Triterpane and sterane biomarkers in the YA13-1 condensates from Qiongdongnan Basin, South China Sea

Triterpanes and steranes in condensates from the YA13-1 gas field, Qiongdongnan Basin, were monitored. The YA13-1 condensates have unusual biomarker distributions dominated by terpanes and steranes derived from higher plants. Anomalously abundant 1 got-oleanane and remarkably abundant bicadinanes are present in the YA13-1 condensates, whereas the 17alpha-hopane contents are extremely low. Taraxastane and significantly abundant 17alpha-diahopanes occur in the condensates. In addition, a number of unknown C-29 and C-30 pentacyclic triterpanes including previously unreported compounds were detected in the condensates, some of which are significantly abundant. The unknown compounds may be terrestrial biomarkers. C-29 homologues are relatively predominant among the regular and rearranged steranes. The diasterane concentrations are markedly higher than those of regular steranes. The maturity of the YA13-1 condensates is relatively high, at the peak to late oil generation stage (corresponding to 0.85-1.10% Ro), based on sterane and terpane and including bicadinane maturity parameters (i.e. T/(T-1 + R) and 2T/R bicadinane ratios). The above maturity assessment result is different from that based on diamondoid maturity parameters (%Ro = 1.60-1.70) [Org. Geochem. 25 (1996) 179], which can be explained by a contribution of hydrocarbons from two sources at different depths. The YA13-1 condensates were probably generated from the Yacheng and Lingshui coal-bearing source rocks buried both in the Qiongdongnan Basin (3400-5000 m) and in the Yinggehai Basin (>5000 m). The possible contribution of lower maturity hydrocarbons from the Yacheng and Lingshui Formations (3400-4100 m) in the Qiongdongnan Basin to the YA13-1 gases and condensates should not be neglected. (C) 2003 Elsevier Science B.V. All rights reserved.

查尔酮异构酶基因过表达对新疆雪莲类黄酮生物合成的调控

新疆雪莲（Saussurea involucrata Kar. et Kir.）是我国名贵中药材，其主要药用活性成分为黄酮类化合物。目前人们对新疆雪莲及它的黄酮类化合物的需求日益增多，但雪莲的人工栽培技术尚未成熟，在野生状态下，新疆雪莲只能生长在海拔4,000到5,000米的雪山上，现在由于过度采挖已濒临灭绝。为解决雪莲资源匮乏，提高雪莲中黄酮类成分的含量，本研究通过基因工程手段利用发根农杆菌将黄酮代谢途径中的关键酶－查尔酮异构酶（CHI）基因导入新疆雪莲，产生转基因新疆雪莲毛状根及再生苗，以期提高新疆雪莲的黄酮类物质含量，进行新疆雪莲黄酮类物质的生产。主要结果如下： 　　1．对克隆到的水母雪莲查尔酮异构酶基因（Smchi）进行功能分析。转Smchi正义、反义烟草的CHI酶活性实验结果表明，转Smchi正义的烟草CHI酶活性比对照提高3-6倍，而转反义Smchi基因的烟草CHI酶活性比对照则显著降低。分析不同株系的转基因烟草和对照烟草的黄酮含量和花色素含量表明，转Smchi正义的烟草积累比对照显著增高水平的总黄酮，其中株系CS-5黄酮含量是对照的6倍，转Smchi反义的烟草则积累较低水平的总黄酮，而且转基因烟草的总黄酮含量与Smchi基因的表达水平和CHI酶活性成正相关。但不论转Smchi基因正义或反义方向的烟草，其花色素含量和对照相比均没有发生显著变化。进一步对转基因烟草的黄酮成分进行分析，发现烟草中的主要黄酮成分芦丁在转Smchi正义烟草中有很高的积累。 　　2．发根农杆菌介导法将Smchi基因导入新疆雪莲，得到转Smchi基因的新疆雪莲毛状根。实验发现，35S-chi转基因对毛状根的生长没有显著影响，但35S-chi转基因毛状根能够合成显著提高水平的芹菜素和总黄酮，其中根系C46经过35 d培养，能产生32.1 mg/L的芹菜素和647.8 mg/L的总黄酮，分别是对照根系的12倍和4倍;不同根系的Smchi基因表达水平、CHI酶活性和芹菜素含量成正相关。本研究为通过基因工程手段提高新疆雪莲毛状根芹菜素和总黄酮含量提供了一个有效方法。 　　3．在1/2MS附加GA1.5 mg/L的培养基上，新疆雪莲毛状根的不定芽再生频率高且不定芽生长健壮。再生苗在MS+BA1.0 mg/L+NAA0.1 mg/L的培养基上继代培养生长量较大，经过20 d的培养，35S-chi转基因新疆雪莲再生苗株系（C17、C27、C46）、对照再生苗（Control-1）和正常试管苗（Control-2）之间生长量差异不显著，增殖倍数都在7倍左右;实验还发现，毛状根再生苗比各自来源的毛状根的芹菜素和总黄酮含量下降了20-30%，但转基因再生苗的芹菜素和总黄酮含量比Control-1和Control-2都有显著提高，其中C46芹菜素和总黄酮含量分别为1.86 mg/g 干重和37.3 mg/g干重，分别是Control-1的12倍和 2.4 倍，Control-2的4 倍和1.6倍。这些结果表明由毛状根诱导出的再生苗可作为增强目标次生代谢产物生产的另外一个有效来源。 　　

东营凹陷幔源流体对原油生物标志化合物的影响研究

A large number of mantle-derived fluid activities occurred in the Dongying Sag. On the basis of the studies on the geochemical characteristics of these fluids in this sag, the spatial distribution of biomarkers in petroleum and their relationships with the parameters of mantle-derived fluids were studied, to reveal the influence of mantle-derived fluids on the biomarkers and to evaluate the reliability of these biomarkers when applied to oil-source correlation and maturity analysis. Most biomarkers used in oil correlation kept the characteristics of their sources during burial thermal evolution. Even some of them were not influenced by mantle derived fluids, such as the relative content of C27-C29 steroid（ααα20R）and C21/C23 tricyclic terpane. However, Pr/Ph and C35/C34 hopanes were sensitive to both heat energy and materiel input by the mantle-derived fluids. γ-waxnae/C30hopanes and C24 tetracyclic terpanes /C26 tricyclic terpanes responded only to thermal influence by mantle-derived fluids. They did not chemically reacted with the mantle-derived fluids. Fluorene series compounds reacted with hydrogen and / or carbon dioxide from the mantle. Mantle-derived fluids affected most maturity index. The huge thermal energy with mantle-devied fluids weakened the relationship between the maturity parameters and depth. Among them, pregnane/C27-29 steroid and Ts/(Ts+Tm) were more sensitive to the heat of the fluids. ααα20S/(20S+20R) took the second place. αββ/(ααα+αββ) and 22S/(22S+22R) were not thermally influenced by the mantle-derived fluid. Besides, the substance of mantle-derived fluids reacted with fragrants, hopanes or moretanoids and thus altered the values of MPI1, MPI2, MPR, C30 hopanes/(C30 hopanes + moretanoids) and alkyl-diben zothiophene/diben zothiophene. The thermal alernation of phenanthrene series and their spatial distribution show that the heat energy carried by mantle-derived fluids was not fierce but spread widely in Dongying Sag, which is favorable to hydrocarbon generation with little destroy. In sum, mantle-derived fluids affected biomarkers through thermal energy and chemical reactions and changed the values of oil-source correlation and maturity parameters in the deep-seated fault belts. Therefore, in the deep-seated fault belts, oil-source correlation should be restudied and the new parameters need to be explored.

黔北早寒武世缺氧事件：生物标志化合物及有机碳同位素特征

本文首次报道了黔北早寒武世黑色岩系的生物标志化合物，并结合有机碳同位素组成特征，探讨该时期所发生的重大地质事件。所有样品均检出了丰富的正构烷烃、类异戊二烯烃、萜类化合物以及甾类化舍物。在GC谱图上，正构烷烃显示明显的单峰型分布特征，碳数分布范围为nC14-nC31主峰碳为nC18、nC19或nC20，nC17／nC31为1．15～50．17，显示轻烃组分占绝对优势，OEP值为0．84～1．11，CPI值为0．92～1．16，接近平衡值1．0，无明显的奇偶碳数优势分布。Pr／Ph值为0．24～0．79，具有明显的植烷优势。萜烷化合物以C30藿烷占优势，其相对丰度三环萜烷〉五环三萜烷〉四环萜烷，并且检出少量的γ-蜡烷。规则甾烷C27-C28-C29呈“V”字型分布，∑（C27＋C28）〉∑C29，其比值为1．25～1．99，∑C27／∑C29为0．78～1．22，重排甾烷C27／规则甾烷C27值为0．21～0．47。4-甲基甾烷普遍存在，但丰度相对较低。在地层剖面上，有机碳含量（TOC（％））从0．05～7．91％，平均为2．52％；有机碳同位素组成（δ^13Corg）从-29．49‰～-34．41‰（PDB），发生负偏移，偏移量达到4．3‰，代表该期海平面处于上升阶段，底层水处于严重缺氧状态，底栖生物缺乏。本文根据生物标志化合物特征参数，结合有机碳同位素组成变化，详细分析了黔北早寒武世这套黑色岩系的有机质来源、成熟度、沉积环境以及古海洋意义，将为我国南方早寒武纪古地理重建、地质事件记录提供可靠依据。

生物标志化合物数据处理及在重建古环境中的应用

本文主要地讨论了生物标志化合物数据的处理方法，并将此应用到古沉积环境的判别方面。全文共分三大部分：第一部分主要讨论是多元统计方法对那些研究的较成熟的烃类生物标志经合物参数在重建古沉积环境方面的应用。对分别采自不同沉积环境的共46个样品中生物标志化合参数的R型因子分析和对应分析的结果表明：生物标志化合物参数可以有效地将所研究的样品分划分为：淡水湖泊相沉积环境、半咸水/淡水环境、膏盐环境和泻湖沉积具火山灰沉积的环境，与此对应，生物标志化合物参数也可以划分为相应的四大类，即：（1）反映淡水湖泊沉积环境的特征为：含较丰富的高等植物输入的生物标志化合物，如：具较高的H/L，C31/C17，C27/C17比值等；（2）半咸水/淡水沉积环境以较高的Pr，Pr/Ph和高的藿/甾比为特征；（3）生物标志化合物参数组合{Ph/nC18，植烷，升藿烷指数，伽玛蜡烷指数， iC25+iC30和Tm/Ts}是反映膏盐环境的一类生物标志化合物参数；（4）可能可以用含较高的C28甾烷和β-胡萝卜烷来区分泻湖具火山灰沉的环境。正构烷烃的分布对于划分沉积环境远不如上述的综合生物标志化合物那样有效，但是，中等链长的正烷烃（C22，C24）在主因子分析的第二主因子轴上有较大的载荷，反晨了含较高中等链长的正烷烃是膏盐环境的一个特征，对其归一化数据的主因子分析的结果还表明，它是一类较好的物源（高等植物或低等生物输入）指示生物标志化合物。第二部分讨论的是膏盐环境中含硫化合物GC/MS数据的处理方法，通过初步确定碳数、碳数的校正、计算TPI（噻吩指数，Thiophene index）和CTPI、确定噻吩代码、建立TPmaster、绘制噻吩化合物分布的“指纹图”等步骤，在大多数噻吩化合物结构不明的情况下，快速地、清楚地、有条理地将这类化合物的所有异构体表达出来，同时还实现了这类化合物在不同样品间的自动对比和直观对比；大大的方便了这类化合物在地质体中应用的研究。在理论上，可以通过TPmater寻找新的异构体和新的噻吩化合物系列。第三部分在计算同位素分子峰发布的基础上，快速地、有效地处理了卟啉的质谱数据，即：卟啉Probe-MS数据的同位素校正。这一计算程序不仅可以计算混合分子同位素峰束中单个化合物的含量（即质谱数据的同位素校正），还可以计算任意化合物和混合物的分子同位素峰的分布类型，通过对比质谱数据，有助于地质体中新的卟啉化合物的发现和鉴定。在用该计算程序处理了采自不同沉积环境的七个样品的Probe-MS数据后，再加上自由基卟啉的HPLC特征，结果表明：可以用卟啉的分布将淡水/半咸水环境的沁阳凹陷、泻湖相具火山灰沉积的准葛尔盆地和四川三叠系海相区分开来。