Comparative analyses of eight complete plastid genomes of two hemiparasitic Cassytha vines in the family Lauraceae.

Cassytha is the sole genus of hemiparasitic vines (ca. 20 spp.) belonging to the Cassytheae tribe of the Lauraceae family. It is extensively distributed in tropical and subtropical regions. In this study, we determined the complete plastid genome sequences of C. filiformis and C. larsenii, which do not possess the typical quadripartite structure. The length of C. filiformis plastomes ranged from 114,215 to 114,618 bp, whereas that of C. larsenii plastomes ranged from 114,900 to 114,988 bp. Comparative genomic analysis revealed 1,013 mutation sites, four large intragenomic deletions, and five highly variable regions in the eight plastome sequences. Phylogenetic analyses based on 61 complete plastomes of Laurales species, 19 ITS sequences, and trnK barcodes from 91 individuals of Cassytha spp. confirmed a non-basal group comprising individuals of C. filiformis, C. larsenii, and C. pubescens in the family Lauraceae and proposed a sister relationship between C. filiformis and C. larsenii. Further morphological comparisons indicated that the presence or absence of hairs on the haustoria and the shape or size of fruits were useful traits for differentiating C. filiformis and C. larsenii.

Two new species of Polyalthiopsis (Annonaceae) based on morphological characters and phylogenetic evidence, with a supplementary description of P. chinensis from China.

Two new species of Polyalthiopsis (Annonaceae), P. nigra Y.H. Tan & Bin Yang from Guangxi and Yunnan Provinces and P. xui Y.H. Tan & Bin Yang from Yunnan Province, are described and illustrated. P. nigra is morphologically similar to P. chinensis in having narrowly elliptic-oblong, lemon to yellowish green petals, but differs by having obovoid monocarps, a higher number of leaf secondary veins, leaf blades usually widest above the middle, and a lower ratio of leaf blade length to width. P. xui is morphologically similar to P. floribunda in having axillary inflorescences, 1-3(-4) flowers, elliptic leaves, and elliptic-ovate petals, but differs in the numbers of carpels per flower and ovules per carpel. The molecular phylogenetic analysis using five plastid markers confirm that the two new species belong to the genus Polyalthiopsis and show clear interspecific divergences between P. nigra and P. xui and between them and other species in the genus. Detailed descriptions, colored photographs, and habitat and distribution data for the two new species are provided. In addition, the fruit morphology of P. chinensis is described for the first time, based on living collections. Geographical distributions and a diagnostic key for all Polyalthiopsis species are also presented.

Splitting one species into 22: an unusual tripling of molecular, morphological, and geographical differentiation in the fern family Didymochlaenaceae (Polypodiales).

The pantropical fern genus Didymochlaena (Didymochlaenaceae) has long been considered to contain one species only. Recent studies have resolved this genus/family as either sister to the rest of eupolypods I or as the second branching lineage of eupolypods I, and have shown that this genus is not monospecific, but the exact species diversity is unknown. In this study, a new phylogeny is reconstructed based on an expanded taxon sampling and six molecular markers. Our major results include: (i) Didymochlaena is moderately or weakly supported as sister to the rest of eupolypods I, highlighting the difficulty in resolving the relationships of this important fern lineage in the polypods; (ii) species in Didymochlaena are resolved into a New World clade and an Old World clade, and the latter further into an African clade and an Asian-Pacific clade; (iii) an unusual tripling of molecular, morphological and geographical differentiation in Didymochlaena is detected, suggesting single vicariance or dispersal events in individual regions and no evidence for reversals at all, followed by allopatric speciation at more or less homogeneous rates; (iv) evolution of 18 morphological characters is inferred and two morphological synapomorphies defining the family are recognized-the elliptical sori and fewer than 10 sori per pinnule, the latter never having been suggested before; (v) based on morphological and molecular variation, 22 species in the genus are recognized contrasting with earlier estimates of between one and a few; and (vi) our biogeographical analysis suggests an origin for Didymochlaena in the latest Jurassic-earliest Cretaceous and the initial diversification of the extant lineages in the Miocene-all but one species diverged from their sisters within the last 27 Myr, in most cases associated with allopatric speciation owing to geologic and climatic events, or dispersal.

Two new species from Sulawesi and Borneo facilitate phylogeny and taxonomic revision of Engelhardia (Juglandaceae).

Engelhardia, a genus of Juglandaceae (the walnut family), is endemic to tropical and subtropical Asia. The rich Cenozoic fossil records and distinctive morphological characters of the living plants have been used to explore the evolutionary history and geographic distribution of Juglandaceae. However, the taxonomy of this genus has been suffered from a lack of in-depth investigation and good specimens across its distribution ranges. Species delimitation of Engelhardia was defined with seven species in 2020, but detailed information on the circumscription of the species still remains poorly understood. In this study, two new species are described from Sulawesi and Borneo, Engelhardia anminiana and E. borneensis. We also revised and reconstructed the phylogeny within Engelhardia using morphological, molecular (plastid and ribosomal), and distribution data. We sampled 787 individuals in 80 populations, and all the samples were genotyped using plastid regions, trnS-trnG, rps16, trnL-trnF, psbA-trnH, and rpl32-trnL; one ribosomal region, nuclear ribosomal internal transcribed spacer (nrITS). The all datasets were used to reconstruct the phylogenetic relationships. Then, the molecular analyses were combined for 738 sheets of specimens with 15 morphological characteristics to further explore the morphological clusters of Engelhardia. Cluster analysis using morphological data confirmed the delimitation of nine Engelhardia species. Also, phylogenetic analysis based on molecular data (i.e., plastid and ribosomal) supported the monophyly of Engelhardia and generated phylogenetic trees that included E. fenzelii, E. roxburghiana, E. borneensis, E. hainanensis, E. anminiana, E. serrata, E. villosa, E. apoensis and the varieties of E. spicata (i.e., E. spicata var. spicata, E. spicata var. rigida, E. spicata var. aceriflora, and E. spicata var. colebrookeana). Our comprehensive taxonomic revision of Engelhardia will provide an insight into understanding the plant diversity in tropical and subtropical Asia.

Plastid genome evolution of a monophyletic group in the subtribe Lauriineae (Laureae, Lauraceae).

Litsea, a non-monophyletic group of the tribe Laureae (Lauraceae), plays important roles in the tropical and subtropical forests of Asia, Australia, Central and North America, and the islands of the Pacific. However, intergeneric relationships between Litsea and Laurus, Lindera, Parasassafras and Sinosassafras of the tribe Laureae remain unresolved. In this study, we present phylogenetic analyses of seven newly sequenced Litsea plastomes, together with 47 Laureae plastomes obtained from public databases, representing six genera of the Laureae. Our results highlight two highly supported monophyletic groups of Litsea taxa. One is composed of 16 Litsea taxa and two Lindera taxa. The 18 plastomes of these taxa were further compared for their gene structure, codon usage, contraction and expansion of inverted repeats, sequence repeats, divergence hotspots, and gene evolution. The complete plastome size of newly sequenced taxa varied between 152,377 bp (Litsea auriculata) and 154,117 bp (Litsea pierrei). Seven of the 16 Litsea plastomes have a pair of insertions in the IRa (trnL-trnH) and IRb (ycf2) regions. The 18 plastomes of Litsea and Lindera taxa exhibit similar gene features, codon usage, oligonucleotide repeats, and inverted repeat dynamics. The codons with the highest frequency among these taxa favored A/T endings and each of these plastomes had nine divergence hotspots, which are located in the same regions. We also identified six protein coding genes (accD, ndhJ, rbcL, rpoC2, ycf1 and ycf2) under positive selection in Litsea; these genes may play important roles in adaptation of Litsea species to various environments.

Opening a door to the spatiotemporal history of plants from the tropical Indochina Peninsula to subtropical China.

The complexity of global biodiversity in the tropical Indochina Peninsula and subtropical China bioregions has fascinated biologists for decades, but little is known about the spatiotemporal patterns in these regions. Accordingly, the aims of present study were to investigate the evolutionary and distribution patterns of Engelhardia in these regions and establish a model for examining biogeographic patterns and geological events throughout the tropical Indochina Peninsula and subtropical China. The effects of geological events occurring in the area between the Indochina Peninsula and subtropical China bioregions on the two trees species (i.e., E. roxburghiana and E. fenzelii) were evaluated. A robust phylogenetic framework of 884 individuals from 79 populations was used to generate time-calibrated cytoplasmic and nuclear phylogenetic frameworks based on cpDNA, nrDNA, and nSSR data, respectively. When considered along with ancestral area reconstructions, the genetic data were also used to assess and reconstruct the species' population genetic structure and diversity. These analyses yielded important information about the (1) historical distribution relationships between the tropical and subtropical flora of China; (2) effects of the East Asian summer monsoon (EASM) on the evolutionary history of Asia's plants; and (3) importance of biogeography in conservation planning. Although cytoplasmic-nuclear discordance indicated cpDNA and nrDNA were subject to distinct evolutionary mechanisms that reflected respective evolutionary histories of the plastid and nuclear genomes of prior demographic and biogeographic events. The tropical elements of Engelhardia occupied the Indochina Peninsula during the early Eocene, whereas the subtropical elements were transformed from the tropical elements during Miocene cooling and the onset of the EASM at the Oligocene-Miocene boundary, intensified during the late Miocene and Pliocene, facilitating the transformation of Engelhardia from the tropical Indochina Peninsula to subtropical China. Demographic history provided insights into prominent planning frameworks in conservation biology, namely that subtropical China functioned as a refugium during past climate oscillations and will continue to serve in this capacity in the future.

A new isotope framework to decipher leaf-root nitrogen allocation and assimilation among plants in a tropical invaded ecosystem.

Exotic plant invasion is an urgent issue occurring in the biosphere, which can be stimulated by environmental nitrogen (N) loading. However, the allocation and assimilation of soil N sources between leaves and roots remain unclear for plants in invaded ecosystems, which hampers the understanding of mechanisms behind the expansion of invasive plants and the co-existence of native plants. This work established a new framework to use N concentrations and isotopes of soils, roots, and leaves to quantitatively decipher intra-plant N allocation and assimilation among plant species under no invasion and under the invasion of Chromolaena odorata and Ageratina adenophora in a tropical ecosystem of SW China. We found that the assimilation of N derived from both soil ammonium (NH4+) and nitrate (NO3-) were higher in leaves than in roots for invasive plants, leading to higher leaf N levels than native plants. Compared with the same species under no invasion, most native plants under invasion showed higher N concentrations and NH4+ assimilations in both leaves and roots, and increases in leaf N were higher than in root N for native plants under invasion. These results inform that preferential N allocation, dominated by NH4+-derived N, to leaves over roots as an important N-use strategy for plant invasion and co-existence in the studied tropical ecosystem.

Oreocharispolyneura, a new species of Gesneriaceae from southern Yunnan, China.

Oreocharispolyneura, a new species from southern Yunnan, China, is described and illustrated. It is morphologically similar to O.rhytidophylla by having more obvious lateral veins forming a crosslinked network on the adaxial surface of leaf blades, but can be distinguished by having more lateral veins (12-15 pairs vs. 7-9 pairs) of leaf blades, shorter corolla tubes (the length ratio of corolla tube to corolla lobes = 1.2-2.9 vs. 3.4-6) and shorter pistils (6-8 mm long vs. 27-30 mm long). In addition, a detailed morphological description, a photographic illustration, the distribution and phenology of the new species are presented.

Rungiafangdingiana (Acanthaceae), a new species from Guangxi, China.

Rungiafangdingiana, a new species of Acanthaceae from Guangxi, China is described and illustrated. This new species belongs to Rungiasect.Rungia, and resembles R.sinothailandica and R.burmanica in the erect perennial herbaceous habit, elliptic leaves and inflorescence form, but differs mainly by the indumentum and the morphology of the bracts and corolla. The pollen and seed micromorphology of this new species are studied, with photographs and a line drawing provided.

Meiogyne oligocarpa (Annonaceae), a new species from Yunnan, China.

Meiogyne oligocarpa sp. nov. (Annonaceae) is described from Yunnan Province in Southwest China. It is easily distinguished from all previously described Meiogyne species by the possession of up to four carpels per flower, its bilobed, sparsely hairy stigma, biseriate ovules and cylindrical monocarps with a beaked apex. A phylogenetic analysis was conducted to confirm the placement of this new species within Meiogyne. Meiogyne oligocarpa represents the second species of Meiogyne in China: a key to the species of Meiogyne in China is provided to distinguish it from Meiogyne hainanensis. Paraffin sectioning was undertaken to study the anatomy of the corrugations on the inner petals of Meiogyne oligocarpa to verify whether they are glandular.

Contributions to the flora of Myanmar from 2000 to 2019.

Myanmar is botanically rich and floristically diverse: one of the world's biodiversity hotspots. However, Myanmar is still very unevenly explored, and until a plant checklist was published in 2003, relatively little work was done on its flora. This checklist included 11,800 species of spermatophytes in 273 families. Since this checklist was published, the botanical exploration of Myanmar has accelerated and there have been many additional publications. We therefore surveyed the literature of taxonomic contributions to Myanmar's vascular flora over the last 20 years (2000-2019) and compiled a list of new and newly described taxa. Our list includes 13 genera, 193 species, 7 subspecies, 19 varieties, and 2 forms new to science; and 3 families, 34 genera, 347 species, 4 subspecies, 7 varieties, and 1 form newly recorded in Myanmar. Altogether, they represent 91 families and 320 genera. Most of the new discoveries belong to 15 families, with more than 25% (146 taxa) belonging to Orchidaceae. These new discoveries are unevenly distributed in the country, with about 41% of the newly discovered species described from Kachin State in northeast Myanmar. This reflects the incompleteness of our current knowledge of the flora of Myanmar and the urgent need for a greatly expanded effort. The completion of the flora of Myanmar requires more fieldwork from north to south, taxonomic studies on new and existing collections, and some mechanism that both coordinates the efforts of various international institutions and initiatives and encourages continued international cooperation. In addition, producing modern taxonomic treatments of the flora of Myanmar requires the participation of experts on all vascular plant families and genera. There is also an urgent need to attract young scientists to plant taxonomy, to work on inventories, identification, nomenclature, herbarium work, and comparative studies.

Shining a light on species delimitation in the tree genus Engelhardia Leschenault ex Blume (Juglandaceae).

Enhanced efficacy in species delimitation is critically important in biology given the pending biodiversity crisis under global warming and anthropogenic activity. In particular, delineation of traditional classifications in view of the complexity of species requires an integrative approach to effectively define species boundaries, and this is a major focus of systematic biology. Here, we explored species delimitation of Engelhardia in tropical and subtropical Asia. In total, 716 individuals in 71 populations were genotyped using five chloroplast regions, one nuclear DNA region (nrITS), and 11 nuclear simple sequence repeats (nSSR). Phylogenetic trees were constructed and relationships among species were assessed. Molecular analyses were then combined with 14 morphological characteristics of 720 specimens to further explore the species boundaries of Engelhardia. Integrating phylogenetic and morphological clusters provided well-resolved relationships to delineate seven species. The results suggested that: first, that E. fenzelii, E. roxburghiana, E. hainanensis, E. apoensis, and E. serrata are distinct species; second, E. spicata var. spicata, E. spicata var. aceriflora, E. spicata var. colebrookeana, and E. rigida should be combined under E. spicata and treated as a species complex; third, E. serrata var. cambodica should be raised to species level and named E. villosa. We illuminated that bias thresholds determining the cluster number for delimiting species boundaries were substantially reduced when morphological data were incorporated. Our results urge caution when using the concepts of subspecies and varieties in order to prevent confusion, particularly with respect to species delimitation for tropical and subtropical species. In some cases, re-ranking or combining subspecies and/or varieties may enable more accurate species delimitation.

From Polyalthia to Polyalthiopsis (Annonaceae): transfer of species enlarges a previously monotypic genus.

The genus Polyalthiopsis Chaowasku (Annonaceae) was a poorly known monotypic genus from Vietnam that was recently segregated from the highly polyphyletic genus Polyalthia s.l. The sister clade relationship between Polyalthiopsis and Miliusa was not well established in previous study. The phylogenetic position of two Polyalthia spp. from China, P. chinensis S.K.Wu ex P.T.Li and P. verrucipes C.Y.Wu ex P.T.Li, remains unresolved and is shown here to be phylogenetically affiliated with Polyalthiopsis. Phylogenetic analyses of six chloroplast regions (matK, ndhF, psbA-trnH, rbcL, trnL-F and ycf1; ca.7.3 kb, 60 accessions) unambiguously placed Polyalthia chinensis and P. verrucipes in the same clade with Polyalthiopsis floribunda (PP = 1, MPBS = 97%); the entire clade is sister to Miliusa with weak to strong support (PP = 1, MPBS = 54%). Polyalthia chinensis and P. verrucipes share several diagnostic characters with Polyalthiopsis floribunda, including the raised midrib on the upper surface of the leaf in vivo, conspicuous foliar glands when dried, petiole with transverse striations when dried and axillary inflorescences. The two species differ from Polyalthiopsis floribunda in having fewer flowers per inflorescence, longer linear petals and two ovules per carpel. On the basis of the combined molecular phylogenetic and morphological data, we propose two new combinations, Polyalthiopsis chinensis (S.K.Wu ex P.T.Li) B.Xue & Y.H.Tan and Polyalthiopsis verrucipes (C.Y.Wu ex P.T.Li) B.Xue & Y.H.Tan. The protologue of Polyalthia verrucipes did not include a description of the flowers, which we provide here. An updated description for the genus Polyalthiopsis and a key to species in the genus Polyalthiopsis is also provided.

Documentation of plant diversity of Southeast Asia: the new role of Belt and Road Initiative.

Editorial of special issue of plant diversity in southeastern Asia.

A new species and two new records of Goniothalamus (Annonaceae) from Lao PDR.

Goniothalamus saccopetaloides Y.H. Tan & Bin Yang, a new species is described and illustrated from Laos. This species shows morphological similarities to G. yunnanensis W.T. Wang, but it differs from the latter by having almost fleshy, involute and saccate outer petals, subglobose monocarps, and single seeded monocarps. Goniothalamus cheliensis, and G. calvicarpus are new records for the Flora of Lao PDR. A key to Goniothalamus species indigenous to Laos is provided here.

Two new species of Alseodaphnopsis (Lauraceae) from southwestern China and northern Myanmar: evidence from morphological and molecular analyses.

Alseodaphnopsis maguanensis and A. putaoensis, two new species of Alseodaphnopsis (Lauraceae) from southwestern China (Yunnan Province) and northern Myanmar (Kachin State), are here described and illustrated based on both morphological and molecular evidence. They are morphologically similar to Alseodaphnopsis rugosa and phylogenetically closely related to A. rugosa and A. hainanensis respectively. Their preliminary conservation status is also estimated according to the IUCN Red List Categories and Criteria.

Notes on the genus Gastrochilus (Orchidaceae) in Myanmar.

Myanmar is known for its high species richness of genus Gastrochilus; however, most of them lack proper information for taxonomic revision. During four years of field investigation in Myanmar, two new distributional records were encountered, namely, G. arunachalensis and G. corymbosus and one species, i.e. G. pechei was rediscovered after its original description. The three species were not easy to interpret from the available original descriptions and types due to severely shrunk or poorly preserved specimens. Therefore, we hereby present more detailed illustrations and updated descriptions for these species, based on freshly collected materials.

Taxonomic studies on Amomum Roxburgh s.l. (Zingiberaceae) in Myanmar II: one new species and five new records for the flora of Myanmar.

In the course of a study of Amomum s.l. (Zingiberaceae) in Myanmar, Amomum schistocalyx Y.H. Tan & H.B. Ding, from Htamanti Wildlife Sanctuary, Sangaing Region of Northern Myanmar is described and illustrated as new to science here. Five taxa: A. yingjiangense S.Q. Tong & Y.M. Xia, A. carnosum V.P. Thomas & M. Sabu, A. tibeticum (T.L. Wu & S.J. Chen) X.E. Ye, L. Bai & N.H. Xia, Lanxangia scarlatina (H.T. Tsai & P.S. Chen) M.F. Newman & Skornick, and Meistera yunnanensis (S.Q. Tong) Skornick. & M.F. Newman, are newly recorded from Myanmar. The photographic illustrations, the distributions, and voucher specimens for each species are provided.

A new species and two new combinations of Monolophus (Zingiberaceae) from Indo-Burma.

Monolophus odontochilus Y.H.Tan & H.B.Ding, a new species from Northern Myanmar, is described and illustrated. The new species is morphologically similar to M. linearis, but differs by having elliptic to oblong leaves (vs. linear-lanceolate to lanceolate), bilobed ligules (vs. entire), purely white corolla (vs. pinkish white), semi-orbicular crenate labellum (vs. trilobed). In addition, a diagnostic key to the new species of Monolophus and its closely related non-yellow flowered species is provided. New combinations are proposed here for Caulokaempferia phokhamii Picheans. & Douangde. and C. wongsuwaniae Picheans. & Douangde. from Laos.