A new species of Saussurea (Asteraceae) from Tibet and its systematic position based on ITS sequence analysis

A new species of Saussurea, S. erecta S. W Liu, J. T Pan A J. Q. Liu sp. nov., is described from Tibet. It resembles S. kingii but may be distinguished by having distinct stems and glabrous achenes. Saussurea kingii was placed in sect. Pseudoeriocoryne of subgen. Eriocoryne; this section was circumscribed by acaulescence and an inflorescence with congested capitula surrounded by a rosette of leaves. The discovery of S. erecta with distinct stems, cauline leaves and corymbose capitula blurred the delimitation of sect. Pseudoeriocoryne and suggested that the section may be polyphyletic. Both the close relationship and the significant difference between S. erecta and S. kingii were confirmed by analyses of nrDNA ITS sequences. The resulting phylogenies based on ITS data further suggest that Saussurea sect. Pseudoeriocoryne, as traditionally defined, does not constitute a monophyletic group. The rapid radiation and speciation of Saussurea in the Qinghai-Tibetan Plateau, as inferred from ITS phylogeny, are discussed. (c) 2005 The Linnean Society of London.

Phylogenetic analyses of Saussurea sect. Pseudoeriocoryne (Asteraceae : Cardueae) based on chloroplast DNA trnL-F sequences

The genus Saussurea is distributed mainly in the temperate and subarctic regions of Eurasia and consists of about 300 species classified into six subgenera and 20 sections. Sect. Pseudoeriocoryne in the subgenus Eriocoryne comprises four species, and is delimited mainly by acaulescence and an inflorescence with congested capitula surrounded by a rosette of leaves. All of these species are endemic to the and Qinghai-Tibet Plateau. Sequences from the chloroplast DNA trnL-F region were obtained for the four species in this section and 26 other species from four subgenera of Saussurea to resolve phylogenetic relationships among these species and to determine whether the shared characters that define sect. Pseudoeriocoryne are synapomorphic or were acquired by convergent evolution. The resulting phylogenies indicated that Saussurea sect. Pseudoeriocoryne as traditionally defined does not constitute a monophyletic group and that each of its species belongs to separate clades. Furthermore, none of these species showed a close relationship with the other species of subgenus Eriocoryne. Our results further indicated that none of the investigated subgenera are monophyletic, and that species from different subgenera clustered together. All these conclusions are provisional and their confirmation would require stronger phylogenetic support. Two possible explanations are suggested for low sequence divergence, poor resolution of internal clades and clustering of species with the rather distinct morphology of Saussurea detected in the present study. The first is rapid radiation and diversification triggered by fast habitat fragmentation due to the recent lifting of the Qinghai-Tibet Plateau and the Quaternary climate oscillations. This could have led to rapid morphological divergence while sequences diverged very little, and also caused the convergent acquisition of similar characteristics in unrelated lineages due to similar selection pressures. The second possible explanation is that both introgressive hybridization and reticulate evolution might have caused the transferring of cpDNA sequences between morphologically dissimilar species, thus leading to homogenization of sequences between lineages. (C) 2004 Elsevier Ltd. All rights reserved.

Uniformity of karyotypes in Ligularia (Asteraceae : Senecioneae), a highly diversified genus of the eastern Qinghai-Tibet Plateau highlands and adjacent areas

Ligularia, a highly diversified genus in the eastern Qinghai-Tibet Plateau and adjacent areas, was chosen as a suitable subject in which to study speciation patterns in this 'hot spot' area at the chromosomal level. Chromosome numbers and karyotypes were studied in 23 populations of 14 species, most of which are endemic to this area. The basic number x = 29 was confirmed for all species. Ligularia virgaurea was found to have diploid and triploid cytotypes, 2n = 58 and 87. Other species are only diploid, with 2n = 58. The karyotypes of all populations within any species, and all species spanning most sections and covering most of the morphological range in Ligularia, are very similar to each other, belonging to type 2A according to Stebbin's classification. This karyotype was also found in its close allies, e.g. Cremanthodium, Ligulariopsis, Parasenecio, and Sinacalia. Aneuploid reduction of chromosome number from 2n = 60 to 58 and karyotypic variation was found in Ligularia and its allies. Such a chromosomal pattern with few polyploids infers that variation of karyotype structure at the diploid level seems to be the predominant feature of chromosomal evolution in this group and sympatric speciation via hybridization and polyploidization has played a minor role in its species diversity. (C) 2004 The Linnean Society of London

Molecular phylogeny and biogeography of the Qinghai-Tibet Plateau endemic Nannoglottis (Asteraceae)

All taxa endemic to the Qinghai-Tibet Plateau are hypothesized to have originated in situ or from immediately adjacent areas because of the relatively recent formation of the plateau since the Pliocene, followed by the large-scaled biota extinction and recession caused by the Quaternary ice sheet. However, identification of specific progenitors remains difficult for some endemics, especially some endemic genera. Nannoglottis, with about eight species endemic to this region, is one such genus. Past taxonomic treatments have suggested its relationships with four different tribes of Asteraceae. We intend to identify the closest relatives of Nannoglottis by evaluating the level of monophyly, tribal delimitation, and systematic position of the genus by using molecular data from ndhF gene, trnL-F, and ITS region sequences. We find that all sampled species of Nannoglottis are a well-defined monophyly. This supports all recent taxonomic treatments of Nannoglottis, in which all sampled species were placed in one broadly re-circumscribed genus. Nannoglottis is most closely related to the Astereae, but stands as an isolated genus as the first diverging lineage of the tribe, without close relatives. A tentative relationship was suggested for Nannoglottis and the next lineage of the tribe was based on the ITS topology, the "basal group," which consists of seven genera from the Southern Hemisphere. Such a relationship is supported by some commonly shared plesiomorphic morphological characters. Despite the very early divergence of Nannoglottis in the Astereae, the tribe must be regarded to have its origin in Southern Hemisphere rather than in Asia, because based on all morphological, molecular, biogeographical, and fossil data, the Asteraceae and its major lineages (tribes) are supposed to have originated in the former area. Long-distance dispersal using Southeast Asia as a steppingstone from Southern Hemisphere to the Qinghai-Tibet Plateau is the most likely explanation for this unusual biogeographic link of Nannoglottis. The 23-32-million-year divergence time between Nannoglottis and the other Astereae estimated by DNA sequences predated the formation of the plateau. This estimation is further favored by the fossil record of the Asteraceae and the possible time of origin of the Astereae. Nannoglottis seems to have reached the Qinghai-Tibet area in the Oligocene-Eocene and then re-diversified with the uplift of the plateau. The molecular infragenetic phylogeny of the genus identifies two distinct clades, which reject the earlier infrageneric classification based on the arrangement of the involucral bracts and the length of the ligules, but agree well with the habits and ecological preferences of its current species. The "alpine shrub" vs. "coniferous forest" divergence within Nannoglottis was estimated at about 3.4 million years ago when the plateau began its first large-scale uplifting and the coniferous vegetation began to appear. Most of the current species at the "coniferous forest" clade of the genus are estimated to have originated from 1.02 to 1.94 million years ago, when the second and third uprisings of the plateau occurred, the climate oscillated and the habitats were strongly changed. The assumed evolution, speciation diversity, and radiation of Nannoglottis based on molecular phylogeny and divergence times agree well with the known geological and paleobotanical histories of the Qinghai-Tibet Plateau. (C) 2002 Elsevier Science (USA). All rights reserved.

Karyological studies on the Sino-Himalayan genus, Cremanthodium (Asteraceae : Senecioneae)

Twenty-two populations of seven species of Cremanthodium from high altitude regions of western China were observed karyologically. C. ellisii, C. microglossum, C. brunneo-pilosum, C. stenoglossum, C. discoideum and C. lineare all had the same chromosome number of 2n=58 whereas C. humile had 2n=60. All chromosome numbers of these species are documented here for the first time. The basic number of x=30 is new for this genus. The karyotypes of all species belong to 2A type according to Stebbins' asymmetry classification of karyotypes. Two basic chromosome numbers, x=30 and x=29 in Cremanthodium, correspond exactly to two branching patterns in this genus, sympodial versus monopodial. The systematic and taxonomic statuses of the sympodial species need further study. The karyomorphological data provide no support to the sectional subdivision in Cremanthodium. (C) 2001 The Linnean Society of London.

东亚千里光族款冬亚族（菊科）的系统学

千里光族是菊科中最大的一个族，约含有120属3400多种，全世界分布。千里光族的族下分类与系统发育一直是菊科研究中的难点。Jeffrey&Chen(1984)和千里光亚族(7属)。自此以后，有关东亚款冬亚族的范畴、亚族内各属的系统位置与亲缘关系就成了千里光族系统学研究中争论的焦点，至今仍未有定论。陈艺(1997)在东亚款冬亚族内发表假橐吾属新属。根据Jeffrey&Chen(1984)的概念，东亚款冬亚族现含13属。东亚款冬亚族目前存在的系统学问题主要有：(1)毛冠菊属是否与款冬亚族内的款冬属及蜂斗菜属近缘;(2)狗舌草亚族所辖3属是否是款冬亚族的自然成员;(3)东亚款冬亚族13属的系统位置及其亲缘关系;(4)东亚款冬亚族的影响范畴。 针对上述问题，本文以发现新的系统学证据为主要目的，较为全面地研究了东亚款冬亚族及其相关类群的外部形态学，首次报道了18属45种植物的花部微观性状，19属64种105个居群的核形态学特征，10属15种植物花粉壁的超薄结构及13属28种植物花粉的外壁纹饰，3种植物的胚胎发育和6种植物的ITS基因序列;并对所获得的证据进行了比较与综合分支分析。根据所获系统学资料，对东亚款冬亚族的上述系统学问题进行了讨论。主要实验结果及结论总结如下。 1. 外部形态 较为全面地观察与分析了东亚款冬亚族及其相关类群外部形态性状及其演化趋势，指出根状茎分枝方式、幼叶出土性状、花部茎基部叶是否宿存、叶柄是否纤维状宿存、花序托空心或实心、花药基部结构等不被以前作者重视或忽视的形态学性状可能具有重要的系统学价值。外部形态上，毛冠菊属与千里光族内的千里光属与款冬属等类群没有亲缘关系，而与紫菀族存在一定的渊源;狗舌草亚族则介于千里光亚族和款冬亚族之间;东亚款冬亚族13属可依据外部形态划分为6个属群。 2. 花部微观性状 东亚款冬亚族及其相关类群的花部微观性状主要包括：花药顶端不育组织、花药药室内壁、花药基部、花药领、花柱分枝顶端、花柱毛被、花柱分枝内表面构型、果柄细胞列数等。花部微观性状研究表明：毛冠菊属似乎应作为紫菀族中的一个孤立属;狗舌草亚族在某些衍征上与千里光亚族接近，而它的原始性状则类似于款冬亚族;根据花部微观性状对东亚款冬亚族的13个属划分出的自然属群与形态性状分析所得结果基本一致。 3. 核形态学 毛冠菊属的染色体基数为x=9，与紫菀族相同：而与千里光族和旋覆花族的染色体x=10存在显著的区别。狗舌草亚族中新发现2n=24、60、72三个染色体数目，亚族内染色体基数之间的演化关系不清楚，利用染色体基数来探讨狗舌草亚族所辖属的系统位置和属间关系仍还比较困难。东亚款冬亚族的染色体基数主要有x=30、29、28、27、26;款冬亚族中普遍存在的染色体基数x=30可能有3种来源;核型可分为2A、3A、2B三大类型，虽然三种类型之间的染色体基数(x=30)有可能是不同起源的，但从整个千里光族来看，3A、2B应是较进化的类型。根据核形态学特征，东亚款冬亚族可分为款冬型、蟹甲草型和大吴风草型三类。在蟹甲草型染色体进化支上，存在染色体的非整倍性下降趋势。东亚款冬亚族存在的种内多倍性可能与不同的生境有关。 4. 花粉学 通过对东亚款冬亚族及其相关类群的花粉壁超微结构与扫描电镜观察，发现花粉壁超微结构存在“千里光型”和“向日葵型”两种类型。扫描电镜下，花粉外壁均为刺状纹饰，但在刺的长短、刺部是否膨大、刺基膨大上的纹饰以及刺基之间的纹饰等方面存在差别。毛冠菊属的花粉性状组合支持将其置于紫菀族中。狗舌草亚族的花粉性状与款冬亚族的有些属相似。款冬亚族各属的花粉生状组合所划分的属群与形态学、核形态学等方面的研究结果较为一致。 5. 胚胎学 款冬、掌裂橐吾和珠毛蟹甲草共同拥有药壁发育双子叶型;绒毡层发育属“The Comos bipinnatus”型;成熟花粉为3细胞型;单珠被，薄珠心，倒生胚珠;具发育相似的珠被绒毡层：胚橐发育4孢子型等胚胎学特征，但它们在胚囊发育方式、反足细胞数目、发殖物候上还存在区别。三属代表植物的胚胎学研究支持将它们作为同一亚族的成员。 6. 分子数据 利用本研究测得的华千里光属4种、狗舌草属1种和大吴风草属的ITS－1和ITS－2基因序列，结合从Genbank中得出的千里光族5属8种植物得ITS序列，用PAUP软件进行分析。研究结果表明，染色体基数、花粉超微结构、叶型和叶脉等性状在ITS分支图上存在趋同进化;华千里光属和狗舌草属不是单系群。并讨论了一些分类群的亲缘关系。 7. 分支分析 选择东亚款冬亚族和狗舌草亚族各属的代表种作为终端代表类群进行分支分析;分支分析表明染色体基数、与头状花序有关的性状、叶型及叶脉等性状存在较多的平行与逆转进化，而根状茎分枝类型、花部微观性状以及花粉性状则表现出较大的一致性。根据分支结果，作者认为应建立包括狗舌草亚族在内的广义东亚款冬亚族。 8. 系统学 毛冠菊属的性状组合支持将其置于紫菀族，排除在千里光族之外。广义东亚款冬亚族可分为7个属群：蟹甲草属群(包括蟹甲草属，华蟹甲草属，小蟹甲草属，兔儿伞属，蟹甲木属)、毛柱菊属群(毛柱菊属)、橐吾属群(包括橐吾属，垂头菊属，假橐吾属)、狗舌草属群(狗舌草属，华千里光属，羽叶千里光属)、多榔菊属群(多榔菊属)、大吴风草属群(大吴风草属)、款冬属群(款冬属，蜂斗菜属)。总结与讨论了各属群的系统位置、亲缘关系与存在的分类学问题。描述了华千里光属1新种。

滇金足草、凋缨菊和长喙吴萸的化学成分研究

本论文对滇金足草（Goldfussia yunnanensis）、凋缨菊（Camchaya loloana）和长喙吴萸（Evodia vestia）的化学成分进行了研究，通过色谱分离得到40个化合物。主要基于波谱数据鉴定了它们的结构，其中10个为新化合物。 1．从滇金足草地上枝叶的95%乙醇提取物中共分离鉴定了16个化合物：泽漆内酯A（1）、18-羟基泽漆内酯A（2）、18-氧代泽漆内酯A（3）、18-羟基-3-O-β-D-吡喃葡萄糖-泽漆内酯A（4）、3-O-β-D-吡喃葡萄糖-泽漆内酯A（5）、3-O-β-D-吡喃半乳糖-泽漆内酯A（6）、6-E-肉桂酰哈巴俄苷（7）、E-哈巴俄苷（8）、5，6-异亚丙二氧基哈巴俄苷（9）、β-谷甾醇（10）、β-胡萝卜苷（11）、齐墩果酸（12）、肉桂酸（13）、麦角固醇（14）、硬脂酸（15）和丁二酸（16）。其中2-7为新化合物。5，6-异亚丙二氧基哈巴俄苷（9）以人工产物形式得到。 2．从凋缨菊地上枝叶的95%乙醇提取物中分离并鉴定了13个化合物：凋缨菊内酯A~C (17-19)、1β-乙酰基凋缨菊内酯C(20)、b-谷甾醇（10）、β-胡萝卜苷（11）、羽扇豆醇（21）、桦木醇（22）、桦木酸（23）、芥子醇（24）、紫丁香苷（25）、咖啡酸（26）和熊果酸（27）。其中化合物17-20为桉叶烷内酯类新化合物。化合物17、18、20对细胞株HepG2的GI50依次为7.80、7.08、4.99 µg/mL。 3．从长喙吴萸（E. vestia）地上枝叶的95%乙醇提取物中分离并鉴定了13个化合物：佛手内酯（28）、花椒毒素（29）、异茴芹内酯（30）、七叶内酯（31）、东莨宕素（32）、瑞香素（33）、异紫花前胡内酯(34)、茵芋碱（35）、山刈碱（36）、白鲜碱（37）、黄柏酮（38）、柠檬苦素（39）和对羟基苯甲醛（40）。 4．综述了1990—2007年期间从菊科植物中发现的桉叶烷-12，6内酯的化学结构、生物活性、生物转化及化学合成方面的研究进展。 Phytochemical investigation on Goldfussia yunnanensis, Camchaya loloana, and Evodia vestia, led to the isolation of 40 compounds, 10 of which were new ones. 1. Six new compounds were isolation from 95% ethanolic extract of the aerial parts of G. yunnanensis, and identified as 18-hydroxyhelioscopinolide A (2), 18-oxohelioscopinolide A (3), 18-hydroxy-3-O-β-D-glucopyranosylhelioscopinolide A (4), 3-O-β-D-glucopyranosylhelioscopinolide A (5),3-O-β-D-Galactopyranosyl helioscopinolide A (6), 6-O-trans-cinnamoyl E-harpagoside (7). The known compounds isolated were helioscopinolide A (1), E-harpagoside A (8), 5,6-isopropylidene E-harpagoside A (9), β-sitosterol (10), β-daucosterol (11), oleanolic acid (12), cinnamic acid (13), ergosterol (14), stearic acid (15) and succinic acid (16). Compound 9 was an artifact. 2. Four new compounds, loloanolides A – C (17 - 19) and 1β-acetoxy-loloanolide C (20), were isolation from 95% ethanolic extract of the aerial parts of C. loloana. The known ones were β-sitosterol (10), β-daucosterol (11), lupeol (21), betulin (22), betulinic acid (23), sinapyl (24), syringin (25), caffeic acid (26) and ursolic acid (27). The GI50 values of compounds 17, 18 and 20 to HepG2 cell line were 7.80, 7.08 and 4.99 µg/mL, respectively. 3. Thirteen were isolated from 95% ethanolic extract of the aerial parts of E. vestia for the first time. They were determined to be bergapten (28), xanthotoxin (29), isopimpinellin (30), esculetin (31), scopoletin (32), daphnetin (33), marmesin (34), skimmianine (35), confusameline (36), dictamine (37), obacunone (38), limonin (39) and p-hydroxy phenyl aldehyde (40). 4. The structures, biological activities, biotransformation and chemical syntheses of eudesmane-12, 6-olides from the Asteraceae during 1990-2007 were reviewed.

Origin of the Qinghai-Tibetan Plateau endemic Milula (Liliaceae): further insights from karyological comparisons with Allium

Milula, a monotypic genus endemic to the Qinghai-Tibetan Plateau, was found to be nested deeply within Allium by the molecular phylogeny despite the aberrant morphology. It remains unknown what had contributed to the rapid evolution of morphology and origin of this exceptional species. In contrast to a previous report of its karyotypes with 2n = 16 = 8M+8SM (2SAT), similar to most species of Allium, a rather different karyotype, 2n = 20 = 4M +10SM+6T (2SAT), was found in examined 31 individuals from 6 populations of M. spicata distributed in the central Tibet. Karyotypes of 7 Allium species occurring in the Qinghai-Tibetan Plateau were further reported. The basic number x = 8 was confirmed for all of them and their karyotypes consist mainly of metacentric and submetacentric chromosomes with rare subterminal and terminal chromosomes. The karyotype of M. spicata is distinctly different from that of most Allium species occurring in the plateau through a complete comparison of all available species in this region and adjacent areas. However, the same chromosome number and similar karyotypic structure were found in A. fasciculatum of Sect. Bromatorrhiza, indicating a possible close relationship between them. But this similarity is contradictory to the preliminary molecular phylogenetic analysis that Milula was closely related to A. cyathophorum of Sect. Bromatorrhiza with x=8, but the other species with x=10 and 11 in this section were clearly placed in the other clade. We therefore suggested that the paralleling evolution from x=8 to x=9, 10 and 11 with increasing asymmetry of karyotype possibly due to the chromosomal Robertsonian translocation might occur separately in the two recognized phylogenetic lineages of Allium. In addition to aneuploidy and following change of the chromosomal structures, the habitat isolation due to the recent uplift of the Qinghai-Tibetan Plateau and the Quaternary climatic oscillation, plays a greater role in origin of Milula and other endemic species (genera) with aberrant morphology from their progenitors.