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Understanding processes that generate and maintain large disjunctions within plant species can provide valuable insights into plant diversity and speciation. The butterfly bush Buddleja alternifolia has an unusual disjunct distribution, occurring in the Himalaya, Hengduan Mountains (HDM) and the Loess Plateau (LP) in China. We generated a high-quality, chromosome-level genome assembly of B. alternifolia, the first within the family Scrophulariaceae. Whole-genome re-sequencing data from 48 populations plus morphological and petal colour reflectance data covering its full distribution range were collected. Three distinct genetic lineages of B. alternifolia were uncovered, corresponding to Himalayan, HDM and LP populations, with the last also differentiated morphologically and phenologically, indicating occurrence of allopatric speciation likely to be facilitated by geographic isolation and divergent adaptation to distinct ecological niches. Moreover, speciation with gene flow between populations from either side of a mountain barrier could be under way within LP. The current disjunctions within B. alternifolia might result from vicariance of a once widespread distribution, followed by several past contraction and expansion events, possibly linked to climate fluctuations promoted by the Kunlun-Yellow river tectonic movement. Several adaptive genes are likely to be either uniformly or diversely selected among regions, providing a footprint of local adaptations. These findings provide new insights into plant biogeography, adaptation and different processes of allopatric speciation.

Rhododendrons are woody plants, famous throughout the world as having high horticultural value. However, many wild species are currently threatened with extinction. Here, we report for the first time a high-quality, chromosome-level genome of Rhododendron griersonianum, which has contributed to approximately 10% of all horticultural rhododendron varieties but which in its wild form has been evaluated as critically endangered. The final genome assembly, which has a contig N50 size of approximately 34 M and a total length of 677 M, is the highest-quality genome sequenced within the genus to date, in part due to its low heterozygosity (0.18%). Identified repeats constitute approximately 57% of the genome, and 38 280 protein-coding genes were predicted with high support. We further resequenced 31 individuals of R. griersonianum as well as 30 individuals of its widespread relative R. delavayi, and performed additional conservation genomic analysis. The results showed that R. griersonianum had lower genetic diversity (θ = 2.58e-3; π = 1.94e-3) when compared not only to R. delavayi (θ = 11.61e-3, π = 12.97e-3), but also to most other woody plants. Furthermore, three severe genetic bottlenecks were detected using both the Stairway plot and fastsimcoal2 analysis, which are thought to have occurred in the late Middle Pleistocene and the Last Glacial Maximum (LGM) period. After these bottlenecks, R. griersonianum recovered and maintained a constant effective population size (>25 000) until now. Intriguingly, R. griersonianum has accumulated significantly more deleterious mutations in the homozygous state than R. delavayi, and several deleterious mutations (e.g., in genes involved in the response to heat stress) are likely to have harmed the adaptation of this plant to its surroundings. This high-quality, chromosome-level genome and the population genomic analysis of the critically endangered R. griersonianum will provide an invaluable resource as well as insights for future study in this species to facilitate conservation and in the genus Rhododendron in general.

Vespicochory, seed dispersal by hornets, is an uncommon seed dispersal pattern in angiosperms. To date, this phenomenon has been recorded in only four families. Because of its rarity, the causes and consequences of vespicochory remain unclear. Hence, this seed dispersal syndrome is often regarded as anecdotal. Through field investigations, chemical analyses, electrophysiological tests, identification of chemosensory proteins from the antennae of hornets, and behavioral assays, we investigated whether olfactory and/or visual cues of the diaspores of Stemona tuberosa mediate the behavior of the social hornets and maintain their mutualism. This study demonstrated that the elaiosome of S. tuberosa emits hydrocarbons, which are attractive to hornets. However, these compounds, which induce responses in the antennae of naive hornets, are ubiquitous substances on insect cuticle surfaces. Innate preference and experienced foraging behavior of hornets can increase their seed dispersal efficiency. This is the first example in which hydrocarbons have been identified as a diaspore odour involved in the attraction of hornets. Given that the ubiquity of hornets, and the communication function of hydrocarbons in insects, we predict that this rare seed dispersal mechanism may be an overlooked mechanism of insect-plant mutualism.

Flowers or inflorescences often deploy various signals, including visual, olfactory, and gustatory cues, that can be detected by their pollinators. In many plants, these cues and their functions are poorly understood. Deciphering the interactions between floral cues and pollinators is crucial for analyzing the reproductive success of flowering plants. In this study, we examined the composition of the fetid floral scents produced by several Stemona species, including nine S. tuberosa populations from across China, using dynamic headspace adsorption, gas chromatography, and mass spectrometry techniques. We compared variations in floral phenotype, including floral longevity, nectar rewards, pollinator behavior, and flower length and color among the Stemona species. Of the 54 scent compounds identified, the major compounds include fetid dimethyl disulfide, dimethyl trisulfide, 1-pyrroline, butyric acid, p-cresol, isoamyl alcohol, and indole. We detected striking differentiation in floral scent at both the species and population level, and even within a population of plants with different colored flowers. Floral characteristics related to sapromyophily and deceptive pollination, including flower color mimicking livor mortis and a lack of nectar, were found in five Stemona species, indicating that Stemona is a typical sapromyophilous taxon. Species of this monocot genus might employ evolutionary tactics to exploit saprophilous flies for pollination.

Vespicochory, seed dispersal by hornets, is a rare seed dispersal mechanism in angiosperms and, to date, there are few records of this phenomenon. Through field investigations and behavioral assays conducted in four populations of Stemona tuberosa from 2011-2016, we demonstrate that hornets are the primary seed dispersers of S. tuberosa and play an important role in "long-distance" seed dispersal in this species. Furthermore, some ant species act as secondary dispersers and may transport the seeds to safe sites. Hornets and ants provide complementary seed dispersal at different spatial scales. This unique example of insect-plant mutualism may be an underestimated but important strategy to ensure "long-distance" seed dispersal in other myrmecochorous plants.

We engaged experts in various fields of study (pollination ecology, chemical ecology, and ethnobotany), invited community participation, and provided environmental education in an effort to conserve an endangered birthwort (Aristolochia delavayi) and a vulnerable pipevine swallowtail (Byasa daemonius). Scientists studied the uptake and sequestration of the secondarymetabolites aristolochic acids from A. delavayi leaves by different stages of pipevine swallowtail as a defense mechanism; low fruit set of the myophilous A. delavayi due to pollinator limitation; and the emission of chemical signals that attract parasitic wasps by the prepupae of B. daemonius. The results of these studies were part of an education program delivered by personnel of non-governmental organizations. The program was devised to deliver information to the public about the health risks of consuming A. delavayi individuals (aristolochic-acid-associated cancers) and to establish a bridge between the public and scientific research. Following delivery of the program, the behavior of residents changed considerably. Community residents were involved in management activities, including participation in a program to promote understanding of ecological interactions between A. delavayi and B. daemonius; designing an in situ conservation site; monitoring A. delavayi and B. daemonius individuals; and promoting the natural fruit set of A. delavayi by scattering animal excrement to attract fly pollinators. The integration of scientific information and community participation appears to have resulted in an increase in abundance of threatened A. delavayi and B. daemonius populations. We believe the involvement of local people in conservation is necessary for successful species conservation.

In this study, floral color, scent composition and emission rate, nectar property, pollinators, and breeding system of dimorphic Buddleja delavayi Gagnep. were investigated. Flower color of B. delavayi was determined using a standard color chart and spectrophotometer, and two distinct color polymorphisms were observed having purple or white flowers. Floral scents of B. delavayi were collected using dynamic headspace adsorption and identified with coupled gas chromatography and mass spectrometry. In total, 28 compounds were identified from the flowers of B. delavayi. The identified scents were divided into three chemical classes based on their biosynthetic origin: terpenes, fatty acid derivatives, and benzenoids. The scent profiles in all individuals were dominated by a few components, such as lilac aldehyde and alcohol, 4-oxoisophorone, benaldehyde, and oxoisophorone oxide. Floral scent composition (benzenoids and terpenes) showed a significant difference between white and purple flower morphs. Flower color-flower scent associations in B. delavayi were identified with two distinct scent profiles in the two color phenotypes. The studies of other floral characteristics (nectar, floral visitors, breeding system, and fruit set) indicated that floral scent emission rate, nectar volume, visitor visitation frequency, and natural fruit set were not significantly different between the two flower color morphs. Bagging experiments revealed that seed production of B. delavayi is dependent mainly on honeybee Apis cerana. Lastly, this study implies that dimorphic floral color in B. delavayi may have been maintained by floral visitors and nectar guide color.

The pollination biology of Rhododendron cyanocarpum (Franch.) W. W. Smith, an alpine species endemic to NW Yunnan, China, was investigated between 2007 and 2008. Floral traits including flowering time, floral morphology, petal color, and floral scents were assessed, and the associated pollinator assemblage and their foraging behavior were recorded. The flowering period of R. cyanocarpum ranges from late March to middle May and a single flower can last 8-10 days in its natural habitat. Flowers generally show marked herkogamy and the protruding style encourages pollinator foraging behavior that counteracts self-pollination. Floral scents comprise mostly aliphatics (64.37%) and terpenoids (29.88%), which make the flowers attractive for several insect groups, but a peak at 430nm in the reflectance spectrum of petals suggest selection for the attraction of bumblebees. The two Bombus species, B. festivus and B. richardsiellus, representing 90% of total recorded visits, were the only insect species that can be considered to effectively pollinate R. cyanocarpum. Pollination treatments indicated the general self-compatibility of R. cyanocarpum; however, outcrossing seemed to be the dominant strategy, with low rates of self-fertilization providing a certain level of reproductive assurance in its alpine habitat.

Natural hybridization was assumed to play an essential role for the diversification of Primula; however, only one study of hybridization in the region of the Himalayas has been undertaken. In the present study, we examined another natural hybrid zone where morphologically putative hybrids as well as P. beesiana Forrest, P. bulleyana Forrest, and P. poissonii Franch. co-occurred. We used molecular data to confirm the parental species of putative hybrids and the unidirectional hybridization pattern between P. beesiana and P. bulleyana. Moreover, with reference to synthetic F(1)s, most hybrids examined are possibly advanced generations, although the possibility of F-1 hybrids currently examined could not be completely excluded. In addition, pollinator observations on experimental arrays of transplanted parental species showed interspecific pollen flows during visitations of shared pollinators, indicating an incomplete pre-zygotic barrier between P. beesiana and P. bulleyana. Seed productions from both flower morphs of putative hybrids were significantly lower than parental species, suggesting lower reproductive success in these hybrids. Combined with the evidence of recent habitat disturbance in the study area, we might witness the early process of hybridization between P. beesiana and P. bulleyana.

Three new clerodane diterpenoids, splendidins A - C (1 - 3, resp.), were isolated from Salvia splendens, together with six known ones. Their structures were elucidated by extensive spectroscopic analysis. Splendidin C (3) was the first diterpenoid glucoside reported from this plant. These compounds were evaluated for their cytotoxic activity; however, none of them were cytotoxic.

Buddleja macrostachya (Buddlejaceae) is a widespread shrub native to the Sino-Himalayan mountains and beyond. It has been found to occur at two ploidy levels, hexaploid, 2n=6x=114 and dodecaploid, 2n=12x=228. To determine if morphological characters might be used as indicators of ploidy levels, we measured floral and fruit length, relative and absolute leaf size, trichome density on both leaf surfaces, and stomatal density and length in different populations of B. macrostachya. In general, flower and fruit length, absolute leaf size, and stomatal length increased with an increase at ploidy level (P 0.05) in different populations. Other characters studied such as trichome type, cuticular membrane and ornamentation of stomata, cell and stomatal shape, and anticlinal wall pattern were quite constant in this species. Thus it appears that flower and fruit length, absolute leaf size, and stomatal frequency and length can be used to distinguish hexaploid from dodecaploid cytotypes either in the field or in herbarium specimens.

Microsatellite primers were developed to assess genetic diversity and population structure in Acer yangbiense, a critically endangered endemic species that occurs in northwestern Yunnan Province, China. •Using the Fast Isolation by AFLP of Sequences Containing repeats (FIASCO) protocol, 34 microsatellite loci were isolated and characterized in A. yangbiense. Polymorphisms were evaluated in 39 individuals from two distinct populations, one of which was naturally occurring and the other an ex situ grouping. Nine of the markers showed polymorphisms with two to five alleles per locus; observed (H(o)) and expected (H(e)) heterozygosities ranged from 0.0000 to 0.8000 and from 0.0000 to 0.6889, respectively. •These microsatellite primers will prove useful in the further investigation of population genetics studies in A. yangbiense and, in addition, will assist related research of other congeneric species.