Monitoring cashew seedlings during interactions with the fungus <Emphasis Type="Italic">Lasiodiplodia theobromae</Emphasis> using chlorophyll fluorescence imaging

The chlorophyll (Chl) fluorescence imaging technique was applied to cashew seedlings inoculated with the fungus Lasiodiplodia theobromae to assess any disturbances in the photosynthetic apparatus of the plants before the onset of visual symptoms. Two-month-old cashew plants were inoculated with mycelium of L. theobromae isolate Lt19 or Lt32. Dark-adapted and light-acclimated whole plants or previously labelled, single, mature leaf from each plant were evaluated weekly for Chl fluorescence parameters. From 21 to 28 days, inoculation with both isolates resulted in the significantly lower maximal photochemical quantum yield of PSII (F_v/F_m) than those for control samples, decreasing from values of 0.78 to 0.62. In contrast, the time response of the measured fluorescence transient curve from dark-acclimated plants increased in both whole plants and single mature leaves in inoculated plants compared with controls. The F_v/F_m images clearly exhibited photosynthetic perturbations 14 days after inoculation before any visual symptoms appeared. Additionally, decays in the effective quantum yield of PSII photochemistry and photochemical quenching coefficient were also observed over time. However, nonphotochemical quenching increased during the evaluation period. We conclude that F_v/F_m images are the effective way of detecting early metabolic perturbations in the photosynthetic apparatus of cashew seedlings caused by gummosis in both whole plants and single leaves and could be potentially employed in larger-scale screening systems.     

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).     

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.     

Chlorophylls <Emphasis Type="Italic">d</Emphasis> and <Emphasis Type="Italic">f</Emphasis> and their role in primary photosynthetic processes of cyanobacteria

The finding of unique Chl d- and Chl f-containing cyanobacteria in the last decade was a discovery in the area of biology of oxygenic photosynthetic organisms. Chl b, Chl c, and Chl f are considered to be accessory pigments found in antennae systems of photosynthetic organisms. They absorb energy and transfer it to the photosynthetic reaction center (RC), but do not participate in electron transport by the photosynthetic electron transport chain. However, Chl d as well as Chl a can operate not only in the light-harvesting complex, but also in the photosynthetic RC. The long-wavelength (Q_y) Chl d and Chl f absorption band is shifted to longer wavelength (to 750 nm) compared to Chl a, which suggests the possibility for oxygenic photosynthesis in this spectral range. Such expansion of the photosynthetically active light range is important for the survival of cyanobacteria when the intensity of light not exceeding 700 nm is attenuated due to absorption by Chl a and other pigments. At the same time, energy storage efficiency in photosystem 2 for cyanobacteria containing Chl d and Chl f is not lower than that of cyanobacteria containing Chl a. Despite great interest in these unique chlorophylls, many questions related to functioning of such pigments in primary photosynthetic processes are still not elucidated. This review describes the latest advances in the field of Chl d and Chl f research and their role in primary photosynthetic processes of cyanobacteria.     

Effect of gamma radiation on mutant induction of <Emphasis Type="Italic">Fagopyrum dibotrys</Emphasis> Hara

Agronomic traits, photosynthetic pigments, gas exchange, and chlorophyll (Chl) fluorescence parameters of red stem buckwheat (Fagopyrum dibotrys Hara) mutants induced by γ-radiation were compared with green control at seedling stage. Plant height, number of first-class branches, and rhizome biomass were inhibited significantly (p<0.01). Chl a, Chl b, and Chl a+b contents decreased with elevated dose of γ-rays, while increasing carotenoid content indicated that buckwheat was capable of adjusting to the radiation damage. Decrease in net photosynthetic rate was the result of both stomatal and non-stomatal limitations. Fluorescence parameters, such as F₀, F_m, F_v/F_m, F_v/F₀, Φ_PS2, electron transport rate, and photochemical quenching declined significantly (p<0.01) as compared with control due to photoinhibition, while non-photochemical quenching increased to enhance thermal dissipation. Lower parameters implied that leaf tissue was damaged significantly by high dose of γ-radiation and therefore leaf senescence was accelerated.     

Fine mapping a major QTL <Emphasis Type="Italic">qFCC7</Emphasis><Subscript><Emphasis Type="Italic">L</Emphasis></Subscript> for chlorophyll content in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) cv. PA64s

Chlorophyll (Chl) content is an important agronomic trait directly affecting the photosynthetic rate. Using a high-density genetic map of 132 recombinant inbred lines (RILs) derived from the cross between 93-11 and PA64s, we detected the quantitative trait loci (QTLs) for Chl content of the top three leaves under two nitrogen (N) conditions at two developmental stages. A total of 32 main-effect QTLs located on chromosomes 1, 4, 5, 6, 7, 8, and 12 were identified, and these QTLs individually accounted for 6.0–20.8?% of the total phenotypic variation. A major QTL qFCC7 _L affecting the Chl content under low N condition was identified, and its positive allele came from PA64s. This QTL might be associated with the ability to tolerate low-N stress in rice. The chromosomal segment substitution line (CSSL) with the corresponding segment from PA64s had a higher SPAD value and photosynthetic rate than 93-11 and showed a lower specific leaf area (SLA). We performed a fine-mapping using a BC₄F₂ population via marker-assisted backcross and finally mapped this QTL to a 124.5 kb interval on the long arm of chromosome 7. Candidate gene analysis showed that there were sequence variations and expression differences in the predicted candidate gene between the two parents. These results suggest that the QTL qFCC7 _L may be useful for breeding the rice varieties with higher photosynthetic rate and grain yield.     

Physiological and biochemical parameters: new tools to screen barley root exudate allelopathic potential (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L. subsp. <Emphasis Type="Italic">vulgare</Emphasis>)

Morphological markers/traits are often used in the detection of allelopathic stress, but optical signals including chlorophyll a fluorescence emission could be useful in developing new screening techniques. In this context, the allelopathic effect of barley (Hordeum vulgare subsp. vulgare) root exudates (three modern varieties and three landraces) were assessed on the morphological (root and shoot length, biomass accumulation), physiological (F_v/F_m and F₀), and biochemical (chlorophyll and protein contents) variables of great brome (Bromus diandrus Roth., syn. Bromus rigidus Roth. subsp. gussonii Parl.). All the measured traits were affected when great brome was grown in a soil substrate in which barley plants had previously developed for 30 days before being removed. The response of receiver plants was affected by treatment with activated charcoal, dependent on barley genotype and on the nature of the growing substrate. The inhibitory effect was lower with the addition of the activated charcoal suggesting the release of putative allelochemicals from barley roots into the soil. The barley landraces were more toxic than modern varieties and their effect was more pronounced in sandy substrate than in silty clay sand substrate. In our investigation, the chlorophyll content and F_v/F_m were the most correlated variables with barley allelopathic potential. These two parameters might be considered as effective tools to quantify susceptibility to allelochemical inhibitors in higher plants.     

Contrasting patterns of sun-red and shade-green leaves of <Emphasis Type="Italic">Buxus microphylla</Emphasis> in response to gradients of excess light during winter acclimation

Leaf reddening in overwintering evergreens largely restricts their application in landscapes and is generally triggered in response to excess light. To explore how leaves respond to excess light and examine the potential relevance of leaf reddening in this process, a comparative field study was conducted on the sun leaves (SUL), shade leaves (SHL) and three levels of artificially shaded sun leaves (SSUL) of Buxus microphylla ‘Wintergreen’. The seasonal changes in leaf colorations, chlorophyll (Chl) and carotenoid contents, leaf absorbance and chlorophyll fluorescence characteristics were investigated. The results showed that SUL upregulated Chl a/b with increased reductions in Chl b compared with Chl a, accumulated red pigments in the upper palisade mesophyll with reduced absorption in blue and red light but increased absorption in green light, and additionally, significantly downregulated photochemical activities through the sustained enhancement of energy dissipation in PSII antenna (ΦD) from fall to midwinter. In the SSUL, as the light intensity decreased, all of the above processes were mitigated except that the SSUL maintained constant absorptions in blue light region and whose levels were similar to those of the SUL and SHL. In contrast, the SHL maintained relatively high levels of Chl a and Chl b, remained completely green and showed regulated ΦD and ΦE (energy dissipation in PSII reaction centers) to maintain relatively high photochemical activity in the winter. We conclude that the sun leaves downregulate Chl contents to reduce the light absorption and simultaneously enhance sustained ΦD to dissipate most of the light energy, whereas shade leaves maintain relatively high Chl contents and demonstrate regulated proportions of ΦD and ΦE to match the extent to which the absorbed light can be utilized through photochemical reactions. The accumulated red pigments in sun phenotypes may provide a shading effect on Chls by directing energy to non-photosynthetic reaction centers in the blue light region where the absorption is offset by the reduced Chls.     

The analysis of determining factors and evaluation of tolerance to photoinhibition in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Photoinhibition is a significant constraint for improvement of radiation-use efficiency and yield potential in cereal crops. In this work, attached fully expanded leaves of seedlings were used to assay the factors determining photoinhibition and for evaluation of tolerance to photoinhibition in wheat (Triticum aestivum L.). Our results showed that even 1 h under PPFD of 600 µmol(photon) m^?2 s^?1 could significantly reduce maximal quantum yield of PSII photochemistry (F_v/F_m) and performance index (PI) compared to low light [300 µmol(photon) m^?2 s^?1]. The decrease of F_v/F_m and PI was more noticeable with the increase of light intensity; irradiance higher than 800 µmol(photon) m^?2 s^?1 resulted in photoinhibition. Compared to 25°C, lower (20°C) or higher temperature (≥ 35°C) aggravated photoinhibition, while slightly high temperature (28°) alleviated photoinhibition. At 25°C, irradiance of 1,000 µmol(photon) m^–2 s^–1 for 1 h was enough to cause photoinhibition and a significant decrease of F_v/F_m, PI, trapped energy flux, electron transport flux, and density of reaction center as well as increase of dissipated energy flux per cross section were observed. In addition, seedlings at 21–32 days after planting showed a relatively stable phenotype, while the younger or older seedlings indicated an increased susceptibility to photoinhibition, especially in senescing leaves. Finally, six wheat varieties with relative tolerance to photoinhibition were identified from 22 Chinese winter wheat varieties by exposing attached leaves of the 25-d old seedlings for 1 h to 1,000 µmol(photon) m^–2 s^–1 at 25°C. Therefore, our work established a possible method for development of new wheat varieties with enhanced tolerance to photoinhibition.     

The eurythermal adaptivity and temperature tolerance of a newly isolated psychrotolerant Arctic <Emphasis Type="Italic">Chlorella</Emphasis> sp.

A new strain of Chlorella sp. (Chlorella-Arc), isolated from Arctic glacier melt water, was found to have high specific growth rates (μ) between 3 and 27 °C, with a maximum specific growth rate of 0.85 day^?1 at 15 °C, indicating that this strain was a eurythermal strain with a broad temperature tolerance range. To understand its acclimation strategies to low and high temperatures, the physiological and biochemical responses of the Chlorella-Arc to temperature were studied and compared with those of a temperate Chlorella pyrenoidosa strain (Chlorella-Temp). As indicated by declining F _v/F _m, photoinhibition occurred in Chlorella-Arc at low temperature. However, Chlorella-Arc reduced the size of the light-harvesting complex (LHC) to alleviate photoinhibition, as indicated by an increasing Chl a/b ratio with decreasing temperatures. Interestingly, Chlorella-Arc tended to secrete soluble sugar into the culture medium with increasing temperature, while its intracellular soluble sugar content did not vary with temperature changes, indicating that the algal cells might suffer from osmotic stress at high temperature, which could be adjusted by excretion of soluble sugar. Chlorella-Arc accumulated protein and lipids under lower temperatures (<15 °C), and its metabolism switched to synthesis of soluble sugar as temperatures rose. This reflects a flexible ability of Chlorella-Arc to regulate carbon and energy distribution when exposed to wide temperature shifts. More saturated fatty acids (SFA) in Chlorella-Arc than Chlorella-Temp also might serve as the energy source for growth in the cold and contribute to its cold tolerance.     

Comparative analyses of physiological assays and chlorophyll <Emphasis Type="Italic">a</Emphasis> variable fluorescence parameters: investigating the importance of phosphorus availability in oligotrophic and eutrophic freshwater systems

Diverse measurements of nutrient status indicators were used to test the severity of physiological phosphorus (P) limitation of phytoplankton among lake systems ranging from oligotrophic to eutrophic, based on P and chlorophyll a (Chl a) concentrations. Metabolic assays and particulate nutrient ratios were used to estimate nutrient status at sites located in Lake Erie, Lake Ontario and Lake Huron. Variable fluorescence ratios (F _v/F _m), relative electron transport rates and their response to irradiance were measured by the pulse-amplitude-modulated fluorometer. Under summer stratified conditions, P deficiency was strongest in the oligotrophic sites and nitrogen (N) status indicators and Chl a variable parameters revealed no severe N deficiency. Nutrient amendment assays showed positive associations with P additions and Chl a fluorescence parameters at P-deficient sites. In the most oligotrophic sites, N additions revealed a modest increase only detected by the Chl a fluorescence parameters. Phytoplankton communities were also associated with nutrient status, where chrysophytes and cryptophytes were important in P-deficient sites and cyanobacteria, phyrrophyta, and diatoms were prevalent in nutrient-rich sites. The results confirmed that Chl a fluorescence parameters can reveal P deficiency and indicate its severity among the range of trophic status in aquatic systems.     

Amino acid composition of leaf,grain and bracts of japonica rice (<Emphasis Type="Italic">Oryza Sativa</Emphasis> ssp. <Emphasis Type="Italic">japonica</Emphasis>) and its response to nitrogen fertilization

Heat stress is one of the main abiotic stresses that limit plant growth. The effects of high temperature on oxidative damage, PSII activity and D1 protein turnover were studied in three wheat varieties with different heat susceptibility (CS, YN949 and AK58). The results showed that heat stress induced lower lipid peroxidation in AK58 and YN949 than CS, which was related to different changes of SOD, CAT, POD and H₂O₂. Similarly, AK58 and YN949 performed better PSII photochemical efficiency (F_v/F_m, ΦPSII and ETR) under high temperature, which was attributed to rapid synthesis and degradation of D1 protein. Moreover, higher expression of D1 protein turnover-related genes (PsbA, STN8, PBCP, Deg1, Deg2, Deg5, Deg8, FtsH1/5 and FtsH2/8) and SOD activity in AK58 and YN949 under normal conditions also established a basis for acclimatizing high temperatures, thereby alleviating PSII photoinhibition and reducing oxidative damage when exposed to heat stress.     

Exogenous proline induces soluble sugar accumulation and alleviates drought stress effects on photosystem II functioning of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> leaves

The effects of exogenous applied proline (Pro), on photosystem II (PSII) photochemistry of drought stressed (DS) 4-week old Arabidopsis thaliana plants, was studied by using chlorophyll (chl) fluorescence imaging. The maximum quantum yield of PSII photochemistry (F _v /F _m) in DS plants decreased significantly to 77% of that of the control value, suggesting that DS plants could not maintain PSII function, possibly due to accelerated photoinhibition of PSII. Free Pro and total soluble sugars (SS) increased, in response to DS. Exogenous foliar application of Pro by spraying, led to a remarkable increase in the accumulation of Pro and surprisingly also of SS. Both of them served to scavenge reactive oxygen species (ROS), as it was evident by the decreased lipid peroxidation level measured as malondialdehyde (MDA). DS plants sprayed with Pro showed a tolerance to photoinhibition, this indicated by F _v/F _m being close to values typical of healthy leaves by maintaining more than 98% of PSII function. Also the higher quantum efficiency of PSII photochemistry (Φ _PSΙΙ ) and the decreased excitation pressure (1 ? q _p ) recorded for stressed leaves with Pro, lead us to conclude that Pro appears to be involved in the protection of chloroplast structures by quenching ROS. The enhanced dissipation of excess light energy of PSII, in part accounts for the observed increased resistance to DS in A. thaliana leaves with Pro. Our data pointed out that Pro signalling interacts with SS signaling pathway and provided a new insight in Pro metabolism.     

Variations in light energy dissipation in <Emphasis Type="Italic">Woodfordia fruticosa</Emphasis> leaves during expansion

Young leaves of tropical trees frequently appear red in color, with the redness disappearing as the leaves mature. During leaf expansion, plants may employ photoprotective mechanisms to cope with high light intensities; however, the variations in anthocyanin contents, nonphotochemical quenching (NPQ), and photorespiration during leaf expansion are poorly understood. Here, we investigated pigment contents, gas exchange, and chlorophyll (Chl) fluorescence in Woodfordia fruticosa leaves during their expansion. Young red leaves had significantly lower Chl content than that of expanding or mature leaves, but they accumulated significantly higher anthocyanins and dissipated more excited light energy through NPQ. As the leaves matured, net photosynthetic rate, total electron flow through PSII, and electron flow for ribulose-1,5-bisphosphate oxygenation gradually increased. Our results provided evidence that photorespiration is of fundamental importance in regulating the photosynthetic electron flow and CO₂ assimilation during leaf expansion.     

Intrapopulation heterogeneity of the fluorescence parameters of the marine plankton alga <Emphasis Type="Italic">Thalassiosira weissflogii</Emphasis> at various nitrogen levels

Fluorescence of the marine alga Thalassiosira weissflogii (Grunow) Fryxell et Hasle with open (F _o ) and closed (F _m ) reaction centers of photosystem 2 (PS 2) and its relative variable fluorescence (F _v/F _m ) were measured at various levels of inorganic nitrogen. A significant heterogeneity of the population in terms of these parameters was revealed. Some cells within the population were more sensitive to nitrogen deficiency, and their photosynthetic apparatus was disrupted to a greater extent. The cells within a population also differed in terms of their ability to recover after incubation at low nitrogen levels. Enhancement of nitrogen deficiency resulted in an increase in the variability of the F _o and F _v/F _m values of the cells. Fluorescence variability decreased at a less pronounced deficiency. Fluorescence variability should be taken into consideration in the studies concerning responses of algae to changes in nutrient contents.     

Gas exchanges of an endangered species <Emphasis Type="Italic">Syringa pinnatifolia</Emphasis> and a widespread congener <Emphasis Type="Italic">S. oblata</Emphasis>

Net photosynthetic rate (P_N), transpiration rate (E), water use efficiency (WUE), stomatal conductance (g_s), and stomatal limitation (L_s) were investigated in two Syringa species. The saturation irradiance (SI) was 400 µmol m^-2s^-1 for S. pinnatifolia and 1 700 µmol m^-2s^-1 for S. oblata. Compared with S. oblata, S. pinnatifolia had extremely low g _s . Unlike S. oblata, the maximal photosynthetic rate (P_max) in S. pinnatifoliaoccurred around 08:00 and then fell down, indicating this species was sensitive to higher temperature and high photosynthetic photon flux density. However, such phenomenon was interrupted by the leaf development rhythms before summer. A relatively lower P_N together with a lower leaf area and shoot growth showed the capacity for carbon assimilation was poorer in S. pinnatifolia.     

Inhibition of root respiration induces leaf senescence in <Emphasis Type="Italic">Alhagi sparsifolia</Emphasis>

Leaf senescence can be induced by numerous factors. In order to explore the relationship between root respiration and leaf senescence, we utilized different types of phloem girdling to control the root respiration of Alhagi sparsifolia and its physiological response. Our results showed that both girdling and inhibition of root respiration led to a decline of stomatal conductance, photosynthesis, transpiration rate, chlorophyll (Chl) a, Chl b, carotenoid (Car) content, Chl a/b, Chl/Car, water potential, and Chl a fluorescence, as well as to an increase of abscisic acid (ABA), proline, and malondialdehyde content in leaves and to upregulation of senescence-associated gene expression. Our present work implied that both inhibition of root respiration and girdling can induce leaf senescence. In comparison with phloem girdling, the leaf senescence caused by inhibition of root respiration was less significant. The reason for girdling-induced senescence was ABA and carbohydrate accumulation. Senescence induced by inhibition of root respiration occurred due to leaf water stress resulting from inhibition of water absorption.     

A rice <Emphasis Type="Italic">White-stripe leaf3</Emphasis> (<Emphasis Type="Italic">wsl3</Emphasis>) mutant lacking an HD domain-containing protein affects chlorophyll biosynthesis and chloroplast development

Leaf-color mutants are ideal genetic materials for understanding the mechanism of chloroplast development and chlorophyll (Chl) biosynthesis. Here we isolated and identified a new leaf-color mutant of rice, named white-stripe leaf3 (wsl3), from a ⁶⁰Co-irradiated mutant pool. The wsl3 mutant displayed a visible white-stripe leaf in both young seedlings and flag leaves of mature plant. Chl content in homozygous wsl3 mutant was approximately 47% of that in the wild type. Besides, chloroplast development in the mutant was severely arrested. By a map-based cloning strategy, the wsl3 gene was finely confined to a 50.8 kb region on chromosome 1. Moreover, a 9-bp deletion was identified in the genomic region of LOC_Os01g01920, which encodes an HD (histidine and aspartic acid) domaincontaining protein. Genetic complementation confirmed that LOC_Os01g01920 could recover the lesion of wsl3 mutation. Real-time PCR analyses showed that the expression levels of WSL3 were the highest in young and flag leaves among various tissues, and most of the genes associated with Chl biosynthesis were significantly down-regulated in the wsl3 mutant. Meanwhile, in contrast to many nuclear gene-encoded phage-type RNA polymerase(s) (NEP) transcribed genes were up-regulated, most of plastid-encoded bacterialtype RNA polymerase (PEP) transcribed genes were downregulated. These results demonstrated that the WSL3 gene, as an HD domain-containing protein, is involved in chl biosynthesis and chloroplast development in rice.