Rubisco small subunits of C4 plants,Napier grass and guinea grass confer C4-like catalytic properties on Rubisco in rice

ABSTRACT

Overexpression of Rubisco small subunit (RbcS) of C₄ plant, sorghum (sorghum bicolor) was shown to enhance the catalytic turnover rate (k _cat) of Rubisco in rice (Oryza sativa). In this study, the effects of other Rubisco small subunits of C₄ plants, Napier grass (Pennisetum purpureum) and guinea grass (Megathyrsus maximus) on kinetic properties of Rubisco in rice were studied. The expression levels of Napier grass RbcS (NgRbcS) and guinea grass RbcS (GgRbcS) proteins accounted for 41% and 45% of total RbcS, respectively in homozygous overexpression lines. The k _cat and K _m for CO₂ (Kc) of Rubisco were increased in all transgenic lines. Interestingly, the k _cat was markedly higher in NgRbcS homozygous line, whereas K _c was notably higher in GgRbcS homozygous line. Although its effects depend on species, these results suggest that the introduction of C₄ RbcS are effective approaches to alter the catalytic properties of Rubisco in rice.     

Production Ecophysiology of Hungarian Green Pepper Under Elevated Air CO2 Concentration

SUMMARY

Production, dry matter (including reproductive) allocation, photosynthesis, transpiration, water use efficiency and carbon and nitrogen responses of a Hungarian sweet pepper (Capsicum annum L.) under continuous elevated CO₂ concentrations are reported. Plants were grown in open top chambers under a temperate-continental climate in Hungary from plantation at ambient (350 μmol mol^?1) and elevated (700 μmol mol^?1) CO₂ concentrations. The CO₂ assimilation responses of the green pepper plants grown in high CO₂ from plantation until final harvest reflected down-regulation of their photosynthesis. The integrated and combined effect of the increased net CO₂ assimilation rate and the unchanged rate of transpiration resulted in higher WUE at elevated CO₂ concentrations in the high CO₂ plants than in the control ones grown at ambientCO₂. However, the improved water use efficiency in the high CO₂ plants was not followed by an acclimation in C-trans-location and C-allocation to the reproductive organs in the required degree. This was reflected in a slightly increased overall plant production and higher reproductive allocation, but was not accompanied by an increased fresh or dry berry mass production. The acclimation discussed may be of advantage for plant growth in a high CO₂ environment with restricted water availability. We did not find worthy statistical difference between the yield mass of the control and elevated CO₂, although the dry matter production parameters of the high CO₂ plants had statistically not significantly higher values.     

Structural and physiological responses of the C4 grass Sorghum bicolor to nitrogen limitation

Nitrogen (N) is one of the major nutrients influencing photosynthesis and productivity of C₄ plants as well as C₃ plants. C₄ photosynthesis operates through close coordination between mesophyll (M) and bundle sheath (BS) cells. However, how the development of structural and physiological traits in leaves of C₄ plants is regulated under N limitation remains uncertain. We investigated structural and physiological responses of leaves of the NADP-ME-type C₄ grass Sorghum bicolor to N limitation. Plants were grown under four levels of N supply (.05 to .6 g N per 5-L pot). Decreasing N supply resulted in decreases in net photosynthetic rate, stomatal conductance, leaf N and chlorophyll contents, and the activity ratio of phosphoenolpyruvate carboxylase to ribulose 1,5-bisphosphate carboxylase/oxygenase and increases in δ¹³C values and photosynthetic N use efficiency. Low-N leaves were thinner and had smaller photosynthetic cells, especially in M, resulting in lower M/BS tissue area ratio, and contained smaller and fewer chloroplasts. The BS chloroplasts in the low-N leaves accumulated abundant starch grains. The number of thylakoids per granal stack was reduced in M chloroplasts but not in BS chloroplasts. The low-N leaves had thicker cell walls, especially in the BS cells, which might be associated with less negative δ¹³C values, and fewer plasmodesmata in the BS cells. These data reveal structural and physiological responses of C₄ plants to N limitation, most of which would be related to cellular N allocation, light use, CO₂ diffusion and leakiness, and metabolite transport under N limitation.     

Analyzing the impact of high temperature and CO2 on net photosynthesis: biochemical mechanisms,models and genomics

Determining plant response at the biochemical/physiological level to a changing global environment is a prerequisite for constructing accurate models to predict plant productivity. High temperature and CO₂ impact plant biomass accumulation by altering the rate of net photosynthesis such that the measured rates differ greatly from the potential rates predicted from commonly used models. Such models are based on assumptions pertaining to biochemical limitations to net photosynthesis by the capacity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), the enzyme that ultimately limits the rate of photosynthetic CO₂ fixation in C3 and C4 plants, or the capacity of electron transport to supply energy for Calvin cycle activity. Here we provide evidence that the impact of high temperature and CO₂ on net photosynthesis can be accurately calculated from predictive models based solely on Rubisco kinetics if the modeled rate of photosynthesis is adjusted for heat and CO₂-induced changes in the activation state of Rubisco. The activation state of Rubisco, which is regulated by the activity of Rubisco activase, thus appears to be the primary limitation to net photosynthesis at high temperature and/or CO₂. This limitation to net photosynthesis has not been incorporated into common biochemical models, thus compromising their effectiveness. The expression of activase mRNA was not indicative of the central role of activase in the response of photosynthesis to high temperature thus revealing limitations of using a broad genomics approach to identify the impact of environmental stress on plant metabolism.     

Mechanism of High Photosynthetic Capacity in BC2F4 LinesDerived from a Cross between Oryza sativa and Wild Relatives O. rufipogon

Abstract

We found that several BC₂F₄ lines had high leaf photosynthetic rates under light-saturated and ambient CO₂ conditions. These lines are progenies of BC₂F₁ plants with high photosynthetic capacities which were generated by backcrossing between Oryza rufipogon (W630) and O. sativa cv. Nipponbare, as a recurrent parent. Some photosynthetic characteristics of the BC₂F₄ lines were investigated to identify the factors increasing photosynthetic rates. Photosynthetic rates of these lines under light-saturated conditions at 50 to 700 ppm CO₂ concentrations were higher than those in Nipponbare. The estimated-maximum photosynthetic rates under light-saturated and CO₂-saturated conditions in BC₂F₄ lines were also higher than that in Nipponbare. The photosynthetic rate under light-saturated and ambient CO₂ conditions was positively correlated with the carboxylation efficiency as an indicator of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity in vivo rather than stomatal conductance. Initial and total Rubisco activities in vitro tended to be higher in the BC₂F₄ lines than in Nipponbare. The content of active Rubisco calculated from the activation state of Rubisco was also higher in the BC₂F₄ lines than in Nipponbare. These results suggest that high photosynthetic capacities of BC₂F₁ plants can be maintained high in their progenies and high photosynthetic rates under light-saturated and ambient CO₂ conditions in the BC₂F₄ lines are achieved mainly by the high activity of Rubisco due to the high active Rubisco content.     

Gene Expression and Accumulation of Rubisco in Bundle Sheath and Mesophyll Cells during Leaf Development and Senescence in Rice,a C3 Plant

Abstract

Gene expression of ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) large subunit (rbcL) and small subunit (rbcS) in bundle sheath and mesophyll cells of rice, a C₃ plant, was examined during leaf development and senescence by in situ hybridization. Localization of Rubisco protein in both cells was also examined by immuno-electron microscopy. Gene expression and accumulation of Rubisco were related with the chlorophyll fluorescence parameters. The chlorophyll fluorescence parameters, such as F_v/F_m and Φ_psii, gradually increased during leaf development with the increase in the accumulation of Rubisco. However, the chlorophyll fluorescence parameters decreased earlier than the Rubisco content during leaf senescence. The expression of rbcS decreased earlier in bundle sheath cells than in mesophyll cells during leaf development, whereas the expression of rbcL in both cells was retained during leaf development and decreased during leaf senescence. On the other hand, Rubisco content of bundle sheath and mesophyll cells increased during leaf development and decreased during leaf senescence. Rubisco was retained even after the disappearance of the expression of rbcS and rbcL detectable by in situ hybridization. The present results suggest that the expression pattern of rbcS in bundle sheath cells was somewhat different from that in mesophyll cells, but this difference was not reflected in Rubisco content.     

Correlation of Chlorophyll Meter Readings with Gas exchange and Chlorophyll Fluorescence in Flag Leaves of Rice (Oryza sativa L.) Plants

Abstract

The objective of this study was to establish the correlation of the chlorophyll meter (SPAD) readings with the contents of chlorophyll (Chl) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), the gross photosynthetic rate (P_g), and the maximum quantum yield of photosystem II (PSII) (F_v/F_m) in flag leaves of rice (Oryza sativa L.) in ripening stage. The SPAD readings significantly correlated with the Chl content, the Rubisco content, P_g and F_v/F_m (R² = 0.848, 0.648, 0.671 and 0.712, respectively), which suggests that the SPAD meter has the potential to estimate the photosynthetic capacity of the flag leaves. However, both P_g and F_v/F_m had a stronger relationship with the Rubisco content than the SPAD readings, indicating that the PSII photochemical and CO₂ assimilation capacities are strongly influenced by the Rubisco content. Therefore, accurate calibration would be indispensable to obtain the physiological information from the SPAD readings of flag leaves.     

Effects of atmospheric CO2 concentration and defoliation on the growth of Themeda triandra

The effects of elevated atmospheric carbon dioxide (CO₂) concentration (700 μmol mol^?1) on defoliated (three clippings at 3‐week intervals) and undefoliated plants were determined for the C₄ grass Themeda triandra, Forsk. The elevated CO₂ concentration significantly increased leaf regrowth following defoliation, and total leaf production was greatest in this treatment. Shoot biomass of undefoliated plants was also increased under the elevated CO₂ concentration treatment. The primary effect of the elevated CO₂ concentration in both defoliated and undefoliated plants was an increase in individual leaf length and mass of dry matter, linked to a higher leaf water content and increased photosynthetic rates at the canopy level. Photosynthetic down‐regulation at the leaf level occurred, but this was compensated for by increased assimilation rates and greater canopy leaf area at the elevated CO₂ concentration. Increases in leaf and sheath growth of defoliated plants in the elevated CO₂ concentration treatment were lost following a final 3‐week reversion to ambient CO₂ concentration, but occurred in plants exposed to the elevated CO₂ concentration for the final 3‐week period only. In conclusion, elevated atmospheric CO₂ concentration increases shoot growth via increased leaf extension, which is directly dependent on stimulation of concurrent photosynthesis. CO₂ responsiveness is sustained following moderate defoliation but is reduced when plants experience reduced vigour as a result of maturation or high frequency of defoliation.     

Evaluation of the Effect of Photosynthesis on Biomass Production with Simultaneous Analysis of Growth and Continuous Monitoring of CO2 Exchange in the Whole Plants of Radish,cv Kosena under Ambient and Elevated CO2

Abstract

The effects of elevated CO₂ (approximate doubling of atmospheric CO₂ concentration) on the rate of photosynthesis estimated from continuous monitoring of CO₂ exchange in whole plants were investigated in radish cv. Kosena accompanied with simultaneous analysis of growth for 6 days from 15 to 21 days after planting (DAP). The elevated CO₂ increased the dry weights of hydroponically grown radish plants by 59% at 21 DAP.

The increase in dry weight was due to a combined effect of increased leaf area and increased photosynthetic rate per unit leaf area. Leaf area and the photosynthetic rate were increased by elevated CO₂ by 18-43% and 9-20%, respectively, during 15 to 21 DAP. Namely, an increase in the rate of photosynthesis is accompanied by an increase in leaf area, both having a significant effect on biomass production.     

玉米磷酸烯醇式丙酮酸羧化酶基因的研究进展

高等植物按照CO₂不同的同化途径,分为C₃、C₄和CAM植物。由于C₄植物具有高光效基因,使其在高温、高光照、高氧分压条件下具有比C₃植物更高的光合作用效率。目前,很多学者致力于研究玉米中的高光效基因-磷酸烯醇式丙酮酸羧化酶基因（PEPC）,并试图将此基因转入到C₃植物中,使C₃植物的光合特性得到一定改善,进而使其产量提高。本文综述将C₄高光效基因转入到C₃植物后,C₃植物的生理生化变化和对其光合作用的影响,对向C₃植物中导入pepc基因能否提高其光合作用效率和产量的潜在可能性进行探讨。     

Characteristics of Gas Exchange and Chlorophyll Fluorescence during Senescence of Flag Leaf in Different Rice (Oryza sativa L.) Cultivars Grown under Nitrogen-Deficient Condition

Abstract

Effects of nitrogen (N) deficiency on photosynthetic gas exchange and photosystem II (PSII) photochemistry of flag leaves during grain-filling stage were investigated in six rice cultivars, Kasalath (a conventional indica), IR36 (an improved indica), Shirobeniya (a conventional japonica), Nipponbare (an improved japonica), Akenohoshi (an improved japonica-indica intermediate type) and BSI429 (an improved tropical japonica, a new plant type line) grown hydroponically in N-sufficient (NS) and N-deficient (ND) solution. From 3 to 24 days after heading (DAH), net photosynthetic rate (P_n), maximum quantum yield of photosystem II (PSII) (F_v/F_m), quantum yield of PSII electron transport ( Φ_psii), and contents of chlorophyll (Chl) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in the flag leaves decreased, particularly under the ND condition in all six cultivars. A substantial difference was observed among the ND plants for the sustainability index (SI, the ratio of the value at 24 DAH to that at 3 DAH) of P_n, F_v/F_m, Φ_psii, Chl content and Rubisco content; SIs of those parameters of Akenohoshi, BSI429, Nipponbare and Shirobeniya were higher than those of IR36 and Kasalath. The SI of P_n showed significant positive correlations with those of F_v/F_m, Φ_psii, and the contents of Chl and Rubisco under the ND condition. It was concluded that the sustainability of photosynthesis in the flag leaves was mainly due to those of PSII photochemistry and electron transport, which was associated with the maintenance of Chl and Rubisco under the ND condition.     

CO2 Enrichment Effects on Forage and Grain Nitrogen Content of Pasture and Cereal Plants

SUMMARY

Increasing atmospheric CO₂ concentrations [CO₂] have the potential to enhance growth and yield of agricultural plants. Con-comitantly plants grown under high [CO₂] show significant changes of the chemical composition of their foliage and of other plant parts. Particularly, high [CO₂] result in a decrease of plant nitrogen (N) concentration, which may have serious consequences for crop quality. This presentation summarizes the results of a variety of CO₂ enrichment studies with pasture plants (Lolium spp., Trifolium repens) and cereal species (Triticum aestivum, Hordeum vulgare) which were conducted at our laboratory under different growth and CO₂ exposure conditions ranging from controlled environment studies to investigations under free air carbon dioxide enrichment (FACE). With the exception of clover in all experiments a CO₂-induced decline of forage and grain N concentration was observed. The magnitude of this reduction differed between species, cultivars, management conditions (N fertilization) and CO₂ exposure conditions. No unambiguous evidence was obtained whether N fertilization can contribute to meet the quality requirements for cereals and grass monocultures with respect to tissue N concentrations in a future high-CO₂ world. As shown in the FACE experiments current application rates of N fertilizers are inadequate to achieve quality standards.     

The Effect of Elevated Atmospheric CO2 Concentration on Two Populus Clones Grown in a FACE System

SUMMARY

Two poplar clones, hybrid Populus deltoides Bartr. Ex Marsh X Populus nigra L. (Populus xeuramericana), clone I-214, and Populus deltoides, clone Lux, were grown from hardwood cuttings for one growing season in either ambient (360 μmol mol^?1) or elevated (560 μmol mol^?1) [CO₂] in FACE-systemrings at Rapolano Terme (Siena, Italy). Both clones I-214 and Lux exhibited a higher aboveground bio-mass, photosynthesis at light saturation and instantaneous transpiration efficiency (ITE) in CO₂-enriched air. The elevated [CO₂]-induced responses of clone I-214 included increased investment in branch and leaf biomass, and enhanced stem volume. The elevated [CO₂]-induced responses of clone Lux included an increase in the number of branches (and foliage area). Indication of photosynthetic acclimation under elevated [CO₂] was found during the early morning, but only in clone I-214. Stomatal conductance decreased under elevated [CO₂] particularly in clone Lux. Clone differences in response to elevated [CO₂] should be taken in account when planning future poplar plantations in forecast warmer and drier Mediterranean sites.     

Effects of Elevated Atmospheric Carbon Dioxide Concentration on Silica Deposition in Rice (Oryza sativa L.) Panicle

Abstract

The effects of elevated carbon dioxide concentration ([CO₂]) on silica deposition on husk epidermis of rice (Oryza sativa L. cv. Akitakomachi) during the flowering stage were investigated in this study. The study was motivated by the concept that the rice yield maybe affected by global warming as a result of elevated [CO₂] environment since sterility of rice is related to the panicle silica content that influences transpiration, and elevated [CO₂] could affect plant transpiration. Silica deposition analysis was focused on the flowering stage of the rice crop grown hydroponically under two [CO₂] conditions: 350 μmol mol-¹ (ambient) and 700 μmol mol-¹ (elevated). Silica deposition on the husk epidermis from three parts of the panicle at four flowering stages were examined using a scanning electron microscope (SEM) combined with an energy dispersive X-ray microanalyzer (EDX). The results demonstrated that elevated [CO₂] significantly suppressed silica deposition on the husk epidermis at the lower part of the panicle, and at the early flowering stage when 1/3 of the panicle emerged from the leaf sheath. In the transverse section analysis of the husk, silica deposition on the husk epidermis under elevated [CO₂] was less than that under ambient [CO₂] at the late flowering stage. The less silica deposition observed on the husks at the late flowering stage under elevated [CO₂] might be related to the suppressed transpiration from the panicle by elevated [CO₂] found in a previous study.     

Effect of carbonic anhydrase-containing endophytic bacteria on growth and physiological attributes of wheat under water-deficit conditions

Drought is one of the major limitations to agricultural productivity, suppressing plant growth and yield of food crops throughout the world particularly in arid and semiarid regions. Drought-tolerant carbonic anhydrase (CA; EC 4.2.1.1)-containing endophytic bacteria may improve plant growth under stressed conditions. In the present study, effect of drought-tolerant CA-containing endophytic bacteria on growth and physiology of wheat under water-deficit conditions was studied. One hundred and fifty isolates were isolated from wheat plants and screened for their ability to tolerate polyethylene glycol (PEG) 6000-induced water-deficit stress (?0.31 to ?3.20 MPa). Fifty isolates exhibiting intrinsic ability to tolerate stress were further screened for CA activity. Ten drought-tolerant isolates with higher CA activity were evaluated for improving wheat growth under water-deficit conditions (?0.04, ?1.09, ?1.23 MPa). Results showed that PEG-mediated water-deficit stress significantly reduced growth of wheat. However, inoculation with isolates WR2, WS11 and WL19 significantly enhanced seedling growth by improving maximum root length, shoot length, root and shoot dry weight under non-stressed as well as stressed conditions. These isolates were identified by 16S rRNA as Bacillus marisflavi (WR2) Bacillus thuringiensis (WS11) and Bacillus subtilis (WL19). Isolate WL19 also improved chlorophyll content, photosynthetic rate, CA activity and relative water content compared to uninoculated control plants. Overall, our findings suggest that endophytic bacterial isolates WR2, WS11 and WL19 with CA activity can enhance photosynthesis and biomass of wheat seedlings under water-deficit conditions.

Abbreviations: CA: Carbonic anhydrase; PEG: Polyethylene glycol; CO₂: Carbon dioxide; HCO^3–: Bicarbonate; TSA: Tryptic Soy Agar; LB: Luria Bertani; A: CO₂ assimilation rate; E: Transpiration rate; g_s: Stomatal conductance; C_i: Substomatal CO₂ concentration; RWC: Relative water content; EL: Electrolyte leakage     

Intracellular position of mitochondria and chloroplasts in bundle sheath and mesophyll cells of C3 grasses in relation to photorespiratory CO2 loss

In C₃ plants, photosynthetic efficiency is reduced by photorespiration. A part of CO₂ fixed during photosynthesis in chloroplasts is lost from mitochondria during photorespiration by decarboxylation of glycine by glycine decarboxylase (GDC). Thus, the intracellular position of mitochondria in photosynthetic cells is critical to the rate of photorespiratory CO₂ loss. We investigated the intracellular position of mitochondria in parenchyma sheath (PS) and mesophyll cells of 10 C₃ grasses from 3 subfamilies (Ehrhartoideae, Panicoideae, and Pooideae) by immunostaining for GDC and light and electron microscopic observation. Immunostaining suggested that many mitochondria were located in the inner half of PS cells and on the vacuole side of chloroplasts in mesophyll cells. Organelle quantification showed that 62–75% of PS mitochondria were located in the inner half of cells, and 62–78% of PS chloroplasts were in the outer half. In mesophyll cells, 61–92% of mitochondria were positioned on the vacuole side of chloroplasts and stromules. In PS cells, such location would reduce the loss of photorespiratory CO₂ by lengthening the path of CO₂ diffusion and allow more efficient fixation of CO₂ from intercellular spaces. In mesophyll cells, it would facilitate scavenging by chloroplasts of photorespiratory CO₂ released from mitochondria. Our data suggest that the PS cells of C₃ grasses have already acquired an initial structure leading to proto-Kranz and further C₃–C₄ intermediate anatomy. We also found that in the Pooideae, organelle positioning in PS cells on the phloem side resembles that in mesophyll cells.     

Identification of Chromosomal Regions Controlling the Leaf Photosynthetic Rate in Rice by Using a Progeny from Japonica and High-yielding Indica Varieties

Abstract

The whole-leaf photosynthetic rate in rice plants is controlled by various physiological processes. In a high-yielding indica rice variety, Habataki, the leaf photosynthetic rate (LPR) of the uppermost fully expanded leaves was approximately 130 to 140% of that in a japonica variety, Sasanishiki, from booting to the early ripening stage. We characterized the difference in the LPR between Habataki and Sasanishiki. Leaves of Habataki contained higher levels of nitrogen and, as a consequence, of Rubisco, and had higher stomatal conductance that was associated with higher hydraulic conductance from roots to leaves than those of Sasanishiki. These features were responsible for the higher LPR of Habataki. An analysis of chromosome segment substitution lines (CSSLs) in which chromosome segments from Habataki were substituted into the genetic background of Sasanishiki showed that three genetic regions on chromosomes 4, 5 and 11 were responsible for the increase in the LPR. Each of these regions was estimated to increase the LPR by 15 to 30%, and we showed that they were associated with higher activity of mesophyll photosynthesis due to higher leaf nitrogen content and greater stomatal conductance. Leaf nitrogen content and stomatal conductance may be useful parameters for further quantitative trait locus analysis of efficient photosynthesis in leaves.     

Variations in structural,biochemical, and physiological traits of photosynthesis and resource use efficiency in Amaranthus species (NAD-ME-type C4)

C₄ plants show higher photosynthetic capacity and productivity than C₃ plants owing to a CO₂-concentrating mechanism in leaves, which reduces photorespiration. However, which traits regulate the photosynthetic capacity of C₄ plants remains unclear. We investigated structural, biochemical, and physiological traits associated with photosynthesis and resource use efficiency in 20 accessions of 12 species of Amaranthus, NAD-malic enzyme-type C₄ dicots. Net photosynthetic rate (P_N) ranged from 19.7 to 40.5 μmol m^?2 s^?1. P_N was positively correlated with stomatal conductance and nitrogen and chlorophyll contents of leaves and was weakly positively correlated with specific leaf weight. P_N was also positively correlated with the activity of the C₃ enzyme ribulose-1,5-bisphoshate carboxylase/oxygenase, but not with the activities of the C₄ enzymes phosphoenolpyruvate carboxylase and NAD-malic enzyme. Structural traits of leaves (stomatal density, guard cell length, leaf thickness, interveinal distance, sizes of mesophyll and bundle sheath cells and the area ratio between these cells) were not significantly correlated with P_N. These data suggest that some of the biochemical and physiological traits are involved in interspecific P_N variation, whereas structural traits are not directly involved. Photosynthetic nitrogen use efficiency ranged between 260 and 458 μmol mol^?1 N s^?1. Photosynthetic water use efficiency ranged between 5.6 and 10.4 mmol mol^?1. When these data were compared with previously published data of C₄ grasses, it is suggested that common mechanisms may determine the variations in resource use efficiency in grasses and this dicot group.     

Interactive Effects of CO2 and O3 on the Growth of Trisetum flavescens and Trifolium pratense Grown in Monoculture or a Bi-Species Mixture

SUMMARY

Golden oat grass (Trisetum flavescens L.) and red clover (Trifolium pratense L.) were grown as monocultures or bi-species mixtures under controlled conditions and exposed to ambient (350 ppm) or elevated (580 ppm) CO₂, with or without addition of O₃ (diel profile with 150 ppb maximum). Shoot biomass measurements after the initial growth and two re-growth periods were used to determine the specific responses of both species, and the difference in the specific response between monocultures and mixtures. T. pratense was much more responsive to CO₂, O₃, and their combination, compared to T. flavescens. In the case of O₃ but not of CO₂, the difference in sensitivity between species was larger in mixture than in monoculture. In contrast to elevated CO₂, O₃ significantly reduced the root:shoot ratio in the mixture, which could explain the increasing negative effect of O₃ on clover with progressing harvests.

The relative CO₂ stimulation of T. pratense and of the cumulative mixture shoot biomass was larger in the presence than in the absence of O₃, which was due to an almost complete protection from O₃ stress by elevated CO₂. In the mixture, the fraction of T. flavescens was small and increased during the experiment; this increase was most pronounced with O3, but any change in mixture biomass was dominated by the response of T. repens.

The results confirm that in grass/legume mixtures legumes are most sensitive to elevated CO₂ and O₃, but the magnitude of specific responses depends on canopy structure and of plant development. Elevated CO₂ minimizes the negative impacts of O₃ stress on above- and below-ground plant growth.     

Effects of temperature on growth and photosynthesis in the seedling stage of the sheath blight-resistant rice genotype 32R

The 32R rice genotype is resistant to sheath blight disease (ShB), with a high-yield potential. We examined effects of temperature on the plant responses of 32R in comparison with those of the ShB-susceptible rice genotype (29S) and Nipponbare (Nb, a Japonica standard cultivar). The seedlings at the 4th leaf stage of rice genotypes were exposed to 14/14, 19/14, 25/20, 31/26, 37/32 and 37/37 °C (day/night) for 5, 10 and 15 days. The dry weight, leaf area, photosynthesis, contents of ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and chlorophyll contents were examined. The dry weight showed lower in 32R than in 29S and Nb at a low temperature, and total dry weight correlated strongly with root dry weight and leaf area. The relative growth rate (RGR) correlated strongly with the net assimilation rate (NAR). Rubisco, chlorophyll contents and the photosynthetic rates were limited at a low temperature and showed lower in 32R than in 29S and Nb. The strong correlations between Rubisco and the rates of maximum photosynthesis and initial slope were found in 32R, but not found in 29S and Nb. In addition, RGR and NAR of 32R correlated positively with Rubisco. These suggest that 32R contains traits of cold-sensitive genotypes that are related to limiting Rubisco at a low temperature, thus diminishing photosynthesis and limiting plant growth. Differences of growth among 32R, 29S and Nb were discussed in the relation of genotypes.