Elevational patterns of tree diversity are well studied worldwide. However, few studies have examined how seedlings respond to elevational gradients and whether their responses vary across climatic zones. In this study, we established three elevational transects in tropical, subtropical and subalpine mountain forests in Yunnan Province, southern China, to examine the responses of tree species and their seedlings to elevational gradients. Within each transect, we calculated species diversity indices and composition of both adult trees and seedlings at different elevations. For both adult trees and seedlings, we found that species diversity decreased with increasing elevation in both tropical and subalpine transects. Species composition showed significant elevational separation within all three transects. Many species had specific elevational preferences, but abundant tree species that occurred at specific elevations tended to have very limited recruitment in the understory. Our results highlight that the major factors that determine elevational distributions of tree species vary across climatic zones. Specifically, we found that the contribution of air temperature to tree species composition increased from tropical to subalpine transects, whereas the contribution of soil moisture decreased across these transects.

In the southern mountain ranges of Yunnan province, China, deep valleys of several large rivers create rain shadows with hot dry summers, and are locally designated tropical; towards the north, notably in the Lancang (Upper Mekong) valley, these regions may experience frost during winter. The woody forest canopy of these valleys is predominantly deciduous, with evergreen elements in the north, where the canopy is open and the forest savanna-like. However, we here present tall forest with a closed deciduous canopy and semi-evergreen subcanopy observed in hot dry valleys of these rivers and their tributaries in the tropical south. The structure and physiognomy of these forests resemble the tall (moist) deciduous forest formation widespread in South Asia and Indo-Burma. Furthermore, these forests are largely composed of tropical elements at both the generic (80%) and the species level (>70%), indicating that these forests are indeed tropical. We originally hypothesized that these isolated forests represent refugia of a pre-Holocene extension of tall (moist) deciduous forest formation of South Asia and Indo-Burma. The sample plot we established to test this hypothesis confirmed that these forests share the structure and physiognomy of the tall (moist) deciduous forest formation; however, the plots also showed that these forests lack the characteristic and dominant species of the formation's Indo-Burmese range. The tree flora, in particular, indicates that both deciduous and evergreen elements are instead mostly derived from the adjacent tropical semi-evergreen forests of tropical southern China; yet they also include an important endemic element, which implies that these forests have survived as refuges possibly since the Pliocene. The exceptional representation of evergreen elements in these forests indicates that they have rarely been subject to hot fires or domestic cattle browsing, adding to the unique nature of the forests and further justifying their strict conservation.

Cymbidium, which includes approximately 80 species, is one of the most ornamental and cultivated orchid genera. However, a lack of markers and sparse sampling have posed great challenges to resolving the phylogenetic relationships within the genus. In the present study, we reconstructed the phylogenetic relationships by utilizing one nuclear DNA (nrITS) and seven plastid genes (rbcL, trnS, trnG, matK, trnL, psbA, and atpI) from 70 species (varieties) in Cymbidium. We also examined the occurrence of phylogenetic conflict between nuclear (nrITS) and plastid loci and investigated how phylogenetic conflict bears on taxonomic classification within the genus. We found that phylogenetic conflict and low support values may be explained by hybridization and a lack of informative characteristics. Our results do not support previous classification of the subgenera and sections within Cymbidium. Discordance between gene trees and network analysis indicate that reticulate evolution occurred in the genus Cymbidium. Overall, our study indicates that Cymbidium has undergone a complex evolution.

The complex interactions of historical, geological and climatic events on plant evolution have been an important research focus for many years. However, the role of desert formation and expansion in shaping the genetic structures and demographic histories of plants occurring in arid areas has not been well explored. In the present study, we investigated the phylogeography of Arnebia szechenyi, a desert herb showing a near-circular distribution surrounding the Tengger Desert in Northwest China. We measured genetic diversity of populations using three maternally inherited chloroplast DNA (cpDNA) fragments and seven bi-paternally inherited nuclear DNA (nDNA) loci that were sequenced from individuals collected from 16 natural populations across its range and modelled current and historical potential habitats of the species. Our data indicated a considerably high level of genetic variation within A. szechenyi and noteworthy asymmetry in historical migration from the east to the west. Moreover, two nuclear genetic groups of populations were revealed, corresponding to the two geographic regions separated by the Tengger Desert. However, analysis of cpDNA data did not show significant geographic structure. The most plausible explanation for the discrepancy between our findings based on cpDNA and nDNA data is that A. szechenyi populations experienced long periods of geographic isolation followed by range expansion, which would have promoted generalized recombination of the nuclear genome. Our findings further highlight the important role that the Tengger Desert, together with the Helan Mountains, has played in the evolution of desert plants and the preservation of biodiversity in arid Northwest China.

Rhododendron meddianum is a critically endangered species with important ornamental value and is also a plant species with extremely small populations. In this study, we used double digest restriction-siteassociated DNA sequencing (ddRAD) technology to assess the genetic diversity, genetic structure and demographic history of the three extant populations of R. meddianum. Analysis of SNPs indicated that R. meddianum populations have a high genetic diversity (π=0.0772 ±0.0024, H_E=0.0742 ±0.002). Both F_ST values (0.1582-0.2388) and AMOVA showed a moderate genetic differentiation among the R. meddianum populations. Meanwhile, STRUCTURE, PCoA and NJ trees indicated that the R. meddianum samples were clustered into three distinct genetic groups. Using the stairway plot, we found that R. meddianum underwent a population bottleneck about 70,000 years ago. Furthermore, demographic models of R. meddianum and its relative, Rhododendron cyanocarpum, revealed that these species diverged about 3.05 (2.21-5.03) million years ago. This divergence may have been caused by environmental changes that occurred after the late Pliocene, e.g., the Asian winter monsoon intensified, leading to a drier climate. Based on these findings, we recommend that R. meddianum be conserved through in situ, ex situ approaches and that its seeds be collected for germplasm.

Determining whether the high-latitude Bering land bridge (BLB) was ecologically suitable for the migration of mesothermal plants is significant for Holarctic phytogeographic inferences. Paleobotanical studies provide a critical source of data on the latitudinal positions of different plant lineages at different times, permitting assessment of the efficacy of the BLB for migration. Here we report exceptionally preserved fossils of Firmiana and Tilia endochrysea from the middle Miocene of South Korea. This represents a new reliable record of Firmiana and the first discovery of the T. endochrysea lineage in the fossil record of Asia. The occurrence of these fossils in South Korea indicates that the two lineages had a distribution that extended much farther north during the middle Miocene, but they were still geographically remote from the BLB. In light of the broader fossil record of Asia, our study shows that, in the middle Miocene, some mesothermal plants apparently inhabited the territory adjacent to the BLB and thus they were possibly capable of utilizing the BLB as a migratory corridor. Some other mesothermal plants, such as Firmiana and the T. endochrysea lineages, however, are restricted to more southern regions relative to the BLB based on current fossil evidence. These lineages may have been ecologically unable to traverse the BLB, which raises questions about the efficacy of the BLB as a universal exchange route for mesothermal plants between Asia and North America during the middle Miocene.

Two new species of Yushania (Poaceae, Bambusoideae, Arundinarieae) are described and illustrated from Hunan, China. Yushania longshanensis D.Z. Li & X.Y. Ye is distinguished from related species (Y. confusa, Y. angustifolia and Y. pachyclada) by having a thinner culm (0.2-0.3 cm in diameter), glabrous sheath scar, no oral setae, a large glabrous leaf blade (10-20×0.9-1.3 cm) and 3-4 pairs of secondary veins. Yushania stoloniforma D.Z. Li & X.Y. Ye has a distinctive scrambling habit, which differs from its putative close allies. Both of these two new species have a solitary branch at the basal nodes and can be assigned to Yushania sect. Yushania based on morphological features. Additionally, we treated Yushania gigantea T.P. Yi & L. Yang as a new synonym of Y. elevata T.P. Yi and renamed Y. microphylla T.P. Yi & L. Yang as Y. weiningensis D.Z. Li & X.Y. Ye.

NAC-LIKE, ACTIVATED BY AP3/PI (NAP) is a NAC transcription factor regulating leaf senescence in Arabidopsis thaliana. In wild tobacco Nicotiana attenuata, a nuclear localized NAC transcription factor NaNAC29 was identified to be highly elicited after inoculation of Alternaria alternata, a notorious necrotic fungus on tobacco species. The NaNAC29 possesses similar tertiary structure to NAP with 60% amino acid identity. However, it remains unknown the role of NaNAC29 in plant defense responses to A. alternata and leaf senescence in N. attenuata. In this paper, Defensin-like protein 1 (NaDLP1) was highly induced in N. attenuata after A. alternata inoculation and bigger lesions were developed in NaDLP1-silenced plants. Interestingly, A. alternata-induced NaDLP1 was reduced by 76% in VIGS NaNAC29 plants and by 61% in JA deficient irAOC plants at 3 days post inoculation. The regulation of NaDLP1 expression by NaNAC29 was clearly independent on JA pathway, since exogenous methyl jasmonate treatment could not complement the induction levels of NaDLP1 in NaNAC29-silenced plants to the levels in WT plants. Otherwise, the expression of NaNAC29 was low expressed in young leaves but highly in senescent leaves and darktreated leaves. NaNAC29-silenced plants, which were generated by virus-induced gene silencing (VIGS NaNAC29), showed delayed senescence phenotype. In addition, constitutive over-expression of NaNAC29 in A. thaliana could rescue the delayed-senescence phenotype of nap and caused precocious leaf senescence of wild-type Col-0 plants. All the data above demonstrate that NaNAC29 is a NAP homolog in N. attenuata participating in the defense responses to A. alternata by regulation of a defensin protein NaDLP1 and promoting leaf senescence.

The bHLH transcription factors play pivotal roles in plant growth and development, production of secondary metabolites and responses to various environmental stresses. Although the bHLH genes have been well studied in model plant species, a comprehensive investigation of the bHLH genes is required for tobacco with newly obtained high-quality genome. In the present study, a total of 309 NtbHLH genes were identified and can be divided into 23 subfamilies. The conserved amino acids which are essential for their function were predicted for the NtbHLH proteins. Moreover, the NtbHLH genes were conserved during evolution through analyzing the gene structures and conserved motifs. A total of 265 NtbHLH genes were localized in the 24 tobacco chromosomes while the remained 44 NtbHLH genes were mapped to the scaffolds due to the complexity of tobacco genome. Moreover, transcripts of NtbHLH genes were obviously tissue-specific expressed from the gene-chip data from 23 tobacco tissues, and expressions of 20 random selected NtbHLH genes were further confirmed by quantitative real-time PCR, indicating their potential functions in the plant growth and development. Importantly, overexpressed NtbHLH86 gene confers improve drought tolerance in tobacco indicating that it might be involved in the regulation of drought stress. Therefore, our findings here provide a valuable information on the characterization of NtbHLH genes and further investigation of their functions in tobacco.