石油污染土壤微生物多样性分析及芘降解菌的鉴定

石油污染是目前最严重的环境问题之一，我国有近千万亩的耕地受到程度不同的石油污染。沈抚灌区位于辽宁省沈阳市和抚顺市之间，全长70多千米，是我国最大的石油污染灌区之一，有四五十年的污灌历史，已对当地的生态环境造成了严重的影响。随着人们对污水灌溉危害的认识，从20世纪80年代起，逐步停止了污水灌溉，开始了清水灌溉和改为旱田耕作，但污染问题仍然存在。目前，由于对该地区污染土壤微生物类群、原位降解菌及它们对环境条件变化的响应等信息缺乏了解，限制了对该地区污染土壤的生物修复。 分子生物学研究手段及稳定同位素分析技术，为污染土壤微生物生态学研究提供了全新的研究手段，为突破当前石油污染土壤生物修复研究止步不前的局面指引了方向。本论文采用变性梯度凝胶电泳（Denaturing Gradient Gel Electrophoresis, DGGE）和磷脂脂肪酸分析（Phospholipid Fatty-acid Analysis, PLFA）分析技术，对沈抚灌区水改旱田石油污染土壤的微生物多样性与环境指标的相关规律进行了分析；以PLFA为生物标示物，结合气相色谱-稳定同位素比率质谱（Gas Chromatography Combustion Isotope Ratio Mass Spectrometry, GC-c-IRMS）分析技术，对石油污染土壤和相对清洁土壤中多环芳烃菲和芘的土著降解菌群进行了鉴定；并以不同的吸附性载体为介质，富集、筛选了芘的降解菌。 研究结果表明，石油污染土壤中的微生物群落结构主要与其相对地理位置有关，当污染物的浓度达到一定程度，土壤中的微生物群落结构会发生明显的改变。以13C标记的稳定同位素菲和芘为代谢底物，对污染土壤中菲和芘的土著降解菌群进行了鉴定。 13C-PLFA- GC-c-IRMS分析结果显示，参与菲降解的微生物有G-、G+细菌和放线菌，其中以G-细菌为主；污染土壤和相对清洁土壤两种土壤都未检出有真菌参与；参与芘降解的微生物有放线菌、G-细菌和真菌，其中以G-细菌为主，同时，也检测到有真菌的参与；污染土壤中的降解菌群较清洁土壤丰富。研究还发现，芘在土壤中的降解可能存在不经过菲为中间产物的代谢途径。采用不同的吸附性载体所富集的优势细菌显著不同。从不同的吸附性载体上最终得到8株芘的降解菌，其中鉴定出的三株菌中Sp-A56，Sp-A21为多食鞘氨醇杆菌（Sphingobacterium multivorum）；Sp-B05氨基杆菌属的一个亚种（Aminobacter sp.），未见有关该菌株芘降解能力的报道，可能是一株新的芘降解菌。

藏东及滇东南新生代钾质岩浆作用及其深部制约

The dissertation focuses on the petrology, geochemistry of the volcanic rocks in east Tibet and southeast Yunnan. It lucubrates the Magmatic process, forming mechanism and the possible tectonic settings of the volcanic rocks. The volcanic rocks of Nangqen basin in east Tibet, Qinghai province are mainly Cenozoic intermediate-acid shoshonites. The rocks are LREE enriched and the LREE/HREE = 3～34; (La/Yb)_N = 18.17-53.59, and ΣREE 222～1260μg/g. There are no Eu anomaly, and Nb, Ta, Zr, Hf, Ti are markedly depleted. The isotopic composition is ~(87)Sr/~(86)Sr = 0.70497～0.70614, ~(206)Pb/~(204)Pb = 18.622～18.974, ~(208)Pb/~(204)Pb = 38.431～38.996, ~(207)Pb/~(204)Pb = 15.511～15.613, respectively. K-Ar age of the whole rocks and the single mineral are between 32.0-36.5Ma. Based on the trace elements and isotopic elements, we get the conclusion that the partial melting is one of the dominated forming mechanisms for the volcanic rocks in Naneqen basin. The magma did not experience the crustal contamination en route to the surface; however, the complex mixture took place in the upper mantle before the melt was formed. There are at least two kinds of mixed sources that can be identified. The basalt in southeast Yunnan province is studied. They are distributed in Maguan, Tongguan, and Pingbian County, which is located on the both sides of the Red River belt, and the ultrabasic xenolith are cursory introduced. The volcanic rocks belongs to the alkali series, which can be subdivided into trachybasalt and basanite(Ol normal molecule >5). The volcanic rocks are characteristics by high Ti and low Mg#. According to the magma calculation model, the original rocks of the basalt in southeast Yunnan province are Spinel Lherzolite in Tongguan, Garnet Lherzolite in Pingbian and Maguan, while Togguan undergoes 2-5 percent and percent of partial melting, whereas volcanism in Maguan and Pingbian was so complex to calculate. The fractional crystallization took place during the magma evoltion in southeast Yunnan. The basalt is enriched in LREE with LREE/HREE=9.23-20.19. All of the trace elements display weak Nb, Ta peak, and the depletion of Zr, Hf and Ti in Maguan and pingbian represent the presence of Garnet in the source. The composition of the isotope ratio are ~(87)Sr/~(86)Sr = 0.70333-0.70427, ~(143)Nd/~(144)Nd = 0.512769-0.512940, ~(206)Pb/~(204)Pb = 18.104-18.424, ~(207)Pb/~(204)Pb = 15.483 -15.527; ~(208)Pb/~(204)Pb = 37.938-38.560, respectively, which shows the characteristics of the HIMU type OIB. The volcanic rocks of the southwest Yunnan are derived from the enriched, OIB type mantle sources by synthesizing all the data from trace and isotope elements. It is similar to that of the volcanic rocks in Hawaii, a typical kind of the mixtures of the recycled oceanic crust plume and depleted asthenosphere. To sum up, the volcanic rocks in southeast Yunnan are formed by the intraplate hotpot volcanism.

中国北方草本植物及表土有机质碳同位素组成

Recently, more and more attention has been paid to stable isotope ratios in terrestrial depositional systems. Among them, δ~(13)C value is mainly determined by the surface vegetation, while vegetation is directly related to climate, therefore, carbon isotope ratio in soil organic matter and pedogenic carbonate has been employed as an important paleoecological indicator. In order to test the paleoecological information extracted from stable isotope ratios in terrestrial depositional systems, it is necessary to study the relationships between δ~(13)C value in standing terrestrial plants and today climate, as well as between δ~(13)C value in modern surface soil organic matter and standing vegetation. Thus, these relationships were studied in this paper by means of analysing δ~(13)C in standing plants and modem surface soil organic matter in North China. The main results and conclusions are presented as following: 1. According to their δ~(13)C values, 40 C-4 species represent about 16% of the 257 plant species sarnpled from the North China. C-4 photosynthesis mainly occurs in Poaceae, Cyperaceae and Chenopidaceae families, and percentage representation of C-4 photosynthesis is up to 56% in Poaceae family. 2. The δ~(13)C values of C-3 plant species in North China vary from -21.7‰ to -32.0‰ with an average of -27.1‰, and 93% focus on the range of -24.0‰ ～ -30.0‰; δ~(13)C values of C-4 plant species in North China are between -10.0‰ ～ -15.5‰ with an average of -12.9‰, and 90% concentrate on the range of -11.0‰ ～ -15.0‰. 3. The δ~(13)C composition of C-3 plant species collected from Beijing, a semi-moist district, mainly vary between -27.0‰ ～ -30.0‰, and the average is -28.7‰; the δ ~(13)C values of plants in the semi-arid district, east and west to the Liu Pan Moutain, focus on the range of-26.0‰ ～ -29.0‰ and -25.0‰ ～ -28.0‰, respectively, with the mean value of -27.6‰ and -26.6‰, respectively; the δ~(13)C composition in the arid district dominantly vary from -24.0‰ to -29.0‰, with the average of -26.2‰, and among them, the δ~(13)C values of C-3 plant species in deserts are often between - 22‰ ～ -24‰; the δ~(13)C values in the cold mountain district concentrate on the range of -24.0‰ to -29.0‰, with the average of -26.3‰. 4. The main range of δ~(13)C composition of C-4 plant species, derived from Beijing, a semi-moist district, are -13.0‰ ～ -15.5‰; the semi-arid district, -11.0‰ ～ -14.0‰; the arid district, -11.0‰ ～ -14.0‰. The mean values of them are -14.0‰, -12.4‰,-12.7‰, respectively. 5. From east to west in North China, δ~(13)C values of C-3 plant species increase with longitude. The correlation between δ~(13)C ratios of C-3 plant species and longitude is linear. Changing temperate and precipitation and changing atmosphere pressure are spossible explanations. 6. Almost all C-3 plant species have the trends that their δ~(13)C values gradually increase with decreasing precipitation, decreasing temperature and increasing altitude. Our results show the increases of the δ~(13)C value by 0.30 ～ 0.45‰, 0.19 ～ 0.27‰ and 1.1 ～ 1.2‰ per 100 mm, I℃ and 1000 m, respectively, for all C-3 plant species together. 7. The δ~(13)C values of all C-3 plant species together and a part of C-3 species show highly significant linear correlation with the mean annual temperature, the mean annual precipitation and the altitude, and the results suggest that they can be used as proxies of these environmental variables, while, those without highly significant correlation, may be not suitable as the proxies. 8. The extent, which of responses of δ~(13)C composition to environmental variables, is different for each C-3 plant specie. 9. The δ~(13)C variations along altitude and longitude may be non-linear for C-4 p1ant species in North China. The mean annual temperature may be not important influential factor, thus, it suggests that the δ~(13)C composition of C-4 plant species may be not suitable as the proxy of the mean annual temperature. The influences of summer temperature on δ~(13)C values are much bigger than that of annual temperature, among them, the influence of September temperature is biggest. The mean annual precipitation may be one of the dominant influential factors, and it shows a highly significant non-linear relationship with δ~(13)C values, and the result indicates that δ~(13) C composition of C-4 plant species can be employed as the proxy of the mean annual precipitation. 10. The variations of δ~(13)C ratios do not show systematic trends along longitude, latitude and altitude for modern surface soil organic in Northwest China. ll. The δ~(13)C ratios of modern surface soil organic do not exhibit systematic patterns with temperature and precipitation in Northwest China, it suggests that, unless soil organic is transferred from pure C-3 or C-4 vegetation, the δ~(13)C composition of soil organic may be not used as proxies of climatic variables. 12. The δ~(13)C values of modem surface soil organic are heavier than that of standing vegetation, and the difference ofrnean δ~(13)C between them is -2.18‰. 13. Without considering the δ~(13)C difference between vegetation and soil organic, as well as the δ~(13)C drift in various enviromnent, we may not obtain the valuable information of C-3, C-4 relative biomass in vegetation. 14. The C-4 biomass contribution in vegetation increase with decreasing latitude, increasing longitude and decreasing altitude in Northwest China. The C-4 biomass almost are zero in those regions north to 38 ° N, or west to 100°E, or above 2400 m. 15. The C-4 relative biomass in vegetation increase with growing temperature and precipitation. and, C-4 plants are rare at those regions where the mean annual temperature is less 4 ℃, or the mean annual precipitation is less 200 mm, and their biomass contribution in vegetation are almost zero. Both the mean annual temperature and the mean annual average precipitation may be the important influential factors of C-4 distribution, but the dominant factors.

碳、氧稳定同位素分析在古环境研究中的应用

A number of proxy records of paleoenvironment using stable isotopes could show the history of past environmental changes. These archives include peat and lake sediments, loess-paleosot sequence, fossil mammals and stalagmite, and so on. The stable isotopic composition of carbonate and organic matter and frequency magnetic susceptibility from Tianshuigou and Yuanlei loess-palesol sequence can be used to give estimates of the paleoenvironmental history of Dali, and even of the whole Chinese Loess Plateau during the last 250ka. Features of the High Temperature and Large Precipitation Event in the Tibet Plateau and its adjacent area during 40~30kaBP had been studied by Professor Shi Y. In this dissertation, its impact on Chinese Loess Plateau has been discussed again. Carbon and oxygen isotopic ratios, magnetic susceptibility and frequency magnetic susceptibility in Tianshuigou and Yuanlei profiles show that the Event in this area is not so stronger as the Tibet Plateau. The carbon isotopic composition of organic matter in Tianshuigou, Yuanlei, dingcun and Jingcun loess-palesol sequences are indicative of major changes in the paleovagetation between terrace and plain of the Chinese Loess Plateau. Water is one of the most important factors adjusting the relative biomass of C4 plant in terrestrial ecosystems. Stable carbon isotope ratio of vertebrate tooth enamel is used increasingly to reconstruct environmental and ecological information modern and ancient ecosystems. The SI3C value of tooth enamel bioapatites can distinguish between browsers and grazers. Data from typical grassland of Inter Mongolia, the Alpine meadow of Qinghai-Tibet Plateau and the Yaluzangbu Great Canyon indicate that diets of mammals could record the relative biomass of C4 plant only in the C4 dominated ecosystem. In a C3 dominated ecosystem, diet of mammals would include more C3 plants than vegetation. According to Professor Cerling, proxy records from North and South America, Africa and Pakistan show that at the end of the Miocene (between 8Ma to 6 Ma) there was a global expansion of CA biomass, probably when atmospheric CO2 levels declined. Thus, "C4 world" and "CO2 starvation" are put forward. In this dissertation, carbon isotopes of fossil tooth such as Equus sanmeniensis and Hipparion chiai from Linxia, China reveal that there is a C3 dominated ecosystem in the late Miocene. Diets of ancient mammals in Linxia are not evidence of global expansion of C4 biomass.