Phylogeography of the Qinghai-Tibetan Plateau endemic Juniperus przewalskii (Cupressaceae) inferred from chloroplast DNA sequence variation

The vegetation of the northeast Qinghai-Tibetan Plateau is dominated by alpine meadow and desert-steppe with sparse forests scattered within it. To obtain a better understanding of the phylogeography of one constituent species of the forests in this region, we examined chloroplast trnT-trnF and trnS-trnG sequence variation within Juniperus przewalskii, a key endemic tree species. Sequence data were obtained from 392 trees in 20 populations covering the entire distribution range of the species. Six cpDNA haplotypes were identified. Significant population subdivision was detected (G(ST) = 0.772, N-ST = 0.834), suggesting low levels of recurrent gene flow among populations and significant phylogeographic structure (N-ST > G(ST), P < 0.05). Eight of the nine disjunct populations surveyed on the high-elevation northeast plateau were fixed for a single haplotype (A), while the remaining, more westerly population, contained the same haplotype at high frequency together with two low frequency haplotypes (C and F). In contrast, most populations that occurred at lower altitudes at the plateau edge were fixed or nearly fixed for one of two haplotypes, A or E. However, two plateau edge populations had haplotype compositions different from the rest. In one, four haplotypes (A, B, D and E) were present at approximately equivalent frequencies, which might reflect a larger refugium in the area of this population during the last glacial period. Phylogenetic analysis indicated that the most widely distributed haplotype A is not ancestral to other haplotypes. The contrasting phylogeographic structures of the haplotype-rich plateau edge area and the almost haplotype-uniform plateau platform region indicate that the plateau platform was recolonized by J. przewalskii during the most recent postglacial period. This is supported by the findings of a nested clade analysis, which inferred that postglacial range expansion from the plateau edge followed by recent fragmentation is largely responsible for the present-day spatial distribution of cpDNA haplotypes within the species.

基于叶绿体DNA trnT-trnF序列研究祁连圆柏的谱系地理学

由于青藏高原的地貌效应,第四纪冰期气候的反复变化应对现今该地区生物的地理分布及其居群遗传结构产生重大影响.本文对这一地区特有分布物种祁连圆柏Juniperus przewalskii Kom.整个分布区内20居群392个个体的trnT-trnF序列变化进行了研究;共发现3种单倍型(haplotype),构成两种地理区域:高原台面上的居群主要固定HapA,而HapA、HapB和HapC在高原边缘居群均有分布.所有居群总的遗传多样性HT=0.511,GST=0.847.在低海拔的高原边缘,Hap A、Hap B和Hap C高频率固定在不同的居群中,表明可能存在多个不同的避难所,居群反复缩小和扩张的瓶颈效应造成了遗传多样性的丢失.而边缘的一个居群含有两种相似单倍型频率则可能是冰期后迁移融合而成或者该居群在冰期经受的瓶颈作用更弱.高原台面东部间断分布的居群只固定Hap A,表明它们可能经历了冰期后共同的回迁过程和由此产生的奠基者效应.我们的研究结果表明祁连圆柏在冰期可能存在多个避难所,瓶颈效应和奠基者效应造成了这些居群现在的遗传多样性分布式样.

Phylogeography of Pinus tabulaeformis Carr. (Pinaceae), a dominant species of coniferous forest in northern China

How coniferous trees in northern China changed their distribution ranges in response to Quaternary climatic oscillations remains largely unknown. Here we report a study of the phylogeography of Pinus tabulaeformis, an endemic and dominant species of coniferous forest in northern China. We examined sequence variation of maternally inherited, seed-dispersed mitochondrial DNA (mtDNA) (nad5 intron 1 and nad4/3-4) and paternally inherited, pollen- and seed-dispersed chloroplast DNA (cpDNA) (rpl16 and trnS-trnG) within and among 30 natural populations across the entire range of the species. Six mitotypes and five chlorotypes were recovered among 291 trees surveyed. Population divergence was high for mtDNA variation (G(ST) = 0.738, N-ST = 0.771) indicating low levels of seed-based gene flow and significant phylogeographical structure (N-ST > G(ST), P < 0.05). The spatial distribution of mitotypes suggests that five distinct population groups exist in the species: one in the west comprising seven populations, a second with a north-central distribution comprising 15 populations, a third with a southern and easterly distribution comprising five populations, a fourth comprising one central and one western population, and a fifth comprising a single population located in the north-central part of the species' range. Each group apart from the fourth group is characterized by a distinct mitotype, with other mitotypes, if present, occurring at low frequency. It is suggested, therefore, that most members of each group apart from Group 4 are derived from ancestors that occupied different isolated refugia in a previous period of range fragmentation of the species, possibly at the time of the Last Glacial Maximum. Possible locations for these refugia are suggested. A comparison of mitotype diversity between northern and southern subgroups within the north-central group of populations (Group 2) showed much greater uniformity in the northern part of the range both within and between populations. This could indicate a northward migration of the species from a southern refugium in this region during the postglacial period, although alternative explanations cannot be ruled out. Two chlorotypes were distributed across the geographical range of the species, resulting in lower levels of among-population chlorotype variation. The geographical pattern of variation for all five chlorotypes provided some indication of the species surviving past glaciations in more than one refugium, although differentiation was much less marked, presumably due to the greater dispersal of cpDNA via pollen.

Estudo filogenético das espécies da seção Torva do gênero Solanum L. (Solanaceae) na região sul do Brasil

O gênero Solanum L. (Solanaceae) compreende mais de 1000 espécies, incluindo táxons de grande interesse econômico por seu valor alimentício e medicinal. Este gênero é dividido em três subgêneros: Bassovia, Solanum e Leptostemonum. O subgênero Leptostemonum é dividido em dez seções, e entre essas destaca-se a seção Torva que possui representantes no sul do Brasil, e cujas espécies têm amplo interesse por apresentarem substâncias ativas de grande utilidade farmacológica. Entretanto, dentro dessa seção existem problemas taxonômicos, inclusive com a presença de indivíduos de morfologia intermediária, que dificultam sua classificação e, conseqüentemente, o seu melhor aproveitamento. Nesse trabalho, foram realizados dois estudos de caráter filogenético a fim de conhecer as relações de parentesco entre as espécies de Solanum seção Torva, presentes no sul do Brasil, e destas com espécies de outras seções do subgênero Leptostemonum. Em ambos os estudos foram utilizados quatro marcadores (genomas nuclear e plastidial): a região ITS (espaçadores internos transcritos do DNA nuclear ribossomal) incluindo ITS1, ITS2 e o gene 5,8S; o íntron trnL e os espaçadores intergênicos trnL-trnF e trnS-trnG do DNA plastidial. O marcador ISSR (Inter Simple Sequence Repeats) foi utilizado para verificar a variabilidade genética entre as espécies de Solanum seção Torva e testar o grau de polimorfismo de quatro “primers” dentro dessa seção. As análises realizadas evidenciaram uma origem monofilética para a seção Torva. Além disso, foi verificada uma relação de parentesco mais acentuado dessa seção com S. melongena, S. jamaicense e S. sisymbriifolium. Dentro da seção Torva foram observados agrupamentos que relacionam a espécie de morfologia intermediária a seus possíveis progenitores S. paniculatum e S. guaraniticum. Os quatro agrupamentos mais freqüentes observados dentro da seção foram: a aproximação de S. guaraniticum, S. bonariense e S. paniculatum X S. guaraniticum; o relacionamento entre S. adspersum e S. tabacifolium; a interação entre S. paniculatum e a espécie de morfologia intermediária; e a aproximação entre S. paniculatum e S. variabile. Este trabalho contribuiu para o conhecimento evolutivo das espécies dessa complexa seção que vem levantando interesse de inúmeros pesquisadores.

崖柏属的系统发育与生物地理学研究－兼论广义柏科4CL基因的进化

第三纪末期和第四纪的气候变冷使广泛分布于北半球的暖温带/亚热带生物区系的分布区破碎化，形成了各种洲际间断分布格局。其中东亚—北美东部间断分布最为常见，自林奈时代起就吸引了植物学家们的关注。为了探讨这一地理分布格局的形成过程，前人开展了大量的研究工作，包括间断类群分布的比较、种间关系的经典分类学和化学分类学研究、地理分布的分支分析、遗传距离的估算等。随着分子系统学的发展，在各级分类阶元水平上探讨植物系统发育关系的研究取得了重大进展，很多植物类群的系统发育得到了重建，这使东亚－北美间断分布类群的历史生物地理学研究变得更为切实可行。同时，生物地理学的理论有了新的发展，新的分析方法不断涌现，化石记录及古气候和古地质资料得到大量积累，为深入探讨东亚—北美间断分布提供了条件。目前，已有20 多个东亚－北美间断分布的植物类群被详细研究，丰富了我们对这一现象的认知。然而，以往的研究多基于单亲遗传的叶绿体基因和（或）PCR 直接测序所得的nrDNA ITS 序列，在探讨杂交和网状进化方面存在较大的局限性。在本研究中，我们选取了崖柏属（Thuja L.）这一典型东亚—北美间断分布类群，用来自叶绿体和细胞核的多个基因序列重建其系统发育，探讨其地理分布格局的形成过程，并讨论不同遗传体系的多基因联合分析在植物生物地理学研究中的应用。此外，我们还初步探讨了低拷贝核基因4CL 在广义柏科的进化式样。 1、崖柏属的系统发育和生物地理学研究 崖柏属共 5 种，其中3 种分布于东亚，2 种分别分布于北美东、西部。我们用5 个叶绿体DNA 片段（rpl16 内含子, atpI-rpoC1、trnS-trnfM 和trnS-trnG 基因间区以及trnT-trnF 区）和3 个核基因片段（ITS，LEAFY，4CL）的序列重建了崖柏属的系统发育。发现：（1）基因树拓扑结构的冲突存在于叶绿体基因和核基因之间，甚至不同的核基因之间，说明崖柏属曾发生多次种间杂交并导致网状进化;（2）崖柏属中存在两个种对，即日本香柏－崖柏和朝鲜崖柏－北美香柏;（3）朝鲜崖柏在叶绿体和核基因树上位于不同的位置，可能因古老的杂交和叶绿体捕获所致;（4）北美乔柏的叶绿体基因存在镶嵌式的变异，可能在物种形成过程中发生了叶绿体重组。根据分子钟度量结果，崖柏属两个种对的分化时间为51.1±3.96 Mya，日本香柏和崖柏的分化时间为23.7±5.04 Mya，朝鲜崖柏和北美乔柏的分化时间为14.7±6.06 Mya。 基于多个基因的系统发育分析、DIVA 分析、化石证据和分子钟度量，我们推测崖柏属在古新世或更早的时候起源于北美高纬度地区，并通过白令陆桥扩散到东亚，然后通过隔离分化形成日本香柏－崖柏这一种对。白令陆桥和阿留申陆桥可能在崖柏属的进一步迁移中起了重要介导作用，使崖柏属内发生了多次种间杂交事件，并导致了崖柏－日本香柏和朝鲜崖柏－北美香柏这两种主要的叶绿体类型间的重组以及朝鲜崖柏对北美香柏叶绿体基因的捕获。携带重组叶绿体DNA 的杂交个体迁入北美西部，产生了北美乔柏。根据分子钟估算结果，该迁移事件可能发生在中新世。 鉴于以往对东亚—北美间断分布植物类群的分子系统学研究多基于单亲遗传的叶绿体基因和（或）PCR 直接测序的ITS 数据，这些类群中的网状进化事件可能被低估。同时，我们的结论也部分解释了为什么东亚、北美东部、北美西部三者间的关系存在很多争议：频繁的杂交和渐渗模糊了种间的系统发育关系。因此，我们建议在生物地理学研究中用来源于多个基因组的多基因分析，特别是用单/低拷贝核基因。 2、广义柏科4CL 基因进化的初步研究 广义柏科是松杉类植物中唯一在南、北半球广泛分布的类群，该科中既有古老的孑遗属和寡种属，也有第三纪起源、呈南北半球间断分布的类群。研究4CL 基因在这一类群中的进化，有助于探讨低拷贝核基因在松杉类植物中的进化式样和规律。4CL 在植物次生化合物的生物合成中起重要作用，它催化激活4-香豆酸和一些相关的底物形成不同的辅酶A，促进各种苯丙烷类的代谢。 我们从广义柏科 17 个属中扩增并克隆到23 条4CL 基因序列。基因结构和系统发育分析表明，4CL 在广义柏科中分为4CLI 和4CL II 两大类，二者间的序列相似性为67-70％，进化速率也有很大差异。RT-PCR 结果证明这两种类型均能转录，推测它们都具有功能，且基因结构的差异和序列之间的高度分化暗示这两大类可能执行不同的功能。在4CL 基因树中，落羽杉亚科、北美红杉亚科、狭义柏科的单系都得到了支持。尽管4CL 在广义柏科中的类型及拷贝数还有待研究，但4CLI 很可能以单拷贝或低拷贝存在，其高变的内含子序列可以用来探讨种间的系统发育关系。

Phylogeny and historical biogeography of the Australian Camphorosmeae (Chenopodiaceae)

Diese Studie befasst sich mit der Phylogenie und Biogeographie der australischen Camphorosmeae, die ein wichtiges Element der Flora arider Gebiete Australiens sind. Die molekularen Phylogenien wurden mit Hilfe Bayes’scher Statistik und „maximum likelihood”berechnet. Um das Alter der Gruppe und interner Linien abzuschätzen, wurden die Methoden „Nonparametric rate smoothing” und “penalized likelihood” benutzt. Morphologische Merkmale wurden nach Kriterien der Parsimonie auf den molekularen Baum aufgetragen. „Brooks parsimony analysis”, „cladistic analysis of distributions and endemism”, „dispersal-vicariance analysis”,„ancestral area analysis” und „weighted ancestral area analysis” wurden angewandt, um Abfolge und Richtungen der Ausbreitung der Gruppe in Australien zu analysieren.Von sieben getesteten Markern hatten nur die nukleären ETS und ITS genügend Variation für die phylogenetische Analyse der Camphorosmeae. Die plastidären Marker trnL-trnF spacer,trnP-psaJ spacer, rpS16 intron, rpL16 intron und trnS-trnG spacer zeigten kein ausreichendes phylogenetisches Signal. Die gefundenen phylogenetischen Hypothesen widersprechen der jetzigen Taxonomie der Gruppe. Neobassia, Threlkeldia, Osteocarpum und Enchylaena sollten den Gattungen Sclerolaena bzw. Maireana zugeordnet werden. Die kladistische Analyse der Fruchtanhängsel unterstützt die taxonomischen Ergebnisse der auf DNA basierenden Phylogenie. Allerdings hat die Behaarung, die bei anderen Gruppen der Chenopodiaceae als wichtiges taxonomisches Merkmal herangezogen wird, die Phylogenie nicht unterstützt. Vorfahren der heutigen Camphorosmeen sind im Miozän, vor ca. 8-14 Millionen Jahren, durch Fernausbreitung vermutlich aus Asien in Australien eingewandert. Anfängliche Diversifizierung fand während des späten Miozäns bis in das frühe Pliozän vor ca. 4-7 Millionen Jahren statt. Am Ende des Pliozäns existierten schon 45% - 72% der Abstammungslinien der jetzigen Camphorosmeen. Dies weist auf eine schnelle Ausbreitung hin. Das Alter stimmt mit dem Einsetzen der Aridisierung Australiens überein, und deutet darauf hin, dass die Ausbreitung der ariden Gebiete eine große Rolle bei der Diversifizierung der Gruppe spielte. Die Vorfahren der australischen Camphorosmeae scheinen die Südküste Australiens zuerst besiedeln zu haben. Dies geschah vor dem Einsetzen der Aridisierung des Kontinents. Die anschließende Ausbreitung erfolgte in verschiedene Richtungen und folgte der fortschreitenden Austrocknung im späten Tertiär und im ganzen Quartär. Durch ihre Anpassung an Trockenheit ist der Erfolg der Camphorosmeae in den ariden Gebieten zu erklären.Die Abwesenheit von klaren phylogenetischen und artspezifischen Signalen zwischen Arten der australischen Camphorosmeae ist auf das junge Alter und die schnelle Diversifizierung der Gruppe zurückzuführen, welche die Häufung von Mutationen und eine starke morphologische Differenzierung nicht zugelassen haben.

Nuclear and plastid markers reveal the persistence of genetic identity: A new perspective on the evolutionary history of Petunia exserta

Recently divergent species that can hybridize are ideal models for investigating the genetic exchanges that can occur while preserving the species boundaries. Petunia exserta is an endemic species from a very limited and specific area that grows exclusively in rocky shelters. These shaded spots are an inhospitable habitat for all other Petunia species, including the closely related and widely distributed species P. axillaris. Individuals with intermediate morphologic characteristics have been found near the rocky shelters and were believed to be putative hybrids between P. exserta and P. axillaris, suggesting a situation where Petunia exserta is losing its genetic identity. In the current study, we analyzed the plastid intergenic spacers trnS/trnG and trnH/psbA and six nuclear CAPS markers in a large sampling design of both species to understand the evolutionary process occurring in this biological system. Bayesian clustering methods, cpDNA haplotype networks, genetic diversity statistics, and coalescence-based analyses support a scenario where hybridization occurs while two genetic clusters corresponding to two species are maintained. Our results reinforce the importance of coupling differentially inherited markers with an extensive geographic sample to assess the evolutionary dynamics of recently diverged species that can hybridize. (C) 2013 Elsevier Inc. All rights reserved.

Evolution of the Neckeraceae (Bryophyta): resolving the backbone phylogeny

Earlier phylogenetic studies, including species belonging to the Neckeraceae, have indicated that this pleurocarpous moss family shares a strongly supported sister group relationship with the Lembophyllaceae, but the family delimitation of the former needs adjustment. To test the monophyly of the Neckeraceae, as well as to redefine the family circumscription and to pinpoint its phylogenetic position in a larger context, a phylogenetic study based on molecular data was carried out. Sequence data were compiled, combining data from all three genomes: nuclear ITS1 and 2, plastid trnS-rps4-trnT-trnL-trnF and rpl16, and mitochondrial nad5 intron. The Neckeraceae have sometimes been divided into the two families, Neckeraceae and Thamnobryaceae, a division rejected here. Both parsimony and Bayesian analyses of molecular data revealed that the family concept of the Neckeraceae needs several further adjustments, such as the exclusion of some individual species and smaller genera as well as the inclusion of the Leptodontaceae. Within the family three well-supported clades (A, B and C) can be distinguished. Members of clade A are mainly non-Asiatic and nontropical. Most species have a weak costa and immersed capsules with reduced peristomes (mainly Neckera spp.) and the teeth at the leaf margins are usually unicellular. Clade B members are also mainly non-Asiatic. They are typically fairly robust, distinctly stipilate, having a single, at least relatively strong costa, long setae (capsules exserted), and the peristomes are well developed or only somewhat reduced. Members of clade C are essentially Asiatic and tropical. The species of this clade usually have a strong costa and a long seta, the seta often being mammillose in its upper part. The peristome types in this clade are mixed, since both reduced and unreduced types are found. Several neckeraceous genera that were recognised on a morphological basis are polyphyletic (e.g. Neckera, Homalia, Thamnobryum, Porotrichum). Ancestral state reconstructions revealed that currently used diagnostic traits, such as the leaf asymmetry and costa strength are highly homoplastic. Similarly, the reconstructions revealed that the 'reduced' sporophyte features have evolved independently in each of the three clades.

藏麻黄与丽江麻黄的形态及遗传变异研究

麻黄属(Ephedra)起源较早，该属植物的形态性状极度退化或简化，可用于经典分类学的有价值的性状非常有限。分布于青藏高原的藏麻黄（E. saxatilis）和丽江麻黄（E. likiangensis）二者间形态相似，性状变异连续，很难分辨，但被《中国植物志》中、英文版作两个种处理。 本文对藏麻黄和丽江麻黄的七个居群、151个个体的叶、雌球花和节间长等形态学性状进行了分析，发现各性状的变异情况在群体间无明显差异。同时我们还对154个个体的叶绿体DNA trnT-trnF和 trnS-trnfM区进行了序列分析，两个片段的联合矩阵长1382bp，共有29个变异位点，其中有9个碱基变异和2个indel，可划分为H1、H2和H3三种单倍型。这3种单倍型在丽江麻黄中均有分布，但藏麻黄仅含H1和H2。 综合来自形态学和分子方面的证据，我们发现藏麻黄和丽江麻黄的关系非常近缘，因此建议予以合并。同时本文还以膜果麻黄（E. przewalskii）为外类群，从谱系生物地理学角度探讨了三种叶绿体单倍型的进化关系，发现H2最原始，分布最广;H1与其它两种单倍型间的序列差异较大，可能是较进化的类型。此外，无性克隆的繁殖方式可能是导致Ephedra单倍型非常简单的重要原因之一。

东亚—北美间断分布类群的分子生物地理学研究—以莲科和菖蒲科为例

在洲际间断生物地理学研究中，东亚—北美间断分布类群的分子生物地理学研究一直是关注和研究的热点。在本论文中我们选取了水生和半水生的植物代表类群，莲科（Nelumbonaceae）和菖蒲科（Acoraceae）作为研究对象，通过来自叶绿体、线粒体和核基因组的DNA 序列分析和微卫星分析，一方面探讨莲科的系统位置、揭示其间断地理格局的形成过程、重建菖蒲科的系统发育及其地理格局的形成过程的同时，另一方面，在总结前人研究成果的基础上，总结东亚—北美间断分布的基本特点。主要成果总结如下。 1. 菖蒲科的系统发育和分子生物地理学 菖蒲科仅含一属，菖蒲属（Acorus），共5 种。其中北美菖蒲（A. americanus） 分布于北美，其余4 种（A. calamus, A. gramineus, A. tatarinowii and A. rumphianus） 分布于亚洲的东部和南部。北美菖蒲和菖蒲（A. calamus）叶片中间具有明显的中肋；其余3 种不具有明显的中肋。本论文的19 份材料包含了4 个种，（不含较狭域分布的长苞菖蒲A. rumphianus），利用4 个叶绿体基因片段（trnL-F, psbA-trnH, rps16-trnK, rbcL）和1 个核基因片段（ITS）的序列重建菖蒲属的系统发育。结果表明（1）具有中肋和不具中肋的物种各自聚为一支；（2）具有中肋的菖蒲和北美菖蒲亲缘关系最近，构成东亚—北美间断种对关系；（3）在不具有中肋的一支内部，来自台湾的材料与其它材料差异最大，其余的材料也明显的分为了两类。基于rbcL 序列，使用松散分子钟模型、贝叶斯算法估算菖蒲属起源时间约为135.17 百万年（mya），菖蒲和北美菖蒲的间断分歧时间约为3.72mya。该结果支持菖蒲属为古老的单子叶植物，但东亚—北美间断物种分化时间较年轻。我们推测间断的种对可能通过白令陆桥，从东亚扩散到了北美。 2. 莲科的系统位置和分子生物地理学 莲科仅含一属，莲属（Nelumbo），两个种莲（N. nucifera）和美洲黄莲（N. lutea），间断分布于东亚、澳大利亚北部和北美东部。莲科的系统位置在形态和分子证据不一致。本论文使用了核基因18S rDNA、26S rDNA，叶绿体基因atpB、rbcL，线粒体基因NAD1 的序列重新构建莲科的系统位置并进行了分化时间推算。结果为：（1）叶绿体和核基因构建的严格一致树的拓扑结构不一致，叶绿体数据支持莲科和山龙眼科、悬铃木科具有较近的亲缘关系，核基因数据显示莲科位于真双子叶植物的基部；（2）5 个基因片段的合并分析结果显示，莲科与山龙眼科、悬铃木科聚为一支但支持率不高；（3）基于核基因、叶绿体和5 个基因的分别合并数据，使用松散分子钟模型、贝叶斯算法估算莲科起源时间分别为，113.13 、109.38 和110.35mya ，两个间断物种的分化时间为，3.77、4.34、5.85mya；（4）根据间断的时间和两个物种的遗传差异程度，现存的两个物种应是来自于东亚或北美的冰期残遗，而不是来自于两个大陆祖先种的独立进化后裔。 3. 莲的分子谱系地理学研究 我们采集了37 份莲的材料，10 份美洲黄莲的材料，代表了两者的主要分布区。我们选取了叶绿体基因（trnL-trnF, trnS-trnG, petB-petD 和psbA-trnH），线粒体基因COX1，以及11 个微卫星位点进行莲的分子谱系地理研究。DNA 序列显示莲和美洲黄莲均具有很低的遗传多样性；微卫星数据揭示了稍高于DNA 序列的遗传多样性。两物种相比，美洲黄莲的多样性较高。基于微卫星数据的遗传结构分析表明，莲存在明显的3 个地理分化区域，这三个区域的遗传分化显著（FST=0.542），说明莲远距离群体间基因交流有限。基于DNA 序列和微卫星数据的单倍型地理分布关系，我们推测东南亚地区是莲的避难所或冰期残遗区，冰期后群体分别向西和向北扩张。 4. 东亚—北美间断分布的一般特点 （1）东亚—北美东部间断分歧时间范围较长，从始新世中期甚至更早一直持续到1mya 左右；东亚—北美西部间断类群分化时间跨度相对小，集中在中新世时期；东亚—整个北美间断分化时间与东亚—北美东部间断类群一样经历较长时间；草本类群晚于木本类群形成间断分布式样，洲际间断分化时间与类群的起源时间并无相关性。（2）东亚与北美间断分布类群的起源地因类群而异。（3） 东亚与北美间断分布类群扩散方向呈不确定性。（4）东亚与北美间断类群扩散有三条可能的路径，即大西洋陆桥、白令陆桥和南半球跨洋长距离传播。

Potential refugium on the Qinghai-Tibet Plateau revealed by the chloroplast DNA phylogeography of the alpine species Metagentiana striata (Gentianaceae)

Metagentiana striata is an alpine annual herbaceous plant endemic to the east of the Qinghai-Tibet (Q-T) Plateau and adjacent areas. The phylogeography of M. striata was studied by sequencing the chloroplast DNA (cpDNA) trnS-trnG intergenic spacer. Ten haplotypes were identified from an investigation of 232 individuals of M. striata from 14 populations covering the entire geographical range of this species. The level of differentiation amongst populations was very high (G(ST) = 0.746; N-ST = 0.774) and a significant phylogeographical structure was observed (P < 0.05). An analysis of molecular variance found a high variation amongst populations (76%), with F-ST = 0.762 (highly significant, P < 0.001), indicating that little gene flow occurred amongst the different regions; this was explained by the isolation of populations by high mountains along the Q-T Plateau and adjacent areas (N-m = 0.156). Only one ancestral haplotype (A) was common and widespread throughout the distributional range of M. striata. The populations of the Hengduan Mountains region of the south-eastern Q-T Plateau showed high diversity and uniqueness of haplotypes. It is suggested that this region was the potential refugium of M. striata during the Quaternary glaciation, and that interglacial and postglacial range expansion occurred from this refugium. This scenario was in good agreement with the results of nested clade analysis, which inferred that the current spatial distribution of cpDNA haplotypes and populations resulted from range expansion, together with past allopatric fragmentation events. (c) 2008 The Linnean Society of London.

Plastid microsatellites for the study of genetic variability in the widespread Cephalanthera longifolia, C. damasonium and C. rubra (Neottieae, Orchidaceae), and cross-amplification in other Cephalanthera species

Plastid microsatellite loci developed for Cephalanthera longifolia were used to examine the level of genetic variation within and between populations of the three widespread Cephalanthera species (C. damasonium, C. longifolia and C. rubra). The most detailed sampling was in C. longifolia (42 localities from Ireland to China; 147 individuals). Eight haplotypes were detected. One was detected in the vast majority of individuals and occurred from Ireland to Iran. Three others were only found in Europe (Ireland to Italy, England to Italy and Austria to Croatia). Two were only found in the Middle East and two only in Asia. In C. damasonium, 21 individuals from 10 populations (England to Turkey) were sampled. Only one haplotype was detected. In C. rubra, 34 individuals from eight populations (England to Turkey) were sampled. Although it was not possible to amplify all loci for all samples of this species, nine haplotypes were detected. Short alleles for the trnS-trnG region found in two populations of C. rubra were characterized by sequencing and were caused by deletions of 26 and 30 base pairs. At this level of sampling, it appears that C. rubra shows the greatest genetic variability. Cephalanthera longifolia, C. rubra and C. damasonium have previously been characterized as outbreeding, outbreeding with facultative vegetative reproduction and inbreeding, respectively. Patterns of genetic variation here are discussed in the light of these reproductive system differences. The primers used in these three species of Cephalanthera were also demonstrated to amplify these loci in another five species (C. austiniae, C. calcarata, C. epipactoides, C. falcata and C. yunnanensis). Although it is sometimes treated as a synonym of C. damasonium, the single sample of C. yunnanensis from China had a markedly different haplotype from that found in C. damasonium. All three loci were successfully amplified in two achlorophyllous, myco-heterotrophic species, C. austinae and C. calcarata. © 2010 The Linnean Society of London.