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.     

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.     

Effects of Salinity Stress on the Structure of Bundle Sheath and Mesophyll Chloroplasts in NAD-Malic Enzyme and PCK Type C4 Plants

Abstract

The effect of NaCl stress on the structure of leaf chloroplasts was investigated in several NAD-Malic enzyme (NAD-ME) and phosphoenolpyruvate carboxykinase (PCK) type C₄ plant species. Seedlings of the monocot species, except Zoysia japonica, grown in 300 mL pots were subjected to salt stress by adding 50 mL of 3% NaCl solution per day to the soil for 5 d after the fourth leaf blades were fully developed. Z. japonica and the dicot species, Amaranthus tricolor, were also treated with 3% NaCl in a similar manner from 5 wk after germination. Salt stress negatively affected the growth, chlorophyll content and chloroplast structure in all the species. At the ultrastructure level, swelling of thylakoids and disruption of envelopes were more or less observed in mesophyll cell (MC) chloroplasts after salt treatment. The structure of bundle sheath cell (BSC) chloroplasts, on the other hand, was hardly damaged under salt condition although stromal and starch areas were considerably decreased. Furthermore, salinity induced granal development in BSC chloroplasts in most species; the number of thylakoids per granum, granal indices and appressed thylakoid density in salt-treated plants were generally higher than those in control. Since the similar responses have also been reported in all NADP-ME type C₄ species investigated in our previous study, the high sensitivity to salt stress in MC chloroplasts and the granal development in BSC chloroplasts by salinity were considered to be common phenomena in all three C₄ subtypes.     

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.     

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.     

Ultrastructure of Fusion Product between Protoplasts from C3 and C4 Species of Amaranthaceae

Summary

This paper describes the ultrastructure of the electric field-induced fusion products of C₃ and C₄ species of Amaranthaceae at the early developmental stage. Protoplasts of C₃ species were isolated from a Ceiosia cristata L. cell suspension and, those of C₄ species were isolated from an Amaranthus tricolor L. cotyledon. Incompatibility occurred in the C₃/C₄ hybrid. The incompatibility reactions were detected in the newly formed hybrid cells accompanied with significant changes in the nucleolus (segregation of nucleolar components) and plastids (cup-like shape or amoeboid plastid enclosing cytoplasmic materials) of C₃ species parent. The structural changes in the organelles of the C₄ partner were less marked. After 5 days of culture, most organelles showed high cellular activity, and a normal dedifferentiation process of mesophyll chloroplasts was observed. At this stage nucleolar segregation was not detected and the C₃ species plastids were difficult to distinguish from the proplastids formed from mesophyll chloroplasts. In addition, some mitochondria showed bursting-like structure. However, under the culture condition used these somatic incompatibility did not seem to impair further growth of fusion products since they were still proliferating well resulting in callus formation.     

Structural and Functional Differentiation of Bundle Sheath and Mesophyll Cells in the Lamina Joint of Rice Compared with that in the Corresponding Region of the Liguleless Genotype

Abstract

The structural and functional characterization of the blade-sheath boundary region of a rice cultivar T65 and its near-isogenic line T65lg were examined by light and electron microscopy and in situ hybridization. Starch accumulation in bundle sheath cells was compared between the lamina joint of T65 and the corresponding region of T65lg and also between the lamina joint and the leaf blade. In the lamina joint of T65, starch grains were predominantly accumulated in bundle sheath cells, and the starch-containing chloroplasts within these cells were spherical in shape. On the other hand, in the blade-sheath transition region of T65lg, little starch accumulation was observed and the chloroplasts were oval in both mesophyll and bundle sheath cells. Furthermore, photosynthesis-related genes, rbcS and cab, were expressed in mesophyll cells within the blade-sheath transition region of T65lg as in the leaf blade and sheath, while no expression of these genes was found within the lamina joint of T65. These facts indicate that T65lg can not develop the lamina joint from either structural or functional aspect. The present results suggest that the control mechanism of starch accumulation in bundle sheath chloroplasts in the lamina joint differs from that in leaf blade in rice.     

玉米C4光合叶不同部位“花环”结构及叶绿素含量的变化

以玉米进行C4光合的全展第5位叶片为材料,分析从叶基部到顶部的解剖结构和叶绿素含量变化,研究玉米C4光合叶片"花环"结构随叶片发育的变化规律。结果表明,玉米第5位叶从基部到顶部都具有完整的典型"花环"结构,维管束鞘细胞(BSC)和叶肉细胞(MC)的体积在叶片发育过程中具有渐变性,从叶基部到顶部BSC和MC均呈先增大后变小的趋势,而且叶绿素a、b和a+b含量呈相同的变化趋势,说明BSC和MC细胞体积与叶绿素含量的变化具有相关性。叶绿素a/b总体呈上升趋势,说明玉米第5叶基部到顶部的光合途径存在C3向C4转变的过程。玉米第5叶不同部位C4光合途径发育的渐变性比前3叶更为明显。     

Differential Sensitivity of Chloroplasts in Mesophyll and Bundle Sheath Cells in Maize,an NADP-Malic Enzyme-Type C4 Plant,to Salinity Stress

Abstract

The changes in chloroplast ultrastructure and the contents of chlorophyll, Na and K in response to salinity stress were investigated in leaves of maize, an NADP-malic enzyme-type C₄ plant species possessing dimorphic chloroplasts. The seedlings were treated with 0, 1, 2 or 3% NaCl for three or five days under a light or dark condition. In both light and dark conditions, the dry weight of salt-treated plants decreased as NaCl concentration increased. Chlorophyll and K contents of the second leaf blade decreased as NaCl concentration increased under the light condition but not under the dark condition. Na content of the second leaf blade was significantly higher at high NaCl concentrations under both light and dark conditions. However, Na content was much lower under the dark condition than light condition. Higher concentrations (2 and 3%) of NaCl significantly increased the size of plastoglobules, decreased the number and size of starch granules and altered the chloroplast ultrastructure. Under the light condition, mesophyll cell (MC) chloroplasts appeared more sensitive to the damaging effect of salinity than the bundle sheath cell (BSC) chloroplasts. MC chloroplasts became more globular in shape and showed swollen and disorganized thylakoids and reduced thickness of grana by salinity. BSC chloroplasts were less affected by salinity than MC chloroplasts. Although chloroplast size and number and size of starch granules were reduced, there was no structural distortion in the thylakoids of BSC chloroplasts. However, the thickness of grana was increased by salinity. Under the dark condition, the chloroplast structure was less affected by salinity. Though the envelope of BSC chloroplasts was occasionally damaged, the thylakoids in both MC and BSC chloroplasts were preserved under salinity stress. The present study suggests that the chloroplast damage caused by salinity is light-dependent and MC chloroplasts are more sensitive to salinity than BSC chloroplasts.     

Photosynthetic Characteristics of an Amphibious C4 Plant,Eleocharis retroflexa ssp. chaetaria

Abstract

Eleocharis retroflexa (Poir.) Urban ssp. chaetaria (Roem. & Schult.) T. Koyama, an amphibious leafless sedge, grows not only under terrestrial conditions but also under completely submerged aquatic conditions. We investigated the photosynthetic traits and structural features of the culms, which are the photosynthetic organs, in the terrestrial and submerged forms of this species and compared them with those of other amphibious species of Eleocharis which are known to change the photosynthetic modes. The culms of the terrestrial form had Kranz anatomy with well-developed Kranz (bundle sheath) cells and high levels of C₄ enzyme activity typical of the NAD-malic enzyme (NAD-ME) subtype of C₄ metabolism. They also had a δ ¹³C value typical of C₄ plants, indicating that the terrestrial form fixes carbon through the C₄ pathway. The culms of the submerged form had not only a Kranz-like anatomy but also revealed anatomical traits typical of leaves of submerged aquatic plants. The activities of the C₄ enzymes in the submerged form were lower than those in the terrestrial form, but were still in the range typical of G4 plants, ¹⁴C pulse-¹²C chase experiments with the submerged form indicated that almost all of the fixed ¹⁴C was incorporated into G4 compounds, and subsequently the raioactivity was transferred into C₃ compounds and sucrose. The submerged form showed no diurnal fluctuation in malate level. These data demonstrate that a C₄ metabolism is operative even in the submerged form. This unique amphibious C₄ plant provides an intriguing example of the physiological and ecological adaptability of C₄plants.     

Production of Raphanus sativus (C3)-Moricandia arvensis (C3-C4 intermediate) Monosomic and Disomic Addition Lines with Each Parentl Cytoplasmic Background and their Photorespiratory Characteristics

Abstract

We are maintaining five Moricandia arvensis monosomic addition lines of Raphanus sativus carrying R. sativus cytoplasm (autoplasmic MALs) and twelve M. arvensis MALs of R. sativus carrying M. arvensis cytoplasm (alloplasmic MALs) from BC₆ to BC₈ generation, and newly produced five M. arvensis disomic addition lines of R. sativus (autoplasmic DALs) and seven M. arvensis DALs of R. sativus carrying M. arvensis cytoplasm (alloplasmic DALs) from selfing and sib-crossing of the MALs and DALs in S₃BC₅ and S₂BC₆ generations. The structural, biochemical and physiological characteristics related to photorespiration of these MALs and DALs were compared to study the genetic mechanisms of the C₃-C₄ intermediate photosynthesis in the individual chromosomes of M. arvensis. The CO₂ compensation point of the autoplasmic and alloplasmic DALs (RMa-b and MaR-b DALs) with one pair of M. arvensis ‘b’ chromosome were 29.4 and 30.1 μmol mol-¹, respectively, which were significantly lower than that of other DALs and MALs as well as R. sativus (34.5 𰂼mol mol-¹). An immunogold electron microscopic study of the P-protein of glycine decarboxylase (GDC) in photosynthetic cells of the RMa-b DAL revealed that the bundle sheath cell (BSC) mitochondria were more intensively labeled for the protein than the mesophyll cell (MC) mitochondria. The ratio of the labeling density of the BSC mitochondria to that of the MC mitochondria was 1.13, which lies between values in M. arvensis (2.66) and R. sativus (0.76). These data suggest that the ‘b’ chromosome of M. arvensis genome controls the expression of C₃-C₄ intermediate characteristics.     

More lightweight and isodiametric seeds for C4 than for C3 grasses are associated with preference for open habitats of C4 grasses in a temperate flora

Seed mass and shape of grasses were analysed in a temperate flora containing 178 and 26 species belonging to C₃ and C₄ photosynthetic types respectively. The weedy character and the annual or perennial status were also considered. On the basis of the seed traits studied, three groups were distinguished: C₄ grasses, annual C₃ grasses of weedy character and perennial non‐weedy C₃ grasses. The C₄ group had more isodiametric (same diameter in all directions) seed shape and lower average seed mass than the C₃ group. To our knowledge, this has not yet been described for temperate C₄ grasses and is certainly associated with their preference for open habitats where competition for light is small. Weedy annual C₃ grasses had heavier and less isodiametric seeds than C₄ grasses did. These species are mostly specialized to establish in the dense cover of perennial vegetation, and this ability distinguishes them from the C₄ group. Non‐weedy perennial C₃ species possessed less isodiametric seeds than did C₄ grasses, but did not differ from weedy annual C₃ grasses. As most alien C₄ grasses naturalized in Hungary are annuals with small, isodiametric seeds, these traits are good candidates to be included in screening for potential future invasives in open habitats.     

外源C4二羧酸对转玉米PEPC基因水稻C4光合途径的促进作用

以转PEPC基因水稻和野生型水稻Kitaake为材料, 研究了不同基因型水稻叶片中的C4光合微循环及其功能。通过测定与光合C4途径有关的关键酶,如磷酸烯醇式丙酮酸羧化酶(PEPC)、NADP苹果酸酶(NADP-ME)、NADP苹果酸脱氢酶(NADP-MDH)和丙酮酸磷酸双激酶(PPDK),说明野生型水稻叶片中具有完整的C4光合酶体系;用外源草酰乙酸(OAA)或苹果酸(MA)饲喂叶切片或叶绿体后明显增加光合速率,证明野生型水稻中具有一个有限的光合C4微循环。将玉米的PEPC基因导入原种水稻后,可大幅度提高光合C4微循环的速率。测定不同基因型的CO2交换速率,看出水稻中C4光合微循环的增强有提高净光合速率(Pn)和降低Pr/Pn比值的作用。叶绿素荧光特性分析表明,C4光合微循环的增强伴随着PSⅡ电子传递效率(Fv/Fm)和光化学猝灭(qp)的增加,以及非光化学猝灭(qN)的降低。      

Response of the leaf photosynthetic rate to available nitrogen in erect panicle-type rice (Oryza sativa L.) cultivar,Shennong265

Increasing the yield of rice per unit area is important because of the demand from the growing human population in Asia. A group of varieties called erect panicle-type rice (EP) achieves very high yields under conditions of high nitrogen availability. Little is known, however, regarding the leaf photosynthetic capacity of EP, which may be one of the physiological causes of high yield. We analyzed the factors contributing to leaf photosynthetic rate (P_n) and leaf mesophyll anatomy of Nipponbare, Takanari, and Shennong265 (a EP type rice cultivar) varieties subjected to different nitrogen treatments. In the field experiment, P_n of Shennong265 was 33.8 μmol m^?2 s^?1 in the high-N treatment, and was higher than that of the other two cultivars because of its high leaf nitrogen content (LNC) and a large number of mesophyll cells between the small vascular bundles per unit length. In Takanari, the relatively high value of P_n (31.5 μmol m^?2 s^?1) was caused by the high stomatal conductance (g_s; .72 mol m^?2 s^?1) in the high-N treatment. In the pot experiment, the ratio of P_n/C_i to LNC, which may reflect mesophyll conductance (g_m), was 20–30% higher in Nipponbare than in Takanari or Shennong265 in the high N availability treatment. The photosynthetic performance of Shennong265 might be improved by introducing the greater ratio of P_n/C_i to LNC found in Nipponbare and greater stomatal conductance found in Takanari.     

Ultrastructure of Hybrid Callus between C3 and C4 Species of Amaranthaceae

Summary

Unlike the parent line, the hybrid calli between C₃ and C₄ species of Amaranthaceae have poor division capability. Therefore, in this study the fine structures of hybrid callus derived from protoplasts of Celosia cristata L. cv. Pink Charm (C₃ species) cell suspension and Amaranthus tricolor L. cv. Perfecta (C₄ species) cotyledon callus were investigated by light and electron microscopy, and compared with the parental. All callus lines were composed of parenchymatous cells possessing a voluminous vacuole. Parental callus lines contained organelles with a relatively normal structure. The peculiar feature characterizing the C₃/C₄ hybrid callus was the presence of highly lobed nucleus with multinucleoli and numerous small vacuoles having autophagic activity scattered in the cytoplasm. The autophagic activity is apparently related to the existence of several inclusions such as cytoplasmic organelles in the central vacuole. It might also be related to the pronounced expansion of central vacuole and the reduced cytoplasm. The failure to sustain proliferation related to some atypical features of the organelles is discussed.     

BS型小麦光温敏雄性不育系光合特性研究

为了解BS型小麦光温敏雄性不育系光合特性的差异,分析其光合特性的影响因子,以指导高光效不育系的选育,以6份BS型小麦光温敏不育系和常规品种京411为研究材料,于北京顺义(不育系繁殖区)和河南南阳(不育系制种区)分期播种,分析不同材料在抽穗期、开花期和花后30 d的净光合速率(P_n)、气孔导度(G_s)、胞间CO_2浓度(C_i)、蒸腾速率(T_r)、叶绿素荧光参数(F_v/F_m)等光合特性指标的差异,以及不同区域不育系的P_n、T_r和结实率与播期、旗叶面积及株高的相关性。结果表明,不育系的P_n、G_s、T_r和F_v/F_m变化趋势均为抽穗期或开花期达到最高值,花后30 d降为最低值,而C_i的变化趋势为花后30 d达到最高值。P_n和T_r较高的材料为BS485,G_s和T_r较高的材料为BS366;P_n较低的材料为BS93,T_r较低的材料为BS1453和BS210。在北京顺义,P_n与结实率、G_s、T_r和F_v/F_m呈正相关,与旗叶面积呈负相关;T_r与播期、P_n和G_s呈正相关,与株高和F_v/F_m呈负相关;结实率与P_n呈正相关,与旗叶面积呈负相关。对P_n影响最主要的因子是播期和T_r,其次是结实率、F_v/F_m、G_s和旗叶面积;对T_r影响最主要的因子是播期、G_s和P_n,其次是株高和F_v/F_m;对结实率影响最主要的因子是P_n,其次是G_s和旗叶面积。在河南南阳,P_n和结实率与各因子相关性均不显著;T_r与播期和G_s呈正相关,与C_i呈负相关;对T_r影响最主要的因子是播期和G_s,其次是C_i。     

Effects of elevated atmospheric CO2 concentration on morphology of leaf blades in Chinese yam

ABSTRACT

The effects of elevated carbon dioxide concentration on the morphology of leaf blades in two Chinese yam lines under different temperature conditions were determined. Plants were grown under two [CO₂] levels, ambient (about 400 µmol mol^?1) and elevated (ambient + 200 µmol mol^?1) in the daytime, and two mean air temperature regimes, approximately ambient temperature (22.2°C) and high temperature (25.6°C). The palisade layer was thicker under elevated [CO₂] than under ambient [CO₂] in both temperature regimes, and the whole yam leaf blade was thicker under elevated [CO₂] than under ambient [CO₂] in the approximately ambient temperature regime. The numbers of chloroplasts per palisade cell and spongy cell as well as per unit profile area of palisade cell, number of starch grains per chloroplast, profile area of the starch grain, and starch-to-chloroplast area ratio in both palisade and spongy cells were higher under elevated [CO₂] than under ambient [CO₂] in both temperature regimes. Furthermore, the stomatal density on the abaxial side of the leaf blade in Chinese yam was greater under elevated [CO₂] than under ambient [CO₂] under both temperature regimes, and stomatal-pore length was higher under elevated [CO₂] than under ambient [CO₂] in the approximately ambient temperature regime. These results indicate that elevated [CO₂] positively affects the photosynthetic apparatus. The results of this study provide information for understanding the response characteristics of the leaf blade under elevated [CO₂] and a possible explanation for the positive photosynthetic responses of Chinese yam to elevated [CO₂] in our previous study.

List of Abbreviations:[CO₂]: carbon dioxide concentration     

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.     

Screening of High kcat Rubisco among Poaceae for Improvement of Photosynthetic CO2 Assimilation in Rice

Abstract

The activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a major limitation of photosynthetic CO₂ assimilation in C₃ plants. In order to find useful Rubisco for improvement of photosynthesis in rice under elevated CO₂, we analyzed the catalytic turnover rate (k_cat) of Rubisco in Poaceae including C₃ alpine plants, C₃ cold-resistant plants and C₄ plants. Rubisco in these plants showed 1.1- to 2.8-fold higher k_cat than that in rice. However, the most of high k_cat Rubisco also showed a higher km for CO₂ (Kc) than that of rice, indicating that increase in k_cat led to decrease in the affinity for CO₂. Rubisco in Festuca ovina, Phleum pratense and Sorghum bicolor showed relatively high k_cat to Kc. Although the k_cat of Rubisco in F. ovina and P. pratense was not so high (1.5-1.6 fold relative to rice), the Kc was comparable to that in rice and the amino acid sequence of RbcL shared higher identity to that in rice than that in S. bicolor. By contrast, Rubisco of S. bicolor showed considerably high k_cat (2.5-fold relative to rice), which is considered to be the most important factor for improvement of photosynthesis. In our estimation, the expression of high k_cat Rubisco of F. ovina and S. bicolor in rice could significantly enhance CO₂ assimilation at Ci of 50 Pa, the level assumed to be reached by the middle of this century.     

Inheritance of C3―C4 Intermediate Photosynthesis in Reciprocal Hybrids Between Moricandia Arvensis (C3―C4) and Brassica Oleracea (C3) That Differ in Their Genome Constitution

Abstract

To Elucidate The Genetic Mechanisms Underlying C₃―C₄ intermediate Photosynthesis, We investigated The Structural and Photosynthetic Characteristics of Leaves of Reciprocal Hybrids Between The C₃―C₄ intermediate Species Moricandia Arvensis (L.) Dc. (Mama) and The C₃ Species Brassica Oleracea L. (Cabbage; Cc), Which Differ in Genome Constitution. Moricandia Arvensis Bundle Sheath (Bs) Cells included Many Centripetally Located Chloroplasts and Mitochondria, Whereas Those of Cabbage Had Few Organelles. Hybrid Leaves Were Structurally intermediate Between Those of The Parents and Showed Stronger intermediate C₃―C₄ Features As The Proportion of The Ma Genome increased. The P-Protein of Glycine Decarboxylase (Gdc) Was Confined Mainly To Bs Mitochondria in M. Arvensis, But Accumulated More in The Mesophyll (M) of Cabbage. in The Hybrids, The Accumulation of Gdc in Bs Cells increased With An increasing Ma:C Ratio. Hybrids Exhibited Gradients in Structural and Biochemical Features, Even in Reciprocal Crosses. The Co₂ Compensation Point of Reciprocal Hybrids With High Ma:C Ratios Was Lower Than That of Cabbage But Higher Than That of M. Arvensis. Thus, The Structural and Biochemical Features in Hybrid Leaves Reduced Photorespiration. Moricandia Arvensis Had A Higher Photosynthetic Rate Than Cabbage, But The Photosynthetic Rates of Hybrids Were intermediate Between Those of The Parents Or Comparable To That of M. Arvensis. Our Results Demonstrate That The C₃―C₄ intermediate Characteristics Are inherited Based On The Ratio of The Parent Genomes, and That There Is No Evidence of Cytoplasmic inheritance in These Characteristics.