Integrating Physiological Features and Proteomic Analyses Provides New Insights in Blue/Red Light-Treated Moso Bamboo (Phyllostachys edulis).

Bamboo is one of the most important nontimber forestry products in the world. Light is not only the most critical source of energy for plant photosynthesis but also involved in regulating the biological processes of plants. However, there are few reports on how blue/red light affects Moso bamboo. This study investigated the growth status and physiological responses of Moso bamboo (Phyllostachys edulis) to blue/red light treatments. The growth status of the bamboo plants was evaluated, revealing that both blue- and red-light treatments promoted plant height and overall growth. Gas exchange parameters, chlorophyll fluorescence, and enzyme activity were measured to assess the photosystem response of Moso bamboo to light treatments. Additionally, the blue light treatment led to a higher chlorophyll content and enzyme activities compared to the red light treatment. A tandem mass tag quantitative proteomics approach identified significant changes in protein abundance under different light conditions with specific response proteins associated with distinct pathways, such as photosynthesis and starch metabolism. Overall, this study provides valuable insights into the physiological and proteomic responses of Moso bamboo to blue/red light treatments, highlighting their potential impact on growth and development.

Poaceae plants transfer cyclobutane pyrimidine dimer photolyase to chloroplasts for ultraviolet-B resistance.

Photoreactivation enzyme that repairs cyclobutane pyrimidine dimer (CPD) induced by ultraviolet-B radiation, commonly called CPD photolyase (PHR) is essential for plants living under sunlight. Rice (Oryza sativa) PHR (OsPHR) is a unique triple-targeting protein. The signal sequences required for its translocation to the nucleus or mitochondria are located in the C-terminal region but have yet to be identified for chloroplasts. Here, we identified sequences located in the N-terminal region, including the serine-phosphorylation site at position 7 of OsPHR, and found that OsPHR is transported/localized to chloroplasts via a vesicle transport system under the control of serine-phosphorylation. However, the sequence identified in this study is only conserved in some Poaceae species, and in many other plants, PHR is not localized to the chloroplasts. Therefore, we reasoned that Poaceae species need the ability to repair CPD in the chloroplast genome to survive under sunlight and have uniquely acquired this mechanism for PHR chloroplast translocation.

Analysis of a radiation-induced dwarf mutant of a warm-season turf grass reveals potential mechanisms involved in the dwarfing mutant.

Zoysia matrella [L.] Merr. is a widely cultivated warm-season turf grass in subtropical and tropical areas. Dwarf varieties of Z. matrella are attractive to growers because they often reduce lawn mowing frequencies. In this study, we describe a dwarf mutant of Z. matrella induced from the 60Co-γ-irradiated calluses. We conducted morphological test and physiological, biochemical and transcriptional analyses to reveal the dwarfing mechanism in the mutant. Phenotypically, the dwarf mutant showed shorter stems, wider leaves, lower canopy height, and a darker green color than the wild type (WT) control under the greenhouse conditions. Physiologically, we found that the phenotypic changes of the dwarf mutant were associated with the physiological responses in catalase, guaiacol peroxidase, superoxide dismutase, soluble protein, lignin, chlorophyll, and electric conductivity. Of the four endogenous hormones measured in leaves, both indole-3-acetic acid and abscisic acid contents were decreased in the mutant, whereas the contents of gibberellin and brassinosteroid showed no difference between the mutant and the WT control. A transcriptomic comparison between the dwarf mutant and the WT leaves revealed 360 differentially-expressed genes (DEGs), including 62 up-regulated and 298 down-regulated unigenes. The major DEGs related to auxin transportation (e.g., PIN-FORMED1) and cell wall development (i.e., CELLULOSE SYNTHASE1) and expansin homologous genes were all down-regulated, indicating their potential contribution to the phenotypic changes observed in the dwarf mutant. Overall, the results provide information to facilitate a better understanding of the dwarfing mechanism in grasses at physiological and transcript levels. In addition, the results suggest that manipulation of auxin biosynthetic pathway genes can be an effective approach for dwarfing breeding of turf grasses.

The transcriptomic responses of C4 grasses to subambient CO2 and low light are largely species specific and only refined by photosynthetic subtype.

Three subtypes of C4 photosynthesis exist (NADP-ME, NAD-ME and PEPCK), each known to be beneficial under specific environmental conditions. However, the influence of photosynthetic subtype on transcriptomic plasticity, as well as the genes underpinning this variability, remain largely unknown. Here, we comprehensively investigate the responses of six C4 grass species, spanning all three C4 subtypes, to two controlled environmental stresses: low light (200 µmol m-2  sec-1 ) and glacial CO2 (subambient; 180 ppm). We identify a susceptibility within NADP-ME species to glacial CO2 . Notably, although glacial CO2 phenotypes could be tied to C4 subtype, biochemical and transcriptomic responses to glacial CO2 were largely species specific. Nevertheless, we were able to identify subtype specific subsets of significantly differentially expressed transcripts which link resource acquisition and allocation to NADP-ME species susceptibility to glacial CO2 . Here, low light phenotypes were comparable across species with no clear subtype response, while again, transcriptomic responses to low light were largely species specific. However, numerous functional similarities were noted within the transcriptomic responses to low light, suggesting these responses are functionally relatively conserved. Additionally, PEPCK species exhibited heightened regulation of transcripts related to metabolism in response to both stresses, likely tied to their C4 metabolic pathway. These results highlight the influence that both species and subtype can have on plant responses to abiotic stress, building on our mechanistic understanding of acclimation within C4 grasses and highlighting avenues for future crop improvements.

A rapid monitoring of NDVI across the wheat growth cycle for grain yield prediction using a multi-spectral UAV platform.

Wheat improvement programs require rapid assessment of large numbers of individual plots across multiple environments. Vegetation indices (VIs) that are mainly associated with yield and yield-related physiological traits, and rapid evaluation of canopy normalized difference vegetation index (NDVI) can assist in-season selection. Multi-spectral imagery using unmanned aerial vehicles (UAV) can readily assess the VIs traits at various crop growth stages. Thirty-two wheat cultivars and breeding lines grown in limited irrigation and full irrigation treatments were investigated to monitor NDVI across the growth cycle using a Sequoia sensor mounted on a UAV. Significant correlations ranging from R2 = 0.38 to 0.90 were observed between NDVI detected from UAV and Greenseeker (GS) during stem elongation (SE) to late grain gilling (LGF) across the treatments. UAV-NDVI also had high heritabilities at SE (h2 = 0.91), flowering (F)(h2 = 0.95), EGF (h2 = 0.79) and mid grain filling (MGF) (h2 = 0.71) under the full irrigation treatment, and at booting (B) (h2 = 0.89), EGF (h2 = 0.75) in the limited irrigation treatment. UAV-NDVI explained significant variation in grain yield (GY) at EGF (R2 = 0.86), MGF (R2 = 0.83) and LGF (R2 = 0.89) stages, and results were consistent with GS-NDVI. Higher correlations between UAV-NDVI and GY were observed under full irrigation at three different grain-filling stages (R2 = 0.40, 0.49 and 0.45) than the limited irrigation treatment (R2 = 0.08, 0.12 and 0.14) and GY was calculated to be 24.4% lower under limited irrigation conditions. Pearson correlations between UAV-NDVI and GY were also low ranging from r = 0.29 to 0.37 during grain-filling under limited irrigation but higher than GS-NDVI data. A similar pattern was observed for normalized difference red-edge (NDRE) and normalized green red difference index (NGRDI) when correlated with GY. Fresh biomass estimated at late flowering stage had significant correlations of r = 0.30 to 0.51 with UAV-NDVI at EGF. Some genotypes Nongda 211, Nongda 5181, Zhongmai 175 and Zhongmai 12 were identified as high yielding genotypes using NDVI during grain-filling. In conclusion, a multispectral sensor mounted on a UAV is a reliable high-throughput platform for NDVI measurement to predict biomass and GY and grain-filling stage seems the best period for selection.

Reactive oxygen species trigger the fast action of glufosinate.

MAIN CONCLUSION: Glufosinate is primarily toxic to plants due to a massive light-dependent generation of reactive oxygen species rather than ammonia accumulation or carbon assimilation inhibition. Glutamine synthetase (GS) plays a key role in plant nitrogen metabolism and photorespiration. Glufosinate (C5H12NO4P) targets GS and causes catastrophic consequences leading to rapid plant cell death, and the causes for phytoxicity have been attributed to ammonia accumulation and carbon assimilation restriction. This study aimed to examine the biochemical and physiological consequences of GS inhibition to identify the actual cause for rapid phytotoxicity. Monocotyledonous and dicotyledonous species with different forms of carbon assimilation (C3 versus C4) were selected as model plants. Glufosinate sensitivity was proportional to the uptake of herbicide between species. Herbicide uptake also correlated with the level of GS inhibition and ammonia accumulation in planta even with all species having the same levels of enzyme sensitivity in vitro. Depletion of both glutamine and glutamate occurred in glufosinate-treated leaves; however, amino acid starvation would be expected to cause a slow plant response. Ammonia accumulation in response to GS inhibition, often reported as the driver of glufosinate phytotoxicity, occurred in all species, but did not correlate with either reductions in carbon assimilation or cell death. This is supported by the fact that plants can accumulate high levels of ammonia but show low inhibition of carbon assimilation and absence of phytotoxicity. Glufosinate-treated plants showed a massive light-dependent generation of reactive oxygen species, followed by malondialdehyde accumulation. Consequently, we propose that glufosinate is toxic to plants not because of ammonia accumulation nor carbon assimilation inhibition, but the production of reactive oxygen species driving the catastrophic lipid peroxidation of the cell membranes and rapid cell death.

Effects of temperature and water availability on light energy utilization in photosynthetic processes of Deschampsia antarctica.

Regional climate change in Antarctica would favor the carbon assimilation of Antarctic vascular plants, since rising temperatures are approaching their photosynthetic optimum (10-19°C). This could be detrimental for photoprotection mechanisms, mainly those associated with thermal dissipation, making plants more susceptible to eventual drought predicted by climate change models. With the purpose to study the effect of temperature and water availability on light energy utilization and putative adjustments in photoprotective mechanisms of Deschampsia antarctica Desv., plants were collected from two Antarctic provenances: King George Island and Lagotellerie Island. Plants were cultivated at 5, 10 and 16°C under well-watered (WW) and water-deficit (WD, at 35% of the field capacity) conditions. Chlorophyll fluorescence, pigment content and de-epoxidation state were evaluated. Regardless of provenances, D. antarctica showed similar morphological, biochemical and functional responses to growth temperature. Higher temperature triggered an increase in photochemical activity (i.e. electron transport rate and photochemical quenching), and a decrease in thermal dissipation capacity (i.e. lower xanthophyll pool, Chl a/b and ß carotene/neoxanthin ratios). Leaf mass per unit area was reduced at higher temperature, and was only affected in plants exposed to WD at 16°C and exhibiting lower electron transport rate and amount of chlorophylls. D. antarctica is adapted to frequent freezing events, which may induce a form of physiological water stress. Photoprotective responses observed under WD contribute to maintain a stable photochemical activity. Thus, it is possible that short-term temperature increases could favor the photochemical activity of this species. However, long-term effects will depend on the magnitude of changes and the plant's ability to adjust to new growth temperature.

Heteropogon contortus BL-1 (Pilli Grass) and Elevated UV-B Radiation: The Role of Growth, Physiological, and Biochemical Traits in Determining Forage Productivity and Quality.

Exposure to elevated UV-B (eUV-B) is well known for its phytotoxicity, although studies made with UV-B exposure and its impact on grasses are limited especially from tropical countries including India. In this study, responses of a valuable grass species, Heteropogon contortus BL-1, were assessed under eUV-B (over ambient UV-B) at different growth stages. Damage caused by eUV-B was observed in the form of membrane damage and loss of pigments at early stages of growth, whereas tannin, phenol, and protein contents showed their increments at all the growth stages, to overcome the imposed stress. Reducing sugar showed its decline at all the growth stages, whereas starch and sucrose contents were higher mostly at later ages of plant growth. eUV-B caused a marked variations in anatomical structures with increase of mesophyll and spongy parenchymatous cells in leaves to reduce severity of irradiation, to maintain the growth and productivity. The study also highlights the significant negative influence of eUV-B on the growth of H. contortus BL-1 and its adaptive strategy to minimize the negative impacts. With the progression of age, plants although adopted to UV-B stress with maintenance of productivity, but palatability of forage was affected due to increment of tannin content.

Optical properties of straw-derived dissolved organic matter and growth inhibition of Microcystis aeruginosa by straw-derived dissolved organic matter via photo-generated hydrogen peroxide.

Recent advances in research on algae inhibition by using low-cost straw proposed a possible mechanism that reactive oxygen species (ROS) generated by the solar irradiation of straw-derived dissolved organic matter (DOM) might contribute to cyanobacteria inhibition. However, this process is not clearly understood. Here, DOM from three types of straw (barley, rice, and wheat) and natural organic matter (NOM) isolates were investigated in terms of their photochemical properties and ROS generating abilities. Results demonstrated that the DOM derived from the aeration decomposition of barley straw (A-DOMbs) yielded the best formation efficiencies of hydrogen peroxide (H2O2) and hydroxyl radicals (•OH) under solar-simulated irradiation in all organic matter samples. Correlation analysis implies that optical parameters and phenolic hydroxyl group contents can signify ROS generating abilities of different DOM solutions. Bioassay results show that A-DOMbs possesses the highest inhibition performance for M. aeruginosa in all DOM samples, much higher than those of NOM isolates. The addition of catalase greatly relieves the inhibition performance, making the loss of chlorophyll a content decreased from 37.14% to 7.83% in 2 h for A-DOMbs, which implies that for cyanobacteria growth inhibition, photochemically-produced H2O2 from SOM is far more important than singlet oxygen (1O2), •OH, and even SOM itself. Our results show that H2O2 photochemically generated from straw-derived DOM is able to result in rapid inhibition of M. aeruginosa in a relatively short period, furthering the understanding of complicated mechanisms of cyanobacteria inhibition by using low-cost straw in eutrophic waters.

Germination ecology of Chloris truncata and its implication for weed management.

Chloris truncata is a significant weed in summer crops in the subtropical region of Australia. A study was conducted to evaluate the effect of environmental factors on germination and emergence of two populations of C. truncata. Overall, germination was not affected by the populations. Seeds germinated at a wide range of alternating day/night temperatures, suggesting that seeds can germinate throughout the spring, winter and autumn seasons. Seed germination was stimulated by the presence of light; however, 51 to 71% of these seeds still germinated in the dark. The sodium chloride concentration and osmotic potential required to inhibit germination of 50% of the population were 179 mM and -0.52 MPa, respectively. A high proportion of seeds germinated over a wide pH range (4 to 10). Seeds placed on the soil surface had greatest germination (67%) and a burial depth of 3 cm resulted in complete inhibition of emergence. The sorghum residue amount required to reduce emergence by 50% was 1.8 t ha-1. The results suggest that, although this weed will be favored in no-till systems, residue retention on the soil surface will help in reducing its infestation. Seed bank buildup can be managed by burying seeds below the depth of emergence.

Inter- and intraspecific variability in physiological traits and post-anoxia recovery of photosynthetic efficiency in grasses under oxygen deprivation.

Low oxygen conditions occur in grass sites due to high and frequent precipitation, poor soil quality, and over-irrigation followed by slow drainage. Three warm-season and one cool-season grass were analyzed at metabolic level during a time-course experiment performed in a controlled anoxic environment. Prolonged oxygen depletion proved detrimental by leading to premature death to all the species, with the exception of seashore paspalum. Moreover, the anoxia tolerance observed in these grasses has been associated with slow use of carbohydrates, rather than with their relative abundance, which was more important than their antioxidant capacity. Further physiological characterization of eight seashore paspalum genotypes to anoxia was also performed, by examining the variation in photosystem II (PSII) efficiency and gas exchange during post-anoxia recovery. Multivariate analysis highlighted the presence of three main clusters of seashore paspalum genotypes, characterized by different ability to restore the PSII photochemistry during recovery after one day of anoxia. Taken together, our data demonstrate that the analysis of post-anoxia recovery of fluorescence and gas exchange parameters can represent a fast and reliable indicator for selecting species and cultivars more able to acclimate their photosynthetic apparatus.

Increased soil nutrition and decreased light intensity drive species loss after eight years grassland enclosures.

Enclosures (fenced, grazing or clipping) within a certain period of years are the most common tools for restoration of degraded grasslands in temperate regions. Short-term enclosures can improve biodiversity and productivity by effectively relieving grazing pressure, while long-term enclosures can reduce species diversity. We therefore carried out a field experiment to investigate the specific causes of the reduced species diversity in Hulunbeier grassland of northern China. After eight years of enclosure, the significantly increased soil available nitrogen (AN) and available phosphorus (AvP) in enclosure community reduced nitrogen (N) limitation but most vegetation was still N limited. Many environmental factors led to decreased species richness, but increased soil AN and decreased light intensity at the community bottom were the most significant ones. Community density decreased independently of soil nutrition but significantly related to decreased species richness. Density of dominant canopy species increased, while dominant understory species decreased during assemblage-level thinning; therefore, the random-loss hypothesis was not supported. The dominant understory species responded to lower light availability by increasing their height, leaf area, and chlorophyll content. Moreover, our results were expected to provide some specific guidance for the restoration mode selection of degraded grasslands in northern China.

Addition of multiple limiting resources reduces grassland diversity.

Niche dimensionality provides a general theoretical explanation for biodiversity-more niches, defined by more limiting factors, allow for more ways that species can coexist. Because plant species compete for the same set of limiting resources, theory predicts that addition of a limiting resource eliminates potential trade-offs, reducing the number of species that can coexist. Multiple nutrient limitation of plant production is common and therefore fertilization may reduce diversity by reducing the number or dimensionality of belowground limiting factors. At the same time, nutrient addition, by increasing biomass, should ultimately shift competition from belowground nutrients towards a one-dimensional competitive trade-off for light. Here we show that plant species diversity decreased when a greater number of limiting nutrients were added across 45 grassland sites from a multi-continent experimental network. The number of added nutrients predicted diversity loss, even after controlling for effects of plant biomass, and even where biomass production was not nutrient-limited. We found that elevated resource supply reduced niche dimensionality and diversity and increased both productivity and compositional turnover. Our results point to the importance of understanding dimensionality in ecological systems that are undergoing diversity loss in response to multiple global change factors.

A self-photoprotection mechanism helps Stipa baicalensis adapt to future climate change.

We examined the photosynthetic responses of Stipa baicalensis to relative long-term exposure (42 days) to the predicted elevated temperature and water availability changes to determine the mechanisms through which the plant would acclimate to future climate change. Two thermal regimes (ambient and +4 °C) and three irrigation levels (partial, normal and excess) were used in environmental control chambers. The gas exchange parameters, light response curves and A/Ci curves were determined. The elevated temperature and partial irrigation reduced the net photosynthetic rate due to a limitation in the photosynthetic capacity instead of the intercellular CO2 concentration. Partial irrigation decreased Rubisco activation and limited RuBP regeneration. The reduction in Vcmax increased with increasing temperature. Excess irrigation offset the negative effect of drought and led to a partial recovery of the photosynthetic capacity. Although its light use efficiency was restricted, the use of light and dark respiration by Stipa baicalensis was unchanged. We concluded that nonstomatal limitation was the primary reason for photosynthesis regulation in Stipa baicalensis under relative long-term climate change conditions. Although climate change caused reductions in the light use efficiency and photosynthetic rate, a self-photoprotection mechanism in Stipa baicalensis resulted in its high ability to maintain normal live activities.

Transcriptome and comparative gene expression analysis of Phyllostachys edulis in response to high light.

BACKGROUND: Photosynthesis plays a vital role as an energy source for plant metabolism, and its efficiency may be drastically reduced owing to abiotic stresses. Moso bamboo (Phyllostachys edulis), is a renewable and versatile resource with significant ecological and economic value, which encounters high light stress with large amplitude in natural environment. However, the gene expression profiles in response to high light were elusive in bamboo. RESULTS: We firstly performed physiological experiments on moso bamboo leaves treated with high light (1200 µmol · m(-2) · s(-1)). Based on the physiological results, three samples of leaves treated with high light for 0 h (CK), 0.5 h (0.5H), and 8 h (8H) were selected to perform further high-throughput RNA sequencing (RNA-Seq), respectively. Then, the transcriptomic result demonstrated that the most genes were expressed at a statistically significant value (FPKM ≥ 1) and the RNA-Seq data were validated via quantitative real time PCR. Moreover, some significant gene expression changes were detected. For instance, 154 differentially expressed genes were detected in 0.5H vs. CK, those in 8H vs. CK were 710, and 429 differentially expressed genes were also identified in 0.5H vs.8 H. Besides, 47 gene annotations closely related to photosynthesis were refined, including 35 genes annotated as light-harvesting chlorophyll a/b-binding (LHC) proteins, 9 LHC-like proteins and 3 PsbSs. Furthermore, the pathway of reactive oxygen species (ROS) in photosynthesis was further analyzed. A total of 171 genes associated with ROS-scavenging were identified. Some up-regulated transcript factors, such as NAC, WRKY, AR2/ERF, and bHLH, mainly concentrated in short-term response, while C2H2, HSF, bZIP, and MYB were largely involved in short and middle terms response to high light. CONCLUSION: Based on the gene expression analysis of moso bamboo in response to high light, we thus identified the global gene expression patterns, refined the annotations of LHC protein, LHC-like protein and PsbS, detected the pathway of ROS as well as identified ROS-scavenging genes and transcription factors in the regulation of photosynthetic and related metabolisms. These findings maybe provide a starting point to interpret the molecular mechanism of photosynthesis in moso bamboo under high light stress.

Use of exotic plants to control Spartina alterniflora invasion and promote mangrove restoration.

In coastal China, the exotic invasive Spartina alterniflora is preventing the establishment of native mangroves. The use of exotic species, control of exotic plant invasion, and restoration of native plant communities are timely research issues. We used exotic Sonneratia apetala Buch.-Ham and S. caseolaris (L.) Engl. to control invasive Spartina alterniflora Loisel through replacement control for five years, which concurrently promoted the restoration of native mangroves. This process includes three stages. I: In a mangrove area invaded by S. alterniflora, exotic S. apetala and S. caseolaris grew rapidly due to their relatively fast-growing character and an allelopathic effect. II: Fast-growing S. apetala and S. caseolaris eradicate S. alterniflora through shading and allelopathy. III: The growth of native mangrove was promoted because exotic plant seedlings cannot regenerate in the understory shade, whereas native mesophytic mangrove plants seedlings can grow; when the area experiences extreme low temperatures in winter or at other times, S. apetala dies, and native mangrove species grow to restore the communities. This model has important implications for addressing the worldwide problems of "how to implement the ecological control of invasion using exotic species" and "how to concurrently promote native community restoration during the control of exotic invasion".

Physical mapping of chromosome 4J of Thinopyrum bessarabicum using gamma radiation-induced aberrations.

KEY MESSAGE: Gamma radiation induced a series of structural aberrations involving Thinopyrum bessarabicum chromosome 4J. The aberrations allowed for deletion mapping of 101 4J-specific markers and fine mapping of blue-grained gene BaThb. Irradiation can induce translocations and deletions to assist physically locating genes and markers on chromosomes. In this study, a 12-Gy dosage of (60)Co-γ was applied to pollen and eggs of a wheat (Triticum aestivum) landrace Chinese Spring (CS)-Thinopyrum bessarabicum chromosome 4J disomic addition line (DA4J), and the gametes from irradiated plants were fertilized with normal CS eggs or pollen to produce M1 seeds. Based on genomic in situ hybridization analysis of 261 M1 plants, we identified 74 lines carrying structural aberrations involving chromosome 4J with the higher aberration rate in treated pollen (31.2 %) than in the treated eggs (21.3 %). We further identified 43 (53.8 %) lines with structural aberrations on chromosome 4J by analyzing another 80 M1 plants with 74 4J-specific markers, indicating that combining molecular and cytological methods was more efficient for detecting chromosome aberrations. Marker analysis thus was performed prior to cytogenetic identification on M2-M4 seeds to detect chromosome structural aberrations. Sixty-eight M3 lines with structural aberrations on chromosome 4J and six previously obtained chromosome 4J alien lines were then analyzed using 101 chromosome 4J-specific markers. After combining marker results with chromosome aberrations in each line, chromosome 4J was physically divided into 24 segmental blocks with 7 in the short arm and 17 in the long arm. The blue-grained gene BaThb was further mapped into the region corresponding to block 4JL-11. The chromosome aberrations and the physical map developed in this research provide useful stocks and tools for introgression of genes on chromosome 4J into wheat.

A high throughput gas exchange screen for determining rates of photorespiration or regulation of C4 activity.

Large-scale research programmes seeking to characterize the C4 pathway have a requirement for a simple, high throughput screen that quantifies photorespiratory activity in C3 and C4 model systems. At present, approaches rely on model-fitting to assimilatory responses (A/C i curves, PSII quantum yield) or real-time carbon isotope discrimination, which are complicated and time-consuming. Here we present a method, and the associated theory, to determine the effectiveness of the C4 carboxylation, carbon concentration mechanism (CCM) by assessing the responsiveness of V O/V C, the ratio of RuBisCO oxygenase to carboxylase activity, upon transfer to low O2. This determination compares concurrent gas exchange and pulse-modulated chlorophyll fluorescence under ambient and low O2, using widely available equipment. Run time for the procedure can take as little as 6 minutes if plants are pre-adapted. The responsiveness of V O/V C is derived for typical C3 (tobacco, rice, wheat) and C4 (maize, Miscanthus, cleome) plants, and compared with full C3 and C4 model systems. We also undertake sensitivity analyses to determine the impact of R LIGHT (respiration in the light) and the effectiveness of the light saturating pulse used by fluorescence systems. The results show that the method can readily resolve variations in photorespiratory activity between C3 and C4 plants and could be used to rapidly screen large numbers of mutants or transformants in high throughput studies.

Transcriptional responses indicate maintenance of photosynthetic proteins as key to the exceptional chilling tolerance of C4 photosynthesis in Miscanthus × giganteus.

Miscanthus × giganteus is exceptional among C4 plants in its ability to acclimate to chilling (≤14 °C) and maintain a high photosynthetic capacity, in sharp contrast to maize, leading to very high productivity even in cool temperate climates. To identify the mechanisms that underlie this acclimation, RNA was isolated from M × giganteus leaves in chilling and nonchilling conditions and hybridized to microarrays developed for its close relative Zea mays. Among 21 000 array probes that yielded robust signals, 723 showed significant expression change under chilling. Approximately half of these were for annotated genes. Thirty genes associated with chloroplast membrane function were all upregulated. Increases in transcripts for the lhcb5 (chlorophyll a/b-binding protein CP26), ndhF (NADH dehydrogenase F, chloroplast), atpA (ATP synthase alpha subunit), psbA (D1), petA (cytochrome f), and lhcb4 (chlorophyll a/b-binding protein CP29), relative to housekeeping genes in M. × giganteus, were confirmed by quantitative reverse-transcription PCR. In contrast, psbo1, lhcb5, psbA, and lhcb4 were all significantly decreased in Z. mays after 14 days of chilling. Western blot analysis of the D1 protein and LHCII type II chlorophyll a/b-binding protein also showed significant increases in M. × giganteus during chilling and significant decreases in Z. mays. Compared to other C4 species, M. × giganteus grown in chilling conditions appears to counteract the loss of photosynthetic proteins and proteins protecting photosystem II typically observed in other species by increasing mRNA levels for their synthesis.