Variations in physiological,biochemical, and structural traits of photosynthesis and resource use efficiency in maize and teosintes (NADP-ME-type C4)

C₄ plants show higher photosynthetic capacity and resource use efficiency than C₃ plants. However, the genetic variations of these traits and their regulatory factors in C₄ plants still remain to be resolved. We investigated physiological, biochemical, and structural traits involved in photosynthesis and photosynthetic water and nitrogen use efficiencies (PWUE and PNUE) in 22 maize lines and four teosinte lines from various regions of the world. Net photosynthetic rate (P_N) ranged from 32.1 to 46.5 μmol m^?2 s^?1. P_N was positively correlated with stomatal conductance, transpiration rate, and chlorophyll, nitrogen and soluble protein contents of leaves, but not with specific leaf weight. P_N was positively correlated with the activities of ribulose-1,5-bisphosphate carboxylase/oxygenase and the C₄-acid decarboxylases, NADP-malic enzyme and phosphoenolpyruvate carboxykinase, but not with the activity of phosphoenolpyruvate carboxylase. Leaf structural traits (stomatal parameters, leaf thickness, and interveinal distance) were not correlated with P_N. These data suggest that physiological and biochemical traits are involved in the genetic variation of P_N, but structural traits are not directly involved. PWUE is in the lower class of values reported for C₄ plants, whereas PNUE is in the highest class of values reported for C₄ plants. PNUE was negatively correlated with leaf nitrogen content but not significantly correlated with P_N. PWUE was not correlated with δ¹³C values of leaves, indicating difficulty in using δ¹³C values as an indicator of PWUE of maize. In general, teosinte lines showed lower P_N but higher PWUE than maize lines.     

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.     

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。     

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.     

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 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.     

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.     

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.     

Effects of partial root-zone drying on yield,tuber size and water use efficiency in potato under field conditions

Water resources are limited for irrigation worldwide; therefore, there is a need for water-saving irrigation practices to be explored. Partial root-zone drying (PRD) is a new water-saving irrigation strategy being tested in many crop species. Experiments were conducted in potato (Solanum tuberosum L. cv. Folva) under open field conditions in 2004 and under a mobile rainout shelter in 2005. Two subsurface irrigation treatments were studied: full irrigation (FI) receiving 100% of evaporative demands, 50.1 and 201 mm of irrigation water in the 2 years, to keep it close to field capacity; and PRD, which received 21.7 and 140 mm of irrigation in 2004 and 2005 respectively. Due to rain in 2004, the PRD treatment was imposed over a short period only during the late tuber filling and maturing stages. In 2005, the PRD treatment was imposed during the whole period of tuber filling and tuber maturation. The PRD treatment was shifted from one side to the other side of potato plants every 5–10 days. Especially in 2005 it was apparent that stomatal conductance was generally lower in the PRD than in the FI plants, whereas leaf water potential tended to be lower in only a few instances. During the treatment period, plants were harvested five times, and no significant difference was found between the treatments in leaf area index, top dry mass and tuber yield. At final harvest, tubers were graded based on size into four classes C₁–C₄, of which the yield of the important marketable class (C₂) was significantly higher (20%) in the PRD than in the FI treatment. Compared with FI, the PRD treatment saved 30% of irrigation water while maintaining tuber yield, leading to a 61% increase of irrigation water use efficiency. The limited data of 2004 support these results. In summary, PRD is a promising water-saving irrigation strategy for potato production in areas with limited water resources.     

Variability of light interception and radiation use efficiency in maize and soybean

Variability of light interception and its derivatives are poorly understood at the field-scale in maize (Zea mays L.) and soybean [Glyine max (L.) Merr.]. Quantifying variability can provide reliable estimates of field-scale processes and reliable methodology. A field study was conducted during the 2005 growing season in a 31 ha maize and 23 ha soybean field rotated annually near Ames, IA to measure variability of cumulatively intercepted photosynthetically active radiation (CI-PAR) and radiation use efficiency (RUE) by deploying eight line quantum sensors in each field. Cumulative mean PAR interception for soybean was 575 MJ m⁻² ending on day of the year (DOY) 249 compared with 687 MJ m⁻² in maize ending on DOY 244. Soybean standard error (s_X) for a single sensor was 4.48% and with six sensors was 1.83% of the final CI-PAR. Maize s_X for a single sensor was 5.29% and with eight sensors was 1.87% of the final CI-PAR. Crop biomass was quantified weekly by collecting four 1 m² samples. Soybean RUE using all sensors was 1.44 ± 0.06 g MJ PAR⁻¹. The highest CI-PAR from a single sensor had RUE of 1.32 and the lowest was 1.55 g MJ PAR⁻¹. Maize RUE using all sensors was 3.35 ± 0.09. The highest CI-PAR from a single sensor had RUE of 2.87 and the lowest was 3.70 g MJ PAR⁻¹. Reliable transmitted PAR and RUE estimates are obtainable at the field-scale in maize and soybean with four and three sensors, respectively, assuming that crop biomass is accurately measured.     

Yield, water use efficiency and nitrogen uptake in potato: influence of drought stress

Summary A lysimeter experiment was performed to study the optimal allocation of limited water supply in potatoes. Irrigation regimes equal to 40, 60 and 80% of maximum evapotranspiration (ET) were evenly applied over the crop cycle. Other treatments involved withholding 80 mm of irrigation, based on ET, beginning at each of three designated growth stages (tuber initiation, early and late tuber growth). An irrigated control treatment, restoring the entire ET, was included for comparison. Continuous drought stress reduced photosynthesis as irrigation volumes were reduced. Plant biomass and tuber yield decreased almost proportionally to water consumption, so that WUE was roughly constant. N uptake was highest in the control and in 80% ET treatment. Withholding water during tuberisation severely hindered plant physiological processes and penalized tuber yield. Reductions in photosynthesis, total biomass and yield were the greatest when drought was imposed during tuber initiation. The earliest stress resulted in the lowest WUE and N uptake. A new crop water stress index (SI) was proposed, which combines atmospheric demand for water and canopy temperature.     

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.     

Radiation use efficiency,N accumulation and biomass production of high-yielding rice in aerobic culture

The concept of aerobic culture is to save water resource while maintaining high productivity in irrigated rice ecosystem. This study compared nitrogen (N) accumulation and radiation use efficiency (RUE) in the biomass production of rice crops in aerobic and flooded cultures. The total water input was 800–1300 mm and 1500–3500 mm in aerobic culture and flooded culture, respectively, and four high-yielding rice cultivars were grown with a high rate of N application (180 kg N ha⁻¹) at two sites (Tokyo and Osaka) in Japan in 2007 and 2008. The aboveground biomass and N accumulation at maturity were significantly higher in aerobic culture (17.2–18.5 t ha⁻¹ and 194–233  kg N ha⁻¹, respectively) than in flooded culture (14.7–15.8 t ha⁻¹ and 142–173 kg N ha⁻¹) except in Tokyo in 2007, where the surface soil moisture content frequently declined. The crop maintained higher N uptake in aerobic culture than in flooded culture, because in aerobic culture there was a higher N accumulation rate in the reproductive stage. RUE in aerobic culture was comparable to, or higher than, that in flooded culture (1.27–1.50 g MJ⁻¹ vs. 1.20–1.37 g MJ⁻¹), except in Tokyo in 2007 (1.30 g MJ⁻¹ vs. 1.37 g MJ⁻¹). These results suggest that higher biomass production in aerobic culture was attributable to greater N accumulation, leading to higher N concentration (N%) than in flooded culture. Cultivar differences in response to water regimes were thought to reflect differences in mainly (1) early vigor and RUE under temporary declines in soil moisture in aerobic culture and (2) the ability to maintain high N% in flooded culture.     

Environmental effects on seed yield determination of irrigated peanut crops: Links with radiation use efficiency and crop growth rate

In Argentina, delayed sowing causes a decrease in seed yield and in radiation use efficiency (RUE) of peanut crops (Arachis hypogaea L.), but it is not known if RUE reduction is mainly due to reduced temperature during late reproductive stages or to a sink limitation promoted by decreased seed number in these conditions. We analyzed seed yield determination and RUE dynamics of two cultivars (Florman and ASEM) in four irrigated field experiments (Exp_n) grown at three sites and five contrasting sowing dates (between 17 October and 21 December) in three growing seasons. An additional field experiment was performed with widely spaced plants (i.e. with no interference among them) to evaluate the effect of peg removal on RUE and leaf carbon exchange rate (CER). Seasonal dynamics of mean air temperature and irradiance, biomass production (total and pods), and intercepted photosynthetically active radiation (IPAR) were followed. Seed yield and seed yield components (pod number, seeds per pod, seed number and seed weight) were determined at final harvest. Crop growth rate (CGR) and pod growth rate (PGR) were computed for growth phases of interest. RUE values for crops sown until 14 November were 1.89–1.98 g MJ⁻¹ IPAR, within the usual range. RUE decreased significantly for cv. Florman in the late sowing of Exp₁ (29 November) and for both cultivars in Exp₃ (21 December sowing). Across experiments, seed yield (4.5-fold variation relative to minimum) was strongly associated (r² = 0.87, P < 0.0001) with variations in seed number (3.5-fold variation relative to minimum), and to a lesser extent (r² ≤ 0.54, P ≤ 0.001) to variations in seed weight (1.9-fold variation relative to minimum). Seed number was positively related (P < 0.01) to CGR (r² = 0.66) and to PGR (r² = 0.72) during the R₃–R_6.5 phase (seed number determination window), while crop growth during the grain-filling phase (i.e. between R_6.5 and final harvest) was positively associated with grain number (r² = 0.80, P < 0.001). No association was found between RUE and mean air temperature, neither for the whole cycle nor for the phase between R_6.5 and final harvest, which showed the largest temperature variation (16.4–22.4 °C) across experiments. Use of mean minimum temperature records (range between 13.8 and 18.5 °C) did no improve the relationship. However, grain-filling phase RUE showed a positive (r² = 0.69, P = 0.003) linear response to seed number across experiments. This apparent sink limitation of source activity was consistent with the reduced RUE (from 2.73 to 1.42 g MJ⁻¹ IPAR) and reduced leaf CER at high irradiance (from ca. 30 to 15 μmol m⁻² s⁻¹) for plants subjected to 75% peg removal.     

Characterizing leaf gas exchange responses of cotton to full and limited irrigation conditions

Plant responses to water deficit need to be monitored for producing a profitable crop as water deficit is a major constraint on crop yield. The objective of this study was to evaluate physiological responses of cotton (Gossypium hirsutum) to various environmental conditions under limited water availability using commercially available varieties grown in South Texas. Soil moisture and variables of leaf gas exchange were measured to monitor water deficit for various varieties under different irrigation treatments. Lint yield and growth variables were also measured and correlations among growth parameters of interest were investigated. Significant differences were found in soil moisture, leaf net assimilation (A_n), stomatal conductance (g), transpiration rate (T_r), and instantaneous water use efficiency (WUE_i) among irrigation treatments in 2006 while no significant differences were found in these parameters in 2007. Some leaf gas exchange parameters, e.g., T_r, and leaf temperature (T_L) have strong correlations with A_n and g. A_n and WUE were increased by 30–35% and 30–40%, respectively, at 600 μmol (CO₂) m⁻² s⁻¹ in comparison with 400 μmol (CO₂) m⁻² s⁻¹. Lint yield was strongly correlated with g, T_r, WUE, and soil moisture at 60 cm depth. Relative A_n, T_r, and T_L started to decrease from FTSW 0.3 at 60 cm and FTSW 0.2 at 40 cm. The results demonstrate that plant water status under limited irrigation management can be qualitatively monitored using the measures of soil moisture as well as leaf gas exchange, which in turn can be useful for describing yield reduction due to water deficit. We found that using normalized A_n, T_r, and T_L is feasible to quantify plant water deficit.