固氮放线菌Frankia与放线菌根植物共生进化的研究进展

固氮放线菌Frankia是能够诱导大范围的放线菌根植物(actinorhizal plants)产生根瘤放线菌.放线菌根植物是植物受Frankia侵染后能够形成根瘤的植物[1].目前,它们之间的进化研究虽然取得了很大进展,但是仍然存在许多不清楚的东西.本文根据最近对植物和Frankia系统进化研究成果、根瘤中放线菌Frankia的多样性研究情况,综述放线菌根植物和共生Frankia进化的研究进展.     

不同宿主植物根瘤Frankia及其生物学特性

对从木麻黄、杨梅、桤木和胡颓子等宿主植物根瘤中获得的19株分离菌的离体培养形态特征、生理特性和交叉侵染特性等进行了比较分析.结果表明,各分离菌均有Frankia属所特有的分枝状菌丝、孢囊、泡囊或串珠状菌丝等形态结构;细胞壁类型多属胞壁Ⅲ型,生理类型多属B型,无氮诱导培养下都具有同氮酶活性,在BAP、JA或S培养液中菌体生长较好,以吐温-80和酪蛋白水解物为最佳碳、氮源.但不同宿主分离菌的形态和培养特征差异明显,木麻黄属分离菌菌丝较粗,孢囊数量较少,在BAP培养液中多旱荔肉白絮状颗粒沉淀;杨梅属菌丝较细,孢囊数量较多,在BAP培养液中多是浅红色颗粒沉淀;桤木属和胡颓子属的菌体形态特征在BAP培养液中与木麻黄属的相似,但桤木属菌丝较细,胡颓子属的菌丝较粗.根据回接及交叉侵染特性可将分离菌分为2个宿主特异类群:能侵染木麻黄苗木的木麻黄类群;只侵染原宿主并能在杨梅属、胡颓子属和桤木属间相互侵染,但不能使木麻黄属苗木结瘤的杨梅-桤木-胡颓子类群.图2表5参19     

云南腾冲热泉极端嗜酸热硫化叶菌多样性研究

硫化叶菌(Sulfolobus)是极端嗜热古菌遗传机制研究的重要模式菌株,广泛分布于酸性热泉中.对分离自云南腾冲酸性热泉的11株极端嗜酸热菌株进行16S rRNA基因序列测定,并通过菌体形态观察、16S rRNA基因可变区序列比较、菌株16S rRNA基因相似性比较及系统发育分析,研究腾冲热泉硫化叶菌的多样性.结果表明,分离自腾冲热泉的11株硫化叶菌的16S rRNA基因序列与分离自世界上其它地区的8个硫化叶菌的标准菌株的相似性为85.8%～94.9%,与同样分离自腾冲热泉的S.tengchongensis标准菌株的相似性为96.6%～97.5%,在分类学上具有形成新种的可能.在16S rRNA基因高可变区序列上,与分离自世界上其它地区的8个硫化叶菌的标准菌株存在明显的差异.系统发育分析表明,分离自腾冲热泉的硫化叶菌在系统发育树上形成两个亲缘关系较为紧密的簇群,可见腾冲热泉硫化叶菌不仅具有一定的多样性,同时具有较明显的地域性特征.     

5种植物根际真菌群落结构与多样性

根际真菌在植物生长和健康方面具有非常重要的作用,然而由于分离和培养技术的限制,对根际真菌群落、多样性及功能仍缺乏认识.为明确不同植物根际真菌群落的结构与差异,揭示其潜在的生态功能,采用Illumina MiSeq技术对马蔺、葡萄、大豆、玉米等5种植物根际真菌ITS rRNA基因的ITS2区进行高通量测序,并利用FUNGuild对其功能进行预测分析.结果显示,5种植物根际中80%以上的真菌序列划分到17-45个OTUs中,优势菌门为子囊菌门,优势菌纲为座囊菌纲和粪壳菌纲等;这5种植物根际真菌群落各自聚群,其香农指数(Shannon)和丰富度(Chao1)从大到小依次为木瓜、马蔺、葡萄、大豆、玉米;在获得的1 290个OTUs中,49.76%的OTUs归属于8个功能类群,其中腐生菌约占总OTUs数的25.89%,是主要功能类群,其次是病原菌/腐生/共生过渡型、植物病原体等,葡萄、木瓜和马蔺根际中腐生菌显著高于大豆和玉米.本研究表明木瓜、马蔺和常见植物根际真菌的群落结构和多样性不同,一年生作物的根际真菌多样性小于多年生植物,进而也表明根际真菌群落及其生态功能受到植物的生活周期或类型的影响.(图4表1参30)     

重金属镉对水田土壤微生物基因多样性的影响

采用16S rDNA DGGE分析技术,探讨了不同处理时间、不同浓度镉胁迫条件下水田土壤微生物种群的基因多样性.研究发现,随着处理时间的不同,各处理间的土壤微生物基因多样性差异存在明显不同,培养初期镉对土壤微生物种群毒害作用非常明显,土壤微生物基因多样性下降,但培养后期不同处理之间DNA基因型差异很小,丰度差异较大.在土壤培养初期、含镉3～10 mg/kg的培养基上出现一条新增条带,为对照及低镉浓度土壤所没有,镉形态稳定后会消失,说明土壤中可能存在适官镉浓度下良好生长的微生物种,经序列测定及与网上比对分析.与一株Uncultured bacterium sp.具有96%的相似率,初步确定为一不可培养细菌种,以其作为淹水条件下镉污染初期的报告基因有一定的可行性.     

黄河三角洲湿地土壤微生物群落结构分析

采用末端限制性片段长度多态性分析(T-RFLP)技术和16S rDNA克隆文库的方法,分析了黄河三角洲滨海湿地土壤不同深度细菌和古菌的群落结构.研究表明,随着深度的增加,细菌群落的多样性下降,而古菌群落多样性则有上升的趋势,且土壤的细菌和古菌群落结构都呈现出规律的层状分布.该土壤包括各种硫酸盐还原菌、产甲烷古菌、光合细菌等丰富的细菌和古菌资源.图5参27     

广州市莲塘村风水林群落特征及植物多样性

风水林是极具南方地域特色的植被景观与物种储存库,是接近地带性植被特性的一种群落类型.研究风水林,对丰富和发展环境科学、生态科学、林业科学和园林科学,具有重要意义.本研究在广州市莲塘村村后风水林设立了样带、标准地和样方,对其乔木层、灌木层和草本层植物进行详细调查,结果表明,该区共有植物73种,隶属于33个科57个属.其植物种类丰富,以茜草科、樟科、大戟科、壳斗科、莎草科、桃金娘科植物占优势.该群落具有南亚热带常绿阔叶林群落的典型群落结构,垂直结构分层现象明显,层次清晰.文章对树种丰富度、Shannon-Winner指数、Simpson多样性指数、种间相遇机率以及均匀度等多样性指标的研究表明:该群落具有较大的物种丰富度,植物多样性较高,物种分布均匀,不同种之间相遇几率较大,其生物多样性各项指标远远大于尾叶桉人工纯林.该风水林群落是目前广州市保存较完好的具有典型地带特色的少数自然林之一,它的研究成果对以后生态公益林造林有极大的参考价值.     

云南稻种磷高效特性相关性状的生态差异

云南不仅是世界稻种(Oryza sativa L.)最大的遗传和生态多样性中心之一,也是中国土壤类型最丰富的地区;中国南方和云南耕地65%以上缺乏有效磷,土壤缺磷已成为稻作生产的主要限制因子.该文中,缺磷(速效磷6.26 mg·kg-1)与适磷(速效磷40 mg·kg-1)酸性红壤进行比较,以相对茎蘖数(RTA)、相对总质量(RTB)、相对地上质量(RDWU)和相对地下质量(RDWG)为参数,研究了来自云南省16个州市5个稻作区577份水稻核心种质的磷高效基因型及其生态差异,结果表明:低磷胁迫下云南稻核心种质的相对茎蘖数、相对总质量、相对地上质量和相对地下质量密切相关,这4个性状是水稻磷高效鉴定较可靠和方便的指标,并筛选出10个磷高效品种;磷高效基因型在籼粳间存在较大差异,但丁颖分类性状中的水陆、粘糯间差异较小;水稻磷高效品种广泛分布于5个稻作区和绝大多数州市,特别是磷高效极强品种主要来自云南稻种的遗传多样性中心(思茅、临沧、西双版纳、德宏和文山州)和基因多样性中心(南部边缘水陆兼作区).这些结果有力地支持了水稻磷高效特性与酸性红壤和遗传多样性中心有关,更好地理解水稻磷高效特性的地理生态和遗传多样性机制.     

基于rpoC1基因的微囊藻属分子系统学研究

测定了广东省9个水库4株铜绿微囊藻Microcystis aeruginosa、5株水华微囊藻M.flos-aquae和4株未定种微囊藻rpoC1基因部分序列,结合GenBank下载的日本5个水体11株铜绿微囊藻、5株绿色微囊藻M.viridis和5株惠氏微囊藻M.wesenbergii间源序列,进行序列分析.结果显示,34株微囊藻存在21种基因型,序列相似性达97.6%～100%.在邻接树上不同形态种和不同地理来源的藻株混杂在一起,没有形成明显的谱系结构和地理结构:同一形态种藻株可能具有不同的基因型,而相同基因型的藻株可能是不同的形态种;从同一水体中分离的微囊藻具有不同的形态种和不同的基因型,而不同水体分离的微囊藻有时又具有相同基因型,表明rpoC1基因序列无法区分形态种和地理株,支持Kondo和Otsuka提出的暂将铜绿微囊藻、水华微囊藻、惠氏微囊藻和绿色微囊藻等归为铜绿微囊藻复合种的分类处理.     

茶园年限对根际土壤真菌群落结构及多样性的影响

为探明不同年限茶园根际土壤真菌菌群结构及多样性的差别,采用MiSeq高通量测序方法分析5年、10年、25年和40年茶园根际土壤真菌的菌落组成、Alpha多样性、Beta多样性和组间差异性.结果显示,随着茶园年限增大,根际土壤pH值显著性降低;而土壤养分含量呈现先增后减趋势,其中25年茶园最高.不同年限茶园根际土壤真菌群落α多样性无显著差异,β多样性则5年与10年茶园相近,而25年和40年茶园相似. 5年和40年茶园土壤中的优势菌目存在明显差异,其中粪壳菌目占5年茶园总菌群的20.93%,糙孢孔目占40年茶园总菌群的14.97%.此外,不同年限茶园根际土壤中差异性显著的菌种主要分布在粪壳菌纲和银耳菌纲.本研究表明,不同年限茶园根际土壤真菌物种组成丰富、结构差异明显;随着茶园年限的增加,茶树根际土壤肥力增加和真菌群落数量增加,有利于茶树生长代谢的真菌种群增多.     

四川省部分豆科植物根瘤菌的遗传多样性

从四川省豆科植物鸡眼草属、合欢属、豇豆属、葛属、金合欢、杭子梢分离获得了103株根瘤菌,采用AFLP、BOXAIR-PCR研究了这些菌株的遗传多样性.结果表明,分离自四川省豆科植物6个属的根瘤菌存在明显的遗传多样性,AFLP分析中,全部供试菌株的相似性为21%,全部供试菌株可分为26个AFLP遗传群;BOXAIR-PCR聚类分析结果表明,103个菌株分在70%相似性水平处,供试菌株被分为18个BOX遗传群.除部分菌株外,多数根瘤菌按宿主类型聚群,同一宿主而地理来源不同的根瘤菌分布于不同的遗传群,表明宿主和地理环境对根瘤菌具有深刻影响.图3表1参9     

Potato Diversity in the Andean Center of Crop Domestication

The diversity and population structure of potato landraces ( Solanum spp.) within their center of domestication was studied using isozyme surveys of four polymorphic loci. The objective in assessing the distribution of genetic diversity was to assist in planning conservation strategies of crop genetic resources that are threatened by genetic erosion. In situ conservation methods depend on this type of analysts. Research was conducted in the region of Cusco, Peru. Eight fields spread among two microregions were randomly sampled, and 610 tubers were studied from this sample. In addition, 503 tubers were collected from markets in seven different meso-regions (provinces) surrounding the regional center of Cusco. Thirty genotypes were identified in the field sample and 82 in the regional sample. The frequency and distribution of genotypes and alleles are described. A high degree of genotype endemism was found at both the field and regional levels. Genotypes were unevenly distributed, and most of the genotypic diversity was between rather than within populations. At the allele level, however, we found that a very high percentage of the diversity was within rather than between populations. The genotype is the key unit for maintaining the population of potato landraces. Our findings suggest that collections need to be both geographically extensive and intensive. Because farmers are able to maintain most alleles on relatively small portions of their land, in situ conservation is a viable strategy.     

The consequences of genetic diversity in competitive communities

Several lines of evidence suggest that the species diversity and composition of communities should depend on genetic diversity within component species, but there has been very little effort to directly assess this possibility. Here I use models of competition among genotypes and species to demonstrate a strong positive effect of the number of genotypes per species on species diversity across a range of conditions. Genetic diversity allows species to respond to selection imposed by competition, resulting in both functional convergence and divergence among species depending on their initial niche positions. This ability to respond to selection promotes species coexistence and contributes to a reduction in variation in species composition among communities. These models suggest that whenever individual fitness depends on the degree of functional similarity between a focal individual and its competitors, genetic diversity should promote species coexistence; this prediction is consistent with the few relevant empirical data collected to date. The results point to the importance of considering the genetic origin and diversity of material used in ecological experiments and in restoration efforts, in addition to highlighting potentially important community consequences of the loss of genetic diversity in natural populations.     

Genetic Diversity and Population Size in Four Rare Southern Appalachian Plant Species

Allozyme diversity was examined in four rare, high-montane plant species from the Appalachian Mountains of southeastern North America. These species may represent relictual members or descendants of an alpine community that was more widespread during the late Pleistocene. We sampled five populations of Geum radiatum (Rosaceae), Carex misera (Cyperaceae), Trichophorum cespitosum (Cyperaceae), and the four known populations of Calamagrostis cainii (Poaceae). Genetic diversity was low for all species but was typical of that found for plant species with limited ranges. Low genetic diversity may reflect historical events associated with changes in the species' biogeography. As the Pleistocene climate warmed, suitable habitat decreased in areal extent and became fragmented, probably resulting in smaller, more-isolated populations. In recent times these species, which co-occur in fragile rock outcrop habitats, have been adversely affected by human activities. Genetic analyses revealed reduced diversity in populations of decreasing size for three species. Estimates of gene flow were low ( Nm < 1.0) in all four species. Positive associations between genetic diversity and population size, evidence of recent population declines, and the low estimates of gene flow suggest that genetic drift may play a prominent role in shaping the present-day genetic composition of these species. Furthermore, these data suggest that the genetically depauperate populations are unlikely to regain genetic variation without human intervention.     

The relative importance of host-plant genetic diversity in structuring the associated herbivore community

Recent studies suggest that intraspecific genetic diversity in one species may leave a substantial imprint on the surrounding community and ecosystem. Here, we test the hypothesis that genetic diversity within host-plant patches translates into consistent and ecologically important changes in the associated herbivore community. More specifically, we use potted, grafted oak saplings to construct 41 patches of four saplings each, with one, two, or four tree genotypes represented among the host plants. These patches were divided among two common gardens. Focusing first at the level of individual trees, we assess how tree-specific genotypic identity, patch-level genetic diversity, garden-level environmental variation, and their interactions affect the structure of the herbivore community. At the level of host-plant patches, we analyze whether the joint responses of herbivore species to environmental variation and genetic diversity result in differences in species diversity among tree quartets. Strikingly, both species-specific abundances and species diversity varied substantially among host-tree genotypes, among common gardens, and among specific locations within individual gardens. In contrast, the genetic diversity of the patch left a detectable imprint on local abundances of only two herbivore taxa. In both cases, the effect of genetic diversity was inconsistent among gardens and among host-plant genotypes. While the insect community differed significantly among individual host-plant genotypes, there were no interactive effects of the number of different genotypes within the patch. Overall, additive effects of intraspecific genetic diversity of the host plant explained a similar or lower proportion (7-10%) of variation in herbivore species diversity than did variation among common gardens. Combined with the few previous studies published to date, our study suggests that the impact of host-plant genetic diversity on the herbivore community can range from none to nonadditive, is generally low, and reaches its most pronounced impact at small spatial scales. Overall, our findings strengthen the emerging view that the impacts of genetic diversity are system, scale, and context dependent. As the next step in community genetics, we should then start asking not only whether genetic diversity matters, but under what circumstances its imprint is accentuated.     

Genotypic richness and phenotypic dissimilarity enhance population performance

Increases in biodiversity can result from an increase in species richness, as well as from a higher genetic diversity within species. Intraspecific genetic diversity, measured as the number of genotypes, can enhance plant primary productivity and have cascading effects at higher trophic levels, such as an increase in herbivore and predator richness. The positive effects of genotypic mixtures are not only determined by additive effects, but also by interactions among genotypes, such as facilitation or inhibition. However, so far there has been no effort to predict the extent of such effects. In this study, we address the question of whether the magnitude of the effect of genotype number on population performance can be explained by the extent of dissimilarity in key traits among genotypes in a mixture. We examine the relative contribution of genotype number and phenotypic dissimilarity among genotypes to population performance of the soil arthropod, Orchesella cincta. Nearly homogeneous genotypes were created from inbred isofemale lines. Phenotypic dissimilarity among genotypes was assessed in terms of three life-history traits that are associated with population growth rate, i.e., egg size, egg development time, and juvenile growth rate. A microcosm experiment with genotype mixtures consisting of one, two, four, and eight genotypes, showed that genotypic richness strongly increased population size and biomass production and was associated with greater net diversity effects. Most importantly, there was a positive log-linear relationship between phenotypic dissimilarity in a mixture and the net diversity effects for juvenile population size and total biomass. In other words, the degree of phenotypic dissimilarity among genotypes determined the magnitude of the genotypic richness effect, although this relationship leveled off at higher values of phenotypic dissimilarity. Although the exact mechanisms responsible for these effects are currently unknown, similar advantages of trait dissimilarity have been found among species. Hence, to better understand population performance, genotype number and phenotypic dissimilarity should be considered collectively.     

The Genetic Diversity of Native, Stocked, and Hybrid Populations of Brook Trout in the Southern Appalachians

Using mitochondrial DNA (mtDNA) restriction analysis and starch-gel electrophoresis of the CK-A2 locus product, we examined genetic variation in 311 brook trout (Salvelinus fontinalis) from 11 native, 5 hatchery-derived, and 8 hybrid populations in the southern Appalachian Mountains. Native southern Appalachian fish were genetically distinct from hatchery-derived fish. Southern and hatchery-derived fish were reliably distinguished based on three mtDNA restriction sites. Native southern haplotypes differed from hatchery-derived haplotypes by an average of 0.84%. Northern hatchery-derived haplotypes varied little in mtDNA compared to native southern haplotypes. Introgression of mtDNA haplotypes and the CK-A2 locus varied among populations, and introgression of allozyme and mtDNA markers was positively correlated. Continued introductions of nonnative strains of brook trout in the southern Appalachians could simplify the genetic structure of native brook trout populations and eliminate unique genotypes.     

植物根瘤内痕量Frankia DNA的提取及鉴定

建立了一种简捷、快速植物根瘤内痕量Frankia DNA的提取方法，即采集自然态下根瘤，剥离瘤瓣，取瘤瓣尖经液氮研主民粉末，以20g/L CTAB（十六烷基三甲基溴化铵）消化胞壁，然后以d=5-10μm的石英粉吸附DNA，再以无菌去离子蒸馏水释放DNA，所获DNA产率及纯度较酚／氯仿法好。经对辽宁沙棘、色赤杨根瘤的瘤内痕量Frankia DNA及体外培养的9种Frankia菌株DNA分纯，并用于酶切及PCR，获满意结果。图3参12