microalgae that enables them to adapt to UVB-induced photosystem alterations during both short-term and long-term exposure.     

The effect of galling aphids feeding on photosynthesis photochemistry of elm trees (<Emphasis Type="Italic">Ulmus</Emphasis> sp.)

Changes of chlorophyll (Chl) a fluorescence and photosynthetic pigment contents were analysed in galled leaves (visibly damaged and undamaged parts) and intact leaves. The values of minimal fluorescence of the dark-adapted state, maximal quantum yield of PSII photochemistry, effective quantum yield of PSII photochemical conversion, and photochemical quenching coefficient decreased in Ulmus pumila L. leaves galled by Tetraneura ulmi (L.) and in U. glabra Huds. galled by Eriosoma ulmi (L.). Colopha compressa (Koch.) feeding affected these parameters only in damaged parts of U. laevis Pall. galled leaves. The increasing number of T. ulmi galls progressively decreased photosynthetic performance. In gall tissues of all analysed aphid species, the lowest photosynthetic pigment content was found, indicating that the photosynthetic capacity must have been low in galls. Significant reduction of Chl and carotenoid contents were observed in damaged and undamaged portions of galled leaves only in the case of T. ulmi feeding.     

Heat shock increases thermotolerance of photosynthetic electron transport and the content of chloroplast membranes and lipids in wheat leaves

Preliminary heating of 15-16-day-old wheat (Triticum aestivum L.) plants for 3 h at 37–38°C (heat shock, HS) increased the tolerance of photosynthetic electron transport (determined as the reduction of 2,6-dichlorophenol indophenol by isolated chloroplasts) toward heating of leaves at 42–48°C in high light (100 klx). At the same time, HS did not affect the activity of the xanthophyll cycle reactions in the 30–48°C temperature range. HS exposure induced an increase in the thylakoid length, the number of grana, and the average number of thylakoids per granum. The volume of the thylakoid system increased 1.4-fold. Such indices as the total content of chlorophylls (a + b), the chlorophyll a/b ratio, as well as the contents of individual carotenoids, chloroplast membrane proteins, and the soluble leaf proteins remained unchanged. The de novo photosynthetic membrane formation was accompanied by the 1.5-fold increase in major chloroplast lipids. It was concluded that, in mature wheat chloroplasts, HS induced the formation of thylakoids characterized by a changed molecular structure and by increased lipid/protein and lipid/chlorophyll ratios.     

Acclimation of shade-tolerant and light-resistant <Emphasis Type="Italic">Tradescantia</Emphasis> species to growth light: chlorophyll <Emphasis Type="Italic">a</Emphasis> fluorescence,electron transport,and xanthophyll content

In this study, we have compared the photosynthetic characteristics of two contrasting species of Tradescantia plants, T. fluminensis (shade-tolerant species), and T. sillamontana (light-resistant species), grown under the low light (LL, 50–125 µmol photons m^?2 s^?1) or high light (HL, 875–1000 µmol photons m^?2 s^?1) conditions during their entire growth period. For monitoring the functional state of photosynthetic apparatus (PSA), we measured chlorophyll (Chl) a emission fluorescence spectra and kinetics of light-induced changes in the heights of fluorescence peaks at 685 and 740 nm (F ₆₈₅ and F ₇₄₀). We also compared the light-induced oxidation of P₇₀₀ and assayed the composition of carotenoids in Tradescantia leaves grown under the LL and HL conditions. The analyses of slow induction of Chl a fluorescence (SIF) uncovered different traits in the LL- and HL-grown plants of ecologically contrasting Tradescantia species, which may have potential ecophysiological significance with respect to their tolerance to HL stress. The fluorometry and EPR studies of induction events in chloroplasts in situ demonstrated that acclimation of both Tradescantia species to HL conditions promoted faster responses of their PSA as compared to LL-grown plants. Acclimation of both species to HL also caused marked changes in the leaf anatomy and carotenoid composition (an increase in Violaxanthin?+?Antheraxantin?+?Zeaxanthin and Lutein pools), suggesting enhanced photoprotective capacity of the carotenoids in the plants grown in nature under high irradiance. Collectively, the results of the present work suggest that the mechanisms of long-term PSA photoprotection in Tradescantia are based predominantly on the light-induced remodeling of pigment-protein complexes in chloroplasts.     

Comparative study of the effects of salinity and UV radiation on metabolism and morphology of the red macroalga <Emphasis Type="Italic">Acanthophora spicifera</Emphasis> (Rhodophyta,Ceramiales)

Increase of harmful radiation to the Earth’s surface due to ozone depletion results in higher exposure to harmful ultraviolet- B radiation (UV), while fluctuations in seawater salinity may alter water density, ionic concentration, nutrient uptake, and osmotic pressure. This study evaluated the effects of salinity and UV on metabolism and morphology of Acanthophora spicifera (M.Vahl) Børgesen. Water with 30 and 37 psu [g(salt) kg^–1(sea water)] was used for experiments during 7 d of exposure to UV (3 h per day). We demonstrated that UV treatment predisposed, irrespective of salinity, A. spicifera to a decrease in its growth rate and cell viability, as well as affected its morphological parameters. After exposure to PAR + UVA + UVB (PAB), samples showed structural changes and damage, such as increasing cell wall thickness and chloroplast disruption. Our results indicate that UV led to dramatic metabolic changes and cellular imbalances, but more remarkable changes were seen in samples exposed to high salinity.     

Activity of the System for Chlorophyll Biosynthesis and Structural and Functional Organization of Chloroplasts in a Plastome <Emphasis Type="Italic">en:chlorina-5</Emphasis> Sunflower Mutant

Chlorophyll (Chl) deficiency in leaves of a plastome sunflower (Helianthus annuus L.) en:chlorina-5 mutant is due to the formation of smaller chloroplasts with a markedly reduced membrane system, as compared to the parent 3629 line. Abnormalities in the structure of the photosynthetic apparatus in the mutant can be mainly attributed to changes in the formation of photosystem I and its light-harvesting complexes. Chl deficiency in en:chlorina-5 correlated with its lower capability of synthesizing the first specific Chl precursor, 5-aminolevulinic acid (ALA) in the light. Light-independent stages of Chl biosynthesis in the mutant had the same efficiency as in leaves of the parent line. ALA formation in darkness and its conversion into protochlorophyllide did not depend on the extent of photosynthetic membrane development and photosynthetic activity.     

Disturbance of chlorophyll biosynthesis at Mg branch affects the chloroplast ROS homeostasis and Ca<Superscript>2+</Superscript> signaling in <Emphasis Type="Italic">Pisum sativum</Emphasis>

Reactive oxygen species (ROS) and calcium (Ca²⁺), two crucial intracellular signaling molecules, have been reported to play important roles in chlorophyll biosynthesis. In this study, we aimed to investigate whether disturbance of chlorophyll synthesis affects chloroplast ROS and Ca²⁺ homeostases. Chlorophyll biosynthesis was inhibited at the Mg branch by virus-induced gene silencing (VIGS) of CHLI gene encoding the Mg chelatase CHLI subunit in pea (Pisum sativum). Subsequently, ROS and intracellular free Ca²⁺ concentration ([Ca²⁺]_i) in these chlorophyll-deficient pea plants were evaluated by histochemical and fluorescent staining assays. The results showed that the superoxide anion and hydrogen peroxide were predominantly generated in chloroplasts of the yellow leaves of pea VIGS-CHLI plants. The expression of genes encoding chloroplast antioxidant enzymes (CuZn-superoxide dismutase, ascorbate peroxidase, glutathione reductase, phospholipid glutathione peroxidase, peroxiredoxin and thioredoxins) were also decreased in the leaves of VIGS-CHLI plants compared with the control plants. Additionally, the [Ca²⁺]_i were significantly reduced in the yellow leaves of VIGS-CHLI plants compared with the green leaves of VIGS-GFP control plants. The expression of genes encoding Ca²⁺ signaling related proteins (thylakoid Ca²⁺ transporter, calmodulins and calcineurin B-like protein) was down-regulated in yellow VIGS-CHLI leaves. These results indicate that inhibition of chlorophyll biosynthesis at the Mg branch by silencing CHLI affects chloroplast ROS homeostasis and Ca²⁺ signaling and down-regulates the expression of ROS scavenging genes and Ca²⁺ signaling related genes.     

Physiological acclimation of <Emphasis Type="Italic">Lessonia spicata</Emphasis> to diurnal changing PAR and UV radiation: differential regulation among down-regulation of photochemistry,ROS scavenging activity and phlorotannins as major photoprotective mechanisms

Intertidal macroalgae are constantly subjected to high variations in the quality and quantity of incident irradiance that can eventually generate detrimental effect on the photosynthetic apparatus. The success of these organisms to colonize the stressful coastal habitat is mainly associated with the complexity of their morphological structures and the efficiency of the anti-stress mechanisms to minimize the physiological stress. Lessonia spicata (Phaeophyceae), a brown macroalga, that inhabits the intertidal zone in central–southern Chile was studied in regard to their physiological (quantum yield, electron transport rate, pigments) and biochemical (phlorotannins content, antioxidant metabolism, oxidative stress) responses during a daily light cycle under natural solar radiation. Major findings were that F _v/F _m, photosynthetic parameters (ETR_max, alpha, E _k) and pigments in L. spicata showed an inverse relationship to the diurnal changes in solar radiation. Phlorotannins levels and antioxidant activity showed their highest values in treatment that included UV radiation. There was an increase in SOD and APX in relation at light stress, with a peak in activity between 5.2 and 10.1 W m^?2 of biologically effective dose. The increase in peroxidative damage was proportional to light dose. These results indicated that different light doses can trigger a series of complementary mechanisms of acclimation in L. spicata based on: (i) down-regulation of photochemistry activity and decrease in concentration of photosynthetic pigments; (ii) induction of phenolic compounds with specific UV-screening functions; and (iii) reactive oxygen species (ROS) scavenging activity via complementary repair of the oxidative damage through increased activity of antioxidant enzymes and potentially increased amounts of phenolic compounds.     

An efficient method for the sequential production of lipid and carotenoids from the Chlorella Growth Factor-extracted biomass of <Emphasis Type="Italic">Chlorella vulgaris</Emphasis>

Efficient methodology for simultaneous extraction of multiple bioactive compounds from microalgae still remains a major challenge. The present study provides a method for the sequential production of three major products: Chlorella Growth Factor (CGF, a nucleotide-peptide complex enriched with vitamins, minerals, and carbohydrates), lipid, and carotenoids from Chlorella vulgaris biomass in an economically feasible manner. After protein-rich CGF was extracted, the spent biomass was found to contain 12% lipid and 3% carotenoids when extracted individually, compared to that of the un-utilized (fresh) biomass (lipid, 14%; carotenoids, 4%). When extracted simultaneously using conventional methods, the yield of lipid from “CGF and carotenoids-extracted biomass,” and carotenoids from “CGF and lipid-extracted biomass” were significantly reduced (50%). However, simultaneous extraction using different solvent mixtures such as hexane:methanol:water and pentane:methanol:water mixture-augmented lipid yield by 38.5% and carotenoids by 14%, and additionally retained chlorophyll and its derivatives. Column chromatographic approach yielded sequential production of lipid (18%), lutein (9%) with better yields as well as without chlorophyll interference. Different geometric isomers of lutein all-E-(trans)-(3R,3′R,6′R)-β,ε-carotene-3,3′diol, 9Z(cis)-(3R,3′R,6′R)-β,ε-carotene-3,3′diol, and 13Z(cis)-(3R,3′R,6′R)-β,ε-carotene-3,3′diol were purified by HPLC and elucidated by CD, UV, NMR, FT-IR, and Mass spectra. In conclusion, the study provides an efficient and economically viable methodology for sequential production of lipid and lutein along with its geometrical isomers without chlorophyll influence and yield loss from the protein-rich CGF-extracted spent biomass of marine microalga, Chlorella vulgaris.     

Changes in photosynthesis of alpine plant <Emphasis Type="Italic">Saussurea superba</Emphasis> during leaf expansion

The native alpine plant Saussurea superba is widely distributed in Qinghai–Tibetan Plateau regions. The leaves of S. superba grow in whorled rosettes, and are horizontally oriented to maximize sunlight exposure. Experiments were conducted in an alpine Kobresia humilis meadow near Haibei Alpine Meadow Ecosystem Research Station (37°29′–37°45′N, 101°12′–101°33′E; alt. 3200 m). Leaf growth, photosynthetic pigments and chlorophyll fluorescence parameters were measured in expanding leaves of S. superba. The results indicate that leaf area increased progressively from inner younger leaves to outside fully expanded ones, and then slightly decreased in nearly senescent leaves, due to early unfavorable environmental conditions, deviating from the ordinary growth pattern. The specific leaf area decreased before leaves were fully expanded, and the leaf thickness was largest in mature leaves. There were no significant changes in the content of chlorophylls (Chl) and carotenoids (Car), but the ratios of Chl a/b and Car/Chl declined after full expansion of the leaves. The variation of Chl a/b coincided well with changes in photochemical quenching (q _P) and the fraction of open PSII reaction centers (q _L). The maximum quantum efficiency of PSII photochemistry after 5 min dark relaxation (F _(v)/F _(m)) continuously increased from younger leaves to fully mature leaves, suggesting that mature leaves could recover more quickly from photoinhibition than younger leaves. The light-harvesting capacity was relatively steady during leaf expansion, as indicated by the maximum quantum efficiency of open PSII centers (\(F_{\text{v}}^{{\prime }}\)/\(F_{\text{m}}^{{\prime }}\)). UV-absorbing compounds could effectively screen harmful solar radiation, and are a main protection way on the photosynthetic apparatus. The decline of q _P and q _L during maturation, together with limitation of quantum efficiency of PSII reaction centers (L _(PFD)), shows a decrease of oxidation state of Q_A in PSII reaction centers under natural sunlight. Furthermore, light-induced (Φ _NPQ) and non-light-induced quenching (Φ _NO) were consistent with variation of L _(PFD). It is concluded that the leaves of S. superba could be classified into four functional groups: young, fully expanded, mature, and senescent. Quick recovery from photoinhibition was correlated with protection by screening pigments, and high level of light energy trapping was correlated with preservation of photosynthetic pigments. Increasing of Φ _NPQ and Φ _NO during leaves maturation indicates that both thermal dissipation of excessive excitation energy in safety and potential threat to photosynthetic apparatus were strengthened due to the declination of q _P and q _L, and enhancement of L _(PFD).     

Concentration of some metals in soil and plant organs and their biochemical profiles in <Emphasis Type="Italic">Tulipa luanica</Emphasis>, <Emphasis Type="Italic">T. kosovarica</Emphasis> and <Emphasis Type="Italic">T. albanica</Emphasis> native plant species

The purpose of this study was to determine the concentration of some metals (Al, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Zn, Ca and Mg) in soil of serpentine and limestone sites, their bioaccumulation and impact on some biochemical parameters in T. luanica, T. kosovarica and T. albanica plants. T. kosovarica and T. albanica grows in serpentine soil, while T. luanica grow in limestone soil. The research showed that concentrations of Cd, Co, Cr, Fe, Mn and Ni were significantly higher at serpentine soil sites in comparison with limestone sites, while concentrations of Pb, Cd, Co and Cr in bulbs, leaves and seeds were under the limit of detection. The concentration of Ni in plant samples of T. kosovarica was significantly higher in comparison with its concentration in T. albanica, but it was under the limit of detection in T. luanica. Moreover, concentrations of Al and Fe in leaves of T. kosovarica and T. albanica were higher in comparison with T. luanica. The concentration of Mg was significantly higher in T. kosovarica and T. albanica than in T. luanica. The δ-aminolevulinic acid dehydratase activity, malondialdehyde and glutathione contents in leaves of T. luanica were higher in comparison with T. kosovarica and T. albanica. In addition, the amounts of total chlorophyll and δ-aminolevulinic acid (ALA) in leaves of T. albanica were higher in comparison with T. kosovarica and T. luanica. Our findings show that target organs of metal accumulation in three Tulip species appears to be leaves?>?seeds?>?bulbs, while the biochemical parameters show that limestone sites represent a less stressful habitat for growing these plant species in comparison with serpentine sites.     

UV-B induces biomass production and nonenzymatic antioxidant compounds in three cyanobacteria

In the present study, the impact of low fluence rate of UV-B (0.045 W.m^?2) on biomass production, photosynthetic pigments (chlorophyll a, carotenoids, and phycobiliproteins), chlorophyll fluorescence, nonenzymatic antioxidants: proline, ascorbate, cysteine, and nonprotein thiols, total phenolic contents, and antioxidant potential (radical scavenging activity) was investigated in three cyanobacteria, viz. Nostoc muscorum, Phormidium foveolarum, and Arthrospira platensis. Selected fluence rate of UV-B caused enhancing effect on these parameters; however, the increased values of these attributes were greater in A. platensis followed by P. foveolarum and N. muscorum. Results indicate that UV-B (at selected fluence rate) could be used as technique that may modify cyanobacterial system for efficient and economic production of natural food supplements and/or natural pharmaceuticals.     

Changes in leaf epicuticular wax,gas exchange and biochemistry metabolism between <Emphasis Type="Italic">Jatropha mollissima</Emphasis> and <Emphasis Type="Italic">Jatropha curcas</Emphasis> under semi-arid conditions

Jatropha curcas and Jatropha mollissima plants were evaluated under conditions of high (HSM) and low (LSM) soil moisture in a semi-arid environment, as changes in the content and concentration of epicuticular wax and the leaf metabolism which could have a relationship with drought tolerance. Besides epicuticular wax, gas exchange, antioxidant system and biochemical parameters of the photosynthetic metabolism were measured. The epicuticular wax content increased only in J. mollissima leaves 95 % under LSM, when compared with HSM conditions. Therefore, J. curcas invested less in the production of long-chain n-alkanes than did J. mollissima under LSM conditions. J. mollissima plants showed the highest CO₂ assimilation rate during the HSM period compared to J. curcas. Both species showed high stability in some leaf biochemistry products, highlighting the highest sugar content, free amino acids, total soluble protein, and photosynthetic pigments in the leaves of J. mollissima plants under both of the soil moisture conditions. Moreover, the stability and performance of the different parameters, such as morphologic variables, seem to allow J. mollissima plants to tolerate semi-arid conditions.     

The responses of an early (<Emphasis Type="Italic">Rhynchospora alba</Emphasis>) and a late (<Emphasis Type="Italic">Molinia japonica</Emphasis>) colonizer to solar radiation in a boreal wetland after peat mining

The responses of seedlings to solar radiation, including ultraviolet (UV), were investigated for Rhynchospora alba, an early colonizer, and Molinia japonica, a late colonizer, in a mined peatland in northern Japan. The solar radiation and rainfall were, respectively, higher and lower in 2008 than in 2009 during the field surveys. The seedlings were transplanted to bare ground, and measurements were made of the biomass, the allocation of biomass to shoots and roots, the absorbance of ultraviolet A and ultraviolet B, and the concentrations of anthocyanin and chlorophyll. R. alba did not change its biomass in response to any solar radiation treatment in 2008 and decreased shoot biomass with low UV and decreased root biomass with shade in 2009. Additionally, M. japonica did not change its biomass in 2008 but decreased its root biomass with low UV in 2009. The chlorophyll concentration of R. alba did not change in 2008 or 2009, whereas the chlorophyll concentration of M. japonica increased with decreased solar radiation, including UV. The UV absorbance of R. alba decreased under shade and with high peat moisture. In contrast, the content of UV-absorbing substances remained unchanged in M. japonica. Therefore, R. alba, the early colonizer, adapted more to strong solar radiation by changing its shoot-root allometry and producing UV-absorbing substances, whereas M. japonica, the late colonizer, tended to respond more to peat moisture. These differing responses to solar radiation and peat moisture may explain the temporal patterns of species replacement from early to late colonizers.     

Accumulation of heavy metals in leaves of submerged hydrophytes (<Emphasis Type="Italic">Elodea canadensis</Emphasis> Michx. and <Emphasis Type="Italic">Potamogeton perfoliatus</Emphasis> L.) and their responses to the effect of the wastewater of a metallurgical plant

The accumulation of heavy metals (Cu, Ni, Zn, Mn, and Fe), parameters of the photosynthetic apparatus, and flavonoid content in leaves of two widespread species of submerged hydrophytes (Elodea canadensis Michx. and Potamogeton perfoliatus L.) from habitats with different levels of pollutions have been studied. The investigations are carried out in the surroundings of the metallurgical plant in the town of Revda, Sverdlovsk oblast, Russia (Revda River, upstream and downstream the confluence of the sewage). It is shown that hydrophytes from polluted habitats (impact site) differ from the plants of a less polluted zone (background site) by a higher accumulation of heavy metals (HMs), greater leaf thickness, and larger cells and chloroplasts. Plants from the polluted site are characterized by a high content of photosynthetic pigments, while flavonoid content in the leaves is lower. The identified trends indicate a high adaptive potential of these species of plants and their ability to inhabit in an aquatic environment polluted with metals and other pollutants.     

Relationship between flavonoids and photoprotection in shade-developed Erigeron breviscapus transferred to sunlight

Flavonoids are thought to participate in protection of the photosynthetic apparatus against photoinhibition under excessive light. Flavone glycoside, scutellarin, is a main active ingredient extracted from Erigeron breviscapus, the plant used in Chinese medicine. Shade-developed leaves of E. breviscapus were transferred from shade to full sunlight to quantify a relationship between the concentration of leaf scutellarin and tolerance to high radiation stress or the recovery from photoinhibition. The maximal quantum yield of PSII photochemistry showed a diurnal fluctuation in both shaded and sunlit leaves throughout the day. It indicated dynamic photoinhibition in the leaves of Erigeron, i.e., higher photoinhibition at solar noon and lower one in the morning and late afternoon. The sun-developed leaves reached the higher scutellarin content and values of nonphotochemical quenching coefficient with a lower degree of photoinhibition than the shade-developed leaves. When the shade-developed leaves were transferred to full sunlight, the content of scutellarin was declining continuously for 10 d and then was increasing for 15 d. After 50 d, all leaves became the sun-developed leaves with their scutellarin contents of about 138.5 ± 5.2 mg g^?1(dry mass, DM) which was significantly higher than that of the shade-developed leaves [107.8 ± 9.8 mg g^?1(DM)]. During acclimatization, the degree of photoinhibition was negatively correlated with the scutellarin content. Our results demonstrated a synchronous fluctuation between the flavonoid content and degree of protection against photoinhibition.     

Genetic analysis and fine-mapping of a new rice mutant, <Emphasis Type="Italic">white and lesion mimic leaf1</Emphasis>

Rice (Oryza sativa L.) leaf color mutants are excellent models for studying chlorophyll biosynthesis and chloroplast development. In this study, we isolated a stable genetic white and lesion mimic leaf1 (wlml1) mutant from an ethyl methanesulfonate (EMS)-mutagenized population of the indica cultivar TN1. Compared with wild-type TN1, the wlml1 mutant had lower contents of chlorophyll and carotenoids, altered chloroplast ultrastructure, and altered regulation of genes associated with chlorophyll metabolism and chloroplast development. In addition, lesions formed on the leaves of wlml1 plants grown at 20 °C and genes related to disease resistance and antioxidant functions were up-regulated; by contrast, the mutant phenotype was partially suppressed at 28 °C. These findings indicated that WLML1 might play a role in chlorophyll metabolism and chloroplast development, as well as in biotic and abiotic stress responses. Genetic analysis showed that WLML1 was controlled by a recessive nuclear gene, and map-based cloning delimited WLML1 to a 159.7-kb region on chromosome 4 that includes 30 putative open reading frames. Based on these findings, the wlml1 mutant will be a good genetic material for further studies on chlorophyll metabolism and stress responses in rice.