Characterization of photosynthetic pathway of plant species growing in the eastern Tibetan plateau using stable carbon isotope composition

The photosynthetic pathway of plant species collected at Menyuan, Henan, and Maduo sites, east of Tibetan Plateau, China, during the growing season were studied using stable carbon isotopes in leaves. The 232 samples leaves analyzed belonged to 161 species, 30 families, and 94 genera. The delta(13)C values (from -24.6 to -29.2 %o) indicated that all the considered species had a photosynthetic C-3 pathway. The absence of plant species with C-4 photosynthetic pathway might be due to the extremely low air temperature characterizing the Tibetan Plateau. The average delta(13)C value was significantly (p < 0.05) different between annuals and perennials at the three considered study sites. Hence the longer-lived species had greater water-use efficiency (WUE) than shorter-lived species, that is, longer-lived species are better adapted to the extreme environmental conditions of the Tibetan Plateau.

Development of near infrared analysis of faeces to estimate non-grass proportions in diets selected by cattle grazing tropical pastures

Grass (monocots) and non-grass (dicots) proportions in ruminant diets are important nutritionally because the non-grasses are usually higher in nutritive value, particularly protein, than the grasses, especially in tropical pastures. For ruminants grazing tropical pastures where the grasses are C-4 species and most non-grasses are C-3 species, the ratio of C-13/C-12 in diet and faeces, measured as delta C-13 parts per thousand, is proportional to dietary non-grass%. This paper describes the development of a faecal near infrared (NIR) spectroscopy calibration equation for predicting faecal delta C-13 from which dietary grass and non-grass proportions can be calculated. Calibration development used cattle faeces derived from diets containing only C-3 non-grass and C-4 grass components, and a series of expansion and validation steps was employed to develop robustness and predictive reliability. The final calibration equation contained 1637 samples and faecal delta C-13 range (parts per thousand) of [12.27]-[27.65]. Calibration statistics were: standard error of calibration (SEC) of 0.78, standard error of cross-validation (SECV) of 0.80, standard deviation (SD) of reference values of 3.11 and R-2 of 0.94. Validation statistics for the final calibration equation applied to 60 samples were: standard error of prediction (SEP) of 0.87, bias of -0.15, R-2 of 0.92 and RPD of 3.16. The calibration equation was also tested on faeces from diets containing C-4 non-grass species or temperate C-3 grass species. Faecal delta C-13 predictions indicated that the spectral basis of the calibration was not related to C-13/C-12 ratios per se but to consistent differences between grasses and non-grasses in chemical composition and that the differences were modified by photosynthetic pathway. Thus, although the calibration equation could not be used to make valid faecal delta C-13 predictions when the diet contained either C-3 grass or C-4 non-grass, it could be used to make useful estimates of dietary non-grass proportions. It could also be ut :sed to make useful estimates of non-grass in mixed C-3 grass/non-grass diets by applying a modified formula to calculate non-grass from predicted faecal delta C-13. The development of a robust faecal-NIR calibration equation for estimating non-grass proportions in the diets of grazing cattle demonstrated a novel and useful application of NIR spectroscopy in agriculture.

浑善达克沙地优势豆科植物 光合生理特性研究

　　氮素是植物光合生产的决定性因素，尤其是在沙地草地生态系统中，氮素贫乏往往限制植物的生长发育。因此，研究沙地植物光合作用与叶片N含量之间的关系，以及不同植物功能型氮素利用效率，有助于理解不同植物资源利用效率的差异。以浑善达克沙地分布的80种植物为研究对象，对不同生境(固定沙丘、丘间低地和湿地)、不同生活型(乔、灌、草)、不同光合途径(C3和C4)以及豆科和非豆科植物等功能型进行研究，结果表明：无论在单位叶面积水平还是单位干重水平上的叶片氮含量，均与光合速率成极显著正相关，但单位氮素的光合利用效率在不同生境以及不同功能型之间差异很大;光合氮素利用效率表现为：湿地植物>沙丘>丘间低地植物;草本植物>灌木>乔木;C4草本>C3草本植物，非豆科植物>豆科植物。 　　为了验证浑善达克沙地豆科植物是否比非豆科植物具有更高的光合潜力，我们比较研究了3种优势豆科植物小叶锦鸡儿(Caragana microphylla)、木岩黄芪(Hedysarum fruticosum var. lignosum)、披针叶黄华(Thermopsis lanceolata)和2种非豆科植物羊草(Leymus chinensis)和黄柳(Salix gordejeviii)，结果表明并非所有豆科植物都比非豆科植物有着显著高的光合速率，仅木岩黄芪表现出较高的光合速率，其它两种豆科植物的光合速率和羊草、黄柳的差异并不显著(P>0.05)，甚至低于后者，这是因为氮素利用效率(PNUE)在其中起关键作用，通过对影响PNUE的几个主要因素进行分析得出：叶绿素对光能的吸收、光化学转换效率和CO2分压并不是构成豆科和非豆科植物PNUE差异的主要因素，而Rubisco羧化效率决定了所实验的5种植物对氮素利用效率的高低。 　　木岩黄芪在浑善达克沙地的沙丘上为优势种，甚至成为流动沙丘的先锋种。除了其显著高的氮含量外，对沙丘胁迫生境的光合适应性是我们关注的另一个重点。通过对木岩黄芪和其伴生种黄柳的光反应曲线以及光合日动态的研究，发现木岩黄芪具有显著高的光合速率、水分利用效率和PSII 光化学效率，其忍受中午强光和高温的能力较强(即“光合午休”现象不明显)。另外，该物种还表现出了显著高的光饱和点和低光补偿点。 对木岩黄芪的模拟降雨试验结果表明：气体交换参数以及叶绿素荧光参数均受到干旱和模拟降雨的影响，其中气孔因素和非气孔因素共同决定了干旱条件下木岩黄芪光合速率的降低;但降雨解除干旱后，气孔导度恢复较快，而PSII 潜在活性和PSII 光能转换效率的恢复却比较缓慢。在0-15mm的降雨量范围内，随降雨量的增加各项生理指标不断升高，但大于15mm的降雨量对木岩黄芪影响不大，因此木岩黄芪可被视为低耗水型植物。 　　对木岩黄芪光合酶的研究结果表明，其C4光合酶的活性很高，磷酸稀醇式丙酮酸羧化酶(PEPcase)、NAD-苹果酸酶(NAD-ME)、NADP-苹果酸酶(NADP-ME)、NAD-苹果酸脱氢酶(NAD-MDH)、NADP-苹果酸脱氢酶(NADP-MDH) 和丙酮酸磷酸双激酶(PPDK)等酶的活性，在整个生育期内为黄柳的5倍以上，但稳定性碳同位素测定结果却表明木岩黄芪为C3植物。因此，我们认为C3豆科植物木岩黄芪体内可能存在着C4光合途径，这种机制使得其对于流动沙丘的胁迫环境有着很强的适应性和很高的资源利用效率。

Natural 13C abundance as a tracer for studies of soil organic matter dynamics

A method for measuring the long- and medium-term turnover of soil organic matter is described. Its principle is based on the variations of 13C natural isotope abundance induced by the repeated cultivations of a plant with a high 13C/12C ratio (C4 photosynthetic pathway) on a soil which has never carried any such plant. The 13C/12C ratio in soil organic matter being about equal to the 13C/12C ratio of plant materials from which it is derived, changing the 13C content of the organic inputs to the soil (by altering vegetation from C3 type into C4 type) is equivalent to a true labelling in situ of the organic matter. Two cases of continuous corn cultivation (Zea mays: δ13C = −12%.) on soils whose initial organic matter average δ13C is −26%. were studied. The quantity of organic carbon originating from corn (that is the quantity which had turned-over since the beginning of continuous cultivation) was estimated using the 13C natural abundance data. After 13 yr, 22% of total organic carbon had turned-over, in the system studied. Particle size fractions coarser than 50μm on the one hand, and finer than 2μm on the other. contained the youngest organic matters. The turnover rate of silt-sized fractions was slower

Effects of afforestation on soil carbon turnover in China's subtropical region

Afforestation in China's subtropics plays an important role in sequestering CO2 from the atmosphere and in storage of soil carbon (C). Compared with natural forests, plantation forests have lower soil organic carbon (SOC) content and great potential to store more C. To better evaluate the effects of afforestation on soil C turnover, we investigated SOC and its stable C isotope (delta C-13) composition in three planted forests at Qianyanzhou Ecological Experimental Station in southern China. Litter and soil samples were collected and analyzed for total organic C, delta C-13 and total nitrogen. Similarly to the vertical distribution of SOC in natural forests, SOC concentrations decrease exponentially with depth. The land cover type (grassland) before plantation had a significant influence on the vertical distribution of SOC. The SOC delta C-13 composition of the upper soil layer of two plantation forests has been mainly affected by the grass biomass C-13 composition. Soil profiles with a change in photosynthetic pathway had a more complex C-13 isotope composition distribution. During the 20 years after plantation establishment, the soil organic matter sources influenced both the delta C-13 distribution with depth, and C replacement. The upper soil layer SOC turnover in masson pine (a mean 34% of replacement in the 10 cm after 20 years) was more than twice as fast as that of slash pine (16% of replacement) under subtropical conditions. The results demonstrate that masson pine and slash pine plantations cannot rapidly sequester SOC into long-term storage pools in subtropical China.

No C_4 Plants Found at the Haibei Alpine Meadow Ecosystem Research Station in Qinghai, China: Evidence from Stable Carbon Isotope Studies

Using the measurement of stable carbon isotopes in leaves as a tool to investigate photosyn-thetic pathway of 102 plant species grown at an alpine meadow ecosystem, at the foot of the Qilian Mountain, Qinghai, China. The results indicate that the δ~(3)C values of plants have a narrow range from -28.24‰ to -24.84‰, which means that none of the species examined belongs to C_4 and crassulaceous acid metabolism (CAM) photosynthetic pathway and all of these species perform photosynthesis through the C_3 pathway. This is likely due to a long-term adaptation to environments at the alpine meadow ecosystem.

Oligocene CO2 decline promoted C4 photosynthesis in grasses.

C4 photosynthesis is an adaptation derived from the more common C3 photosynthetic pathway that confers a higher productivity under warm temperature and low atmospheric CO2 concentration [1, 2]. C4 evolution has been seen as a consequence of past atmospheric CO2 decline, such as the abrupt CO2 fall 32-25 million years ago (Mya) [3-6]. This relationship has never been tested rigorously, mainly because of a lack of accurate estimates of divergence times for the different C4 lineages [3]. In this study, we inferred a large phylogenetic tree for the grass family and estimated, through Bayesian molecular dating, the ages of the 17 to 18 independent grass C4 lineages. The first transition from C3 to C4 photosynthesis occurred in the Chloridoideae subfamily, 32.0-25.0 Mya. The link between CO2 decrease and transition to C4 photosynthesis was tested by a novel maximum likelihood approach. We showed that the model incorporating the atmospheric CO2 levels was significantly better than the null model, supporting the importance of CO2 decline on C4 photosynthesis evolvability. This finding is relevant for understanding the origin of C4 photosynthesis in grasses, which is one of the most successful ecological and evolutionary innovations in plant history.

Nitrogen washing from C3 and C4 cover grasses residues by rain

Crop species with the C-4 photosynthetic pathway are more efficient in assimilating N than C-3 plants, which results in different N amounts prone to be washed from its straw by rain water. Such differences may affect N recycling in agricultural systems where these species are grown as cover crops. In this experiment, phytomass production and N leaching from the straw of grasses with different photosynthetic pathways were studied in response to N application. Pearl millet (Pennisetum glaucum) and congo grass (Brachiaria ruziziensis) with the C-4 photosynthetic pathway, and black oat (Arena Strigosa) and triticale (X Triticosecale), with the C-3 photosynthetic pathway, were grown for 47 days. After determining dry matter yields and N and C contents, a 30 mm rainfall was simulated over 8 t ha(-1) of dry matter of each plant residue and the leached amounts of ammonium and nitrate were determined. C-4 grasses responded to higher fertilizer rates, whereas N contents in plant tissue were lower. The amount of N leached from C-4 grass residues was lower, probably because the C/N ratio is higher and N is more tightly bound to organic compounds. When planning a crop rotation system it is important to take into account the difference in N release of different plant residues which may affect N nutrition of the subsequent crop.

Phylogenetic Relationships in Eleocharis (Cyperaceae): C-4 Photosynthesis Origins and Patterns of Diversification in the Spikerushes

Eleocharis (Cyperaceae) is a morphologically and physiologically diverse lineage of 250 + species with a cosmopolitan distribution. We here explore phylogenetic relationships in this lineage using maximum parsimony and Bayesian inference analyses of nrDNA ITS and cpDNA trnC-ycf6 and ycf6-psbM sequence data with the goals of comparing our phylogenetic hypotheses to previous classifications, morphological variation, and photosynthetic pathway variation. Our results suggest that in Eleocharis C, photosynthesis has been derived at least three times, with several cases of possible reversion to C-3-like or intermediate pathways and several additional origins of C-3-C-4 intermediate photosynthetic pathways, as inferred by carbon isotope ratio measurements. Many classification units currently recognized in Eleocharis are not monophyletic, however, E. subgenus Limnochloa and E. subgenus Scirpidium are monophyletic. Other classification units largely corresponding to clades include E. subgenus Zinserlingia, E. subseries Chaetariae, and E. series Maculosae. Problems with species circumscription and morphological variation in several groups are discussed in light of the phylogeny, particularly in the context of species membership of seven focal clades found in the analyses.

Carbon isotope composition as a tool to control the quality of herbs and medicinal plants

Isotope screening is a simple test for determining the photosynthetic pathway used by plants. The scope of this work was to classify the photosynthetic type of some herbs and medicinal plants through studies of the carbon isotope composition (δ13C). Also, we propose the use of carbon isotope composition as a tool to control the quality of herbs and medicinal plants. For studies of δ13C, δ 13C‰ = [R (sample)/R (standard) - 1] × 10-3, dry leaves powdered in cryogenic mill were analyzed in a mass spectrometer coupled with an elemental analyzer for determining the ratio R = 13CO2/12CO2. In investigation of δ13C of 55 species, 23 botanical families, and 44 species possessed a C3 photosynthetic type. Six species found among the botanical families Euphorbiaceae and Poaceae were C4 plants, and 5 species found among the botanical families Agavaceae, Euphorbiaceae, and Liliaceae possessed CAM-type photosynthesis. Carbon isotope composition of plants can be used as quality control of herbs and medicinal plants, allowing the identification of frauds or contaminations. Also, the information about the photosynthetic type found for these plants can help in introducing and cultivating exotic and wild herbs and medicinal plants.

Carbon isotope composition as a tool for quality control of herbs, spices and medicinal plants

Isotope screening is a simple and cheap test for determining the photosynthetic pathway used by plants. The scope of this work was to classify the photosynthetic type of Melissa officinalis L. and Cymbopogon citratus [DC.] Stapf, through studies of the carbon isotope composition (δ13 C), and we are proposing the use of carbon isotope composition results as a tool to control the quality of medicinal plants. For studies of δ 13C (13 C% = [R (sample)/R (standard) - 1] × 10 -3), dried, powdered leaves were analyzed in a mass spectrometer coupled with an elemental analyzer for determining the ratio R (R = 13CO2/12CO2). As results, M. officinalis presented a C3 photosynthetic type, and C. citratus presented a C4 photosynthetic type. The carbon isotope composition from this study can be used as quality control of M. officinalis adulterants.

Análise Isotópica de Carbono-13 e Nitrogênio-15 da Ipomoea violacea para Determinação de sua Origem Geográfica

Along the Earth globe we can find many types of psychoactive plants. Among them is the Ipomoea violacea, popularly known as Morning Glory. There are ergotalkaloids producer associated-fungus in its leaves and seeds. One of these alkaloids that can be found is the ergine (or LSA), a homologous substance of the lysergic acid diethylamide (LSD). There are many discussions around the world about the inclusion of LSA in the list of controlled substances. In Brazil, this was recently prohibited. One of the most important point of view in the study of isotopic composition of 13C and 15N of this plant is the fact that there is a total alkaloid variation in function of its geographic origin like was verified in 1960’s, besides to aggregate knowledge about it. This work was made to verify if the isotopic ratio can be used as a tool in tracing this illegal Brazilian plant. We could conclude that this plant presents a C3 photosynthetic pathway, its parts has different isotopic carbon and nitrogen composition and that stable isotope analysis can be successfully used as a tool to detect its geographic origin

Turnover do δ¹³C na substituição de dietas com ingredientes de diferentes ciclos fotossintéticos em ovos de codornas japonesas (Coturnix coturnix japonica)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Thermoperiod affects the diurnal cycle of nitrate reductase expression and activity in pineapple plants by modulating the endogenous levels of cytokinins

Nitrate reductase (NR, EC 1.6.6.1) activity in higher plants is regulated by a variety of environmental factors and oscillates with a characteristic diurnal rhythm. In this study, we have demonstrated that the diurnal cycle of NR expression and activity in pineapple (Ananas comosus, cv. Smooth Cayenne) can be strongly modified by changes in the day/night temperature regime. Plants grown under constant temperature (28 degrees C light/dark) showed a marked increase in the shoot NR activity (NRA) during the first half of the light period, whereas under thermoperiodic conditions (28 degrees C light/15 degrees C dark) significant elevations in the NRA were detected only in the root tissues at night. Under both conditions, increases in NR transcript levels occurred synchronically about 4 h prior to the corresponding elevation of the NRA. Diurnal analysis of endogenous cytokinins indicated that transitory increases in the levels of zeatin, zeatin riboside and isopentenyladenine riboside coincided with the accumulation of NR transcripts and preceded the rise of NRA in the shoot during the day and in the root at night, suggesting these hormones as mediators of the temperature-induced modifications of the NR cycle. Moreover, these cytokinins also induced NRA in pineapple when applied exogenously. Altogether, these results provide evidence that thermoperiodism can modify the diurnal cycle of NR expression and activity in pineapple both temporally and spatially, possibly by modulating the day/night changes in the cytokinin levels. A potential relationship between the day/night NR cycle and the photosynthetic pathway performed by the pineapple plants (C(3) or CAM) is also discussed.