塔里木沙漠公路防护林对极端环境胁迫的生理生态响应

论文对塔里木沙漠公路防护林对环境胁迫的生理生态响应及稳定性进行了研究。研究内容包括防护林不同树种对环境胁迫的响应、防护林的环境效应及防护林系统的稳定性初步研究。其中，在塔里木沙漠公路防护林建设方面，主要开展了防风固沙植物种的选择和防沙固沙植物灌溉技术研究；在防护林与环境之间的相互关系方面，通过塔里木沙漠公路防护林树种对环境胁迫如：高温、干旱、风沙和盐分等的响应研究，阐明植物在逆境胁迫下的生理生态特性和适应逆境的内在机理，为沙漠腹地的防护林树种选择和人工绿地建设提供科学依据；同时，通过对防护林地固定样方的长期监测，探讨了防护林对恶劣环境条件的改善作用；通过对防护林及其周边环境的长期监测，发现了影响防风固沙林稳定性的限制因素，初步建立了沙漠公路防护林稳定性评价的综合指标体系。在对防护林引种植物进行野外栽培试验、生态生理试验、种源试验、大规模造林示范基础上，应用SPSS统计软件包进行判别分析，评价89种引种植物的适应性，将引种植物划分为完全适应、一般适应和不适应三类。采用主成分分析法（PCA）研究了防护林树种对干旱环境的生理响应并对植物的抗早性进行排序，在此基础上提出了防护林树种应以灌木树种梭梭归改勿砂lon anllnodendronB岁）、白梭梭（Haloxylon Perscuyn Bge���、多枝柽柳（Tamarix ramosissima Ldb．）和头状沙拐枣（Callugonum caput-medusae Schrenk）等为主。在实验的基础上，发现沙漠极端环境下高温胁迫可导致植物细胞膜透性、膜脂过氧化作用的增强及植物叶片或同化枝的失水，而树木体内渗透调节物质积累和保护酶活性的提高是植物抵御这种胁迫的主要机制。在长期定位监测和样方调查的基础上，应用系统生态学系统模型分析了咸水灌溉条件下防护林地土壤的水盐动态和平衡，分析了植物各营养器官盐分离子含量的变化，认为经过筛选的防护林树种基本上不存在盐害，植物能够生存的主要原因在于植物本身耐盐、避盐和泌盐的生理特性及沙地水、盐的强烈渗漏。通过防护林区内外的风沙观测，发现防护林有效改变了风沙流的结构，降低了风速，增加了沙丘的稳定程度。同时，对防护林地风沙土的理化特性进行定点的长期监测，应用通径系数分析等方法分析了土壤酶和微生物的活性与土壤质地的关系，发现防护林的建立改善了风沙土的质地，提高了土壤酶和微生物的活性，使风沙土向有利于提高其肥力质量的方向发展。本文首次把层次分析法（AHP）应用到防护林系统的稳定性初步研究，结果初步表明：塔里木沙漠公路防护林系统处于相对脆弱状态，恶劣的气候条件是系统稳定的主要干扰因素，它的剧烈波动使生态系统常常偏离稳定状态，使系统的组成结构发生变化，影响生态系统功能的发挥。人工措施可在一定程度改善沙漠公路防护林系统的环境质量，有助于提高生态系统的稳定程度。

几种中药材的化学成分及其定性定量检测方法研究

本论文由四部分组成。第一部分报道了佛手参提取物的化学成分研究，建立了活性成分含量测定的高效液相测定和指纹图谱研究，采用液质联用技术鉴定了主要色谱峰；第二部分报道了丹参及其复方制剂的特征图谱研究；第三部分探讨了两面针生物碱的电喷雾质谱裂解规律，并采用液质联用技术分离鉴定了提取物中的多种生物碱。第四部分概述了液质联用在药物代谢研究中的运用。 第一部分包括第一、第二和第三章。第一章针对佛手参（Gymnadeniaconopsea）块茎的甲醇提取物，采用大孔树脂和反相硅胶柱层析等各种分离方法，共分离鉴定出4 个化合物，通过波谱分析将它们的结构确定为dactylorhin B (1)、loroglossin (2)、dactylorhin A (3)和militarine (4)。这4 个化合物均是首次从佛手参中分离得到的琥珀酸葡萄糖苷类成分。第二章采用高效液相色谱法对西藏、四川、河北、青海和尼泊尔等不同地区产的十个佛手参样品进行腺嘌呤核苷和对羟基苯甲醇的定量分析，结果表明这2 个成份可视为佛手参的特征成分，但也注意到产地不同该2 个特征成分的含量也有所不同。第三章采用标准中药指纹图谱相似度计算软件，以10 个佛手参样品HPLC 图谱的平均值为相似性评价对照模板，对10 个样品进行了相似度评价，并经液质联用分析指认了7 个共有峰，分别为腺嘌呤核苷(1)、对羟基苯甲醇(2)、对羟基苯甲醛(3) 、dactylorhin B(4) 、loroglossin(5)、dactylorhin A(6)和militarine(7)。 第二部分包括第四、第五、第六和第七章。第四章运用电喷雾质谱检测了对照药材和五个不同产地的丹参药材中脂溶性和水溶性成分，系统地探讨了多种成分的电喷雾质谱规律，并以对照药材为标准建立了特征指纹图谱。五个产地的药II材通过与对照药材相对比，采用聚类分析的方法，得到了定性的鉴别与判断。并采用液质联用技术对丹参药材提取液中的化学成份进行分析，推测了九个特征峰，并对六样品的液相色谱图进行了聚类分析。第五章探讨了三七皂苷的电喷雾质谱电离和裂解规律，并采用电喷雾质谱法对三七标准药材，血通片中的皂苷成分进行了分析。第六章运用电喷雾质谱研究复方丹参片提取液的特征图谱，并和单味药材丹参和三七的特征图谱进行了对比研究。并运用HPLC-ESI MSn 分析鉴定了复方丹参片提取液中的化学成分，推测了12 个色谱峰。第七章总结了电喷雾质谱和液质联用技术在丹参药材，三七药材及复方丹参制剂中的运用的优势和局限性。 第三部分（第八章）研究了两面针生物碱中二氢白屈菜红碱(1)、二氢两面针碱(2)、8-酮基二氢白屈菜红碱(3)、8-丙酮基二氢两面针碱(4)、两面针碱(5)、和1,3-二(8-二氢两面针碱)丙酮(6)等六个苯并菲啶型生物碱的电喷雾质谱裂解规律，其中二氢两面针碱和二氢白屈菜红碱，8-丙酮基二氢两面针碱和8-酮基二氢白屈菜红碱是两对二个甲氧基分别在C-9 和C-10，C-10 和C-11 的同分异构体。实验结果表明，在相同的碰撞能下，这类位置异构体的ESI MS2 质谱二级碎片离子的相对峰度存在很大差异，这可以用于区分该类同分异构体，采用液-质联用可以对两面针的总生物碱提取物中的这些同分异构体加于区分。同时在本实验采用的液相色谱条件下，多种生物碱得到较好的分离，通过和对照品的保留时间，紫外吸收光谱及电喷雾质谱图对照，鉴定了11 个主要色谱峰。 第四部分（第九章）对液质联用技术在药物代谢中的运用进行了综述。 This dissertation consisted of four sections. The first two sections elaborated thephytochemical investigation of the rhizomes Gymnadenia conopsea R. Br., methoddevelopment for rapid identifying and qutifying the chemical condtituent of thistibetant medicine, and the chemical fingerprint analysis of rhizomes of G. conopsea,Salviae miltiorrhiza and P. notoginseng. The third section studied the fragmentationmechanism of six alkaloids from Zanthoxylum nitidium and method development forrapid identifying varieties of alkaloids from the extract of this herbal medicine. Thefourth section reviewed HPLC- MS method in drug metabolism studies. The first section consisted of chapters 1, 2, 3. Chapter 1 elaborated the phytochemicalinvestigation of Gymnadenia conopsea R. Br. Four succinate derivative esters wereisolated from the methanol extract of the rhizomes of G. conopsea through repeatedcolumn chromatography on normal and reversed phase silica gel, their structures weredetermined by ESI-MS, 1D and 2D NMR evidence. They were firstly discoveredfrom this species. In chapter 2, a high-performance liquid chromatography.diodearray detection (HPLC-DAD) method has been firstly developed for quantitation oftwo characteristic constituents, adenosine and 4-hydroxybenzyl alcohol, from theextract of rhizomes of G. conopsea. All 10 samples of G. conopsea contained differentamount of adenosine and 4-hydroxybenzyl alcohol. Adenosine and the4-hydroxybenzyl alcohol can be applied in identification and quality control for theroots of G. conopsea. In chapter 3, a high-performance liquid chromatography.diodearray detection.tandem mass spectrometry (HPLC-DAD-MSn) method has been firstly developed for chemical fingerprint analysis of rhizomes of G. conopsea andrapid identification of major compounds in the fingerprints. Comparing the UV andMS spectra with those of authentic compounds, seven main peaks in the fingerprintswere identified as adenosine, 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde,dactylorhin B, loroglossin, dactylorhin A and militarine. The Computer AidedSimilarity Evaluation System for Chromatographic Fingerprint of TraditionalChinese Medicine (CASES) was employed to evaluate the similarities of 10 samplesof the rhizomes of G. conopsea collected from Sichuan, Qinghai and Hebei provincesand Tibet autonomous region of China, and Nepal. These samples from differentsources had similar chemical fingerprints to each other. The second section consisted of chapters 4, 5, 6 and 7. In chapter 4，both thecharacteristic spectra of liposoluble tanshinones and aqueous-soluble salvianolic acidswere established by the electrospray ionization mass spectrometry (ESI MS)technique and the differences between standard and crude rhizomes of Salviaemiltiorrhiza Bge. from 5 sources were analyzed. The law of electrospray ion trap mass(ESI ITMS) of typical tanshinones and salvianolic acids is studied.The analysis of the chemical constituent of rhizomes of Salviae miltiorrhiza Bge. byliquid chromatography coupled with mass spectrum (LC/MS) technique wasestablished，and the distances among standard herb and crude herb from 5 sourceswere calculated by clustering analysis. According the DAD spectra and MS2 data，9tanshinones could be speculated. In chapter 5, the character spectra of total saponinsin P. notoginseng extracts were established by ESI ITMS and selective ion monitoring(SIM) technology. The law of notoginsenosides by ESI MS2 was studied. In chapter 6,the characteristic spectra of Compound Danshen Tablet established and compared byESI-MS and HPLC/DAD/MS, 6 known tanshinones and 3 saponins were speculated.In chapter 7, the advantage and disadvantage of the strategy, using the ESI ITMS andLC/MS techniques for study of characteristic spetra of danshen and Compound Danshen Tablet, were summerized. The third section (chapter 8) studied the fragmentation mechanism of six alkaloids,dihydronitidine, dihydrochelerythrine, 8-acetonyl dihydronitidine,8-acetonyldrochelerythrine, nitidine and 1,3-bis (8-dihydronitidinyl)-acetone, by ESIMSn. Tandem mass spectrometry experiments indicated that different substitutionsites of the methoxyl groups at C-9 and C-10 or at C-10 and C-11 determined thedifferent abundances of the MS2 fragmentation ions using the same collision energy.According to the different abundances of MS2 product ions, positional isomericbenzo[c] phenanthridine alkaloids can be differentiated. Moreover, ten constituents inthe crude alkaloids extract from the roots of Zanthoxylum nitidium were rapidlyidentified by high-performance liquid chromatography coupled with tandem massspectrometry (HPLC-MSn), through comparing the retention times and ESI MSn spectra with the authentic standards. The fourth section (chapter 9) is a review on HPLC-MS method development in drug metabolism studies.

瑞香狼毒（Stellera chamaejasme L.）对高寒草甸系统生长和非生长季节碳氮循环的影响及其花粉化感效应

瑞香狼毒（Stellera chamaejasme L.）是瑞香科（Thymelaeaceae）狼毒属的一种多年生野草，有毒。据调查，从20 世纪60 年代开始至今，狼毒在青藏高原东缘的高寒草甸上不断蔓延、密度不断变大，在一些地段甚至成为优势物种。有关狼毒在高寒草甸蔓延的生态系统效应的研究尚未见报道。本文从系统碳、氮循环的角度，分别研究狼毒在生长和非生长季节对高寒草甸生态系统的影响。同时，从花粉化感的角度，深入研究狼毒对当地同花期物种有性繁殖的影响。系统地研究高寒草甸生态系统物质循环过程，特别是非生长季节微生物和土壤碳氮库的动态变化，有助于揭示狼毒在系统物质循环方面的“物种效应”以及这种效应的季节变化，为丰富有关高海拔生态系统，特别是其非生长季的物质循环的科学理论做出贡献。同时，碳氮循环和花粉化感的研究还有助于深刻地理解狼毒作为一种入侵性很强的杂草的特殊的蔓延机制，从而为狼毒的有效防治、高寒草甸的科学管理提供依据。 针对狼毒在青藏高原高寒草甸上蔓延的生态系统碳氮循环方面的影响，开展以下2 方面的研究：（1）在生长季，研究松潘县尕米寺附近（北纬32°53'，东经103°40'，海拔3190 m）的两种地形（平地和阳坡）条件下狼毒对土壤碳氮循环影响及可能的原因。狼毒和其它几个主要物种（圆穗蓼（Polygonummacrophyllum D. Don var. Macrophyllum），草地早熟禾（Poa pretensis L.），四川嵩草（Kobresia setchwanensis Hand.-Maizz.），鹅绒委陵菜（Potentilla anserina L.var. anserine）和鳞叶龙胆（Gentiana squarrosa Ledeb.）的地上凋落物产量以及地上凋落物和根的化学组成被测量。在有-无狼毒斑块下，各种土壤的库（比如，铵态氮、硝态氮、无机磷和微生物生物量）和周转率（包括净矿化、净硝化、总硝化、反硝化和微生物呼吸速率）被测量和比较。（2）在非生长季节，尤其是春季冻融交替期，选取了两个研究地点——尕米寺和卡卡沟（北纬32°59'，东经103°41'，海拔3400 m），分别测定有狼毒和无狼毒斑块下土壤微生物生物量碳和氮、可溶性有机碳和氮以及铵态氮和硝态氮的动态变化。同时，分别在上述两个地点有-无狼毒的样地上，研究6 个主要物种（狼毒、圆穗蓼、草地早熟禾、四川嵩草、鹅绒委陵菜和鳞叶龙胆）从秋季开始、为期1 年的凋落物分解过程。 针对狼毒花粉化感对同花期其它物种可能的花粉化感作用开展以下工作：在实验室中，用一系列浓度的狼毒花粉水浸提液对与它同花期的其它物种以及自身花粉进行测试，测定花粉萌发率；在野外自然条件下的其它物种的柱头上施用上述浓度的狼毒花粉水浸提液，观测种子结实率，同时，观察狼毒花粉的种间花粉散布数量。 生长季节的研究结果表明，狼毒地上凋落物氮含量比其它几个主要物种更高，而木质素-总氮比更低。狼毒显著地增加其斑块下表层土壤中有机质的含量，而有-无狼毒的亚表层土壤在有机碳和总磷方面没有显著差异。狼毒表土中硝态氮含量在平地和阳坡比无狼毒土壤分别高113%和90%。狼毒表土中微生物生物量碳和氮量显著高于无狼毒表土。无论是平地还是阳坡，狼毒土壤的总硝化和微生物呼吸速率显著高于无狼毒土壤；而它们的反硝化速率只在平地有显著的差异。狼毒与其它物种间地上凋落物的产量和质量的差异可能是导致有-无狼毒土壤碳氮循环差异的原因。我们假设，狼毒可能通过增加贫氮生态系统土壤中的有效氮含量提高其入侵能力。 非生长季的研究结果表明，在青藏高原东缘的高寒草甸上，土壤微生物生3物量在11 月的秋-冬过渡期达到第一个峰值；在春季的冻融交替期，微生物生物量达到第二个峰值后又迅速降低。无机氮以及可溶性有机碳氮与微生物生物量有相似的变化过程。微生物碳氮比呈现显著的季节性变化。隆冬季节的微生物生物量碳氮比显著高于生长旺季的微生物碳氮比。这种变化可能暗示冬、夏季微生物的群落组成和对资源的利用有所不同。有-无狼毒斑块下土壤微生物和土壤碳、氮库一般只在秋-冬过渡期有显著差异，有狼毒土壤微生物生物量和土壤碳、氮库显著高于无狼毒土壤；而在之后的冬季和春季没有显著差异。所有6 个物种凋落物在非生长季分解率为24%-50%，均高于生长季的10%到30%。其中在秋-冬过渡期，凋落物开始埋藏的两周时间内，分解最快，达10%-20%。不同物种凋落物全年的分解率和分解过程有显著差异。圆穗蓼在全年的分解都较缓慢（非生长季26%，生长季15%），草地早熟禾和四川嵩草等全年的分解速率比较均匀（非生长季和生长季均为30%，非生长季略高），而狼毒在非生长季分解较快（约50%），而在接下来的生长季分解变得缓慢（约12%）。所有物种的凋落物氮含量在非生长季下降，而在随后的生长季上升。 实验室的花粉萌发试验证明，狼毒花粉对自身花粉萌发没有自毒作用，而其它受试的所有物种（圆穗蓼，秦艽（Gentiana macrophylla Pall. var. fetissowii），湿生扁蕾（Gentianopsis paludosa (Hook. f.) Ma var. paludosa），鳞叶龙胆，椭圆叶花锚（Halenia elliptica D. Don var. elliptica），蓝钟花（Cyananthus hookeri C. B.Cl. var. grandiflorus Marq.），小米草（Euphrasia pectinata Ten.），川西翠雀花（Delphinium tongolense Franch.），高原毛茛（Ranunculus tanguticus (Maxim.)Ovcz. var. tanguticus）和鹅绒委陵菜）的花粉萌发率随着狼毒花粉浸提液浓度的增加呈显著的非线性降低。大约3 个狼毒花粉的浸提液就可以抑制受试的多数物种的50%的花粉萌发。在鳞叶龙胆和小米草柱头上狼毒花粉的数量分别为5.76 个和3.35 个。狼毒花粉散布数量的差异最可能的原因在于是否有共同的传粉昆虫。花的形状（辐射对称VS 左右对称）、植株或花的密度以及花期重叠性可以部分解释这种差异。在野外试验中，我们发现6 个物种（秦艽、湿生扁蕾、鳞叶龙胆、椭圆叶花锚、蓝钟花和小米草）的种子结实率随狼毒花粉浸提液浓度的增加呈显著的非线性降低。鳞叶龙胆和小米草柱头上狼毒花粉的数量（分别是5.76 个和3.35 个）分别达到了抑制它们63%和55%种子结实率的水平。因此，狼毒对鳞叶龙胆和小米草可能存在明显的花粉化感抑制作用。狼毒周围的物种可能通过花期在季节或昼夜上的分异避免受到狼毒花粉化感的影响或者通过无性繁殖来维持种群繁衍，因此狼毒通过花粉化感作用对其周围物种繁殖的影响程度还需要进一步地研究。如果狼毒的花粉化感抑制作用确实存在，那么它可能成为一种自然选择压力，进而影响物种的进化。 Stellera chamaejasme L., a perennial toxic weed, has emerged and quicklydominated and spread in the high-frigid meadow on the eastern Tibetan Plateau ofChina since the 1960s. In the present study, effects of S. chamaejasme on carbon andnitrogen cycles on the high-frigid meadow on the eastern Qinghai-Tibetan Plateau ingrowing and non-growing season, and its pollen allelopathic effects on the sympatricspecies were determined. The present study that focused on carbon and nitrogencycles, especially on microbial biomass and pools of carbon and nitrogen innon-growing season, could profoundly illuminate plant-species effects on carbon andnutrient cycles and its seasonal pattern and help to understand spread mechanism ofS. chamaejasme as an aggressive weed. The present study also contributed to furtherunderstand carbon and nutrient cycles on alpine regions in non-growing season andprovide a basis on weed control of S. chamaejasme and scientific management in thehigh-frigid ecosystem. Effects of S. chamaejasme on carbon and nitrogen cycles on the high-frigidmeadow on the eastern Qinghai-Tibetan Plateau were determined. The study couldbe divided into two parts. (1) In the growing season, we quantified the effects of S.chamaejasme on carbon and nitrogen cycles in two types of topographic habitats, theflat valley and the south-facing slope, where S. chamaejasme was favored to spreadlitter and root were measured to explain the likely effects of S. chamaejasme on soilcarbon and nutrient cycles. The sizes of various soil pools, e.g. nitrite, ammonium,inorganic phosphorus and microbial biomass, and turnover rates including netmineralization, gross nitrification, denitrification and microbial respiration weredetermined. (2) In the non-growing season study, microbial biomass carbon andnitrogen, soluble organic carbon and nitrogen, ammonium and nitrate weredetermined through the non-growing season, especially in the processes offreeze-thaw of spring in two high-frigid sites, i.e. Kaka valley and Gami temple, onthe eastern Qinghai-Tibetan Plateau. Meanwhile, litter decomposition of six commonspecies, including Stellera chamaejasme L., Polygonum macrophyllum D. Don var.Macrophyllum, Poa pretensis L., Kobresia setchwanensis L., Potentilla anserina L.var. anserine and Gentiana squarrosa Ledeb., were also examined under theabove-mentioned experimental design through one whole-year, which began in theautumn in 2006. In the study of pollen allelopathy, several work, including in vitro study oneffects of extract of pollen from S. chamaejasme on sympatric species and pollenfrom itself, field experiments on effects of pollen extract with the same regime ofconcentrations on seed set and field observation on heterospecific pollen transfer ofS. chamaejasme to six of those sympatric species has been done. The results in the growing season showed that aboveground litter of S.chamaejasme had higher tissue nitrogen and lower lignin: nitrogen ratio than thoseco-occurring species. S. chamaejasme significantly increased topsoil organic matter,whereas no significant differences were found for organic C and total P in subsoilbetween under-Stellera and away-Stellera locations. The nitrate in Stellera topsoilwas 113% and 90% higher on the flat valley and on the south-facing slope,respectively. Both microbial biomass C and N were significantly higher in Stelleratopsoil. Gross nitrification and microbial respiration were significantly higher inStellera topsoil both on the flat valley and on the south-facing slope, whereassignificant differences of denitrification were found only on the flat valley. Thedifferences in the quantity and quality of aboveground litter are a likely mechanismresponsible for the changes of soil variables. We assumed that S. chamaejasme couldenhance their spread by increasing nutrient availability in N-deficient ecosystems. The results in the non-growing season showed that microbial biomass achievedthe first summit in late autumn and early winter on the eastern Qinghai-TibetanPlateau. In the stages of freeze-thaw of spring, microbial biomass firstly achieved thesecond summit and subsequently sharply decreased. Inorganic nitrogen, solubleorganic carbon and nitrogen had a similar dynamics with that of microbial biomass.Ratio of microbial biomass carbon and nitrogen had an obviously seasonal pattern.The highest microbial C: N were in the non-growing season, which weresignificantly higher than those in the growing season. The seasonal pattern inmicrobial biomass C: N suggested that large changes in composition of microbialpopulation and in resources those used by microbes between summer and winter.Generally, microbial biomass and pools size of carbon and nitrogen in Stellera soilwere significantly higher than those under adjacent locations in late autumn andearly winter, but there were not significant differences in winter and in spring. Litterof all the focal species (Stellera chamaejasme L., Polygonum macrophyllum D. Donvar. Macrophyllum, Poa pretensis L., Kobresia setchwanensis Hand.-Maizz.,Potentilla anserina L. var. anserine and G. squarrosa Ledeb.) decomposed about24%-50% in the non-growing season, which were higher than those in the growingseason (ranged from 10% to 30%). Litter decomposed 10%-20% within the first twoweeks in late autumn and early winter. Significant differences in the whole-yeardecomposition rate and in the processes of decomposition were found among species.Polygonum macrophyllum decomposed slowly through the whole year (26% and15% in the non-growing season and in the growing season, respectively). Certainspecies, such as P. pretensis L. and K. setchwanensis, decomposed at a similar rate(30% both in the non-growing and in the growing season, slightly higher in the8growing season than those in the growing season), whereas S. chamaejasmedecomposed more rapidly (about 50%) in the non-growing season and subsequentlydecomposition became slow (about 12%) in the growing season. Litter nitrogencontents of all the focal species firstly decreased in the non-growing season and thenincreased in the growing season. In vitro experiments of pollen allelopathy, the results showed that pollen from S.chamaejasme was not autotoxic, whereas pollen germination in all the sympatricspecies (Polygonum macrophyllum D. Don var. Macrophyllum, Gentianamacrophylla Pall. var. fetissowii, Gentianopsis paludosa (Hook. f.) Ma var. paludosa,Gentiana squarrosa Ledeb., Halenia elliptica D. Don var. elliptica, Cyananthushookeri C. B. Cl. var. grandiflorus Marq., Euphrasia pectinata Ten., Delphiniumtongolense Franch., Ranunculus tanguticus (Maxim.) Ovcz. var. tanguticus andPotentilla anserina L. var. anserina) decreased nonlinearly as the increasingconcentrations of extract of pollen from S. chamaejasme. Pollen Extract of threepollens from S. chamaejasme generally inhibited 50% pollen germination of most ofthe focal species. 5.76 and 3.35 pollens from S. chamaejasme were observed in fieldon stigmas of G. squarrosa and E. pectinata, respectively. Differences inheterospecific pollen transfer of S. chamaejasme could be attributed to the primaryreason whether they shared common pollinators. Flower morphology (e.g.zygomorphic or actinomorphic), plant or floral density and concurrence in floweringphonologies could explain, in part, the differences in heterospecific pollen transfer.In field experiments, the results showed that seed set in six sympatric species(Gentiana macrophylla Pall. var. fetissowii, Gentianopsis paludosa (Hook. f.) Mavar. paludosa, Gentiana squarrosa Ledeb., Halenia elliptica D. Don var. elliptica,Cyananthus hookeri C. B. Cl. var. grandiflorus Marq. and Euphrasia pectinata Ten.)decreased nonlinearly as the increasing concentrations of extract of pollen from S.chamaejasme. According to the nonlinear curves, the amounts of pollens from S.chamaejasme on stigmas of G. squarrosa and of E. pectinata (i.e. 5.76 grains and3.35 grains, respectively) could reduce 63% and 55% seed set of G. squarrosa and ofE. pectinata, respectively. Thus, allelopathic effects of S. chamaejasme on G.squarrosa and E. pectinata could be realistic. The sympatric species of S.chamaejasme could avoid pollen allelopathy of S. chamaejasme to sustainthemselves. This highlights the need to study how much pollen allelopathy in S.chamaejasme influences the sympatric species through divergence in seasonal ordiurnal flowering phonologies or through asexual reproduction. If pollen allelopathyin S. chamaejasme was confirmed, it could be as a pressure of natural selection andthus play an important role in species evolution.

短毛柽柳(TamarixkareliniiBge)的分类学地位探讨

短毛柽柳的分类学地位一直是柽柳属中争论的焦点之一。由于所持划分依据不同 ,认为短毛柽柳为刚毛柽柳杂种、变种或一独立的种。本文系统回顾了短毛柽柳的研究历史 ,对前人所做的有关刚毛柽柳和短毛柽柳形态学、解剖学、孢粉学、植物化学及分子生物学的研究结果进行了分析 ,并查阅了大量的标本 ,认为短毛柽柳是刚毛柽柳的一个变种 ,是从刚毛柽柳进化到另一个极端种的中间过渡阶段 ,同意Baum的意见 ,将其学名恢复为Tamarixhispidavarkarelinii (Bge)Baum .。

黄芪幼苗N,P,K营养缺乏症状和生理特性研究

黄芪是我国重要的传统大宗药材,以豆科紫云英属植物膜荚黄芪Astragalus membranaceus(Fisch.)或蒙古黄芪A.membranaceus(Fisch.)Bge.Var.mongolicusBge.Hsiao.的干燥根系入药[1],具有补中益气、降压利尿、排脓消痈、脱毒生肌、滋养肌肤等多种功效,现代临床及药理研究证明,黄芪还有增强肌体免疫功能,促进抗体合成等作用,有关黄芪的药理活性及其有效成分已得到了较为深入的研究[2-4],随着黄芪市场需求量的增加,野生黄芪资源已难以满足需求,人工栽培黄芪受到重视,并且初具规模。田间栽培黄芪的营养特性和施肥管理是人工栽培质量和产量的理论基础。有关氮磷钾化肥用量及其比例对黄芪生长及产量的影响已有一些研究[5,6],但由于黄芪生长中的营养元素比例不平衡现象时常发生,至于某一种营养元素缺乏时黄芪的外观特征表现,目前尚未见报道,出现缺素症状时难以判断,限制对黄芪及时进行营养调控,对人工栽培黄芪的质量与产量产生影响。本研究的目的是研究黄芪生长中的营养缺乏时的外观表现特征和生理特性变化。本试验采用溶液培养的方法,在氮、磷、钾营养元素缺乏条件下,对黄芪的生长发育状况、营...

CE coupling with end-column electrochemiluminescence detection for chiral separation of disopyramide

CE with electrochemiluminescence, (ECL) detection technique was successfully applied for the chiral separation of a kind of class IA antiarrhythmic racemic drug. To the best of our knowledge, this is the first report of ECL detection used in chiral CE. To get better detection sensitivity and good enantioresolution at the same time, the conditions of capillary inlet and outlet buffer were systematically optimized. Unlike the traditional chiral separation method, the buffers we used in the capillary inlet and outlet differed from each other in terms of buffer pH, ionic strength, type of BGE as well as buffer composition. Under the optimum conditions, baseline enantioseparation and highly sensitive detection of the enantiomers were achieved. Wide linear relationship of each enantiomer was achieved in the range of 5 x 10(-7) to 2 x 10(-5) mol/L with relative coefficients of 0.996 and 0.997, respectively. The detection limits were estimated to be 8 x 10(-8) and 1.0 X 10(-7) mol/L (S/N = 3) for the enantiomers, respectively. In addition, a successful application of this new method to the chiral separation of the racemic drug in spiked plasma samples confirmed the validity and applicability of the chiral CE-ECL method.

围封对退化矮嵩草草甸群落结构和主要种群空间分布格局的影响

通过研究围栏封育1年后中度退化的矮嵩草草甸（夏季牧场）群落结构的变化,探讨主要植物种群在小尺度（50 cm×50 cm）上的空间分布格局,并从生活史特征和生态适应对策等角度探讨产生和维持这些格局的机理。结果表明,围封1年显著降低了群落中主要种群矮嵩草（Kobresia humilis）、高山唐松草（Thalictrum alpinum）和雪白委陵菜（Potentilla nivea）的重要值,而增加了线叶龙胆（Gentiana farreri）的重要值;显著增加了群落的地上生物量和总生物量,但对地下生物量和群落多样性的影响不显著。围封使退化矮嵩草草甸主要种群矮嵩草、高山唐松草、珠芽蓼（Polygonum viviparum）、线叶嵩草（Kobresia capillifolia）和金露梅（Potentilla fruticosa）等空间分布格局从放牧后的随机分布向聚集分布发展,而雪白委陵菜、重齿风毛菊（S.katochaeteMaxim）、矮火绒草（Leonto-podium nanum）和美丽风毛菊（Saussurea pulchra）等种群的空间分布格局没有发生改变;但黑褐苔草（Carex atro-fusca）的空间格局从放牧后的聚集分布转向随机分布。因此,退化的矮嵩草草甸在围封的初始阶段,由于避免了家畜的选择性采食及其践踏作用,首先可能是使主要种群的空间分布格局有从随机分布向聚集分布变化的趋势,从而使小尺度的种间隔离来降低种间的竞争强度,从而改变了不同物种对资源和空间的竞争能力,进而推动群落物种组成和结构的恢复演替。

管花秦艽的传粉生态学研究——兼与同域分布近缘种的比较

在青藏高原东北部连续两年观察了晚秋开花植物管花秦艽Gentiana siphonantha的传粉生态学特征,并在此基础上进一步比较分析了与该物种同域分布且亲缘关系较近、但开花较早的麻花艽G.straminea之间的传粉生态学特征。管花秦艽的花发育过程表现出雌雄异熟和雌雄异位的特点,不存在花内的自花传粉,套袋隔离的花不结实也支持这一结论;株内自交的高结实率表明该物种是自交亲和的。盛花期每植株平均有15朵开放的花,雄性和雌性阶段的花比例为1.2：1;自然条件下产生种子必须依赖传粉媒介;苏氏熊蜂是最有效的传粉昆虫,且访花过程中对雄性和雌性阶段花不具明显的偏向性;株内连续访花的频率高达87.8%,从而导致同株异花传粉自交的广泛存在。与同域分布的麻花艽相比,管花秦艽的单花花期、雄性和雌性期持续时间缩短。但盛花期开花数量明显增加。令人感兴趣的是尽管两个近缘种的花形态特征存在显著差异,但都是由同一种熊蜂传粉。这一特点与过去认为花颜色和花管长度是物种分化过程中与不同传粉昆虫协同进化导致生殖隔离的假说不相符合。管花秦艽单花的访花频率和同株异花连续访花的比例都明显高于麻花艽。两个物种不同花序设计导致访花昆虫行为的改变可能是造成这一差异的主要原因。两个物种具有不同的开花时间,但仍然存在一定的花期重叠,表现出不完全的传粉生殖隔离状态。

高寒草甸植物抗氧化系统对长期增强UV-B辐射的响应

通过研究自然条件下模拟平流层臭氧破坏5%时近地表面增加的太阳UV-B辐射对高寒草甸4种典型植物（矮嵩草Kobresia humilis、垂穗披碱草Elymus nutans、麻花艽Gentiana straminea和鹅绒委陵菜Po-tentilla anserina）的抗氧化系统的影响表明,尽管各植物的抗氧化系统组分变化不同,但4种植物的膜脂过氧化程度没有加剧,长期增强UV-B辐射没有对膜系统造成损伤。在自然长期增强UV-B条件下,4种植物的膜脂过氧化产物——丙二醛（MDA）含量与对照相比无显著差异。垂穗披碱草、鹅绒委陵菜的谷胱甘肽（GSH）含量增加,麻花艽的超氧化物歧化酶（SOD）活性与鹅绒委陵菜的过氧化氢酶（CAT）活性上升,同时麻花艽的类胡萝卜素（Car）含量亦显著增加。可见这些植物已能很好地适应UV-B强辐射,其抗氧化能力除了与抗氧化系统各组分的协同作用有关外,也可能与种的适应性有关。

粗茎秦艽与西藏秦艽(龙胆科)种间的形态学、染色体与分子界定

粗茎秦艽Gentiana crassicaulis Duthie ex Burk和西藏秦艽G. tibetica King ex Hook. f.的主要区别是前者花冠小、檐部蓝色, 后者花大、檐部紫褐色; 但二者在西藏东南部地区的形态界限不清楚, 具有较多的中间形态过渡类型.本文以该地区居群作为主要研究对象, 并以云南和西藏其他地区的典型形态类群为对照, 比较研究了两个物种间的形态学、染色体和分子标记界定.共研究了10个居群69个个体的trnS-G和ITS序列、6个居群26株植株的染色体倍性和54份标本的形态变异.结果表明, 花冠大小与染色体倍性密切相关.云南等地分布的粗茎秦艽均为二倍体(2n=26), 花冠较小; 西藏西部的西藏秦艽均为四倍体(2n=52), 花冠较大.而西藏东南部的形态过渡类型也为四倍体, 花冠较大, 且其中的部分植株花颜色变浅, 类似西藏秦艽.但母系与双亲遗传背景的分子标记都表明西藏东南部居群与典型形态特征上的粗茎秦艽聚为一支, 应属于粗茎秦艽; ITS序列上也未发现杂合位点.这些分子证据都不支持近期杂交渗入造成形态过渡性的假设.综合分析表明多倍化造成了粗茎秦艽种内形态特征的变异, 其四倍体个体与四倍体西藏秦艽在形态特征上的趋同, 最终导致了二者在西藏东南部地区形态界限的模糊.但是, 分子证据可以很好地区分这种多倍化造成的形态趋同.我们的研究结果还表明, 综合利用分子、染色体和广义形态证据来解决近缘种之间种范围的界定具有更大的优势.此外, 还讨论了粗茎秦艽不同染色体倍性的分布以及该高山物种可能的起源与扩散途径.

藏药麻花艽愈伤组织的诱导及植株再生

1植物名称麻花艽(Gentiana straminea Maxim.),又名麻花秦艽,藏药中称为"解吉嘎保".2材料类别幼嫩叶子.3培养条件愈伤组织诱导培养基:(1)MS+2,4-D3.0 mg•L-1(单位下同);(2)MS+2,4-D 3.0+KT 1.0.继代培养基:(3)MS+2,4-D 1.0+6-BA 0.5+NAA 0.5.分化培养基:(4)MS+NAA 1.0+6-BA 2.0+ZT 3.0.以上培养基均附加CH(水解酪蛋白)300、肌醇200、3%蔗糖、5 g•L-1琼脂粉,pH 5.8.培养温度为(25±1)℃,光源为日光灯,光强为30～60μmol•m-2•s-1,光照时间12 h•d-1.

不同生长季节下藏药麻花秦艽活性成分含量研究

采用高效液相色谱法测定了野生与栽培藏药麻花秦艽（Gentiana straminea）根中龙胆苦苷、落干酸、獐牙菜苦苷和獐牙菜苷四种环烯醚萜苷类化学成分的含量及其在不同生长季节的变化趋势。结果表明，4种环烯醚萜苷类成分的含量随植物的生长季节而波动，其活性成分含量在野生种与栽培种之间发生了一定的差异。但是，栽培根中的龙胆苦甙已达到药典的标准，可供药用。

青藏高原几种典型高山植物的光合特性比较

选用西宁地区人工栽培的高山植物唐古特大黄（Rheum tanguticum）、山莨菪（Anisodus tanguticus）和麻花艽（Gentiana straminea），比较了3种高山植物之间光合作用的光响应和CO2响应特性，叶片光合色素以及UV—B吸收物质的差异；并以低海拔植物菘蓝（Isatis indigotica）为对比，分析了高山植物与低海拔植物的差异。结果表明：与低海拔植物菘蓝相比，3种高山植物光合作用的表观量子效率（AQY）都偏低；唐古特大黄叶片的AQY、羧化效率（CE）和光呼吸速率（Rp）都很低，净光合速率（Pn）的光响应曲线在全日照光辐射范围内并没有达到完全饱和，这与单位面积叶片具有较高的光合色素以及UV—B吸收物质有关；麻花艽植物与唐古特大黄一样，具有较高的UV-B吸收物质和光合色素含量，但其Rp较高，加之Pn受气孔限制较为明显，故其光合作用的饱和光强很低，Pn相对于其它3种植物也较低；山莨菪与低海拔植物菘蓝的光合特性很相似，都具有较高的AQY和CE。这些结果表明，3种高山植物的光合特性有较大差异，但并没有一致的相对于低海拔植物的共性。4种植物Pn的胞间CO2浓度（G）响应曲线在CO2饱和点以后都表现为无机磷（Pi）再生限制，其吃的变化与CO2饱和点以后的最大Pn的变化基本一致。

藏药麻花秦艽的研究进展

麻花秦艽（Gentiana straminea Maxim．）具有清热利胆、舒筋止痛之功效，是一种治疗风湿性关节炎、肺结核、低热盗汗、黄疸型肝炎等的珍稀藏药草本植物。笔者系统阐述了近年来该植物的研究进展，并对其研究方向进行了展望，为该物种资源的进一步有效开发利用提供参考。

用nrDNA ITS序列探讨狭蕊龙胆属及其近缘属(龙胆科)的系统发育

运用核糖体DNA内转录间隔区ITS序列对狭蕊龙胆属Metagentiana10种及其近缘属22种植物进行了系统发育分析.ITS分析结果表明狭蕊龙胆属是一个多系群:在系统发育树上,双蝴蝶属Tripterospermum和蔓龙胆属Crawfurdia的种类位于狭蕊龙胆属各分支中,而且双蝴蝶属和蔓龙胆属的种类也相互交叉;这一结果不支持将3个属各自独立为属.但是,在所有分析中,3个属共同形成一单系分支,是龙胆属Gentiana的姊妹群;这一结果与形态学、花部解剖学、细胞学、孢粉学和胚胎学等证据基本一致,狭蕊龙胆属应该从龙胆属中分离出来,它与双蝴蝶属和蔓龙胆属有更为密切的亲缘关系.根据分支图,在狭蕊龙胆属、双蝴蝶属和蔓龙胆属组成的复合群中,现已知的染色体基数x=17、21和23可能存在网状和平行进化.