染色体数据的挖掘及其在植物多样性进化研究中的利用

多倍化(或全基因组加倍)是植物物种形成的重要途径,现存的被子植物可能都发生过一次甚至多次多倍化事件。多倍化传统的定义是染色体数目相对于祖先类群呈整倍性增加。其中最常用的研究方法是核型分析,核型能够提供物种的基本细胞学参数,包括染色体数目、倍性水平、核型不对称性、核型变异系数等。目前核型研究的趋势表现出从物种基本核型参数分析逐渐演化到多类群、多学科交叉融合的特点:一方面植物核型分析从种群、物种、科属的类群到生命之树,探讨染色体核型在各支系的进化特征、趋势以及驱动植物系统进化的细胞学机制;另一方面探讨和分析区域或生态系统植物区系的染色体谱或倍性等细胞学特征,可以探究区域地质环境变化或生态环境对染色体倍性等的影响,或通过区域染色体谱的构建,分析区域植物区系的形成和进化历史。因而,植物核型研究为系统发育、分子系统进化、生命之树以及植物区系地理的起源和演化研究提供了新思路。越来越多的新方法、新手段在植物核型分析与多倍化研究中得到运用,从而揭示了植物类群或植物区系的染色体进化以及细胞地理特征。今后植物细胞学研究趋势会向多学科交叉融合,整合各研究领域证据,从不同水平角度综合分析植物核型多样性形成的原因及意义,从而更加全面地认识和理解植物物种多样化与物种形成原因。     

天门冬科广义铃兰族的染色体数目与核型进化研究

广义铃兰族包括6属200余种单子叶植物,具有较为稳定的染色体数目以及明显的核型变化,是研究植物染色体进化非常理想的类群。该研究采用常规压片法对铃兰族5个物种植物的染色体形态、数目及其核型进行了观察分析,并全面收集和整理了铃兰族所有已报道的共81种253条的染色体记录,通过综合统计、重建系统发育树,探讨铃兰族属间以及各属下种间的系统关系,以揭示物种间的亲缘关系和起源演化趋势。结果表明:(1)细胞学实验观察发现,铃兰族5个物种植物的染色体数目除蜘蛛抱蛋为2n=36外,其余4种均为2n=38,且均为二倍体,不对称性核型均为Stebbins’-2C型,核型不对称系数As.K%的变化范围为65%～69%,但染色体核型公式有差别。(2)所收集的铃兰族染色体报道记录中,绝大多数物种的染色体数目为2n=38,只在蜘蛛抱蛋属中有部分物种为2n=36;根据染色体长度变化核型主要分为单型和三型;统计结果显示,染色体2n=36的类群比2n=38的类群通常多1对中型染色体,少2对小型染色体,推测可能是由于2对小的染色体融合成为1对中型染色体。(3)基于4个叶绿体基因片段(rbcL、matK、psbA-trnH和trnL-F)和核基因ITS构建的铃兰族最新分子系统发育树显示,铃兰族分为三支,第Ⅰ支位于整个族的基部,为首先分化出来的铃兰-白穗花分支,均以x=19为基数;第Ⅱ支为万年青-吉祥草分支,染色体基数为x=19;第Ⅲ支为蜘蛛抱蛋-长柱开口箭分支,染色体基数为x=18或19;结合铃兰族染色体基数和核型分析发现,铃兰族染色体低基数的x=18是由x=19通过染色体融合或丢失进化而来,推测核型则是由比较对称的单型向不对称的三型演化而来。该研究结果为铃兰族属间关系及物种进化关系提供了细胞学支持证据。     

横断山区六种八居群鼠尾草属植物的核型分析

鼠尾草属(Salvia)是唇形科(Lamiaceae)最大的属,属下多种为民间常用草药,亦有供观赏的种类。为探究横断山区物种在细胞学水平的进化方式,讨论形态分类学与分子系统学之间的分类关系,该研究通过广泛收集染色体文献资料,采用植物常规压片法对采集自横断山地区6种8居群鼠尾草属植物进行核型分析,并构建了中国地区分布的鼠尾草属植物叶绿体系统发育树。统计结果表明:(1)全世界范围内报道了约23%的鼠尾草属植物染色体数据,其中分布在中国地区的鼠尾草属植物染色体报道率为32.10%,分布在横断山地区的鼠尾草属植物报道率为40.54%,(2)鼠尾草属植物染色体基数以x=8和x=11为主,分布在中国地区的鼠尾草属植物染色体基数均为x=8。实验结果表明:(1)西藏鼠尾草(S. wardii)核型数据为首次报道。(2)雪山鼠尾草(S. evansiana)首次在云南德钦地区发现二倍体居群。将细胞学数据结合叶绿体进化树开展染色体进化关联分析,论证多倍化可能不是鼠尾草属物种适应高海拔环境的主要机制,表明多倍体不是该属物种形成的主要进化途径而是以二倍体水平为主,推测染色体组的加倍可能是物种在形态学与分子系统学上分类关系不一致的原因之一。该研究丰富了横断山区鼠尾草属植物的染色体核型数据,结合区域分子系统树探讨染色体特征的进化关系,为今后深入研究该属物种的核型进化做出了探索,为开展祖先物种染色体基数推演分析补充了基础数据。     

中国葱属根茎组植物15种25居群的核型研究

采用常规压片法,对15种25居群葱属根茎组植物进行核型研究,首次报道荒漠韭Allium tekesicola、蓝花韭A.beesianum的染色体数目和核型,并增补了一些根茎组植物的细胞学资料。结果表明：供试类群为二倍体或四倍体,核型类型为1A、2A或2B;高山韭A.sikkimense壤塘居群和荒漠韭具短臂随体,宽苞韭A.platyspathum的和布克赛尔居群、奇台居群具居间随体;野黄韭A.rude雅江居群和荒漠韭中各存在1条B染色体。结合前人研究结果,我们讨论了中国根茎组植物的染色体基数、随体染色体的多型性、B染色体的进化意义和该组植物的进化方式,探讨了天蓝韭A.cyaneum、齿丝山韭A.nutans的地理分布成因,得到了如下推论：（1）中国根茎组植物的染色体基数x=8;（2）中国根茎组植物的核型进化趋势为：1A→2A→2B→2C;（3）中国根茎组植物的随体染色体具多型性;（4）多倍化和结构变异是中国根茎组植物进化的两种重要方式;（5）天蓝韭和齿丝山韭以多倍化和无性生殖来克服扩大新的生存空间遇到的困难。     

中国西南地区鹿药属4种15居群核型研究

对产于中国西南部的鹿药属（Maianthemum）4种植物进行了细胞学研究,包括染色体数目,多倍化,非整倍性和随体染色体,以及核型不对称性和核型进化。结果表明：1）除了在云南丽江采集的Maianthemum tatsienensis染色体数目为2n=72之外,其余的居群全为2n=36;2）核型在居群间存在变异,特别是在具中部染色体和近中部染色体的数目以及随体染色体的数目和位置上。此外,M．nanchuanense和M．szechuanicum的核型是首次报道,B染色体也是首次在该属中发现。我们推测鹿药属的进化方式包括频繁的染色体畸变以及不同水平上的多倍化,而中国西南部是该属的分化中心。     

13种21居群葱属植物的细胞分类学研究

采用常规压片法,对13种（含1变种）21居群葱属（Allium L.）植物进行了细胞分类学研究。结果表明：所研究类群的染色体基数为7、8和11,核型的类型为2A、2B和3A型,并且存在2倍体、3倍体和4倍体。主要讨论了葱属根茎组（Sect.Rhizirdium G.Don）部分类群的核型分化和进化机制,高山韭（A.sikkimense Baker）和多星韭（A.wallichii Kunth）的细胞地理学,以及B染色体的多态性及其在生境上的适应意义。最后在本研究的基础上,结合前人的细胞分类学研究结果,对葱属植物的核型进化形成了如下认识：（1）该属植物的原始染色体基数为8,其他基数的类群是由基数为8的类群进化而来的;（2）葱属植物在核型类型的进化上存在两条路线：基数为7的类群核型进化趋势为2A→2B→2C,而基数为8的类群的核型进化趋势为1A→2A→2B→2C;（3）多倍化是葱属植物进化的重要机制之一。     

铁线莲属21个类群的染色体核型分析

铁线莲属(Clematis L.)为毛茛科大属之一,具有重要的园艺与药用价值。为探讨铁线莲属植物染色体组演化规律,揭示属下组间及种间的亲缘关系,该研究采用常规压片法对铁线莲属21个类群的根尖进行处理、压片,对染色体的形态特征进行观察及核型分析,同时利用Ward联接法进行聚类分析研究。狭裂太行铁线莲、毛果扬子铁线莲、卷萼铁线莲、中印铁线莲、钝萼铁线莲的染色体形态为首次报道。结果表明:铁线莲属21个类群均为二倍体,染色体数均为16(2n=2x=16),除中印铁线莲外其他类群均具随体;长瓣铁线莲、钝萼铁线莲、芹叶铁线莲、褐毛铁线莲、C.flammula、毛果扬子铁线莲的染色体为“2B”型,其他类群染色体为“2A”型;铁线莲属核型不对称系数在60.29%~63.79%之间;铁线莲属植物染色体组表现的较为原始,种间核型存在广泛变异。综上结果表明,铁线莲属植物染色体数目应由二倍体向多倍体演化后通过多倍体的二倍化过程产生非整倍体方向演化。铁线莲属染色体的演化主要在二倍体水平上进行,通过产生染色体结构变异的方式实现,通过产生杂合染色体、加强核型不对称性、染色体类型改变以及随体染色体的变化四种途径进化。同时,核型特征在分组水平与物种水平上的划分与传统分类基本一致,说明核型分析可为铁线莲属下组一级分类提供一定的线索。该研究结果为铁线莲属植物系统分类、遗传演化与资源利用等研究领域提供了新的参考资料。     

葱属根茎组8种21居群植物的核型研究

对葱属根茎组Allium sect.Rhiziridium的8种21个地方居群的核型进行研究,以期为解决该组的种间亲缘关系和物种进化机制提供依据。贺兰韭A.eduardii和阿拉善韭A．flavovirens2个种的核型以及辉韭A．strictum的六倍体核型均属首次报道。研究结果表明：贺兰韭A.eduardii、阿拉善韭A.tlavovirens、北韭A.lineare、蒙古韭A.mongolicum和滩地韭A．oreoprasum的各居群均为二倍体,核型类型为Stebbins的2A型;韭A．tuberosum和野韭A.ramosum的各个居群均为四倍体,核型类型为2A型：辉韭A．Jtrictum的4个居群均为六倍体,核型类型为2B型。通过研究可以得出如下推论：（1）该组植物中存在着大量的多倍体或多倍体系列,染色体数目变化与物种进化具有密切相关性,多倍化可能是根茎组植物核型进化的重要机制之一;（2）随体染色体多为st或t染色体,均位于短臂末端;（3）可以认为辉韭是以增加倍性来克服该物种扩大新的生存空间所带来的困难;（4）现今栽培的韭可能是由野生的二倍体韭和四倍体韭经过长期人工驯化而来的,现今栽培的三倍体韭可能是二倍体韭和四倍体韭杂交而来,并且以无性繁殖方式保存三倍体类群的存在。     

国产13种鸢尾属植物的核型研究

对中国产13种鸢尾属Iris植物进行了核型研究。其中中甸鸢尾I.subdichotoma、长葶鸢尾I.delavayi、大锐果鸢尾I.cuniculiformis为中国特有。大锐果鸢尾的染色体数目及核型为首次报道,核型公式为2n=22=4m 6sm 12st(2SAT)。长管鸢尾I.dolichosiphon的核型为首次报道,核型公式为2n=22=4m 12sm 6st。中甸鸢尾的染色体数目为新报道,核型公式为2n=42=20m 22sm。矮紫苞鸢尾I.ruthenicavar.nana的染色体数目为新报道,3个居群的染色体数目均为2n=42,核型公式分别为中甸居群2n=42=30m 12sm(2SAT),丽江甘海子居群2n=42=28m 14sm(2SAT),中甸尼西居群2n=42=36m 6sm(4SAT)。结合以往的细胞学研究结果,显示尼泊尔鸢尾亚属subgen.Nepalensis是一个染色体数目变化较大的类群,其中的中甸鸢尾可能是联系野鸢尾属Pardanthopsis与尼泊尔鸢尾亚属的重要类群。已报道的紫苞鸢尾I.ruthenica染色体数目为2n=84,与我们所研究的变种矮紫苞鸢尾(2n=42)呈倍性关系,通过与相邻类群的分析比较,认为紫苞鸢尾应是由二倍体类群演化而来。还对鸢尾属内染色体数目的变化和核型进化的趋势进行了探讨。     

植物单倍体

在植物界细胞染色体的数目因种而异。在植物进化过程中,出现多倍化使染色体数增加;也存在一种相反的情况,即染色体倍性逐渐降低,使染色体数目减少。如在被子植物内系统发育较进化的草本植物的染色体基数比较衰老的的木本植物低;种子植物与蕨类相比,前者又少得多。减少多倍性的一个重要途径是成倍地减少细胞中的染色体数目或单倍化。研究单倍性和单倍体在理论上对于种的细胞遗传结构和进化途径的分析,对数量性状遗传学有重要意义。     

中国蕨类植物细胞分类学研究概况

中国蕨类植物类群十分丰富,由63科230属约2600种组成,其中约有10％的种类为中国特有。迄今为止,中国蕨类植物进行过染色体计数的种类约有395种,仅占中国蕨类植物总数的15％,其中48％为多倍体。本文对中国蕨类植物细胞分类学研究的历史和现状进行了综合评述。文中论述了蕨类植物染色体数目和染色体基数在系统学上的应用以及蕨类植物染色体组型分析、多倍化、无融合生殖、多倍体复合体及网状进化的研究概况。中国20多年的蕨类植物细胞分类学研究表明,蕨类植物的染色体数目、基数、组型分析、多倍化及其繁殖方式对蕨类植物的分类、起源和演化的研究都有重要作用,对种及种复合体的鉴定、科属的划分也有重要的参考价值。并对中国蕨类植物细胞分类学研究提出了一些建议。     

Species diversity and ecological range in relation to ploidy level in the flora of the Pyrenees

Polyploidy is a major process in plant evolution. Surprisingly, no study has examined its role in species diversification and ecological distribution in relation to other life history traits. In this study, we examine to what extent polyploidy and the other traditionally examined biological traits (pollination mode, dispersal mode and growth form) account for ecological and taxonomic diversity in the flora of the Pyrenees. Fifty genera (in 22 angiosperm families) were classified according to ploidy level, growth form, pollination mode and dispersal mode, and 451 species and/or subspecies in these 50 genera were classified according to ploidy level and growth form. We examined the contribution of ploidy level, pollination and dispersal modes and growth form to (i) the ecological range of species and genera, i.e., the number of natural habitats (defined by a combination of ecological characteristics) where they occur, and (ii) the taxonomic diversity of the 50 genera. Ploidy level and dispersal mode had significant effects on the taxonomic diversity of the 50 genera. Taxonomic diversity, but not polyploidy per se, was significantly correlated with ecological range of genera. For individual species, diploids had a larger ecological range than polyploids, and herbaceous growth forms had wider ecological distributions than other growth forms. Our results indicate that polyploidisation may be a source of ecological diversification of genera, not by increasing the ecological range of particular polyploid species compared to diploids, but rather by creating taxonomic diversity that leads in some genera to a diversification of the habitats occupied by different ploidy levels. This observation is consistent with previous observations of ecological divergence of chromosomal races in some species in the Alps and in the Pyrenees. As found in other studies, species diversification in the studied flora appears to be greatly influenced by the occurrence of multiple dispersal modes, while ecological range of species or subspecies is significantly increased by the presence of herbaceous species.     

Chromosome and genome size variation in Luzula (Juncaceae), a genus with holocentric chromosomes

The present study examines chromosome and genome size evolution in Luzula (woodrush; Juncaceae), a monocot genus with holocentric chromosomes. Detailed karyotypes and genome size estimates were obtained for seven Luzula spp., and these were combined with additional data from the literature to enable a comprehensive cytological analysis of the genus. So that the direction of karyotype and genome size changes could be determined, the cytological data were superimposed onto a phylogenetic tree based on the trnL‐F and internal transcribed spacer (ITS) DNA regions. Overall, Luzula shows considerable cytological variation both in terms of chromosome number (2n = 6–66) and genome size (15‐fold variation; 2C = 0.56–8.51 pg; 547.7–8322.8 Mb). In addition, there is considerable diversity in the genomic mechanisms responsible, with the range of karyotypes arising via agmatoploidy (chromosome fission), symploidy (chromosome fusion) and/or polyploidy accompanied, in some cases, by the amplification or elimination of DNA. Viewed in an evolutionary framework, no broad trend in karyotype or genome evolution was apparent across the genus; instead, different mechanisms of karyotype evolution appear to be operating in different clades. It is clear that Luzula exhibits considerable genomic flexibility and tolerance to large, genome‐scale changes. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 170 , 529–541.     

Karyotype diversity and genome size variation in Neotropical Maxillariinae orchids

• Orchidaceae is a widely distributed plant family with very diverse vegetative and floral morphology, and such variability is also reflected in their karyotypes. However, since only a low proportion of Orchidaceae has been analysed for chromosome data, greater diversity may await to be unveiled. Here we analyse both genome size (GS) and karyotype in two subtribes recently included in the broadened Maxillariinea to detect how much chromosome and GS variation there is in these groups and to evaluate which genome rearrangements are involved in the species evolution.
• To do so, the GS (14 species), the karyotype – based on chromosome number, heterochromatic banding and 5S and 45S rDNA localisation (18 species) – was characterised and analysed along with published data using phylogenetic approaches.
• The GS presented a high phylogenetic correlation and it was related to morphological groups in Bifrenaria (larger plants – higher GS). The two largest GS found among genera were caused by different mechanisms: polyploidy in Bifrenaria tyrianthina and accumulation of repetitive DNA in Scuticaria hadwenii. The chromosome number variability was caused mainly through descending dysploidy, and x=20 was estimated as the base chromosome number.
• Combining GS and karyotype data with molecular phylogeny, our data provide a more complete scenario of the karyotype evolution in Maxillariinae orchids, allowing us to suggest, besides dysploidy, that inversions and transposable elements as two mechanisms involved in the karyotype evolution. Such karyotype modifications could be associated with niche changes that occurred during species evolution.

     

The evolutionary origin of a novel karyotype in Timarcha (Coleoptera, Chrysomelidae) and general trends of chromosome evolution in the genus

In this work, we have analysed the karyotypes of six species of Timarcha for the first time and updated the cytological information for two additional taxa, for one of them confirming previous results ( Timarcha erosa vermiculata ), but not for the other ( T. scabripennis ). We describe the remarkable karyotype of T. aurichalcea , the lowest chromosome number in the genus (2 n  = 18), distinctive as well for the presence of an unusual chiasmatic sexual bivalent hitherto unreported for Timarcha . This study increases the number of species studied cytologically in this genus to forty. Additional cytogenetic analyses are performed on several species, including Ag-NOR staining and fluorescent in situ hybridization (FISH) studies with ribosomal DNA probes. Karyotype evolution is analysed by tracing different karyotype coding strategies on a published independent phylogenetic hypothesis for Timarcha based on the study of three genetic markers. The implementation of a likelihood model of character change optimized onto the phylogeny is tentatively used to detect possible drifts in chromosome changes. These analyses show that karyotype is conservative in the evolution of the genus and that there is an apparent trend to reducing chromosome number. Cytological and phylogenetic data are used to explain the evolutionary origin of the karyotype of T. aurichalcea by two centric fusions involving one pair of acrocentric autosomes and the sexual chromosomes.     

青藏高原4种无心菜属植物的细胞学研究

以四川康定和稻城两个居群的无心菜属（Arenaria）雪灵芝亚属（A. subgen.Eremogoneastrum）的雪灵芝（A.brevipetala Y.W.Tsui et L.H.Zhou）、西藏八宿业拉山和安久拉山两个居群的八宿雪灵芝（A.baxoiensis L.H.Zhou）、西藏当雄和工布江达两个居群的瘦叶雪灵芝（A.ischnophylla Williams）以及西藏拉萨居群的藓状雪灵芝（A.bryophylla Fernald）为研究材料,采用植物根尖常规压片法,对它们的细胞分类学特征进行研究。结果显示：7个居群中,4种植物的染色体数目均为2n=22,为二倍体,核型公式为2n=2x=22=22 m,核型不对称性属于Stebbins's-1A型,AI值在1.75~0.32范围内变化。结合无心菜属已有细胞学资料,推断雪灵芝亚属核型特征原始,染色体数目及倍性稳定。本研究中4种植物的染色体数目和核型资料均为首次报道,补充了青藏高原无心菜属的细胞学资料。     

Tempo and mode in karyotype evolution revealed by a probabilistic model incorporating both chromosome number and morphology

Karyotype, including the chromosome and arm numbers, is a fundamental genetic characteristic of all organisms and has long been used as a species-diagnostic character. Additionally, karyotype evolution plays an important role in divergent adaptation and speciation. Centric fusion and fission change chromosome numbers, whereas the intra-chromosomal movement of the centromere, such as pericentric inversion, changes arm numbers. A probabilistic model simultaneously incorporating both chromosome and arm numbers has not been established. Here, we built a probabilistic model of karyotype evolution based on the “karyograph”, which treats karyotype evolution as a walk on the two-dimensional space representing the chromosome and arm numbers. This model enables analysis of the stationary distribution with a stable karyotype for any given parameter. After evaluating their performance using simulated data, we applied our model to two large taxonomic groups of fish, Eurypterygii and series Otophysi, to perform maximum likelihood estimation of the transition rates and reconstruct the evolutionary history of karyotypes. The two taxa significantly differed in the evolution of arm number. The inclusion of speciation and extinction rates demonstrated possibly high extinction rates in species with karyotypes other than the most typical karyotype in both groups. Finally, we made a model including polyploidization rates and applied it to a small plant group. Thus, the use of this probabilistic model can contribute to a better understanding of tempo and mode in karyotype evolution and its possible role in speciation and extinction.