Allozyme variation and population differentiation of the Aconitum delavayi complex (Ranunculaceae) in the Hengduan Mountains of China

The Aconitum delavayi complex is a group of four climbing species with trisect-leaves occurring in the Hengduan Mountains. The species of this complex are highly localized on very narrow regions with quite small population sizes. Because of rapid environmental changes recently in the Hengduan Mountains, this complex shows complicated morphological variability, which makes it difficult to delimit species. In the present study, 10 enzyme systems coding for 14 putative loci were employed to detect the interspecific and intraspecific genetic variation of the complex. In addition to low genetic diversity within all eight populations surveyed, the results indicate that A. episcopale is a distinct species because of high genetic identities among its three populations. Very low genetic divergence among populations of A. stapfianum and A. delavayi suggests that the two species should be treated as a single one.     

紫乌头复合体nrDNA的ITS序列与系统发育分析

对乌头属 Aconitum L. 27个类群的nrDNA ITS序列进行了简约法与邻接法分析,两种方法得到的系统发育树基本一致。乌头亚属subgen. Aconitum 的蔓乌头系 ser. Volubilia 不是一个单系类群,显柱乌头系 ser. Stylosa 与兴安乌头系ser. Ambigua 各自作为单系类群也没有得到支持。特产于云南西北部横断山区的一些种之间存在非常近的系统发育关系,说明这些种可能是近期物种形成的产物。紫乌头 A. delavayi 复合体的不同类群在系统发育树上位     

Molecular markers reveal little genetic differentiation among Aconitum noveboracense and A. columbianum (Ranunculaceae) populations

Aconitum noveboracense, a rare, herbaceous perennial, is restricted to recently unglaciated areas in Iowa, Wisconsin, Ohio, and New York, and federally classified as a threatened species. These populations may be disjuncts of the common congener, A. columbianum Nutt., which occurs in the mountains of the western United States. Morphological characters do not reliably separate these taxa. The identity of Black Hills populations, located between the ranges of the rare and common species, is also uncertain. We characterized genetic variation within and among the Aconitum populations in question using isozymes and randomly amplified polymorphic DNA (RAPDs). Isozymes indicate a high degree of similarity among all populations and a high level of genetic diversity in Black Hills populations. Of 97 scorable RAPD loci, 89.7% are polymorphic and clearly resolve most populations. Like isozymes, RAPDs indicate high levels of genetic diversity in the Black Hills and very strong similarity of these populations to A. columbianum from the Bighorn Mountains. Aconitum noveboracense populations show >80% similarity to A. columbianum populations. A population of A. uncinatum from Ohio shows the greatest differentiation from other populations. Therefore, both isozyme and RAPD data concur with the recent treatment of A. noveboracense and A. columbianum as a single species.     

五种齿突蟾在横断山南潜在地理分布预测

由于栖息地质量下降,近年来齿突蟾属物种种群数量急剧减少,明确齿突蟾属物种空间分布,是监测、管理、保护齿突蟾属物种的基础。横断山区可能是齿突蟾属的起源中心和分化中心,但齿突蟾属在横断山区的地理分布格局尚不明确。利用优化后Maxent模型,首次预测西藏齿突蟾Scutiger boulengeri、刺胸齿突蟾Scutiger mammatus、胸腺齿突蟾Scutiger glandulatus、圆疣齿突蟾Scutiger tuberculatus、贡山齿突蟾Scutiger gongshanensis 5种高海拔齿突蟾属物种在横断山南生物多样性保护优先区域的潜在地理分布,并分析其与环境因子的关系。结果显示,5种齿突蟾属物种在横断山南的潜在地理分布格局存在差异,西藏齿突蟾主要分布在横断山南的北部,圆疣齿突蟾主要分布在横断山南东北部的四川省境内,贡山齿突蟾主要分布在横断山南的西南部,刺胸齿突蟾和胸腺齿突蟾的潜在分布格局较为相似,在横断山南的中部、西北部地区都有较多分布,但胸腺齿突蟾潜在分布区更为碎片化。另外,横断山南北部地区的齿突蟾属丰富度明显高于南部地区。环境变量贡献率和刀切法结果显示温度因子和降水因子是决定横断山南齿突蟾属潜在分布的主要因素,最冷季降水量对西藏齿突蟾、贡山齿突蟾、圆疣齿突蟾潜在分布有重要影响,但它们对最冷季降水量的偏好存在差异。此外,研究也显示,通过评估潜在的Maxent参数组合,选择最佳的Maxent模型是有效且必要的。     

Karyotype and cytogeography of the genus Heracleum (Apiaceae) in the Hengduan Mountains

Abstract In the present study, the karyotypes of 34 populations belonging to 11 species and one variety of Heracleum from the Hengduan Mountains in China were examined. Chromosome numbers and the karyotypes of three species (H. souliei, H. kingdoni, and H. wenchuanense) are reported for the first time, as are the karyotypes of H. moellendorffii and H. henryi (tetraploid). Populations of H. candicans, H. franchetii, and H. kingdoni in the Hengduan Mountains were found to consist of a mixture of diploid and tetraploid plants. Except for four species of Heracleum, namely H. candicans, H. franchetii, H. henryi, and H. kingdoni, which have both diploid and tetraploid karyotypes, all other species of Heracleum are were found to be diploid. All karyotypes were found to belong to the 2A type of Stebbins, with the exception of H. candicans var. obtusifolium, which belongs to 2B, and H. hemsleyanum and H. franchetii (Mt. Dujuan, Daocheng, Sichuan, China), which belong to 1A. There was only a slight difference in the karyotype asymmetry index, which suggests a close kinship for species of Heracleum and that the entire phylogenetic development of Heracleum is relatively primitive. Species that exhibited advanced morphological features were also more advanced in karyotype structure, with the order of karyotype evolution being 1A→2A→2B. This phenomenon indicates that the species distributed in the Hengduan Mountains have not diverged completely and that the Hengduan Mountains are a relatively young and active area for the evolution of Heracleum. Polyploidization in Heracleum may be an important evolutionary mechanisms for some species, generating diversity. The biological attributes, distribution range, and the geological history of the genus have all played a part in accelerating the evolution through polyploidization or aneuploidization. It is known that as the distribution latitude of Heracleum decreases from north to south, the chromosome number, ploidy level, and asymmetry structure appear to increase. In the Hengduan Mountains, these tendencies are also evident. Finally, based on all the available cytogeographic data, we speculate that the more advanced tetraplont or aneuploid species of Heracleum in India may be derived from early diplont species that were distributed in the Caucasus region and Hengduan Mountains. The dispersal of Heracleum was from Eurasia to India, because this correlates with the emergence of the Himalayan Mountains through tectonic movement. Thus, the Hengduan Mountains are not only a center of diversity for Heracleum, but also a center of active speciation in modern times.     

Molecular and morphological analyses of EuropeanAconitum species (Ranunculaceae)

The systematics of the yellow-floweredAconitum lycoctonum species complex (Ranunculaceae) has long been considered difficult because of high morphological variability and hypothesized hybridization. To elucidate the systematic value of these taxa, and to study their phylogeny in a broader context, we studied all species of the subgenusLycoctonum from Europe and the Caucasus Mountains and the European species of the subgen.Aconitum. We sequenced the ITS region and the chloroplast intergenic spacerpsbA-trnH and conducted parsimony analyses. We also measured morphological characters from 19A. lycoctonum populations. The two subgenera in Europe are clearly separated but hybridization between subgenera might have taken place in the past. Sequence variation withinA. lycoctonum and among all taxa of subgenusLycoctonum was very low and probably the result of recent speciation, leading to one morphologically highly variable species in which color morphs could be seen as subspecies. There was high morphological variability within and among populations and the morphological characters have no value as systematic characters.     

Genetic Diversity Analysis of Starchy Curcuma Species Using RAPD Markers

The genetic variability in starchy Curcuma species was assessed using Random Amplified Polymorphic DNA (RAPD) technique. The RAPD pattern generated by 20 primers revealed a high degree of polymorphism. A total of 274 bands were generated of which 264 were polymorphic. All the species were separated into 3 clusters using UPGMA. C. aromatica, C. leucorrhiza, and C. brog formed a cluster within which C. longa and C. zedoaria formed a subgroup. C. harita was genetically distinct from all the other Curcuma species. Since it is difficult to distinguish different species by leaf morphology, the RAPD pattern has high utility in identification of starchy Curcuma species.     

Evaluation of genetic relationship in Typhonium species through random amplified polymorphic DNA markers

Studies were undertaken to identify genetic relationships in three species of Typhonium and to evaluate the genetic variance within populations of Typhonium trilobatum, Typhonium roxburghii and Typhonium flagelliforme by using random amplified polymorphic DNA (RAPD) markers. A total of 193 distinct DNA fragments ranging from 0.2 to 3.2 kb, were amplified using 22 selected random decamer primers. The cluster analysis indicated that the three species of Typhonium formed two clusters: the first one consisted of T. trilobatum and T. roxburghii, the second one was represented by T. flagelliforme. A maximum similarity of 63 % was observed in T. trilobatum and T. roxburghii. T. flagelliforme shared up to 43 % similarity with T. trilobatum and T. roxburghii. The closest genetic distance was obtained within populations of different Typhonium species.     

Genetic relationships among NE Turkish Lilium L. (Liliaceae) species based on a random amplified polymorphic DNA analysis

Random amplified polymorphic DNA (RAPD) fingerprinting was used to study species boundaries in six closely related NE Turkish Lilium (Liliaceae) taxa of the section Liriotypus. The investigated taxa were L. ciliatum, L. akkusianum, L. ponticum, L. kesselringianum, L. armenum, and L. szovitsianum. Of the 108 primers screened, 11 provided polymorphic and reproducible bands. A total of 93 polymorphic bands were scored for 122 individuals from 18 populations of the six Lilium taxa and principle coordinate analysis and neighbour-joining cluster analysis based on these RAPD profiles were performed. The results demonstrate a clear distinction between the two species L. ciliatum and L. akkusianum, and the other four species. While populations of the two species groups are found to be allopatrically distributed, the two species groups overlap in their geographical ranges. Analysis of molecular variance (AMOVA) indicated that nearly half of the total molecular variance is found within the individual populations and that the molecular variance among species is as high as the variance within the individual species, indicating that genetic differentiation of the species is rather weak.     

云南高山姬鼠头骨的几何形态学研究

表型可塑性是指同一基因型在不同环境条件下而产生多种不同表现型的反应能力。哺乳动物头骨形态的变化是在进化过程中出现的重要表型特征之一。云南省地势西高东低、海拔变化剧烈、是古北界寒带物种南迁,中南半岛热带物种北移的交汇地,自然环境的地带性和非地带性变化明显,栖息在该地区的动物为了适应多样的环境,可能出现不同的表型分化。高山姬鼠(Apodemus chevrieri)为我国特有种,主要分布在中国西南的横断山及其附近地区,是研究表型与生态适应之间关系的理想物种。为研究在云南特殊生态环境下物种的微进化,采用几何形态学的方法测量云南不同地区高山姬鼠头骨形态的变异,采用线粒体细胞色素b(Cytochrome b,Cyt b)基因和线粒体控制区(Mitochondrial control region,D-loop)探讨基因型的变化。结果显示:昆明种群与横断山种群(中甸、剑川、丽江)间发生了明显的形态变异和遗传分化,高山姬鼠头骨背面和腹面,横断山种群和滇中昆明种群很少重叠,这两个种群变异较大;但头骨的侧面和下颌侧面变异不明显。因此,分布于云南地区的高山姬鼠可能正处于生态物种形成的早期阶段。     

Intraspecific molecular variation inHieracium sect.Alpina (Asteraceae), an apomictic group

Intraspecific variation of four agamospecies ofHieracium sect.Alpina was studied using RAPD and isozyme techniques. No variation in either multiprimer RAPD or multi-enzyme phenotypes was observed withinH. holosericeum, suggesting that this widespread species consists of only a single genotype. A low level of within-population isozyme variation was seen inH. tenuifrons andH. calenduliflorum, the origin of which appears to be consistent with somatic mutation. Most isozyme and all RAPD variation in these two species was partitioned between populations. A strong correlation with geography suggests that its cause may be due to polytopic (-polyphyletic?) origin or perhaps to mutation and dispersal. The most variable species wasH. alpinum, in which isozyme variation occurred mostly within populations rather than between them, suggesting occasional sexual events or that the parents ofH. alpinum were heterozygous. RAPD variation in this species, in contrast, was partitioned between Scottish and Swiss populations, suggesting the existence of geographical races.     

中国假百合属亲缘关系及假百合遗传多样性的ISSR分析

利用7条ISSR引物对中国假百合属植物24个居群255个个体的遗传多样性进行了初步探讨。结果表明:(1)UPGMA聚类结果显示,假百合(Notholirion bulbuliferum)、钟花假百合(N.campanulatum)与大叶假百合(N.macrophyllum)分别聚为三支,在分子水平上出现了明显的分化,揭示三者是独立物种。(2)N.bulbuliferum×N.campanulatum与钟花假百合聚为一支,这可能与其母系遗传和极强的无性繁殖体系有关,但两者间基因流(N_m=0.216 0)很低,假百合与N.bulbuliferum×N.campanulatum的基因流(N_m=0.144 9)也极低,说明N.bulbuliferum×N.campanulatum正在分化。(3)假百合种下各居群间按照地理结构可明显分为4支,经Mantel检测表明存在显著的谱系地理结构(r=0.410,P=0.01)。(4)AMOVA分析显示,假百合居群间遗传变异为77.12%(P<0.01),而居群内为22.88%(P<0.01),同样表明居群间遗传分化大于居群内。研究结果揭示了横断山区假百合属植物的亲缘关系,从分子水平上为其鉴定提供了依据;并从遗传结构上为药用植物假百合的可持续利用和开发奠定了理论基础。     

Morphological variation pattern of Aquilegia ecalcarata and its relatives

Aquilegia ecalcarata Maxim. is the only spurless species within the genus Aquilegia and comprises a monophyletic clade with A. yabeana Kitag., A. kansuensis Brühl, and A. rockii Munz. Our previous study on the genetic diversity of those four species revealed that the populations of A. ecalcarata can be divided into two groups, indicating possible genetic difference within A. ecalcarata. However, it is not clear whether the genetic difference is related to the morphological variation among species and groups of A. ecalcarata populations. To answer that question, the morphological variation patterns based on 22 floral and 19 vegetative traits from 42 populations, covering the entire distribution of A. ecalcarata and its relatives, were analyzed in the present study. The result showed that: (i) the differences among the four species were reflected in the floral rather than the vegetative traits; (ii) populations of A. yabeana and A. rockii fell into one cluster each, and each of the six clusters occupied its own distribution range; (iii) one of two A. ecalcarata clusters fell into a subgroup and shared common floral traits with A. rockii; (iv) the individuals of A. ecalcarata form. ecalcarata and form. semicalcarata were often mixed in the same population; and (v) the populations of A. kansuensis were split into two clusters, which differed obviously in floral traits. These results will provide an important morphological basis for the redefinition of species and lay a foundation for the further exploration of the “spurless” A. ecalcarata.     

云南蚤类区系及分布特征

【目的】在收集汇总以往野外调查资料以及已经发表的历史文献资料基础上,归纳云南蚤类的分布特点,分析云南地理小区对蚤类的影响。【方法】在实地采集和补充文献记录基础上,对收集到的蚤类进行系统分类整理;运用SPSS13.0的主成分分析及系统聚类对数据进行处理。【结果】本研究共记录云南蚤类9科45属144种,其中古北种15种,东洋种 108种,东洋、古北两界兼有种15种,广布种6种。144种蚤中,属于云南特有蚤类有75种;横断山中部小区记载蚤类122种,横断山南部小区26种,滇东高原小区41种,滇西高原小区28种,滇南山地小区25种。主成分分析和聚类分析结果基本一致,均可聚为两类：一类为横断山南部小区、滇南山地小区、滇西高原小区及滇东高原小区; 另一类为横断山中部小区。【结论】云南省蚤类分布明显受到地理小区环境的影响。云南省蚤类昆虫种类繁多,蚤类的自然分布受到云南省特定地形地貌影响。     

Species reassessment congruent with the phylogeographical study of the Biston falcata species group

The Himalaya, Hengduan, and Qinling mountains are very important mountain systems in China. Because they are geographically adjacent, species distributed in these three mountains usually have close relationships, but species/populations distributed in the first two mountain systems have higher genetic diversity and more complex phylogenetic structures than those found in the Qinling Mountains. In this study, we used molecular and morphological methods to explore the phylogeographical structure, species differentiation, and evolutionary history of the Biston falcata species group, which is widely distributed in the Himalaya, Hengduan, and Qinling mountains. Five species were identified; B. quercii is distributed in the Qinling Mountains and its adjacent areas, and the other four species are mainly distributed in the Himalaya and Hengduan Mountains. Four gradual species differentiation events occurred at approximately 3.07, 1.75, 1.50, and 1.30 Ma, corresponding to the stage with strong geological movements of the Qinghai‐Tibet Plateau. The low land between the Qinling and the northern Hengduan Mountains, the Sichuan Basin, the low land between the eastern and central Himalaya Mountains and the Mekong‐Salween Divide are four geographical barriers that resulted in multiple speciations. In addition, long‐range dispersal as another mechanism for speciation is non‐negligible. In spite of failing to confirm the ancestral area, our study provided direct evidence of the speciation induced by the geographical barriers and adaptation to the plateau.     

四川西部濒危植物桃儿七遗传多样性的RAPD分析

桃儿七是一种具有重要药用价值的珍稀濒危植物。采用RAPD分子标记技术,对在四川西部地区的桃儿七7个自然种群的遗传多样性水平和遗传结构进行了分析。用12个RAPD引物对7个种群共140个样品进行了扩增,共得到111条清晰带,其中32条具有多态性,在物种水平上多态位点百分率(PPB)为28.83%,在种群内的多态位点百分率变动幅度为4.50%至16.22%。同其它一些濒危植物相比,桃儿七种群具有较低的遗传多样性(He=0.0622,Ho=0.0987)。7个自然种群间出现了很强的遗传分化,分化指数接近70%。种群间的基因流低(Nm=0.2240)。造成上述结果的可能原因是与桃儿七的繁育方式和有限的基因流等因素有关。应将遗传多样性相对较高的松潘县牟尼沟种群作为原位保护的核心种群进行保护,尽量保护所有现有种群。     

Chromosome counts and karyotypes in Chaetoseris and Stenoseris (Asteraceae‐Cichorieae) from the Hengduan Mountains of SW China

Abstract Chaetoseris and Stenoseris are two morphologically close genera from the tribe Cichorieae of the sunflower family and they are endemic in alpine eastern Himalayas to the Hengduan Mountains of SW China. Mitotic chromosome numbers and karyotypes are reported for 12 populations representing eight species of Chaetoseris and two species of Stenoseris from the Hengduan Mountains region. Eight species are new and the other two provide confirmation of previous reference. All Chaetoseris and Stenoseris taxa are diploidy with 2n= 16 and their basic number is tentatively suggested as x= 8. Karyotypes of Chaetoseris and Stenoseris are similar to each other with 2A and 2B for the former and 2A for the latter. Cytological data of chromosomal numbers and karyotypes support a close relationship of the two genera. Currently no polyploids are found for these two genera and it seems that polyploidization has played a minor role in their evolutionary speciation in the Hengduan Mountains region.     

Speciation slowing down in widespread and long-living tree taxa: insights from the tropical timber tree genus Milicia (Moraceae)

The long generation time and large effective size of widespread forest tree species can result in slow evolutionary rate and incomplete lineage sorting, complicating species delimitation. We addressed this issue with the African timber tree genus Milicia that comprises two morphologically similar and often confounded species: M. excelsa, widespread from West to East Africa, and M. regia, endemic to West Africa. We combined information from nuclear microsatellites (nSSRs), nuclear and plastid DNA sequences, and morphological systematics to identify significant evolutionary units and infer their evolutionary and biogeographical history. We detected five geographically coherent genetic clusters using nSSRs and three levels of genetic differentiation. First, one West African cluster matched perfectly with the morphospecies M. regia that formed a monophyletic clade at both DNA sequences. Second, a West African M. excelsa cluster formed a monophyletic group at plastid DNA and was more related to M. regia than to Central African M. excelsa, but shared many haplotypes with the latter at nuclear DNA. Third, three Central African clusters appeared little differentiated and shared most of their haplotypes. Although gene tree paraphyly could suggest a single species in Milicia following the phylogenetic species concept, the existence of mutual haplotypic exclusivity and nonadmixed genetic clusters in the contact area of the two taxa indicate strong reproductive isolation and, thus, two species following the biological species concept. Molecular dating of the first divergence events showed that speciation in Milicia is ancient (Tertiary), indicating that long-living tree taxa exhibiting genetic speciation may remain similar morphologically.     

Two new species of Panorpa (Mecoptera,Panorpidae) from the Hengduan Mountains in Yunnan,China

Two new species of the genus Panorpa Linnaeus, 1758 are described from the Hengduan Mountains in Yunnan, China. Panorpa diqingensis sp. n. can be distinguished from its congeners by vertex dark brown, and occiput yellow; pleura dark brown; gonocoxites bearing a cluster of stout black setae on inner apex; parameres S-shaped in male genitalia. Panorpa dispergens sp. n. can be readily differentiated from other Panorpa species by apical band with a hyaline spot in wings; gonocoxites with a cluster of stout dark brown setae and 3–5 long setae on inner apex; female medigynium with a pair of basal plates. The number of Panorpa species in Yunnan is raised to six. A key to Yunnan species of Panorpa is presented.     

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。