Rosidae, a clade of approximately 90 000 species of angiosperms, exhibits remarkable morphological diversity and extraordinary heterogeneity in habitats and life forms. Resolving phylogenetic relationships within Rosidae has been difficult, in large part due to nested radiations and the enormous size of the clade. Current estimates of phylogeny contain areas of poor resolution and/or support, and there have been few attempts to synthesize the available data into a comprehensive view of Rosidae phylogeny. We aim to improve understanding of the phylogeny of Rosidae with a dense sampling scheme using both newly generated sequences and data from GenBank of the chloroplast rbcL, atpB, and matK genes and the mitochondrial matR gene. We combined sequences from 9300 species, representing 2775 genera, 138 families, and 17 orders into a supermatrix. Although 59.26% of the cells in the supermatrix have no data, our results generally agree with previous estimates of Rosidae phylogeny and provide greater resolution and support in several areas of the topology. Several noteworthy phylogenetic relationships are recovered, including some novel relationships. Two families (Euphorbiaceae and Salvadoraceae) and 467 genera are recovered as non-monophyletic in our sampling, suggesting the need for future systematic studies of these groups. Our study shows the value of a botanically informed bioinformatics approach and dense taxonomic sampling for resolving rosid relationships. The resulting tree provides a starting point for large-scale analyses of the evolutionary patterns within Rosidae.Key words: phylogeny, rapid radiation, Rosidae, supermatrix.With approximately 90 000 species (estimated from Hinchliff et al., 2015), 135-140 families, and 17 orders (Soltis et al., 2005;APG III, 2009;APG IV, 2016), Rosidae contains at least one quarter of all angiosperm species and approximately 39% of eudicot species diversity (Magall on et al., 1999;Wang et al., 2009). Molecular dating indicates that Rosidae originated in the Early to Late Cretaceous, between 115 and 93 million years ago (Mya), followed by rapid diversification resulting in the Fabidae and Malvidae crown groups approximately 112 to 91 Mya (Albian to Coniacian) and 109 to 83 Mya (Cenomanian to Santonian), respectively (Wang et al., 2009;Bell et al., 2010), with major lineages diversifying in perhaps as little as 4 to 5 million years (Wang et al., 2009;Soltis et al., 2010). The radiations in Rosidae also represent the rapid rise of angiosperm-dominated forests and associated co-diversification events that have profoundly shaped much of the current terrestrial biodiversity (Wang et al., 2009).This extraordinarily diverse clade exhibits enormous heterogeneity in habitats and life forms, including herbs, shrubs, trees, vines, aquatics, succulents, and parasites. Species of Rosidae generally have bitegmic, crassinucellate ovules, distinguishing them from Asteridae, which are generally characterized by unitegmic, tenuinucellate ovules. Moreover, some members possess novel bio...

We reconstructed a phylogenetic tree of Chinese vascular plants (Tracheophyta) using sequences of the chloroplast genes atpB, matK, ndhF, and rbcL and mitochondrial matR. We produced a matrix comprising 6098 species and including 13 695 DNA sequences, of which 1803 were newly generated. Our taxonomic sampling spanned 3114 genera representing 323 families of Chinese vascular plants, covering more than 93% of all genera known from China. The comprehensive large phylogeny supports most relationships among and within families recognized by recent molecular phylogenetic studies for lycophytes, ferns (monilophytes), gymnosperms, and angiosperms. For angiosperms, most families in Angiosperm Phylogeny Group IV are supported as monophyletic, except for a paraphyletic Dipterocarpaceae and Santalaceae. The infrafamilial relationships of several large families and monophyly of some large genera are well supported by our dense taxonomic sampling. Our results showed that two species of Eberhardtia are sister to a clade formed by all other taxa of Sapotaceae, except Sarcosperma. We have made our phylogeny of Chinese vascular plants publically available for the creation of subtrees via SoTree (http://www.darwintree.cn/flora/index.shtml), an automated phylogeny assembly tool for ecologists.

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Capparaceae (Brassicales) as traditionally circumscribed is heterogeneous, and several genera have been segregated from it based on molecular and/or morphological data. However, Borthwickia and Stixis, two Southeast Asian endemic genera of Capparaceae with controversial positions, have not previously been evaluated in a molecular phylogenetic study. Here, we used four plastid DNA regions (matK, ndhF, rbcL, trnL–trnF) and pollen data to determine their phylogenetic relationships within core Brassicales. Our results showed that neither Borthwickia nor Stixis is a member of Capparaceae. The two genera, together with Forchhammeria, Gyrostemonaceae, Resedaceae, and Tirania, formed a clade with strong support. Stixis is closely related to Tirania, a relationship that is also supported by morphological characters, such as six sepals and three–or four–locular ovaries. Most interestingly, Borthwickia was resolved as sister to the Forchhammeria–Resedaceae–Stixis–Tirania clade with moderate to strong support. However, Borthwickia differs markedly from its sister group in having opposite leaves, one indistinct stigma, more than four carpels and locules, a linear ovary with ridges, and pollen grains with perforate exine sculpturing. Thus, we describe a new family, Borthwickiaceae, for the genus.

DNA barcoding is a biological technique that uses short and standardized genes or DNA regions to facilitate species identification. DNA barcoding has been used successfully in several animal and plant groups. Ligustrum (Oleaceae) species occur widely throughout the world and are used as medicinal plants in China. Therefore, the accurate identification of species in this genus is necessary. Four potential DNA barcodes, namely the nuclear ribosomal internal transcribed spacer (ITS) and three chloroplast (cp) DNA regions (rbcL, matK, and trnH-psbA), were used to differentiate species within Ligustrum. BLAST, character-based method, tree-based methods and TAXONDNA analysis were used to investigate the molecular identification capabilities of the chosen markers for discriminating 92 samples representing 20 species of this genus. The results showed that the ITS sequences have the most variable information, followed by trnH-psbA, matK, and rbcL. All sequences of the four regions correctly identified the species at the genus level using BLAST alignment. At the species level, the discriminating power of rbcL, matK, trnH-psbA, and ITS based on neighbor-joining (NJ) trees was 36.8%, 38.9%, 77.8%, and 80%, respectively. Using character-based and maximum parsimony (MP) tree methods together, the discriminating ability of trnH-psbA increased to 88.9%. All species could be differentiated using ITS when combining the NJ tree method with character-based or MP tree methods. Overall, the results indicate that DNA barcoding is an effective molecular identification method for Ligustrum species. We propose the nuclear ribosomal ITS as a plant barcode for plant identification and trnH-psbA as a candidate barcode sequence.

Species concept and delimitation are fundamental to taxonomic and evolutionary studies. Both inadequate informative sites in the molecular data and limited taxon sampling have often led to poor phylogenetic resolution and incorrect species delineation. Recently, the whole chloroplast genome sequences from extensive herbarium specimen samples have been shown to be effective to amend the problem. Stachyuraceae are a small family consisting of only one genus Stachyurus of six to 16 species. However, species delimitation in Stachyurus has been highly controversial because of few and generally unstable morphological characters used for classification. In this study, we sampled 69 individuals of seven species (each with at least three individuals) covering the entire taxonomic diversity, geographic range, and morphological variation of Stachyurus from herbarium specimens for genome‐wide plastid gene sequencing to address species delineation in the genus. We obtained high‐quality DNAs from specimens using a recently developed DNA reconstruction technique. We first assembled four whole chloroplast genome sequences. Based on the chloroplast genome and one nuclear ribosomal DNA sequence of Stachyurus, we designed primers for multiplex polymerase chain reaction and high throughput sequencing of 44 plastid loci for species of Stachyurus. Data of these chloroplast DNA and nuclear ribosomal DNA internal transcribed spacer sequences were used for phylogenetic analyses. The phylogenetic results showed that the Japanese species Stachyurus praecox Siebold & Zucc. was sister to the rest in mainland China, which indicated a typical Sino‐Japanese distribution pattern. Based on diagnostic morphological characters, distinct distributional range, and monophyly of each clade, we redefined seven species for Stachyurus following an integrative species concept, and revised the taxonomy of the family based on previous reports and specimens, in particular the type specimens. Furthermore, our divergence time estimation results suggested that Stachyuraceae split from its sister group Crossosomataceae from the New World at ca. 54.29 Mya, but extant species of Stachyuraceae started their diversification only recently at ca. 6.85 Mya. Diversification time of Stachyurus in mainland China was estimated to be ca. 4.45 Mya. This research has provided an example of using the herbarium specimen‐based phylogenomic approach in resolving species boundaries in a taxonomically difficult genus.

There has been increasing interest in integrating a regional tree of life with community assembly rules in the ecological research. This raises questions regarding the impacts of taxon sampling strategies at the regional versus global scales on the topology. To address this concern, we constructed two trees for the nitrogen-fixing clade: (i) a genus-level global tree including 1023 genera; and (ii) a regional tree comprising 303 genera, with taxon sampling limited to China. We used the supermatrix approach and performed maximum likelihood analyses on combined matK, rbcL, and trnL-F plastid sequences. We found that the topology of the global and the regional tree of the N-fixing clade were generally congruent. However, whereas relationships among the four orders obtained with the global tree agreed with the accepted topology obtained in focused analyses with more genes, the regional topology obtained different relationships, albeit weakly supported. At a finer scale, the phylogenetic position of the family Myricaceae was found to be sensitive to sampling density. We expect that internal support throughout the phylogeny could be improved with denser taxon sampling. The taxon sampling approach (global vs. regional) did not have a major impact on fine-level branching patterns of the N-fixing clade. Thus, a well-resolved phylogeny with relatively dense taxon sampling strategy at the regional scale appears, in this case, to be a good representation of the overall phylogenetic pattern and could be used in ecological research. Otherwise, the regional tree should be adjusted according to the correspondingly reliable global tree.

The evolutionary history of herbaceous species in the Northern Hemisphere remains poorly understood. The genus Cerastium is one of the most species-rich herbaceous genera in the Alsineae (Caryophyllaceae), and is mainly distributed in the Northern Hemisphere. Thus, it provides an ideal opportunity to explore the biogeographical history of herbs in the Northern Hemisphere. Here, we present a comprehensive phylogenetic tree for Cerastium with 75 species based on one nuclear and five plastid DNA markers. We then use an integrated phylogenetic, molecular dating, biogeographical, and diversification rate method to examine the spatiotemporal evolution of Cerastium. Cerastium is strongly supported as monophyletic and contains three major clades (Orthodon, Strephodon, and C. fragillimum). Cerastium originated from Europe in the middle Miocene (c. 13.45 million years ago), and subsequently colonized the New World, Africa, Asia, and Australasia through multiple intercontinental dispersal events. The dispersal of Cerastium from Europe (to both the East and West) and subsequent diversifications contributed to the formation of its contemporary Holarctic distribution pattern. The genus experienced rapid lineage accumulation during the late Miocene, possibly coinciding with a global decrease in temperature. These findings highlight the importance of the Northern Hemisphere in herbaceous species diversification in the late Miocene and Pliocene, and will deepen our understanding of the evolution of herbaceous plants.