角蟾科（Megophryidae; Anura）蝌蚪的形态多样性和生态适应

角蟾科（Megophryidae）是以角蟾属（Megophrys Kuhl and Van Hasselt, 1822）为模式属而建立的，隶于无尾目（Anura），变凹型亚目（Anomocoela）。角蟾科包括2 亚科11 属142 种，分布于东洋界，从巴基斯坦、中国西部向东直到菲律宾和苏达群岛；中国有9 属75 种分布于华中和华南地区。角蟾科被认为是原始的两栖动物之一，其分类学、系统学、生态学、动物地理学的研究均深受中外科学家的瞩目。近年来，通过形态学、古生物学、细胞学、生态学、支序系统学的研究，角蟾科的分类与系统学研究取得了较大进展。与成体形态和分子系统学研究结果相比较，蝌蚪的研究存在更多的问题和挑战，尚需深入研究：（1）角蟾科蝌蚪的形态多样性分析；（2）角蟾科的系统发育关系与蝌蚪的演化，以及口漏斗的起源；（3）角蟾科蝌蚪表型分化与栖息环境和觅食行为的适应演化。针对上述问题，本文对角蟾科9 属30 种蝌蚪的形态特征，包括外部宏观形态和口器外部结构特征、口器内部显微结构、唇齿和角质颌的亚显微结构作了深入细致、多层次的比较研究；通过12s rRNA 和cytochrome b 基因构建最大简约树，采用贝叶斯系统发育进行分析，蝌蚪型的演化采用祖先性状的重建方法分析；得到如下结论：1）初步将角蟾科蝌蚪分为4 种类型；并且建立了2 种新的角蟾科蝌蚪类型。A 型：拟髭蟾型蝌蚪，该型蝌蚪包括拟髭蟾属、髭蟾属、齿蟾属和齿突蟾属的物种；B 型：新类型，掌突蟾型蝌蚪，该型蝌蚪在本文中包括掌突蟾属、小臂蟾属的物种；C 型：新类型，短腿蟾型蝌蚪，一种特化类型，该型蝌蚪在本文中仅包括短腿蟾属的物种；D 型：角蟾型蝌蚪，该型蝌蚪在本文中包括无耳蟾属、小口拟角蟾属和异角蟾属的物种。2）对角蟾科的分类进行了修订：（1）支持角蟾科两个亚科的分类系统；（2）角蟾亚科包括拟角蟾属、异角蟾属、无耳蟾属和短腿蟾属；该亚科形态差异小，系统学关系比较复杂，暂不作族级分类的再划分；（3）拟髭蟾亚科分为2 个族：拟髭蟾族，该族物种具有类型A 的蝌蚪，包括4 个属：拟髭蟾属、髭蟾属、齿蟾属、齿突蟾属；掌突蟾族，该族物种具有类型B 的蝌蚪，包括2 个属：掌突蟾属和小臂蟾属。3）结合分子系统进化关系探讨了4 种蝌蚪类型的演化。（1）角蟾科蝌蚪的最近共同祖先来自于一类具有拟髭蟾型蝌蚪性状的蝌蚪；（2）掌突蟾型蝌蚪和角蟾亚科的蝌蚪是由具有拟髭蟾型蝌蚪性状的祖先蝌蚪分别演化而来；（3）短腿蟾型蝌蚪是角蟾型蝌蚪的一种特化类型；（4）外群蝌蚪具有与拟髭蟾型蝌蚪相似的性状，进一步印证了类拟髭蟾型蝌蚪是角蟾科蝌蚪的最近共同祖先的假说；（5）具有口漏斗的蝌蚪类型是由不具口漏斗的蝌蚪类型演化而来，在角蟾科中口漏斗是一种衍生性状。4）分析了角蟾科四种蝌蚪类型与栖息环境的适应演化。（1）角蟾科蝌蚪的口部和体形的变化反映了该科蝌蚪由缓流向类似静水生境的回水凼的渐变式适应，角蟾科蝌蚪的形态显示了多方面的适应变化；（2）随着蝌蚪类型由A 向D的演化，当水速较大时，拟髭蟾型的蝌蚪营流水攀吸型生活方式；当水速递减时，掌突蟾型蝌蚪营流水附着型生活方式；当水速进一步递减时，具有较小口漏斗的短腿蟾型蝌蚪和具有大漏斗的角蟾型蝌蚪营流水浮泳型生活。角蟾科蝌蚪对于水流递减的适应演化说明蝌蚪的生态学适应是具有进化意义的；（3）蝌蚪口器内部结构的分化揭示了蝌蚪和食性的适应关系，蝌蚪以口部的唇齿与角质颌刮取或吞吸水中的物质，然后，通过口乳突有选择地过滤进入口腔中食物。拟髭蟾亚科蝌蚪的唇齿多而窄，唇齿间距宽，颌鞘粗而稀，反映了其植食性为主的特点；它们的舌前乳突一般为指状，在口腔入口处所占面积小，其机械过滤的作用很多被唇齿和角质颌分担了；而角蟾亚科的蝌蚪，其角质颌弱，其舌前乳突一般为匙状，几乎填满了口腔入口处，因此舌前乳突起了主要的机械过滤作用。The family Megophryidae is the largest and most diverse families inArchaeobatrachia, and most of its species occur in India, Pakistan, and eastward intoChina, Southeast Asia, Borneo and the Philippines to the Sunda Islands. Currently thefamily includes 142 species have been grouped into two subfamilies, Megophryinaeand Leptobrachiinae. The mountains of central and southern China are rich in speciesof Megophryidae, 75 species belong to 9 genera and two subfamilies.The family was supposed to be ideal materials of studies in many fields of biology,such as taxonomy, evolution, systematics, ecology, and biogeography. Recently, therehave a great development in taxonomy and systematics of megophryids throughstudied by morphology, paleontology, cytology, ecology, and cladistics. However,larvae of megophryids were generally unknown, although the tadpoles might be veryimportant for above studies.In this paper, we examined the evolutionary scenario of the tadpoles’ morphologyin the context of a phylogenetic framework. Our objectives are (1) to evaluate thedivergence of larval body shape and oral discs in the family Megophryidae, (2) toexplore the evolutionary trends of the larvae in megophryidae, and test if thefunnel-shaped oral disc is apomorphic, and (3) to explore the relationship of the larvalstructure, diet and microhabitat.We examined larval morphology of 30 megophryid species, the larval body shape,oral discs, the buccopharyngeal cavity, and jaw sheaths and denticles of the Chinesemegophryid frogs were re-examined. We constructed a phylogeny of the species on thebasis of published mitochondrial cytochrome b and 16S rRNA gene segments usingpartitioned Bayesian analyses. Furthermore, hypothetical changes of larval morphologywere inferred using parsimony principle on the phylogeny. The results showed that:1) Four tadpole types in Megophryidae. The larval morphological charactersseries in Chinese megophryids fall into four general categories according to the bodyshape and oral discs: (A) Leptobrachiini type, species from genera Leptobrachium,Oreolalax, Scutiger and, Vibrissaphora share this type of tadpoles. (B) Leptolalax type,species of genus Leptolalax have this type of tadpoles. (C) Brachytarsophrys type,species of the genus Brachytarsophrys have this type of tadpoles. (D) Megophryinitype, species of the genera Atympanophrys, Ophryophryne, and Xenophrys share this type of tadpoles. Of which B and C are two novel types.2）Taxonomic implications. The present study leads us to reconsider the generalclassification of tribes attributed to members of Megophryidae. More specifically,concerning the phylogenetic relationships and the two novel tadpole types describedherein, we propose a provisional taxonomy for the family but suggest that further taxasampling of other megophryids be performed to confirm this taxonomic change. TheMegophryidae is composed of two subfamilies (Leptobrachiinae and Megophryinae).The Leptobrachiinae was recogonized the two tribes: (1) tribe Leptobrachiini sensuDubois, corresponding to the tadpole of type A, including four genera, i.e.,Leptobrachium, Oreolalax, Scutiger and, Vibrissaphora; (2) tribe Leptolalaxini,corresponding to the tadpole of novel type B, including two genera, i.e., Leptolalaxand Leptobrachella. However, the relationships among the genera of Megophryinaewere largely unresolved, they recognized no monophyletic groups above the generalevel. A more thorough sampling will likely foster a better taxonomic solution.3) The larval evolutionary scenario in Megophryidae.Type A is characteristicof normal-mouthed with multiple tooth rows, representing the tadpole type of theMRCA of Chinese megophryids. Type B is characteristic of normal-mouthed withreduced tooth rows, prolonging labium, and integumetary glands. Type C ischaracteristic of no labial teeth and smaller umbeliform oral disc. Type D ischaracteristic of no labial teeth, enlarged umbeliform oral disc, representing the tadpoleof the MRCA of subfamily Megophryinae. A previous hypothesis, referring tofunnel-shaped oral discs as an apomorphy, is supported.4) The larval adaptation to habitats in Megophryidae. Tadpoles generallyadhere to substrates using their mouths, and the microhabitat that the tadpoles occupyreflects the degree of adhesion and oral complexity. The morphological changes inmegophryid tadpoles virtually allow a progressive adaptation to a changing habitatfrom faster water to slower water. Within the tadpoles of Type A to type D, the TOTbecomes smaller and smaller, and the oral disc orientates from anteroventral toumbelliform upturned, and eye position orientates from dorsal to lateral, and the trunkis more and more depressed and tail becomes relatively longer and slender. Within therunning water, the normal-mouthed with multiple tooth rows of Leptobrachiini tadpoles are correlated with lotic-suctorial, benthic feeders with anteroventral oraldisc and the largest body. With the water’s velocity decreasing, the lotic-adherentfeeders of Leptolalax tadpoles have tube-shaped labium with reduced tooth rows andintegumetary glands. And then, the smaller umbeliform in Brachytarsophrys tadpolesand the enlarged umbeliform oral disc in the Megophryini tadpoles are inhabitmicrohabitats of non-flowing backwaters of rivers, indicative of adaptive traits oflotic-neustonic surface feeders. The scheme of megophryid tadpoles andmicrohabitats provided the first clear evidence which congruent with the hypothesis ofAltig and Johnston (1989). The ecological divergence plays a general role in thedivergence and evolution of megophrid larvae. There is a definite correlation amongthe buccopharyngeal cavity, diet and feeding mechanisms, the tadpole graze orswallow the food particles, then through papillae which like a sieve and sort out foodparticles to the oesophagus. The tadpole of Leptobrachiinae possess multiple toothrows, wide intertooth distance as well as thick and sparse jaw sheath, these tadpolesinhabit bottom of the streams and graze on epiphyton or major detritus of organicmatter on the substrates, their prelingual papillae like single finger, the mechanicalpurpose of papillae served share in by tooth and jaw. The tadpoles of Megophryinaeoccur near the water surface of small streams and are the filter feeder, their dietincludes plankton and organic debris floating on the water surface, those tadpolepossess weak jaw, their prelingual papillae like spoon, the mechanical purpose ofpapillae served mostly for sieve.

大熊猫“盛林1号”放归监测

大熊猫(Ailuropoda melanoleuca)是我国特有的珍稀濒危物种，国家Ⅰ级重点保护野生动物，被称为“国宝”。目前，大熊猫被局限在我国中西部的岷山、邛崃、大相岭、小相岭、凉山和秦岭6大山系中。对大熊猫的保护和研究，我国政府、保护生物学科研人员、社会各界及国际保护组织都做了大量的工作。根据全国三次大熊猫调查结果显示，大熊猫栖息地片段化现象依然存在，形成多个隔离的大熊猫小种群。尤其在小相岭、大相岭、岷山B和岷山C种群，大熊猫数量较少，且栖息地破碎，面临较大威胁。有的山系大熊猫种群数量些已低于最小可存活大熊猫种群的数量，如果不采取人工措施，这些种群的大熊猫存在灭绝的危险。 将圈养大熊猫放归野外，以补充野外大熊猫种群数量，增加其遗传多样性，复壮和扩大野生大熊猫种群，是大熊猫人工繁育的最终目标。为降低放归的风险性，在放归人工繁育大熊猫前，将救护存活的野生大熊猫先有计划放归野外，并对其进行跟踪监测，对积累大熊猫放归经验，进一步研究大熊猫野外生物学习性，丰富放归地大熊猫种群遗传多样性，为人工繁育大熊猫放归野外夯实基础，具有十分重要的意义。2005年8月8日，国家林业局和四川省人民政府联合将救护野生大熊猫“盛林1号”放归于龙溪－虹口国家级自然保护区内岷山B大熊猫种群栖息地，并进行系统监测研究。成功的积累了一些放归经验和放归大熊猫的生物学资料，为人工繁育大熊猫的放归奠定了一定基础。 2005年8月至2007年6月期间，我们采用GPS无线电项圈、粪便DNA检测和红外线自动触发相机陷阱的方法，对大熊猫“盛林１号”进行了追踪监测，获得了以下成果： 1.通过分析“盛林1号”放归后了活动趋势和采用两种贝叶斯方法，利用目前五大山系的已有微卫星遗传数据，检测“盛林１号”与五大山系的遗传关系的远近，推测其来源于邛崃山系的可能性较大。 2.收集了大量“盛林1号”野外生境选择数据。我们认为“盛林1号”放归后经历了应急期、初步稳定期、长途迁徙期三个阶段（这可能是今后放归大熊猫都必经的三个时期），并与当地大熊猫种群已发生交流。目前“盛林１号”仍在寻找适合的巢域。 3.结合过去监测数据分析，在放归区域大熊猫和羚牛尽管同域分布，但由于食性不同，对微生境选择还是有着很大差异，因此保护管理对策要有针对性。 4.“盛林1号”的放归是成功的。救护大熊猫异地放归工作应继续开展，但要改进放归后的监测技术。要改进现有对人工饲养大熊猫野化培训方法和放归方式，才能真正将人工繁殖个体放归野外。 Giant Panda (Ailuropoda melanoleuca) is an endangered species endemic to China. It was listed as National Protected I Class Species and is crowned as “National treasure” of China. The populations of Giant Panda are limited in 6 mountain system in Center-West of China, i.e. Mingshan, Mt. Qionglai, Mt. Daxiangling，Mt. Xiaoxiangling, Mt. Liangshan and Mt. Qinling. The results of the Third National Survey on Giant Panda showed that the habitats of Giant Panda is still fracted and Giant Panda population is divided into several isolated small populations. Population B from Mt. Daxiangling, Mt. Xiaoxiangling and Mt. Mingshan and Population C from Mt. Mingshan are very small with very fracted habitat and are more endangered. Several populations in those mountain systems are smaller than Minimum Viable Population of Giant Panda. It is very possible that those populations will be extinct without artificial help. The ultimate Goal of Reintroduction caged Giant Panda to wild is to increase wild population size and genetics diversity and rebuild and expand wild Giant Panda population. It is of significant to return rescued wild Giant Panda to wild and monitor their behavior before reintroduction artificial reproduced Giant Panda. It will increase our knowledge on reintroduction of Giant Panda. Aug 8th, 2005, “Shenglin 1”, a rescued wild Giant Panda was returned to Longxi-Hongkou National Nature Reservoir, which is habitat of Giant Panda Population B of Mt. Mingshan. A systematic monitor was carried out on “Shenglin 1”, and the successful return enriched our biological knowledge on Giant Panda reintroduction. It will be very help for future conservation work on reintroduce artificial reproduced Giant Panda. “Shenglin 1” was tracked with GPS collar, DNA in feces and infrared-trigged camera from Aug 2005 to Jun 2007. 1. Locomotion behavior and microsatellites comparison with Giant Panda from the 5 mountain systems indicated that “Shenglin 1” is possibly from Mt. Qionglai. 2. Habitat usage of “Shenglin 1” was studied. It was suggested that there were 3 phases after return, i.e. emergency response, preliminary stable phase and long distance locomotion, which could be a general process for other returned Giant Panda. It was indicated that there was some interaction between “Shenglin 1” and local population. “Shenglin 1” is seeking for suitable home range now. 3. Monitor data also indicated that microhabitat preference of Giant Panda and takin (Budorcas taxicolor) are different because of different diet, though they are sympatric. It was suggested that conservation management for the two species should be plan in particular. 4. The reintroduction of “Shenglin 1” is a successful case. The program of return rescued Giant Panda to other habitats is of value and should be continued. However, more improvement is needed for the monitor technique. More improvement is need for feralization and returning before we return artificial reproduced Giant Panda to wild.

青藏高原栽培青稞遗传多样性研究

青稞，是我国藏区居民对裸大麦的称谓，它不仅是藏民的主要食粮、燃料和牲畜饲料，而且也是啤酒、医药和保健品生产的原料；青稞不仅为藏区人民的健康和经济发展做出了很大的贡献，而且对人类健康和社会经济的可持续发展都有重要的意义。青藏高原是我国及世界上青稞分布和种植面积最大的地区，资源极其丰富。虽然从经典遗传直到分子标记对我国大麦遗传多样性都有研究，但研究手段、数量仍然不够深入，对我国大麦资源遗传多样性研究的信息非常有限，不能很好地满足大麦遗传研究和育种应用的需要，尤其是对西藏栽培大麦的遗传多样性的研究还只是刚刚开始，关于栽培青稞多态性的研究报道很少。本研究采用SSR标记和蛋白质电泳两类技术，从SSR标记位点、单体醇溶蛋白、B组醇溶蛋白和淀粉粒结合蛋白（SGP）等四个方面对我国青藏高原栽培青稞的遗传多样性进行了综合评价。 SSR标记具有基因组分布广泛、数量丰富、多态性高、容易检测、共显性、结果稳定可靠、实验重现性好、操作简单、经济、易于高通量分析等许多优点，被认为是用于遗传多样性、品种鉴定、物种的系统发育、亲缘关系及起源等研究的非常有效的分子标记。本研究采用SSR标记分析了64份青藏高原栽培青稞的遗传多样性，同时评估SSR标记在我国大麦育种和品种鉴定中的应用潜力。选择了30个已知作图位点SSR标记，其中25个标记与重要性状的控制位点连锁紧密。选择的30个SSR标记，5个未得到很好的扩增产物，3个无多态性。22个多态性SSR标记位点中，每位点检测出等位基因2~15个，共检测出等位基因132个，平均每位点6.0 个。各多态位点检测出基因型为2~11种，位点HVM33的基因型最多。各多态位点的多态信息指数为0.16~0.91, 平均为0.65。根据PIC值选择了13个SSR标记用于我国青藏高原栽培青稞基因型鉴定，这些标记的PIC值为0.6以上。结合PIC值和基因型差异，选择了8个多态信息含量高的SSR标记，构建了高效指纹图谱，此图谱能把64份材料完全区分。 贮藏蛋白电泳分析是研究相关编码蛋白基因多态性的非常有效的方法。大麦单体蛋白与小麦醇溶蛋白相对应，具有丰富的多态性，可用于大麦遗传多样性、品种鉴定和群体进化等研究。本研究通过A-PAGE电泳技术研究了84份青藏高原栽培青稞的单体醇溶蛋白多态性。大麦单体醇溶蛋白图谱与小麦醇溶蛋白电泳图谱类似，所分离的蛋白清晰地分为ω－，γ－，β－和α－四个部分。青藏高原栽培青稞单体醇溶蛋白具有丰富的多态性，84份青稞材料中存在43条不同的蛋白带，75种组合带谱；其中67种为单一材料所独有，另8种则分别包含了2-3份材料。每份材料中拥有醇溶蛋白带为6－16条，含有6-10条单体醇溶蛋白带材料较多。西藏和四川材料群体单体醇溶蛋白多态性不同，具有区域特异性。西藏材料中发现了40条不同蛋白带，3条特异带，46 种蛋白组合；四川材料中出现了40种不同蛋白带，26种条带组合, 3条特异带。基于单体蛋白多态性的聚类与材料的来源有一定的相关性。A-PAGE单体蛋白具有丰富的多态性，可作为遗传研究和品种鉴定的标记。 大麦醇溶蛋白（hordein）是大麦籽粒的主要贮藏蛋白，与大麦的营养品质和加工品质密切相关，而且具有丰富的多态性，广泛用于品种鉴定、种质筛选、遗传多样性和亲缘关系研究。B组醇溶蛋白是主要的醇溶蛋白组份，约占总醇溶蛋白的80%，而且具有丰富的多态性。本研究采用SDS-PAGE分析了72份青藏高原栽培青稞B组醇溶蛋白的遗传多样性。青藏高原栽培青稞B组醇溶蛋白具有丰富的多态性，72份青稞材料中存在15种蛋白带，30种组合带谱，其中15种为单一材料所独有，另15种则分别包含了2-10份材料。每份材料中B组醇溶蛋白条带数为4-8条，含5、6条的材料较常见。不同来源的群体材料间B组醇溶蛋白组成存在差异，西藏青稞含有26种蛋白组合带谱，其中有19种特异带谱；四川群体中共发现11种蛋白组合带型，其中有4种特有带谱。两群体中都存在稀有条带。聚类分析将材料分成三组，材料聚类与材料来源地没有明显的相关性。 淀粉粒蛋白（Starch granule proteins, SGPs）是一类与淀粉粒结合的微量蛋白，一些淀粉粒蛋白具有淀粉生化合成中主要的酶蛋白功能，其变异会影响淀粉含量和特性，从而影响淀粉的应用。关于我国大麦淀粉粒组成研究还未见报道。本实验首次开创了我国大麦淀粉粒结合蛋白的研究工作。采用SDS-PAGE电泳技术研究了青藏高原栽培青稞的SGP组成，并分析了不同SGP组合间淀粉含量的差异，初步探索了所分离的SGP蛋白与淀粉合成的关系。66份青稞材料中分离了10种主要的SGP，其表观分子量为40-100KD，低于60KD的SGP带有7条，共有16种组合带谱；各SGP蛋白和组合带谱出现的频率存在差异，青藏高原青稞的SGP组成存在多态性。西藏青稞和四川青稞的SGP组成有很大差异，SGP组成具有地域差异性，西藏青稞含有12种蛋白组合带谱，其中有9种特异带谱；四川群体中共发现7种蛋白组合带型，其中有4种特有带谱；两群体中仅有3种共同的蛋白组合带谱。SGP蛋白特性将66份青稞分为三组, 即Ⅰ、Ⅱ、Ⅲ，材料聚类与材料来源具有一定的相关性。不同组合带谱材料间淀粉含量差异显著性检验结果显示，不同带谱间材料的总淀粉含量、直链淀粉含量和支链淀粉含量有差异，带谱2（SGP1+3+7+9+10）和8（SGP1+2+4+6+8）的总淀粉含量及支链淀粉含量显著大于组合带谱3（SGP1+3+7+10）的总淀粉含量。组合带谱7（SGP1+2+6+8）的直链淀粉含量显著低于带谱11（SGP1+5+8）的直链淀粉。带谱SGP2、3、4、5、6、7、8、9、10可能参与淀粉合成，SGP9可能与高支链淀粉的合成相关。 SSR标记位点、单体醇溶蛋白、B组醇溶蛋白、淀粉结合蛋白等四个方面的研究结果表明青藏高原SSR标记多态性、单体醇溶蛋白多态性、B组醇溶蛋白多态性和SGP多态性都非常丰富，与青藏高原是栽培青稞的多样性分布中心的观点一致。 青藏高原栽培青稞的SSR标记、单体醇溶蛋白、B组醇溶蛋白和SGP多态性表现出很大差异。SSR标记覆盖了整个基因组，多态性非常高。单体蛋白、B组醇溶蛋白、SGP蛋白是育种中非常关注的性状，他们只是代表基因组中的某一区域或位点，多态性相对较低。但单体蛋白多态性很高，84份材料中检测出43条不同蛋白带，75种不同的组合带谱。SSR标记技术和单体蛋白技术都是遗传多样性研究的有力工具，但单体蛋白技术不仅多态性高，而且经济、操作简便，是种质鉴定的理想方法。 对不同标记的多态性材料数据进行聚类，聚类图能为我们提供各材料间的遗传相似信息，为材料选择提供参考。但材料聚类与材料来源的地理区域的相关性表现不一致。SSR聚类和B组醇溶蛋白聚类与材料的来源地无相关性，而单体醇溶蛋白和SGP聚类与材料来源地有一定相关性，即西藏群体和四川群体分别有集中类群，这可能是人为选择的附加效应。 不同来源的群体材料的遗传多样性不同，具有区域特异稀有基因，加强不同地区间资源的交换和配合使用，有利于增加群体遗传多样性和新品种培育。 青藏高原栽培青稞的麦芽浸提性状、淀粉性状、病虫及裸粒等重要农艺性状控制位点存在丰富的变异，遗传基础宽广，可能蕴藏着多种不同的等位基因，是研究重要性状遗传特性、基因资源挖掘和遗传育种的宝贵资源库。 Hulless barley, due to its favorable attributes such as high feed value, good human nutrition，rich dietary fiber and ease processing, attracts people,s attention . Hulless barley plays a very important role in Tibetan life, used as essential food crop, main animal feed and important fuel. In addition to tsampa (roasted barley flour), a main food for Tibetan, hulless barley is also made into cake, soup, porridge, recent naked barley liquor and cornmeal. Qinghai-Tibet Plateau is one of a few areas which plant naked barley widely in the world and also has a long growing history. Genetic diversity of the cultivated hulless barley in this region , however, has not been documented. The study of genetic diversity existing within this population is of particular interest in germplasm identification, preservation, and new cultivar development. This study analyzed the genetic diversity of the cultivated naked barley from Qinghai-Tibet plateau through the study of SSR marker loci and monomeric prolamins, B-horden and starch granule proteins. SSRs are present abundantly in genomes of higher organisms and have become a popular marker system in plant studies. SSRs offer a number of advantages, such as the high level of polymorphisms, locus specificity, co-dominance, reproducibility, ease of use through PCRand random distribution throughout the genome. In barley, several hundred SSRs have been developed and genetically mapped and can therefore be selected from specific genomic regions. The genetic diversity of 64 cultivated naked barley from Tibet and Sichuan was studied with 30 SSRs of known map location.Among the selected SSR markers, PCR products of 5 SSR markers were not obtained and 3 SSR marker loci were monomeric. A total of 132 alleles were identified at 22 polyomeric SSR loci. The number of alleles per locus ranged from 2 to 15, with an average of 6.0. The polymorphism information content values for the SSRs ranged from 0.08 to 0.94, with an average of 0.65. 13 SSR markers with the PIC value >0.6 have been selected for discrimination of Qinghai-Tibet naked barley genotypews. A finger Print map was developed through 7 SSR markers with the high PIC value. It could be used as an efficient tool for gene discovery and identification of gernplasm. Hordeins, the main storage proteins of the barley seed, are composed of momomeric and polymeric prolamins and divided into -A, B, C and D groups in order of decreasing electrophoretic mobility. Hordeins show high inter-genotypic variation and have been extensively used as markers for cultivar identification and analyzing the genetic diversity. This study analyzed the genetic diversity of B-hordein in 72 naked barley from Qinqhai-Tibet Plateau. Extensive diversity was observed. A total of 15 different bands and 30 distinct patterns were found. Jaccard's coefficient of similarity was calculated, and the accessions were divided into three　main groups by cluster analysis using UPGMA. Differentiation among the populations from different collecting regions based on the polymorphism of B-hordein was investigated. Monomeric prolamins show high inter-genotypic variation and have been used as molecular markers for cultivar identification, analyzing the genetic diversity in collections and investigating the evolution processes and structure of populations However, the cultivated hulless accessions from Qinghai-Tibet Pateau in China have never been examined with respect to monomeric prolamins. This study analyzed the genetic diversity of monomeric prolamins (protein fraction corresponding to wheat gliadins) using the Acid -PAGE technique in eighty-four cultivated hulless barley from Qinqhai-Tibet Plateau in China. Extensive diversity was observed. A total of 43 different bands were found, of which 21 different bands were in the region of ω group, 8 in the region of γ, 8 in the region of β, and 6 in the region of α group. Among the 86 accessions, 75 distinct patterns were identified. The number of bands ranged from 6 to 16, depending on the variety. Jaccard’s coefficient of similarity was calculated, and the lines were grouped by cluster analysis using UPGMA. A dendrogram was obtained from the analysis of the groups and five main clusters were identified. No relationship between the distribution in the dendrogram and growth habits and origins of the cultivars could be detected. Starch is the major constituent of the cereal endosperm, comprising approximately 65% of the dry weight of the mature wheat grain. The starch formed in all organs of plants is packaged into starch granules, which vary widely between species and cultivars in size and shape. Wheat endosperm starch granules contain about corresponding to the main biosynthase of starch. This report firstly dealed with intraspecific variation of the major SGPs in cultivated naked barley from Qinghai-Tibet plateau. A total of 10 major SGPs were observed in the range of 40KD-100KD and 16 types of patterns were found. Based on the variation of SGPs, accessions studied were classified into 3 groups. A geographical cline of electrophoregram was observed. In addition, significance test of the difference of starch content among groups and types of patterns were done, and the results indicated those SGPs could be related to the content of starch. Diagram obtained through cluster analysis exhibited a structuration of diversity and genetic relationship among cultivated hulless accessions. In breeding program, parents with genetically distant relationship for hybridization will increase genetic diversity of progenies. In conclusion, cultivated naked barley from Qinghai-Tibet Plateau in China presents a high variability with respect to monomeric prolamins，SSR markers , B- hordeins and SGPs. The result of this study supports Qinghai-Tibet Plateau is the center of cultivated hulless barley and the cultivated naked barley is considered to be a gene pool with large diversity and could be applied to breeding for cereal.

影响沼气甲基产孢弧菌生长及其甲烷单加氧酶活性的若干因素研究

本文叙述了影响甲烷氧化细菌沼气甲基产孢弧菌81Z菌株生长和甲烷单加氧酶(MMO)活性的若干因素。沼气甲基产孢弧菌81Z菌株细胞生长被高浓度PO43-(>8mM),NH4+([NH4cl]>500mg/l)抑制；[CuSO4·5H2O]在0～4mg/l范围内。生长随[Cu2+]升高而加强，低[Cu2+](0.1mg/l CuSO4·5H2O)培养基中，添加Cocl2·6H2O(0.238mg/l)；促进菌体细胞生长。发酵罐分批培养过程中，生长延迟期过后，沼气甲基产孢弧菌81Z菌株细胞MMO比活很快达到最高，并稳定至对数生长中后期，随即急剧下降至初始水平。发现沼气甲基产孢弧菌81Z细胞中存在一种MMO活性，它不同于已报道过的两种MMO,MMOL最适PH6.2～6.4，4℃相对稳定，其产生不受培养基中[Cu2+]调控能与甲醇-甲醇脱氢酶系统相偶联，在无细胞抽提液中其活性被400μM[Cu2+]抑制。在低[Cu2+]发酵罐培养条件下，沼气甲基产孢弧菌81Z菌株产生可溶性MMC，其最适PH7.0，4℃不稳定，可被DE-52分离为三组分：A、B、C。为了获得沼气甲基产孢弧菌81Z细胞MMO的最佳催化活性，①采用高[Cu2+]培养基进行发酵罐培养，收集对数生长中期的细胞；②选择反应缓冲液PH6.3；③反应体系中添加5mM甲醇或甲酸是有效的方法。在本研究所采取过的最佳条件下，测得MMO活性为15.9nmol/min·mg干细胞，是以前报道的该菌株活性0.97nmol/min·mg干细胞的十六倍。Some factors which influence growth and MMO activity of Methylosporovibrio methanica 81Z were described. The growth of Methylosporovibrio methanica 81Z is inhibited by high concentration of PO43-(8mM)or NH4+(500mg/lNH4cl). The growth of Methylosporovibrio methanica 81Z increased with rising of copper concentration up to 4mg/l CuSO4·5H2O. At low copper concentration(0.1mg/lCuSO4·5H2O),adding Cocl2·6H2O(0.238mg/l)could enhance the growth of Methylosporovibrio methanica 81Z.With batch culture of Methylosporovibrio methanica 81Z in a fermentor, after lag phase, the activity of MMO reached the highest level rapidly and steady until later log phase, then falled to initial level.MMOL activity differenct from that of two types of MMO reported before was found from Methylosporovibrio methanica 81Z with optimum PH value from 6.2 to 6.4 and relative stabilty at 4℃. Synthsis of the MMOL was not regulated by copper concentaration in medium. Its activity could couple with methane-l-methanoldehydrogenase system, and in cell-free extract, were inhibited by 400μm copper ion. At low copper concentration(0.1mg/lCuSO4·5H2O) and in a fermentor, Methylosporovibrio methanica 81Z could syntheis soluble MMO similar to solble MMO reported before by Palton and Patel. Its optimum PH value was 7.0. It was unstable at 4℃. It could be resoluted into three components: A, B, and C. It was effentive for obtaining the maxtmum MMO with Methylosporovibrio methanica 81Z that (1) to keep high copper concentration(4mg/lCuSO4·5H2O) in a fermentor and harvest cell at middlel lag phase;(2) to choose 6.3 as the PH value of reaction buffer;(3)and to add 5mM methanol or formate into reaction system. In this dy, the MMO activity of cells of Methylosporovibrio methanica 81Z was reached 15.9 nmol/min.mg, dry weight, sixteen times as high as the value(0.97nmol/min.mg, dry weight) reported with the same strain.

~(19)F+~(27)Al耗散反应产物激发函数的长程关联

在中国原子能科学研究院的HI-13串列静电加速器上完成了19F+27Al耗散反应产物的激发函数测量。束流19F的入射能量从110.25MeV到118.75MeV,步长250keV,测量了实验室系+57°,+31°,-15°和-29°出射的类弹产物B,C,N,O,F和Ne的激发函数。观测到激发函数具有耗散反应的特征涨落结构,分析了激发函数之间的长程角度关联以及强烈的电荷数关联。

~(19)F+~(27)Al耗散反应中双核系统的转动与形变

测量了19F+27Al耗散反应产物B,C,N,O,F和Ne的激发函数,入射束流的能量从110．25MeV 到118．75MeV,能量步长为250keV．从产物的能量自关联函数中提取了反应中所形成的中间双核系统的转动惯量,与相粘模型计算的刚体转动惯量相比较,结果表明形成的双核系统有大的形变．

~(19)F+~(27)Al耗散反应产物截面测量中的不重复性

在中国原子能科学研究院的HI-13串列静电加速器上完成了114MeV和118．75MeV的19F+27Al 耗散反应产物B,C,N,O,F,Ne,Na和Mg的微分截面测量．条形27Al同位素靶的大小尺寸为 10mm×50mm,平均厚度约67μg／cm2．固定入射束流19F的能量,通过移动靶位,先后轰击了靶上20个不同的位置．实验结果表明,在弹靶系统、入射能量以及探测系统都相同的多次独立的微分截面测量中,截面的涨落不呈现高斯型几率分布．截面测量的这种不重复现象难以用有限计数率的统计性质来解释．

(19)~F+(27)~Al耗散反应中双核系统的转动与衰变

测量了19F+27Al耗散反应产物B,C,N,O,F和Ne的激发函数,入射束流的能量从110.25MeV 到118.75MeV,能量步长为250keV.用能量自关联函数方法从激发函数中提取了各反应产物的平均衰变宽度,利用同时考虑反应中所形成的中间双核系统的转动特性和衰变特性而发展了的Ericson核反应统计理论,讨论了19F+27Al耗散反应中双核系统随时间的演化过程.

~(19)F+~(27)Al反应产物的角分布、角分散与耗散机制

完成了114MeV19F+27Al深部非弹性碰撞产物的角分布测量.分析了反应产物B,C,N,O,F,Ne,Na,Mg和Al的实验室系角分布,讨论了反应形成的中间双核系统随时间演化过程中角分布所表现出的耗散特点.从实验上提取了反应产物各元素的角分散参数并作了角分散参数的理论拟合,讨论了角分散参数的物理意义以及角分散参数与耗散反应产物的电荷数Z之间的依赖关系.

~(19)F+~(27)Al深部非弹性碰撞产物的角分布

完成了19F+27Al深部非弹性碰撞产物的角分布测量.初步分析了反应产物B,C,N,O,F,Ne,Na,Mg和Al的实验室系角分布,展现出深部非弹性反应机制的特点,显示了反应系统随时间的演化过程.

Rotation and deformation of the dinuclear system formed in dissipative reaction of F-19+Al-27

Excitation functions of the reaction products B, C, N, O, F and Ne emitted from the dissipative reaction of (19) F+(27) Al have been measured at incident energies from 110.25MeV to 118.75MeV in steps of 250keV. The moments of inertia of the intermediate dinuclear system formed in the reaction are extracted from the energy autocorrelation functions of the products. Comparing the moment of inertia extracted from the experimental data with the calculated one by using the sticking limit, it indicates that the formed dinuclear system has a large deformation in the reaction process.

同中子素核9C、8B、7Be和6Li在中能区与28Si反应总截面的测量

放射性束流(RIB)装置拓广了实验核物理在同位旋(T_z)自由度上从稳定核直到滴线核的广袤空间。通常，位于β-稳定线及其附近的核，N／Z在1－1.5范围变化，其分离能E_s无论对于质子还是中子，总是在6-8 MeV之间；对于远离稳定线的非稳定核，N／Z可在0.5-4范围变化，如~9C的N／Z = 0.5，~(10)He的N／Z = 4，而且分离能E_s是在0-40 MeV之间变化的，开展对这些远离β-稳定线非稳定核性质、结构的研究是目前核物理的前沿之一。核反应总截面σ_R是表征原子核性质特征的一个基本物理量，从实验测得的核反应总截面中可以得到有关核结构和核内核子分布的信息。在由放射性束流所产生奇异核的结构与各种反应机制研究中，反应总截面的测量更是有其特殊的重要性，具有奇异核结构如晕核的一个典型的物理特征就是其反应总截面要比稳定核大得多，Tanihata等人最早就是通过对放射性束流的相互作用截面的测量发现了具有奇异结构的核，即中子晕核。由于反应总截面的测量对探测器的要求不高，而且数据分析过程相对较为简单，因而反应总截面的测量已经成为放射性束核物理的研究的一个非常重要实验手段。中子晕核以及中子皮核的发现促使人们去寻找质子晕核和质子皮核，由于最后一个质子的结合能非常小只有136，keV，并且有较大的电四极矩，因而使得~8B成为质子晕的最大热门候选核，有关~8B是否具有质子晕核结构的问题，许多实验科学家得出了相互矛盾的回答；而目前有关另外一个质子晕候选核~9C的实验数据非常少，目前还没有人从实验上对~9C是否为质子晕核这一问题进行肯定或否定的回答。因此非常有必要测量~9C和~8B的反应总截面。对反应总截面进行研究的一个非常有用理论就是Glauber模型，该模型考虑了库伦效应的多次散射理论。它是一种基于自由核子-核子(N-N)碰撞的与核物质密度相关的理论，因而能够从实验测量到的反应总截面中提取核物质分布的信息。该理论对中低能区的反应总截面描述却有一个缺憾：理论值比实验值都要小。本论文主要描述了利用透射法测量了中能区同中子素核~9C、~8B、~7Be及~6Li与~(28)Si的反应总截面，并介绍了重离子碰撞以及描述重离子性质的几种常用理论。在论文里对实验测量得到的结果进行了理论分析，这些理论包括半经验的Shen公式、Glauber模型、BUU模型以及SHF理论。如果将~9C和~8B当成具有正常核结构来处理，半经验的Shen公式和Glauber模型(HO密度分布)的理论计算值总是比实验值要小得多；对于Glauber模型的理论计算值和实验值的差异，Ozawa等定义了一个差值因子d，方德清等人对轻核系统的d值进行了详细的分析。一般认为，正常核的d值在20％以内，而对于具有晕或皮奇异结构的核，其d值则超过30％，甚至可达50％，因此可根据一个核的d值是否超过30％而且比相邻核的d值明显大这种半经验的方法来判断一个核是否具有奇异结构；利用d值的分析结果，我们认为：~9C和~8B都具有奇异核结构；对于BUU模型用同样的方法引进差值因子d值，对于~9C和~8B有相同的结论。用SHF理论计算得到B和C同位素的密度分布结果显示，~9C和~8B的密度分布比相邻的同位素扩展都要大得多。为减小Glauber模型计算的反应截面与实验值的差别，本论文还对Glauber模型的输入密度形式进行了修改，在原单一HO分布基础上加一个高斯分布的尾巴，并对丰质子的同中子素核~9C、~8B、~7Be及~6Li与~(28)Si靶以及~(12)C和丰中子的C同位素核~(13-16)C与~(12)C靶的反应截面重新进行了计算，结果显示在中能区的计算值比原来单一密度分布的计算结构有明显改善。

黄河源区退化高寒草地土壤种子库:种子萌发的数量和动态

对青藏高原黄河源区不同退化程度高寒草地的土壤种子库土样用土壤分析筛进行浓缩,并以萌发法分析土壤种子库萌发种子数量和动态.结果表明,孔径0.25~2 mm的土壤分析筛分离土样中萌发种子可达萌发种子总量的85%~97%,而小于0.25 mm的土样中未发现种子.因此,用0.25 mm孔径大小的土壤筛对高寒草地土壤种子库土样进行大规模浓缩是一种方便、可靠的方法.4种不同退化程度高寒草地(A:未退化草甸;B:轻度退化草甸;C:中度退化草甸;D:重度退化草甸)的土壤种子库在实验室条件下萌发的种子数量分别为:A 1 194~3 744粒/m2,平均2 421.3粒/m2;B 5 376~1 0912粒/m2,平均7 786.7粒/m2;C 2 304~1 3216粒/m2,平均8 695.5粒/m2;D 4 768~12 352粒/m2,平均8 125.9粒/m2.除样地A外,其它3个样地的可萌发种子数量差异不大.单子叶植物种子在培养到d 10左右开始萌发,双子叶植物在5~7 d内开始萌发,前者3 wk后基本不再萌发,后者5 wk左右停止萌发.4个样地土壤种子库种子萌发主要集中在第2~3周,并表现出近似单峰型格局.图1表3参39