Evolution of C4 photosynthetic genes and overexpression of maize C4 genes in rice

C3 plants including many agronomically important crops exhibit a lower photosynthetic efficiency due to inhibition of photosynthesis by O₂ and the associated photorespiration. C4 plants had evolved the C4 pathway to overcome low CO₂ and photorespiration. This review first focuses on the generation of a system for high level expression of the C4-specific gene for pyruvate, orthophosphate dikinase (Pdk), one of the key enzyme in C4 photosynthesis. Based on the results with transgenic rice plants, we have demonstrated that the regulatory system controlling thePdk expression in maize is not unique to C4 plants but rice (C3 plant) posses a similar system. Second, we discussed the possibility of the high level expression of maize C4-specific genes in transgenic rice plants. Introduction of the maize intact phosphoenolpyruvate carboxylase gene (Ppc) caused 30–100 fold higher PEPC activities than non-transgenic rice. These results demonstrated that intact C4-type genes are available for high level expression of C4 enzymes in rice plants. The extended abstract of a paper presented at the 13th International Symposium in Conjugation with Award of the International Prize for Biology “Frontier of Plant Biology”     

Restoration of C4 grasses with seasonal fires in a C3/C4 grassland invaded by Prosopis glandulosa,a fire‐resistant shrub

Questions: Can prescribed fire restore C₄ perennial grasses in grassland ecosystems that have become dominated by fire‐resistant C₃ shrubs (Prosopis glandulosa) and C₃ grasses? Do fires in different seasons alter the direction of change in grass composition? Location: Texas, USA. Methods: We quantified short‐ and long‐term (12 yr post‐fire) herbaceous functional group cover and diversity responses to replicated seasonal fire treatments: (1) repeated‐winter fires (three in 5 yr), (2) repeated‐summer fires (two in 3 yr), and (3) alternate‐season fires (two winter and one summer in 4 yr), compared with a no‐fire control. Results: Summer fires were more intense than winter fires, but all fire treatments temporarily decreased Prosopis and C₃ annual grass cover. The alternate‐season fire treatment caused a long‐term increase in C₄ mid‐grass cover and functional group diversity. The repeated‐summer fire treatment increased C₄ short‐grass cover but also caused a long‐term increase in bare ground. The repeated winter fire treatment had no long‐term effects on perennial grass cover. Mesquite post‐fire regrowth had increasingly negative impacts on herbaceous cover in all fire treatments. Conclusions: Summer fire was necessary to shift herbaceous composition toward C₄ mid‐grasses. However, the repeated‐summer fire treatment may have been too extreme and caused post‐fire herbaceous composition to “over‐shift” toward less productive C₄ short‐grasses rather than C₄ mid‐grasses. This study provides some of the first long‐term data showing a possible benefit of mixing seasonal fires (i.e., the alternate‐season fire treatment) in a prescribed burning management plan to restore C₄ mid‐grass cover and enhance overall herbaceous diversity.     

Invasion promotes invasion: Facilitation of C3 perennial grass dominance in mixed C3/C4 grassland by an invasive C3 woody sprouter (Prosopis glandulosa)

相似文献   

Neotropical C3/C4 grass distributions – present,past and future

Changes in C₄ grass distribution and abundance are frequently observed in Quaternary, Holocene and future environmental‐change scenarios. However, the factors driving these dynamics are not fully understood, and conflicting theories have been reported. In this paper, we present a very large dataset of modern altitudinal distribution profiles of C₃ and C₄ grasses covering the entire Neotropical Andes, which was compared with actual climate data. The results of multivariate analysis demonstrate that, in the Neotropical Andes, mean annual temperature is the main factor governing the modern altitudinal distribution of C₃ and C₄ grass species. The C₃ and C₄ grass distributions were compared with simulations based on the Lund‐Potsdam‐Jena dynamic global vegetation model (LPJ‐DGVM), which allowed the present grass distribution to be estimated. Finally, the DGVM was employed to simulate past and future scenarios, using the IPCC's climate projections for 2100 and PMIP2 models for the Holocene Optimum (HO, 6000 years bp ) and the Last Glacial Maximum (LGM, 21 000 years bp ). The results were found to be significantly different from those obtained using a simple photosynthetic model. According to LPJ forced with the PMIP2 models for the LGM, during the LGM, the C₄ grasses would not have reached higher altitudes than found in the present day.     

Comparative analysis of thylakoid protein complexes in the mesophyll and bundle sheath cells from C3, C4 and C3–C4 Paniceae grasses

To better understand the coordination between dark and light reactions during the transition from C₃ to C₄ photosynthesis, we optimized a method for separating thylakoids from mesophyll (MC) and bundle sheath cells (BSCs) across different plant species. We grew six Paniceae grasses including representatives from the C₃, C₃–C₄ and C₄ photosynthetic types and all three C₄ biochemical subtypes [nicotinamide adenine dinucleotide phosphate‐dependent malic enzyme (NADP‐ME), nicotinamide adenine dinucleotide‐dependent malic enzyme (NAD‐ME) and phosphoenolpyruvate carboxykinase (PEPCK)] in addition to Zea mays under control conditions (1000 μmol quanta m^?2 s^?1 and 400 ppm of CO₂). Proteomics analysis of thylakoids under native conditions, using blue native polyacrylamide gel electrophoresis followed by liquid chromatography‐mass spectrometry (LC‐MS), demonstrated the presence of subunits of all light‐reaction‐related complexes in all species and cell types. C₄ NADP‐ME species showed a higher photosystems I/II ratio and a clear accumulation of the NADH dehydrogenase‐like complexes in BSCs, while Cytb₆f was more abundant in BSCs of C₄ NAD‐ME species. The C₄ PEPCK species showed no clear differences between cell types. Our study presents, for the first time, a good separation between BSC and MC for a C₃–C₄ intermediate grass which did not show noticeable differences in the distribution of the thylakoid complexes. For the NADP‐ME species Panicum antidotale, growth at glacial CO₂ (180 ppm of CO₂) had no effect on the distribution of the light‐reaction complexes, while growth at low light (200 μmol quanta m^?2 s^?1) promoted the accumulation of light‐harvesting proteins in both cell types. These results add to our understanding of thylakoid distribution across photosynthetic types and subtypes, and introduce thylakoid distribution between the MC and BSC of a C₃–C₄ intermediate species.     

A role for differential Rubisco activase isoform expression in C4 bioenergy grasses at high temperature

Rubisco activase (Rca) facilitates the release of sugar‐phosphate inhibitors at Rubisco catalytic sites during CO₂ fixation. Most plant species express two Rca isoforms, the larger Rca‐α and the shorter Rca‐β, either by alternative splicing from a single gene or expression from separate genes. The mechanism of Rubisco activation by Rca isoforms has been intensively studied in C₃ plants. However, the functional role of Rca in C₄ plants where Rubisco and Rca are located in a much higher [CO₂] compartment is less clear. In this study, we selected four C₄ bioenergy grasses and the model C₄ grass setaria (Setaria viridis) to investigate the role of Rca in C₄ photosynthesis. All five C₄ grass species contained two Rca genes, one encoding Rca‐α and the other Rca‐β, which were positioned closely together in the genomes. A variety of abiotic stress‐related motifs were identified in the Rca‐α promoter of each grass, and while the Rca‐β gene was constantly highly expressed at ambient temperature, Rca‐α isoforms were expressed only at high temperature but never surpassed 30% of Rca‐β content. The pattern of Rca‐α induction on transition to high temperature and reduction on return to ambient temperature was the same in all five C₄ grasses. In sorghum (Sorghum bicolor), sugarcane (Saccharum officinarum), and setaria, the induction rate of Rca‐α was similar to the recovery rate of photosynthesis and Rubisco activation at high temperature. This association between Rca‐α isoform expression and maintenance of Rubisco activation at high temperature suggests that Rca‐α has a functional thermo‐protective role in carbon fixation in C₄ grasses by sustaining Rubisco activation at high temperature.     

Interaction between C4 barnyard grass and C3 upland rice under elevated CO2: Impact of mycorrhizae

Atmospheric CO₂ enrichment may impact arbuscular mycorrhizae (AM) development and function, which could have subsequent effects on host plant species interactions by differentially affecting plant nutrient acquisition. However, direct evidence illustrating this scenario is limited. We examined how elevated CO₂ affects plant growth and whether mycorrhizae mediate interactions between C₄ barnyard grass (Echinochloa crusgalli (L.) Beauv.) and C₃ upland rice (Oryza sativa L.) in a low nutrient soil. The monocultures and combinations with or without mycorrhizal inoculation were grown at ambient (400 ± 20 μmol mol^?1) and elevated CO₂ (700 ± 20 μmol mol^?1) levels. The ¹⁵N isotope tracer was introduced to quantify the mycorrhizally mediated N acquisition of plants. Elevated CO₂ stimulated the growth of C₃ upland rice but not that of C₄ barnyard grass under monoculture. Elevated CO₂ also increased mycorrhizal colonization of C₄ barnyard grass but did not affect mycorrhizal colonization of C₃ upland rice. Mycorrhizal inoculation increased the shoot biomass ratio of C₄ barnyard grass to C₃ upland rice under both CO₂ concentrations but had a greater impact under the elevated than ambient CO₂ level. Mycorrhizae decreased relative interaction index (RII) of C₃ plants under both ambient and elevated CO₂, but mycorrhizae increased RII of C₄ plants only under elevated CO₂. Elevated CO₂ and mycorrhizal inoculation enhanced ¹⁵N and total N and P uptake of C₄ barnyard grass in mixture but had no effects on N and P acquisition of C₃ upland rice, thus altering the distribution of N and P between the species in mixture. These results implied that CO₂ stimulation of mycorrhizae and their nutrient acquisition may impact competitive interaction of C₄ barnyard grass and C₃ upland rice under future CO₂ scenarios.     

Germination sensitivities to water potential among co‐existing C3 and C4 grasses of cool semi‐arid prairie grasslands

An untested theory states that C₄ grass seeds could germinate under lower water potentials (Ψ) than C₃ grass seeds. We used hydrotime modelling to study seed water relations of C₄ and C₃ Canadian prairie grasses to address Ψ divergent sensitivities and germination strategies along a risk‐spreading continuum of responses to limited water. C₄ grasses were Bouteloua gracilis, Calamovilfa longifolia and Schizachyrium scoparium; C₃ grasses were Bromus carinatus, Elymus trachycaulus, Festuca hallii and Koeleria macrantha. Hydrotime parameters were obtained after incubation of non‐dormant seeds under different Ψ PEG 6000 solutions. A t‐test between C₃ and C₄ grasses did not find statistical differences in population mean base Ψ (Ψ_b(50)). We found idiosyncratic responses of C₄ grasses along the risk‐spreading continuum. B. gracilis showed a risk‐taker strategy of a species able to quickly germinate in a dry soil due to its low Ψ_b(50) and hydrotime (θ_H). The high Ψ_b(50) of S. scoparium indicates it follows the risk‐averse strategy so it can only germinate in wet soils. C. longifolia showed an intermediate strategy: the lowest Ψ_b(50) yet the highest θ_H. K. macrantha, a C₃ grass which thrives in dry habitats, had the highest Ψ_b(50), suggesting a risk‐averse strategy for a C₃ species. Other C₃ species showed intermediate germination patterns in response to Ψ relative to C₄ species. Our results indicate that grasses display germination sensitivities to Ψ across the risk‐spreading continuum of responses. Thus seed water relations may be poor predictors to explain differential recruitment and distribution of C₃ and C₄ grasses in the Canadian prairies.     

Grasshoppers efficiently process C4 grass leaf tissues: implications for patterns of host-plant utilization

Leaf‐chewing insects are commonly believed to be unable to crush the nutrient‐rich bundle sheath cells (BSC) of C₄ grasses. This physical constraint on digestion is thought to reduce the nutritional quality of these grasses substantially. However, recent evidence suggests that BSC are digested by grasshoppers. To directly assess the ability of grasshoppers to digest C₄ grass BSC, leaf particles of Bouteloua curtipendula (Poaceae) were examined from the digestive tracts of two grasshopper species: Camnula pellucida (Scudder) (primarily a grass feeder) and Melanoplus sanguinipes (Fabricius) (a forb and grass generalist) (Orthoptera: Acrididae). Transmission electron microscopy was used to make the first observations of BSC crushing by herbivorous insects. Camnula pellucida and M. sanguinipes crushed over 58% and 24%, respectively, of the BSC in ingested leaf tissues. In addition, chloroplast and cell membranes were commonly disrupted in uncrushed BSC, permitting soluble nutrients to be extracted, even when BSC walls remain intact. The greater efficiency with which C. pellucida crushes BSC is consistent with the idea that grass‐feeding species are better adapted for handling grass leaf tissues than are generalist species. By demonstrating the effectiveness with which the BSC of B. curtipendula can be crushed and extracted by both species of grasshoppers, this study suggests one reason why C₄ grasses are not generally avoided by grasshoppers: at least some C₄ grasses can be more easily digested than has been hypothesized.     

Influence of salt stress on C4 photosynthesis in Miscanthus sinensis Anderss.

• Miscanthus sinensis Anderss. is a good candidate for C₄ bioenergy crop development for marginal lands. As one of the characteristics of marginal lands, salinization is a major limitation to agricultural production. The present work aimed to investigate the possible factors involved in the tolerance of M. sinensis C₄ photosynthesis to salinity stress.
• Seedlings of two accessions (salt‐tolerant ‘JM0119’ and salt‐sensitive ‘JM0099’) were subjected to 0 mm NaCl (control) or 250 mm NaCl (salt stress treatment) for 2 weeks. The chlorophyll content, parameters of photosynthesis and chlorophyll a fluorescence, activity of C₄ enzymes and expression of C₄ genes were measured.
• The results showed that photosynthesis rate, transpiration rate, chlorophyll content, PSII operating efficiency, coefficient of photochemical quenching, activity of phosphoenolpyruvate carboxylase (PEPC) and pyruvate, orthophosphate dikinase (PPDK) and gene expression of PEPC and PPDK under salinity were higher after long‐term salinity exposure in ‘JM0119’ than in ‘JM0099’, while activity of NADP‐malate dehydrogenase (NADP‐MDH) and NADP‐malic enzyme (NADP‐ME), together with expression of NADP‐MDH and NADP‐ME, were much higher in ‘JM0099’ than in ‘JM0119’.
• In conclusion, the increased photosynthetic capacity under long‐term salt stress in the salt‐tolerant relative to the salt‐sensitive M. sinensis accession was mainly associated with non‐stomatal factors, such as reduced chlorophyll loss, higher PSII operating efficiency, enhanced activity of PEPC and PPDK and relatively lower activity of NADP‐ME.

     

C4 anatomy can evolve via a single developmental change

C₄ photosynthesis is a complex trait that boosts productivity in warm environments. Paradoxically, it evolved independently in numerous plant lineages, despite requiring specialised leaf anatomy. The anatomical modifications underlying C₄ evolution have previously been evaluated through interspecific comparisons, which capture numerous changes besides those needed for C₄ functionality. Here, we quantify the anatomical changes accompanying the transition between non‐C₄ and C₄ phenotypes by sampling widely across the continuum of leaf anatomical traits in the grass Alloteropsis semialata. Within this species, the only trait that is shared among and specific to C₄ individuals is an increase in vein density, driven specifically by minor vein development that yields multiple secondary effects facilitating C₄ function. For species with the necessary anatomical preconditions, developmental proliferation of veins can therefore be sufficient to produce a functional C₄ leaf anatomy, creating an evolutionary entry point to complex C₄ syndromes that can become more specialised.     

Shifting dominance from native C4 to non‐native C3 grasses: relationships to community diversity

Many field studies have examined how site fertility, soil differences and site history influence the diversity of a plant community. However, only a few studies have examined how the identity of the dominant species influences the diversity in grasslands. Plant species differ widely in phenology, growth form and resource uses; thus, communities dominated by different species are also likely to strongly differ in the environment that they create and in which the subdominant species exist. We examined the correlation between the four most dominant species and community diversity in 2100 plots, located in 21 abandoned agricultural fields in central Minnesota over a 23‐year period. The four most common species were two non‐native C₃ cool season species, Poa pratensis and Agropyron repens, and two native C₄ warm season species, Schizachyrium scoparium and Andropogon gerardii. We found that the differences in the dominants explained up to 27% of the community diversity. Thus, the identity of the dominant species can have a strong influence on community diversity and studies examining factors that influence plant community diversity need to incorporate the effect of the dominants. Secondly, we found that the non‐native C₃ grass dominated communities had lower overall and lower native species richness relative to the native C₄ grass dominated communities. Therefore, a shift in dominants from C₄ to C₃ may lead to a large community diversity decline. We found that Poa pratensis, the most abundant non‐native C₃ grass increased in abundance over the 23 years; thus, the negative influence of non‐natives on the community diversity is not decreasing over time and active management is required to restore native grassland plant communities.     

The age of the grasses and clusters of origins of C4 photosynthesis

At high temperatures and relatively low CO₂ concentrations, plants can most efficiently fix carbon to form carbohydrates through C₄ photosynthesis rather than through the ancestral and more widespread C₃ pathway. Because most C₄ plants are grasses, studies of the origin of C₄ are intimately tied to studies of the origin of the grasses. We present here a phylogeny of the grass family, based on nuclear and chloroplast genes, and calibrated with six fossils. We find that the earliest origins of C₄ likely occurred about 32 million years ago (Ma) in the Oligocene, coinciding with a reduction in global CO₂ levels. After the initial appearance of C₄ species, photosynthetic pathway changed at least 15 more times; we estimate nine total origins of C₄ from C₃ ancestors, at least two changes of C₄ subtype, and five reversals to C₃. We find a cluster of C₄ to C₃ reversals in the Early Miocene correlating with a drop in global temperatures, and a subsequent cluster of C₄ origins in the Mid‐Miocene, correlating with the rise in temperature at the Mid‐Miocene climatic optimum. In the process of dating the origins of C₄, we were also able to provide estimated times for other major events in grass evolution. We find that the common ancestor of the grasses (the crown node) originated in the upper Cretaceous. The common ancestor of maize and rice lived at 52 ± 8 Ma.     

C4 photosynthesis in C3 rice: a theoretical analysis of biochemical and anatomical factors

Engineering C₄ photosynthesis into rice has been considered a promising strategy to increase photosynthesis and yield. A question that remains to be answered is whether expressing a C₄ metabolic cycle into a C₃ leaf structure and without removing the C₃ background metabolism improves photosynthetic efficiency. To explore this question, we developed a 3D reaction diffusion model of bundle‐sheath and connected mesophyll cells in a C₃ rice leaf. Our results show that integrating a C₄ metabolic pathway into rice leaves with a C₃ metabolism and mesophyll structure may lead to an improved photosynthesis under current ambient CO₂ concentration. We analysed a number of physiological factors that influence the CO₂ uptake rate, which include the chloroplast surface area exposed to intercellular air space, bundle‐sheath cell wall thickness, bundle‐sheath chloroplast envelope permeability, Rubisco concentration and the energy partitioning between C₃ and C₄ cycles. Among these, partitioning of energy between C₃ and C₄ photosynthesis and the partitioning of Rubisco between mesophyll and bundle‐sheath cells are decisive factors controlling photosynthetic efficiency in an engineered C₃–C₄ leaf. The implications of the results for the sequence of C₄ evolution are also discussed.     

The efficiency of the CO2-concentrating mechanism during single-cell C4 photosynthesis

The photosynthetic efficiency of the CO₂‐concentrating mechanism in two forms of single‐cell C₄ photosynthesis in the family Chenopodiaceae was characterized. The Bienertioid‐type single‐cell C₄ uses peripheral and central cytoplasmic compartments (Bienertia sinuspersici), while the Borszczowioid single‐cell C₄ uses distal and proximal compartments of the cell (Suaeda aralocaspica). C₄ photosynthesis within a single‐cell raises questions about the efficiency of this type of CO₂‐concentrating mechanism compared with the Kranz‐type. We used measurements of leaf CO₂ isotope exchange (Δ¹³C) to compare the efficiency of the single‐cell and Kranz‐type forms of C₄ photosynthesis under various temperature and light conditions. Comparisons were made between the single‐cell C₄ and a sister Kranz form, S. eltonica[NAD malic enzyme (NAD ME) type], and with Flaveria bidentis[NADP malic enzyme (NADP‐ME) type with Kranz Atriplicoid anatomy]. There were similar levels of Δ¹³C discrimination and CO₂ leakiness (?) in the single‐cell species compared with the Kranz‐type. Increasing leaf temperature (25 to 30 °C) and light intensity caused a decrease in Δ¹³C and ? across all C₄ types. Notably, B. sinuspersici had higher Δ¹³C and ? than S. aralocaspica under lower light. These results demonstrate that rates of photosynthesis and efficiency of the CO₂‐concentrating mechanisms in single‐cell C₄ plants are similar to those in Kranz‐type.