Carbon isotope discrimination in photosynthetic bark

We developed and tested a theoretical model describing carbon isotope discrimination during photosynthesis in tree bark. Bark photosynthesis reduces losses of respired CO₂ from the underlying stem. As a consequence, the isotopic composition of source CO₂ and the CO₂ concentration around the chloroplasts are quite different from those of photosynthesizing leaves. We found three lines of evidence that bark photosynthesis discriminates against ¹³C. First, in bark of Populus tremuloides, the '¹³C of CO₂ efflux increased from -24.2‰ in darkness to -15.8‰ in the light. In Pinus monticola, the '¹³C of CO₂ efflux increased from -27.7‰ in darkness to -10.2‰ in the light. Observed increases in '¹³C were generally in good agreement with predictions from the theoretical model. Second, we found that '¹³C of dark-respired CO₂ decreased following 2-3 h of illumination (P<0.01 for Populus tremuloides, P<0.001 for Pinus monticola). These decreases suggest that refixed photosynthate rapidly mixes into the respiratory substrate pool. Third, a field experiment demonstrated that bark photosynthesis influenced whole-tissue '¹³C. Long-term light exclusion caused a localized increase in the '¹³C of whole bark and current-year wood in branches of P. monticola (P<0.001 and P<0.0001, respectively). Thus bark photosynthesis was shown to discriminate against ¹³C and create a pool of photosynthate isotopically lighter than the dark respiratory pool in all three experiments. Failure to account for discrimination during bark photosynthesis could interfere with interpretation of the '¹³C in woody tissues or in woody-tissue respiration.     

Tracing carbon and oxygen isotope signals from newly assimilated sugars in the leaves to the tree-ring archive

The analysis of δ ¹³C and δ ¹⁸O in tree-ring archives offers retrospective insights into environmental conditions and ecophysiological processes. While photosynthetic carbon isotope discrimination and evaporative oxygen isotope enrichment are well understood, we lack information on how the isotope signal is altered by downstream metabolic processes.
In Pinus sylvestris , we traced the isotopic signals from their origin in the leaf water ( δ ¹⁸O) or the newly assimilated carbon ( δ ¹³C), via phloem sugars to the tree-ring, over a time-scale that ranges from hours to a growing season.
Seasonally, variable ¹³C enrichment of sugars related to phloem loading and transport did lead to uncoupling between δ ¹³C in the tree-ring, and the c _i/ c _a ratio at the leaf level. In contrast, the oxygen isotope signal was transferred from the leaf water to the tree-ring with an expected enrichment of 27‰, with time-lags of approximately 2 weeks and with a 40% exchange between organic oxygen and xylem water oxygen during cellulose synthesis.
This integrated overview of the fate of carbon and oxygen isotope signals within the model tree species P. sylvestris provides a novel physiological basis for the interpretation of δ ¹³C and δ ¹⁸O in tree-ring ecology.     

Hydrogen isotope discrimination in higher plants: Correlations with photosynthetic pathway and environment

Summary The ratio of deuterium to hydrogen (expressed as D) in hydrogen released as water during the combustion of dried plant material was examined. The D value (metabolic hydrogen) determined on plant materials grown under controlled conditions is correlated with pathways of photosynthetic carbon metabolism. C₃ plants show mean D values of-132 for shoots and -117 for roots; C₄ plants show mean D values of -91 for shoots and-77 for roots and CAM plants a D value of-75 for roots and shoots. The difference between the D value of shoot material from C₃ and C₄ plants was confirmed in species growing under a range of glasshouse conditions. This difference in D value between C₃ and C₄ species does not appear to be due to differences in the D value (tissue water) in the plants as a result of physical fractionation of hydrogen isotopes during transpiration. In C₃ and C₄ plants the hydrogen isotope discrimination is in the same direction as the carbon isotope discrimination and factors contributing to the difference in D values are discussed. In CAM plants grown in the laboratory or collected from the field D values range from-75 to +50 and are correlated with ¹³C values. When deprived of water, the D value (metabolic hydrogen) in both soluble and insoluble material in leaves of Kalanchoe daigremontiana Hamet et Perr., becomes less negative. These changes may reflect the deuterium enrichment of tissue water during transpiration, or in field conditions, may reflect the different D value of available water in areas of increasing aridity. Whatever the origin of the variable D value in CAM plants, this parameter may be a useful index of the water relations of these plants under natural conditions.     

土壤水分与遮荫水平对棉花叶片光全特性的影响研究

以盆栽棉花（中棉２３号）做为供试材料,通过人工遮荫,设计了３种遮荫水平（不遮光（ＣＫ）,遮光７５％（ＤＮ）,遮光４０％（ＳＮ）和高、中、低３种水分组合处理,其土壤含水量分别为田间持水量的８５％￣１００％（高水分,ＨＷ）、６５％￣８５％（中水分,ＭＷ）和４５％￣６５％（低水分,ＬＷ）。探讨了土壤水分和遮荫水平共同作用对嘿期至蕾期中午的气孔寻度、净江合速率、蒸腾速率和叶水势等的影响。高水分下ＤＮ和ＳＮ     

Genetic variation in photosynthetic capacity, carbon isotope discrimination and mesophyll conductance in provenances of Castanea sativa adapted to different environments

1. Provenances of Castanea sativa from populations adapted to different climatic areas of Turkey were grown in a field trial in Italy. Carbon isotope discrimination (Δ) in leaf dry matter and in leaf soluble sugar, were measured, along with photosynthesis, stomatal conductance and mesophyll conductance, to study the variability of primary productivity and its ecological significance in European Chestnut.
2. Genetic variations were found in RuBP carboxylase, chlorophyll, leaf soluble protein and leaf thickness.
3. Carbon isotope discrimination (Δ) in leaf dry matter was greater in drought-adapted than in wet-adapted provenances. A similar variation of Δ was observed in leaf soluble carbohydrates either under watered or drought conditions. Possible environmental effects of variables such as vapour pressure difference, on the relationship between transpiration efficiency and carbon isotope discrimination are discussed, on the basis of short-term and long-term results.
4. Generally low values of Δ encountered among provenances were explained not only by low values of intercellular CO₂ partial pressure but also by consistently low values of mesophyll conductance leading to reduced chloroplastic CO₂ partial pressure. A decrease in mesophyll conductance was induced by water shortage. Co-ordination was found between stomatal and mesophyll conductance, with the drought-adapted provenances showing much higher mesophyll conductance than the wet-adapted provenances. Variations in mesophyll conductance were related to differences in leaf protein content.
5. Possible ecophysiological adaptive mechanisms are discussed taking into account stomatal sensitivity, modulation of photosynthetic capacity and water-use efficiency under drought conditions.     

Effects of soil strength on the relation of water-use efficiency and growth to carbon isotope discrimination in wheat seedlings

The ratio of carbon accumulation to transpiration, W, of wheat (Triticum aestivum L.) seedlings increased with increasing soil strength, measured as soil penetrometer resistance, and this was already apparent at the two leaf stage. The ratio was negatively correlated with carbon isotope discrimination, in accord with theory. This means that decrease in intercellular partial pressure of CO₂ accounted for an important part of the increase in W with increasing soil strength. Despite a lower CO₂ concentration in the leaves at high soil strength, assimilation rate per unit leaf area was enhanced. Greater ribulose 1,5-bisphosphate carboxylase activity confirmed that photosynthetic capacity was actually increased. This pattern of opposite variation of assimilation rate and of stomatal conductance is unusual. The ratio of plant carbon mass to leaf area increased markedly with increasing soil strength, mainly because of a greater investment of carbon into roots than into shoots. A strong negative correlation was found between this ratio and carbon isotope discrimination. For a given increase in discrimination, decrease in carbon mass per leaf area was proportionally larger than decrease in assimilation rate, so that relative growth rate was positively correlated to carbon isotope discrimination.     

Decarboxylation of glycine contributes to carbon isotope fractionation in photosynthetic organisms

Carbon isotope effects were investigated for the reaction catalyzed by the glycine decarboxylase complex (GDC; EC 2.1.2.10). Mitochondria isolated from leaves of pea (Pisum sativum L.) and spinach (Spinacia oleracea L.) were incubated with glycine, and the CO₂ evolved was analyzed for the carbon isotope ratio (δ¹³C). Within the range of parameters tested (temperature, pH, combination of cofactors NAD⁺, ADP, pyridoxal 5-phosphate), carbon isotope shifts of CO₂ relative to the C₁-carboxyl carbon of glycine varied from +14‰ to −7‰. The maximum effect of cofactors was observed for NAD⁺, the removal of which resulted in a strong ¹²C enrichment of the CO₂ evolved. This indicates the possibility of isotope effects with both positive and negative signs in the GDC reaction. The measurement of δ¹³C in the leaves of the GDC-deficient barley (Hordeum vulgare L.) mutant (LaPr 87/30) plants indicated that photorespiratory carbon isotope fractionation, opposite in sign when compared to the carbon isotope effect during CO₂ photoassimilation, takes place in vivo. Thus the key reaction of photorespiration catalyzed by GDC, together with the key reaction of CO₂ fixation catalyzed by ribulose-1,5-bisphosphate carboxylase, both contribute to carbon isotope fractionation in photosynthesis. This revised version was published online in June 2006 with corrections to the Cover Date.     

Short-term variation in the isotopic composition of organic matter allocated from the leaves to the stem of Pinus sylvestris: effects of photosynthetic and postphotosynthetic carbon isotope fractionation

We aimed to quantify the separate effects of photosynthetic and postphotosynthetic carbon isotope discrimination on δ¹³C of the fast‐turn‐over carbon pool (water soluble organic carbon and CO₂ emitted from heterotrophic tissues), including their diel variation, along the pathway of carbon transport from the foliage to the base of the stem. For that purpose, we determined δ¹³C in total and water‐soluble organic matter of the foliage plus δ¹³C and δ¹⁸O in phloem organic matter of twigs and at three heights along the stem of Pinus sylvestris over a nine‐day period, including four measurements per day. These data were related to meteorological and photosynthesis parameters and to the δ¹³C of stem‐emitted CO₂. In the canopy (foliage and twigs), the δ¹³C of soluble organic matter varied diurnally with amplitudes of up to 1.9‰. The greatest ¹³C enrichment was recorded during the night/early morning, indicating a strong influence of starch storage and remobilization on the carbon isotope signatures of sugars exported from the leaves. ¹³C enrichment of soluble organic matter from the leaves to the twig phloem and further on to the phloem of the stem was supposed to be a result of carbon isotope fractionation associated with metabolic processes in the source and sink tissues. CO₂ emitted from the stem was enriched by 2.3–5.2‰ compared with phloem organic matter. When day‐to‐day variation was addressed, water‐soluble leaf δ¹³C and twig phloem δ¹⁸O were strongly influenced by c_i/c_a and stomatal conductance (G_s), respectively. These results show that both photosynthetic and postphotosynthetic carbon isotope fractionation influence δ¹³C of organic matter over time, and over the length of the basipetal transport pathway. Clearly, these influences on the δ¹³C of respired CO₂ must be considered when using the latter for partitioning of ecosystem CO₂ fluxes or when the assessment of δ¹³C in organic matter is applied to estimate environmental effects in c_i/c_a.     

黄瓜叶片光合电子传递对水分胁迫的响应

黄瓜叶片在水分胁迫下叶片相对含水量减少,类囊体室温吸收光谱的吸收峰降低,同时其NADP光还原活性、Ca^2 -ATPase活性也相应降低,全链电子传递明显受阻。类囊体膜蛋白电泳分析结果显示：类囊体膜色素蛋白复合体含量有不同程度的降低,其中PSⅡ色素蛋白复合体含量下降较多,试验结果表明水分胁迫通过限制光能的吸收,传递双及转换效率,抑制了光合电子传递过程。     

Rapid isolation of mesophyll cells from leaves of soybean for photosynthetic studies

Mesophyll cells were rapidly isolated from soybean (Glycine max [L.]) leaves using a combined Macerase enzyme-stirring technique. About 50% to 70% of the leaf cells on a chlorophyll basis from 3 grams of leaves could be isolated in 15 minutes. The cells obtained by this method were capable of high rates of photosynthesis even after storage in the dark for periods of up to 9 hours. The CO₂-saturated rate of photosynthesis increased from 5 μm CO₂/mg Chl·hour at 5 C to 170 μm CO₂/mg Chl·hour at 40 C. At atmospheric CO₂ concentration, the rate varied from 5 to 55 μm CO₂/mg Chl·hour over this temperature range. The reduced temperature response of photosynthesis at low CO₂ concentration was due to an increased Km(CO₂) of the cells with increasing temperature. The products of photosynthesis in the isolated cells were similar to the products of leaf photosynthesis.     

A global survey of carbon isotope discrimination in plants from high altitude

Summary Carbon 13/12 isotope ratios have been determined from leaves of a hundred C₃ plant species (or ecotypes) from all major mountain ranges of the globe, avoiding drought stressed areas. A general increase in ¹³C content was found with increasing altitude, i.e. overall discrimination against the heavy isotope is reduced at high elevation. The steepest decline of discrimination is observed in taxa typically ranging to highest elevations (e.g. the genus Ranunculus). Mean ¹³C for all samples collected between 2500 and 5600 m altitude is-26.15 compared to the lowland average of-28.80 (P<0.001). Forbs from highest elevations reach-24. According to theory of ¹³C discrimination this indicates decreasing relative limitation of carbon uptake by carboxylation. In other words, we estimate that the ratio of internal to external partial pressure of CO₂ (p _i /p _a )in leaves of high elevation plants is lower than in leaves of low altitude. These results confirm recent gas exchange analyses in high and low elevation plants.     

Determination of C distribution in photosynthetic serine and phosphoglycerate from grape leaves

Serine and phosphoglyceric acid are the classical marker intermediates of photorespiration and reductive carbon assimilation in C₃ plants. The present paper introduces a new and fast method for the determination of ¹⁴C distribution in these compounds by selective elimination of C-3 (NaIO₄) or C-1 (ninhydrin/ceric sulfate). Reproducibility of the procedure was found to be better than ±1% upon degradation of [U-¹⁴C]serine and [U-¹⁴C]glycerate standards.     

花生幼苗光合特性对弱光的响应

在苗期用黑色遮阳网对丰花1号和丰花2号进行不同遮光处理(不遮光,遮光27%、43%和77%),研究了苗期遮光及恢复对花生叶片光合特性的影响.结果表明： 遮光后,随遮光程度增强,叶片叶绿素含量显著增加,实际光化学效率(Ф_PSⅡ)和最大光化学效率(F_v/F_m)升高,叶绿素a/b和净光合速率(P_n)降低.恢复自然光照后1 d,在高光强下测定时,随前期遮光程度增强,P_n、气孔导度(G_s)下降,细胞间隙CO₂浓度(C_i)升高;在低光强下测定时,P_n显著升高,G_s和C_i下降;低光强与高光强下测定的P_n比值显著升高;恢复自然光照后,随前期遮光程度增强,光补偿点、光饱和点、CO₂补偿点、CO₂饱和点和羧化效率显著降低,表观量子效率显著升高.恢复自然光照后,P_n、Ф_PSⅡ和F_v/F_m先迅速下降,3～5 d后逐渐回升;恢复15 d后,遮光27%处理的各项指标恢复到对照水平,其他处理的恢复程度则因遮光程度和品种而异.丰花1号各处理叶绿素含量、P_n、Ф_PSⅡ均高于丰花2号.苗期遮光提高了花生利用弱光的能力,降低了其利用强光的能力.     

茶树叶片净光合速率对生态因子的响应

试验以铁观音品种为材料 ,研究盆栽茶树叶片净光合速率对生态因子的响应。结果表明 :茶树叶片净光合速率 ( Pn)随着光合有效辐射 ( PAR)增加而迅速升高。成龄叶片光补偿点和光饱和点分别在 50～ 1 1 0 μmol/m2· s和 1 80 0～2 0 0 0 μmol/m2· s左右 ,光补偿点和光饱和点随叶龄、环境温度不同而异。茶树叶片净光合速率对叶温的响应曲线类似抛物线型 ,环境温度为 2 4℃和 2 7℃时光合最适温度分别为 2 7± 2℃和 3 0± 2℃。光合最适温度也随叶龄不同而不同。当空气 CO2 浓度在 2 90～ 3 60μl/L时 ,茶树叶片净光合速率 ( Pn)随空气 CO2 浓度提高而提高 ,但当浓度高于 3 70μl/L或低于2 80μl/L时 ,Pn的增大或减小都很急剧 ,CO2 补偿点在 2 83μl/L左右。茶树叶片净光合速率 ( Pn)随着土壤水势逐渐下降而下降 ,临时性萎蔫点在 -50 k Pa左右 ,永久性萎蔫点在 -62 k Pa左右 ,同时复胁迫茶树的抗旱性明显提高。     

Isolation and photosynthetic characteristics of mesophyll cells from developing leaves of soybean

Mesophyll cells were isolated from developing sink leaves (25 to 30 mm in length) of soybean, Glycine max (L.) Merr. cv. Will. Leaf strips were incubated for two h in a buffered medium containing osmoticum and 0.2% Pectolyase Y-23. Gently stirring the leaf strips released from 7 to 16% of the total leaf mesophyll cells. Other pectinase enzymes, effective in releasing cells from mature source leaves (70 to 75 mm in length), did not release cells from sink leaves. Sink and source cell preparations were about 50 and 95% intact, respectively, based on the exclusion of Evans Blue dye. Intact cells could not be separated from broken cells on Ficoll or metrizamide density gradients. Total protein and catalase, glyceraldehyde-3-phosphate dehydrogenase, glycolate oxidase, phosphoenolpyruvate carboxylase, and ribulose 1,5-bisphosphate carboxylase activities on a chlorophyll basis were about 50% lower in sink mesophyll cells than in sink leaf homogenates indicating that broken sink cells lost soluble protein to the medium. Source cells and source leaf homogenates had comparable amounts of protein and enzymatic activities. Enzymatic activities on a chlorophyll basis were similar in source and sink leaves with the exception of phosphenolpyruvate carboxylase, which was two times higher in sink leaves. This enzyme was also exceptionally low in source and sink cells being only 61 and 23%, respectively, of whole leaf activities. Sink cell rates of ¹⁴CO₂ fixation were only 7% of source cell rates and sink cells did not show light-dependent O₂ evolution. Both cell preparations had photosystem II activiteis which were comparable to rates of ¹⁴CO₂ fixation at satuarating light and CO₂ concentration. It was concluded that the reduced photosynthetic rate of sink cells was limited by the low photochemical capacity rather than a limitation of Calvin cycle enzymes.     

Modelling of carbon isotope discrimination by vegetation

The paper presents a simple box model simulating the temporal variation of atmospheric ¹³CO₂ concentration, atmospheric CO₂ mixing ratio and ¹³C content of plant material. The model is driven by observed meteorological and measured biosphere-atmosphere CO₂ exchange data. The model was calibrated and validated using measurements from a Hungarian atmospheric monitoring station. The simulated atmospheric stable carbon isotope ratio data agreed well with the measured ratios considering both the magnitude and the seasonal dynamics. Observed deviations between the measured and simulated δ¹³C_air values were systematically negative in winters, while deviations were random in sign and smaller by an order of magnitude during periods when the vegetation was photosynthetically active. This difference, supported by a significant correlation between the deviation and modeled fossil fuel contributions to CO₂ concentration, suggests the increased contribution of ¹³C-depleted fossil fuel CO₂ from heating and the lower boundary layer heights during winter.     

Differences in photosynthetic apparatus of leaves from different sides of the chestnut canopy

In crowns of chestnut trees the absorption of radiant energy is not homogeneous; leaves from the south (S) side are the most irradiated, but leaves from the east (E) and west (W) sides receive around 70 % and those from north (N) face less than 20 % of the S irradiation. Compared to the S leaves, those from the N side were 10 % smaller, their stomata density was 14 % smaller, and their laminae were 21 % thinner. N leaves had 0.63 g(Chl) m⁻², corresponding to 93 % of total chlorophyll (Chl) amount in leaves of S side. The ratios of Chl a/b were 2.9 and 3.1 and of Chl/carotenoids (Car) 5.2 and 4.8, respectively, in N and S leaves. Net photosynthetic rate (P _N) was 3.9 μmol(CO₂) m⁻² s⁻¹ in S leaves, in the E, W, and N leaves 81, 77, and 38 % of that value, respectively. Morning time (10:00 h) was the period of highest P _N in the whole crown, followed by 13:00 h (85 % of S) and 16:00 h with 59 %. Below 500 μmol m⁻² s⁻¹ of photosynthetic photon flux density (PPFD), N leaves produced the highest P _N, while at higher PPFD, the S leaves were most active. In addition, the fruits from S side were 10 % larger than those from the N side.     

Topography of photosynthetic activity of leaves obtained from video images of chlorophyll fluorescence

The distribution of photosynthetic activity over the area of a leaf and its change with time was determined (at low partial pressure of O₂) by recording images of chlorophyll fluorescence during saturating light flashes. Simultaneously, the gas exchange was being measured. Reductions of local fluorescence intensity quantitatively displayed the extent of nonphotochemical quenching; quench coefficients, q_N, were computed pixel by pixel. Because rates of photosynthetic electron transport are positively correlated with (1 − q_N), computed images of (1 − q_N) represented topographies of photosynthetic activity. Following application of abscisic acid to the heterobaric leaves of Xanthium strumarium L., clearly delineated regions varying in nonphotochemical quenching appeared that coincided with areoles formed by minor veins and indicated stomatal closure in groups.