Independence of alpine meadow plants from soil organic matter for nitrogen supply: evidence from stable nitrogen isotopes

Stable nitrogen isotope signatures of major sources of mineral nitrogen ( mineralization of soil organic nitrogen, biological N-2 fixation by legumes, annual precipitation and plant litter decomposition) were measured to relatively define their individual contribution to grass assimilation at the Haibei Alpine Meadow Ecosystem, Qinghai, China. The results indicated that delta N-15 values (- 2.40 parts per thousand to 0.97 parts per thousand) of all grasses were much lower than those of soil organic matter (3.4 +/- 0.18 parts per thousand) and mineral nitrogen ( ammonium and nitrate together,7.8 +/- 0.57 parts per thousand). Based on the patterns of stable nitrogen isotopes, soil organic matter (3.4 +/- 0.18 parts per thousand), biological N-2 fixation (0 parts per thousand), and precipitation (- 6.34 +/- 0.24 parts per thousand) only contributed to a small fraction of nitrogen requirements of grasses, but plant litter decomposition (- 1.31 +/- 1.01 parts per thousand) accounted for 67%.

The effect of land management on plant community composition, species diversity, and productivity of alpine Kobersia steppe meadow

Grassland degradation is widespread and severe on the Tibet Plateau. To explore management approaches for sustainable development of degraded and restored ecosystems, we studied the effect of land degradation on species composition, species diversity, and vegetation productivity, and examined the relative influence of various rehabilitation practices (two seeding treatments and a non-seeded natural recovery treatment) on community structure and vegetation productivity in early secondary succession. The results showed: (1) All sedge and grass species of the natural steppe meadow had disappeared from the severely degraded land. The above-ground and root biomass of severely degraded land were only 38 and 14.7%, respectively, of those of the control. So, the original ecosystem has been dramatically altered by land degradation on alpine steppe meadow. (2) Seeding measures may promote above-ground biomass, particularly grass biomass, and ground cover. Except for the grasses seeded, however, other grass and sedge species did not occur after seeding treatments in the sixth year of seeding. Establishment of grasses during natural recovery treatment progressed slowly compared with during seeding treatments. Many annual forbs invaded and established during the 6 years of natural recovery. In addition, there was greater diversity after natural recovery treatment than after seeding treatments. (3) The above-ground biomass after seeding treatment and natural recovery treatment were 114 and 55%, respectively, of that of the control. No significant differences in root biomass occurred among the natural recovery and seeded treatments. Root biomass after rehabilitation treatment was 23-31% that of the control.

Development of a sensitive fluorescent derivatization reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) and its application for determination of amino acids from seeds and bryophyte plants using high-performance liquid chromatography with fluorescence detection and identification with electrospray ionization mass spectrometry

A pre-column derivatization method for the sensitive determination of amino acids and peptides using the tagging reagent 1,2-benzo-3,4dihydrocarbazole-9-ethyl chloroformate (BCEOC) followed by high-performance liquid chromatography with fluorescence detection has been developed. Identification of derivatives was carried out by liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS/MS). The chromophore of 2-(9-carbazole)-ethyl chloroformate (CEOC) reagent was replaced by 1,2-benzo-3,4-dihydrocarbazole functional group, which resulted in a sensitive fluorescence derivatizing reagent BCEOC. BCEOC can easily and quickly label peptides and amino acids. Derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography. The derivatives showed an intense protonated molecular ion corresponding m/z (M + H)(+) under electrospray ionization (ESI) positive-ion mode with an exception being Tyr detected at negative mode. The collision-induced dissociation of protonated molecular ion formed a product at m/z 246.2 corresponding to the cleavage of C-O bond of BCEOC molecule. Studies on derivatization demonstrate excellent derivative yields over the pH 9.0-10.0. Maximal yields close to 100% are observed with a 3-4-fold molar reagent excess. Derivatives exhibit strong fluorescence and extracted detzvatization solution with n-hexane/ethyl acetate (10:1, v/v) allows for the direct injection with no significant interference from the major fluorescent reagent degradation by-products, such as 1,2-benzo-3,4-dihydrocarbazole-9-ethanol (BDC-OH) (a major by-product), mono- 1,2-benzo-3,4-dihydrocarbazole-9-ethyl carbonate (BCEOC-OH) and bis-(1,2-benzo-3,4-dihydrocarbazole-9-ethyl) carbonate (BCEOC)(2). In addition, the detection responses for BCEOC derivatives are compared to those obtained with previously synthesized 2-(9-carbazole)-ethyl chloroformate (CEOC) in our laboratory. The ratios AC(BCEOC)/AC(CEOC) = 2.05-6.51 for fluorescence responses are observed (here, AC is relative fluorescence response). Separation of the derivatized peptides and amino acids had been optimized on Hypersil BDS C-18 column. Detection limits were calculated from 1.0 pmol injection at a signal-to-noise ratio of 3, and were 6.3 (Lys)-177.6 (His) fmol. The mean interday accuracy ranged from 92 to 106% for fluorescence detection with mean %CV < 7.5. The mean interday precision for all standards was < 10% of the expected concentration. Excellent linear responses were observed with coefficients of > 0.9999. Good compositional data could be obtained from the analysis of derivatized protein hydrolysates containing as little as 50.5 ng of sample. Therefore, the facile BCEOC derivatization coupled with mass spectrometry allowed the development of a highly sensitive and specific method for the quantitative analysis of trace levels of amino acids and peptides from biological and natural environmental samples. (c) 2005 Elsevier B.V. All rights reserved.

陆生蜗牛壳体同位素组成及其对生长季节的响应

Stable isotope compositions of land snail shells have a great potential as an indicator of paleoclimatic and paleoenvironmental changes. However, some key issues, such as the relationship of carbon isotope between snail food and local vegetation, and the uncertainty of the dominant factors about snail body fluid changes in oxygen isotope composition, remain less well known, strongly limiting shell isotopic application. In this study, we measure the stable isotope compositions on the shells of both live snails and fossils collected from the Chinese Loess Plateau and a loess sequence at Mangshan, Xingyang, respectively. Based on the analyses, the association of the stable isotope compositions of land snail shells with their growing seasons is investigated. In addition, the climatic and environmental significances of isotopic differences among several snail species are discussed. The main results and conclusions are presented as follows: 1. δ18O values for the shell lip samples of Bradybaena ravida redfieldi range from -6.79‰ to -1.92‰, and parallels to the monthly changes of local rain water δ18O, temperature and humidity. The compatibility of shell lip δ18O with monthly modeled shell δ18O indicates that the shell lip δ18O changes are mainly resulted from the 18O variations of rain-water. The shells of a land snail growing in spring could be enriched in 18O, and those growing in summer depleted in 18O. 2. Carbon isotope compositions of snail shells are controlled by their diet, which is affected by the relative proportion of C3 to C4. There are some differences in carbon isotopic compositions among different snail species, especially between P. orphana and V. tenera or P. aeoli. Shell δ13C for P. orphana is the most positive with an average of -5.88 ± 2.54 ‰. The C4 plant fraction of the food for “cold-aridiphilous” taxa, P. aeoli and V. tenera, is distinctly lower than that for “thermo-humidiphilous” taxa, P. orphana, indicating that summer is likely to be the main active season of P. orphana and spring of P. aeoli and V. tenera. Therefore, some discrepancy of carbon isotopic compositions among different species may be related to snail active season. 3. δ13C values among different species have a certain degree of positive correlation, which may be influenced by local vegetation ecosystem. δ13C value of the snail shells (especially P. orphana) shows an eastward increasing trend and consists with the variations of C4 plants biomass in Loess Plateau. The result shows that the carbon isotope in local vegetation ecosystem is one of the main factors influencing δ13C values of snail food. Therefore, both carbon isotopes of local vegetation ecosystem and snail active season contribute to the carbon isotopic differences among different snail species and in different areas. 4. δ13C values of living snail shells and soil organic matter have a positive correlation with each other, which further supports the view that carbon isotope in local vegetation ecosystem is one of the main factors influencing δ13C values of snail food. However, the range of δ13C values of snail food for various species in response to carbon isotope in local vegetation ecosystem is different. It is suggested that 13C enrichment of snail shells relative to local vegetation ecosystem has a potential to indicate snail active season and the degree of climate temperature and humidity. 5. There is a significant negative correlation between carbon and oxygen isotopic compositions of living snail shells in Loess Plateau. This result further supports that snail active season can be inferred based on the shell carbon and oxygen isotopic compositions. Moreover, there are some positive correlations between mean annual temperature and differences of shell δ13C values ( 13CV. tenera-P. orphana) and that of δ18O values ( 18OV. tenera-P. orphana) for P. orphana, a typical “thermo-humidiphilous” taxa, and V. tenera, a typical “cold-aridiphilous” taxa, respectively. It shows that  13CV. tenera-P. orphana and  18OV. tenera-P. orphana may have a potential to indicate mean annual temperature or the length of biological growing season. 6. Stable isotopes of land snail shell in the Mangshan loess sequence show that the shell δ18O value of “cold-aridiphilous” taxa V. tenera is more positive than “thermo-humidiphilous” taxa P. orphana and δ13C value of the former is more negative than the latter. In addition, the shell δ18O value of V. tenera varies significantly in different period. During the last glacial maximum, its δ18O value with an average of -7.89 ‰ is more negative than that (-5.88 ‰) from the last deglaciation to the early Holocene. This phenomenon indicates that its growing season during different period is significantly different. It tends to grow in summer in last glacial maximum. With climate warming, it prefers growing in spring with relatively low temperature. While the shell δ18O value of P. orphana varies in a little range, which shows that its activity season is shorter and mainly in summer. These results further support that the change of the snail growing season is one of the main factors of differences of carbon isotopic compositions among different snail species and varies with time. Furthermore, it is consistent that changes in magnetic susceptibility and trend of differences of shell δ18O values and δ13C values respectively between the two snail fossils. It is further testified that 13CV. tenera-P. orphana and  18OV. tenera-P. orphana may have a potential to indicate mean annual temperature or the length of biological growing season.

末次盛冰期以来中国陆地生态系统碳储量变化研究

Carbon is an essential element for life, food and energy. It is also a key element in the greenhouse gases and therefore plays a vital role in climatic changes. The rapid increase in atmospheric concentration of CO_2 over the past 150 years, reaching current concentrations of about 370 ppmv, corresponds with combustion of fossii fuels since the beginning of the industrial age. Conversion of forested land to agricultural use has also redistributed carbon from plants and soils to the atmosphere. These human activities have significantly altered the global carbon cycle. Understanding the consequences of these activities in the coming decades is critical for formulating economic, energy, technology, trade, and security policies that will affect civilization for generations. Under the auspices of the International Geosphere-Biosphere Programme (IGBP), several large international scientific efforts are focused on elucidating the various aspects of the global carbon cycle of the past decade. It is only possible to balance the global carbon cycle for the 1990s if there is net carbon uptake by terrestrial ecosystems of around 2 Pg C/a. There are now some independent, direct evidences for the existence of such a sink. Policymarkers involved in the UN Framework Convention on Climate Change (UN-FCCC) are striving to reach consensuses on a 'safe path' for future emissions, the credible predictions on where and how long the terrestrial sink will either persist at its current level, or grow/decline in the future, are important to advice the policy process. The changes of terrestrial carbon storage depend not only on human activities, but also on biogeochemical and climatological processes and their interaction with the carbon cycles. In this thesis, the climate-induced changes and human-induced changes of carbon storage in China since the past 20,000 years are examined. Based on the data of the soil profiles investigated during China's Second National Soil Survey (1979-1989), the forest biomass measured during China's Fourth National Forest Resource Inventory (1989-1993), the grass biomass investigated during the First National Grassland Resource Survey (1980-1991), and the data collected from a collection of published literatures, the current terrestrial carbon storage in China is estimated to -144.1 Pg C, including -136.8 Pg C in soil and -7.3 Pg C in vegetation. The soil organic (SOC) and inorganic carbon (SIC) storage are -78.2 Pg C and -58.6 Pg C, respectively. In the vegetation reservoir, the forest carbon storage is -5.3 Pg C, and the other of-1.4 Pg C is in the grassland. Under the natural conditions, the SOC, SIC, forest and grassland carbon storage are -85.3 Pg C, -62.6 Pg C, -24.5 Pg C and -5.3 Pg C, respectively. Thus, -29.6 Pg C organic carbon has been lost due to land use with a decrease of -20.6%. At the same time, the SIC storage also has been decreased by -4.0 Pg C (-6.4%). These suggest that human activity has caused significant carbon loss in terrestrial carbon storage of China, especially in the forest ecosystem (-76% loss). Using the Paleocarbon Model (PCM) developed by Wu et al. in this paper, total terrestrial organic carbon storage in China in the Last Glacial Maximum (LGM) was -114.8 Pg C, including -23.1 Pg C in vegetation and -86.7 Pg C in soil. At the Middle Holocene (MH), the vegetation, soil and total carbon were -37.3 Pg C, -93.9 Pg C and -136.0 Pg C, respectively. This implies a gain of-21.2 Pg C in the terrestrial carbon storage from LGM to HM mainly due to the temperature increase. However, a loss of-14.4 Pg C of terrestrial organic carbon occurred in China under the current condition (before 1850) compared with the MH time, mainly due to the precipitation decrease associated with the weakening of the Asian summer monsoon. These results also suggest that the terrestrial ecosystem in China has a substantial potential in the restoration of carbon storage. This might be expected to provide an efficient way to mitigate the greenhouse warming through land management practices. Assuming that half of the carbon loss in the degraded terrestrial ecosystem in current forest and grass areas are restored during the next 50 years or so, the terrestrial ecosystem in China may sequestrate -12.0 Pg of organic carbon from the atmosphere, which represents a considerable offset to the industry's CO2 emission. If the ' Anthropocene' Era will be another climate optimum like MH due to the greenhouse effect, the sequestration would be increased again by -4.3 - 9.0 Pg C in China.