Dysfunction of CYC2g is responsible for the evolutionary shift from radiate to disciform flowerheads in the Chrysanthemum group (Asteraceae: Anthemideae)

Evolutionary shifts among radiate, disciform and discoid flowerheads have occurred repeatedly in a number of major lineages across the Asteraceae phylogeny; such transitions may also appear within evolutionarily young groups. Although several studies have demonstrated that CYC2 genes partake in regulating floral morphogenesis in Asteraceae, the evolution of capitulum forms within a recently diverging lineage has remained poorly understood. Here, we study the molecular regulation of the shift from a radiate to a disciform capitulum within the Chrysanthemum group. This is a recently radiating group mainly comprising two genera, Chrysanthemum and Ajania, that are phylogenetically intermingled but distinct in flowerhead morphology: Chrysanthemum spp. with radiate capitula and Ajania spp. with disciform capitula. We found that the morphogenesis of zygomorphy in the marginal floret in Ajania was disrupted soon after floral primordium emergence; CYC2g, one of the CYC2 copies that was expressed prominently in the ray floret of Chrysanthemum was not expressed in flowerheads of Ajania. Weakening the expression of ClCYC2g in Chrysanthemum lavandulifolium led to the gradual transition of a ray flower toward the disc-like form. Molecular evolutionary analyses indicated that the disciform capitulum might have evolved only once, approximately 8 Mya, arising from dysfunction of the CYC2g orthologs. A 20-nt deletion, including a putative TATA-box of the Ajania-type CYC2g promoter, appeared to inhibit the expression of the gene. Considering the divergent habitats of Chrysanthemum and Ajania, we propose that the shift from radiate to disciform capitulum must have been related to changes in pollination strategies under selective pressure.     

Localization of 45S and 5S rDNA sites and karyotype of Chrysanthemum and its related genera by fluorescent in situ hybridization

The phylogenetic relationships among Chrysanthemum and its related genera (Anthemideae, Asteraceae) is poorly understood. In the present study, these relationships were investigated using 45S and 5S ribosomal DNA (rDNA)-targeted fluorescent in situ hybridization. The results showed that there were two 45S rDNA signals present in Crossostephium chinense, four 45S rDNA signals in Cercidiphyllum japonicum, Artemisia sieversiana, Artemisia annua and Artemisia absinthium, six 45S rDNA signals in Chrysanthemum boreale and Pyrethrum parthenium, eight 45S rDNA signals in Chrysanthemum nankingense, Chrysanthemum dichrum, Chrysanthemum lavandulifolium and Tanacetum vulgare, and ten 45S rDNA signals in Ajania przewalskii. For the 5S rDNA locus, two 5S rDNA signals were present in C. nankingense, C. dichrum, C. lavandulifolium, C. boreale, C. japonicum, C. chinense and P. parthenium, four in A. sieversiana, A. annua, A. absinthium and A. przewalskii, and six 5S in T. vulgare. In addition, karyotypes of the 12 species were investigated. From this study, we inferred that Chrysanthemum was closely related to Ajania, and that Chrysanthemum species originating from China and Japan may have evolved differently. These findings add a new level to the understanding of the phylogenetic relationships of Chrysanthemum and related genera.     

Phylogeny of the Genus Chrysanthemum L.: Evidence from Single-Copy Nuclear Gene and Chloroplast DNA Sequences

Chrysanthemum L. (Asteraceae-Anthemideae) is a genus with rapid speciation. It comprises about 40 species, most of which are distributed in East Asia. Many of these are narrowly distributed and habitat-specific. Considerable variations in morphology and ploidy are found in this genus. Some species have been the subjects of many studies, but the relationships between Chrysanthemum and its allies and the phylogeny of this genus remain poorly understood. In the present study, 32 species/varieties from Chrysanthemum and 11 from the allied genera were analyzed using DNA sequences of the single-copy nuclear CDS gene and seven cpDNA loci (psbA-trnH, trnC-ycf6, ycf6-psbM, trnY-rpoB, rpS4-trnT, trnL-F, and rpL16). The cpDNA and nuclear CDS gene trees both suggest that 1) Chrysanthemum is not a monophyletic taxon, and the affinity between Chrysanthemum and Ajania is so close that these two genera should be incorporated taxonomically; 2) Phaeostigma is more closely related to the Chrysanthemum+Ajania than other generic allies. According to pollen morphology and to the present cpDNA and CDS data, Ajania purpurea is a member of Phaeostigma. Species differentiation in Chrysanthemum appears to be correlated with geographic and environmental conditions. The Chinese Chrysanthemum species can be divided into two groups, the C. zawadskii group and the C. indicum group. The former is distributed in northern China and the latter in southern China. Many polyploid species, such as C. argyrophyllum, may have originated from allopolyploidization involving divergent progenitors. Considering all the evidence from present and previous studies, we conclude that geographic and ecological factors as well as hybridization and polyploidy play important roles in the divergence and speciation of the genus Chrysanthemum.     

Phylogenetic relationship and taxonomic position of Xylanthemum tianschanicum (Krasch.) Muradyan (Compositae,Anthemideae) as inferred from nrDNA ITS data

The central Asian endemic Xylanthemum tianschanicum (Krasch.) Muradyan (Compositae, Anthemideae) is the only species of Xylanthemum Tzvelev with radiate capitula. While the species was formerly found to be closely related to members of the subtribe Handeliinae, other representatives of the genus were considered to be similar to members of Tanacetum. In order to assess its phylogenetic relationships and taxonomic classification, a molecular analysis based on ITS sequences of 13 representatives of the subtribe Handeliinae and 17 species of three other subtribes (Anthemidinae, Artemisiinae and Matricariinae) was performed. Due to the close relationship of the X. tianschanicum to the type species of Richteria (Richteria pyrethroides) in a well supported clade (PP = 1, BS = 100%), which is supported by morphological features concerning the indumentum, capitulum morphology, and pappus structure, the combination Richteria tianschanica (Krasch.) Sonboli & Oberpr. is proposed based on the basionym Pyrethrum tianschanicum Krasch.     

Chloroplast genome sequences of Artemisia maritima and Artemisia absinthium: Comparative analyses,mutational hotspots in genus Artemisia and phylogeny in family Asteraceae

Artemisia L. is a complex genus of medicinal importance. Publicly available chloroplast genomes of few Artemisia species are insufficient to resolve taxonomic discrepancies at species level. We report chloroplast genome sequences of two further Artemisia species: A. maritima (151,061 bp) and A. absinthium (151,193 bp). Both genomes possess typical quadripartite structure comprising of a large single copy, a small single copy and a pair of long inverted repeats. The two genomes exhibited high similarities in genome sizes, gene synteny, GC content, synonymous and non-synonymous substitutions, codon usage, amino acids frequencies, RNA editing sites, microsatellites, and oligonucleotide repeats. Transition to transversion ratio was <1. Maximum likelihood tree showed Artemisia a monophyletic genus, sister to genus Chrysanthemum. We also identified 20 highly polymorphic regions including rpoC2-rps2, trnR-UCU-trnG-UCC, rps18-rpl20, and trnL-UAG-rpl32 that could be used to develop authentic and cost-effective markers to resolve taxonomic discrepancies and infer phylogenetic relationships among Artemisia species.     

Systematic status and phylogenetic relationships of the enigmatic Tanacetum paradoxum Bornm. (Asteraceae, Anthemideae): evidences from nrDNA ITS, micromorphological, and cytological data

In order to evaluate the phylogenetic relationships and taxonomic classification of the Iranian endemic Tanacetum paradoxum Bornm. (Asteraceae, Anthemideae), a molecular analysis based on nrDNA ITS sequences of 16 species of Tanacetum together with representatives of 27 genera of subtribes Artemisiinae and Handeliinae as well as a micromorphological study of achenes and pollen grains was performed. The chromosome number of 2n?=?2x?=?16 and the genome size of T. paradoxum (2C?=?8.81?±?0.19?pg) are reported here for the first time. Due to the well-supported close phylogenetic relationship of T.?paradoxum to the species of the genus Artemisia (Bayesian inference, posterior probability PP?=?1), which is supported by cytological (x?=?8) and micromorphological features concerning the achene shape and pollen type, the combination Artemisia paradoxa (Bornm.) Sonboli is proposed.     

Genetic polymorphism of ferula mushroom growing on Ferula sinkiangensis

Mating tests, internal transcribed spacer (ITS) sequence analysis, intergenic spacer 1–restriction fragment length polymorphism (IGS1-RFLP), IGS1 sequence analysis, and IGS2-RFLP analysis were carried out on isolates of 17 morphologically different Pleurotus mushrooms collected on Ferula sinkiangensis. The isolates were divided, based on mating tests and ITS sequence analysis, into two groups identical to P. eryngii var. ferulae and P. nebrodensis, respectively. Single spores from these two groups were incompatible, but those from P. eryngii var. ferulae and P. eryngii were compatible and combined to produce 56.25% dikaryon mycelia with clamp connections. The ITS of P. eryngii var. ferulae and P. nebrodensis (GenBank accession no. AY311408) were both 638 bp in size but differed by 3% in sequence. P. eryngii var. ferulae and P. eryngii (GenBank accession no. AY368658) were identical in ITS size and sequence. P. nebrodensis was the dominant population of Pleurotus mushroom growing on F. sinkiangensis. It exhibited genetic diversity. The two species could also be distinguished by IGSI-RFLP, similar to identification by mating tests and ITS sequence analysis. Difference in IGS1-RFLP existed between P. eryngii var. ferulae and P. nebrodensis. The sequence difference reached 2.28%. Both IGS1 size and IGS1-RFLP were similar among the different samples of P. nebrodensis. The 17 isolates were separated into five types based on IGS2 size and IGS2-RFLP, with both interspecies and extraspecies differences. P. nebrodensis exhibited polymorphism and was divided into four types. These results agreed with macroscopic differences. IGS2 might be the effective domain of genetically polymorphic ribosomal DNA in P. nebrodensis mushrooms found in Xinjiang, China.     

Phylogenetic relationships among Artemisia species based on nuclear ITS and chloroplast psbA-trnH DNA markers

The taxonomic and phylogenetic relationships within the genus Artemisia s.l. (Asteraceae) are controversial, and it has been considered 1 to 8 different genera. This work re-investigated the phylogenetic relationships in Artemisia using nuclear ribosomal (ITS) and chloroplast psbA-trnH DNA sequences using three sections of Artemisia, Dracunculus, and Serphidium. Three phylogenetic trees were conducted separately on the basis of ITS, psbA-trnH and combined sequences using maximum parsimony. The results showed that the three sections were clearly separated from each other, and that the heterogamous Dracunculus and Artemisia are closely related to each other than either to homogamous Serphidium. This may suggest the taxonomic importance of capitulum morphology in Artemisia s.l. Our data also cast doubt on the use of cytogenetic similarity e.g., basic chromosome number in grouping Serphidium and Artemisia s.s. Furthermore, AMOVA analysis showed a higher level of ITS (55.29%) and combined ITS+cppsbA-trnH (55.63%) variations among sections. This provides further evidence for separation of these three sections and supports the phylogenetic results. The higher ITS nucleotide differences detected in Artemisia (30.4737) compared to very low value in Dracunculus (2.3333) and Serphidium (1.23077) may propose that the Artemisia comprises of several incipient sections. This supports the previous suggestion that Artemisia is a complex group.     

The Genus Artemisia and its Allies: Phylogeny of the Subtribe Artemisiinae (Asteraceae, Anthemideae) Based on Nucleotide Sequences of Nuclear Ribosomal DNA Internal Transcribed Spacers (ITS)

Abstract: Sequences of the internal transcribed spacers (ITS1 and ITS2) of nuclear ribosomal DNA were analysed for 44 Artemisia species (46 populations) representing all the five classical subgenera and the geographical range of the genus, 11 species from 10 genera closely related to Artemisia, and six outgroup species from five other genera of the Anthemideae. The results definitely support the monophyly of the genus Artemisia in its broadest sense (including some taxa segregated as independent genera, like Oligosporus and Seriphidium ). Eight main clades are established in this molecular phylogeny within Artemisia; they agree in part with the classical subdivision of the genus, but they also suggest that some infrageneric groups must be redefined, especially the subgenus Artemisia. The subgenera Tridentatae and Seriphidium are independent from each other. Some of the satellite genera are clearly placed within Artemisia ( Artemisiastrum, Filifolium, Mausolea, Picrothamnus, Sphaeromeria, Turaniphytum ), whereas some others fall outside the large clade formed by this genus (Brachanthemum, Elachanthemum, Hippolytia, Kaschgaria). Our results, correlated to other data such as pollen morphology, allow us to conclude that the subtribe Artemisiinae as currently defined is a very heterogeneous group. Affinities of the largest genus of the subtribe and tribe, Artemisia, and of other genera of the subtribe to some genera from other subtribes of the Anthemideae strongly suggest that subtribe Artemisiinae needs a deep revision and redefinition. Phylogenetic utility of region trnL-F of the plastid DNA in the genus Artemisia and allies was also evaluated: sequences of the trnL-F region in Artemisia do not provide phylogenetic information.     

Phylogenetic relationships among Strobilanthes s.l. (Acanthaceae): evidence from ITS nrDNA, trnL-F cpDNA, and morphology

Chloroplast trnL-F sequence data, nuclear ribosomal internal transcribed spacer (ITS) sequence data, and morphology were used to analyze phylogenetic relationships among members of the subtribe Strobilanthinae. Parsimony and maximum likelihood analyses of trnL-F indicate that the Strobilanthinae are a monophyletic group. While parsimony analysis of ITS recovers a nonmonophyletic subtribe, maximum likelihood analysis of ITS corroborates results from trnL-F and suggests that systematic error is impacting on ITS parsimony analysis. A combined ITS and trnL-F analysis strengthens the signal and also recovers a monophyletic subtribe. All analyses indicate that Hemigraphis, Sericocalyx, and Strobilanthes are nonmonophyletic. With one exception, all morphological characters included in a combined ITS and morphological analysis are homoplastic. The prospect for a new informative generic classification of the Strobilanthinae aiming to recognize and diagnose only monophyletic groups is considered. While some groups can be diagnosed, adequate diagnosis of the majority of groups remains problematic. Consequently, a single expanded genus Strobilanthes sensu lato is proposed at the level of the well-supported and monophyletic Strobilanthinae.     

菊花近缘种属植物幼苗耐阴特性分析及其评价指标的确定

以龙脑菊、菊花脑、野菊等15个菊花近缘种属植物幼苗为材料,对其进行不同梯度遮荫处理(全光照,遮光率60%,遮光率78%,遮光率95%),从形态和生理等方面的22个指标进行测定,以各项指标的耐阴系数作为衡量耐阴性的指标,利用主成分分析、回归分析和聚类分析法对其耐阴性进行综合评价。结果表明:遮光率78%时的植物茎粗(X2)、叶片厚度(X10)、叶绿素含量(X16),遮光率60%时的植物叶绿素含量(X15),以及遮光率95%时的植物叶面积(X13)、相对含水量(X14)和胞间二氧化碳浓度(X21)8个指标可作为菊花近缘种属植物耐阴性评价指标,建立菊花近缘种属植物耐阴性评价的数学模型:Y=82.876-0.153X2+0.094X10+0.741X13+0.084X14+0.054X15-0.087X16-0.472X2,(R2=0.998),预测精度大于0.97。13份材料的耐阴性极强,矶菊的耐阴性较差,即多数菊花近缘种属植物具有较好的耐阴能力。     

Palynological Features as a Systematic Marker in Artemisia L. and Related Genera (Asteraceae, Anthemideae) - II: Implications for Subtribe Artemisiinae Delimitation

Abstract: Using optical and scanning electron microscopy, we completed a palynological study of the subtribe Artemisiinae (Asteraceae, Anthemideae), which we started in a previous paper. This subtribe contains different genera with a systematic position that, in many cases, has been controversial. There is a group of genera closely related to Artemisia, and another one more related to Chrysanthemum sensu lato. We confirm the existence of two pollen morphological patterns - concerning exine ornamentation - in the tribe Anthemideae and in the subtribe Artemisiinae as currently considered: one with long spines ( Anthemis type) and the other with short spinules ( Artemisia type). This feature is a good taxonomic marker, well correlated with other morphological and with molecular characters. This enables a new delimitation of the subtribe Artemisiinae, which is characterized by pollen grain ornamentation constituted by short spinules (microechinate pollen), and should be restricted to Artemisia and the closely related genera that share this trait.     

Polyphyly of the grass tribe Hainardieae (Poaceae: Pooideae): identification of its different lineages based on molecular phylogenetics, including morphological and cytogenetic characteristics

The small pooid grass tribe Hainardieae comprises six genera with approximately ten species; however, this tribe was not accepted by all previous taxonomic treatments. To study the relationships among these genera and to infer the phylogeny and evolutionary patterns, we used sequence variation of the internal transcribed spacers (ITS) of nuclear ribosomal and chloroplast (cp) matK DNA and morphology. Many genera of the Aveneae/Poeae tribe complex additionally were included. Both molecular datasets showed Hainardieae to be highly polyphyletic, and its genera to branch with different groups of the Aveneae/Poeae. Parapholis and Hainardia are corroborated as being closely related, and belonging to a firmly supported Eurasian clade together with Catapodium incl. Scleropoa, Cutandia, Desmazeria, Sphenopus, Vulpiella (subtribe Parapholiinae) and with Cynosurus as sister to this assemblage. The other genera of traditionally recognised Hainardieae are positioned phylogenetically distant: Mediterranean Narduroides is verified as more or less related to Festuca and relatives (subtribe Loliinae), whereas the west Eurasian Pholiurus is close to the lineage of Poa and relatives (subtribe Poinae). North American Scribneria is sister to Deschampsia and both genera should be unified under a common subtribe (Aristaveninae or Holcinae). The phylogenetic position of the Algerian genus Agropyropsis (close to Narduroides and within the Loliinae) is suggested on morphology only, because no molecular data was obtained for it. Considering classification, we support the abandonment of tribe Hainardieae and argue to abandon Poeae subtribe Scribneriinae. Poeae subtribe Parapholiinae is redefined with a novel genus content, due to the exclusion of Agropyropsis and Pholiurus and the inclusion of Vulpiella.     

Phylogenetic relationships among diploid species of Symphyotrichum (Asteraceae: Astereae) based on two nuclear markers, ITS and GAPDH

The mostly North American subtribe Symphyotrichinae (Asteraceae: Astereae) comprises Canadanthus, Ampelaster, Psilactis, Almutaster, and Symphyotrichum. Intergeneric and interspecific relationships within the subtribe have been investigated in the past, particularly by Nesom [Nesom, G.L., 1994. Review of the taxonomy of Aster sensu lato (Asteraceae: Astereae), emphasizing the new world species, Phytologia 77, 141–297] and Semple [Semple, J.C., 2005. Classification of Symphyotrichum. Available from: <http://www.jcsemple.uwaterloo.ca/Symphyotrichumclassification.htm/>], using morphological and cytological approaches. Symphyotrichum is the largest and most complex genus within the subtribe and includes four subgenera: Symphyotrichum (x = 7, 8), Virgulus (x = 4, 5), Astropolium (x = 5), and Chapmaniani (x = 7). In this study we used two nuclear markers, the nrDNA internal transcribed spacer (ITS) and the low-copy nuclear gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH), to resolve intergeneric and interspecific relationships within the subtribe at the diploid level, and to determine whether our phylogenies validate the classifications of Nesom or Semple. Our results confirm the distinct generic status of Canadanthus and Ampelaster, whereas Psilactis and Almutaster form a polytomy with Symphyotrichum. Within Symphyotrichum, subg. Virgulus is monophyletic based on ITS and appears polyphyletic based on GAPDH. Neither the ITS nor the GAPDH analyses support a distinct status for subg. Astropolium, which groups within subg. Symphyotrichum. In general, interspecific relationships within Symphyotrichum are unresolved. Lack of resolution may be interpreted as a case of recent and rapid evolutionary radiation.     

Generic Delimitation and Phylogeny of the Subtribe Centaureinae (Asteraceae): A Combined Nuclear and Chloroplast DNA Analysis

Taxonomic complexity has hindered the partitioning of the artificial genus Centaurea, even though it has long been recognized as a polyphyletic assemblage. On the basis of morphology, pollen type, karyology and DNA sequence analysis, previous workers have defined five informal groups in the genus (Acrocentron,Centaureasensu stricto , Cyanus, Jacea and Psephellus). However, the precise delimitation of these groups and their relationships remain largely unknown. Moreover, although some informal groups have been established among the rest of the subtribe (Amberboa, Carthamus and Stemmacantha), many genera cannot be classified in any group. Newer molecular approaches are essential for resolving these problems. Sequences of the internal transcribed spacers (ITS) of nuclear ribosomal DNA and the chloroplast gene mat K were analysed for a comprehensive sample of the whole subtribe, with the aim of clarifying the delimitation and the phylogeny of the groups of the Centaureinae. Results largely confirm the suggested informal entities as natural groups, with some interesting changes of placement of some genera, especially in the Acrocentron and the Stemmacantha groups. Our results confirm that the sections of Centaurea with Dealbata pollen type should be classified as a different genus, Psephellus. In addition to morphology and suggested pollen type evolution, the ITS and mat K phylogenies are also supported by karyological evidence. Our results confirm that the natural delimitation of Centaurea that minimizes nomenclatural changes is possible only if a new type of the genus is designated.