The complete chloroplast genome of Hemiboea pterocaulis (Gesneriaceae) exclusively distributed in Guilin, Guangxi, China.

Hemiboea pterocaulis is a unique species only found in Guilin, Guangxi, China. In this study, we sequenced and assembled the complete chloroplast genome of H. pterocaulis and revealed its phylogenetic relationship with other Hemiboea species. The chloroplast genome sequence of H. pterocaulis is 153,159 bp in length and comprises a large single-copy (LSC) region of 84,178 bp, a small single-copy (SSC) region of 18,087 bp, and a pair of inverted repeat (IR) regions, each with a length of 25,447 bp. It has a total GC content of 37.6% and encodes 132 genes, including 87 protein-coding genes, 37 tRNA genes, and eight rRNA genes. The phylogenetic relationships based on the complete chloroplast genome sequences of Hemiboea taxa indicate that H. pterocaulis is most closely related to H. suiyangensis, indicating that H. pterocaulis is an independent species and is separated from the H. subcapitata complex. These results provide valuable insights into the phylogeny, species divergence, and delimitation of the Hemiboea genus.

Evolution of intraspecific floral variation in a generalist-specialist pollination system.

Intraspecific processes impact macroevolutionary patterns through individual variation, selection, and ecological specialisation. According to the niche variation hypothesis, the broader ecological niche of gen- eralist species results in an increased morphological variation among individuals, either because they are constituted of diversified specialised individuals each exploiting a fraction of the species' niche, or because they are constituted of true generalist individuals that experience relaxed selection. To test this hypoth- esis, we surveyed the individual floral morphology of species of Antillean Gesneriaceae, a group that has transitioned between specialisation for hummingbird pollination and generalisation multiple times throughout its evolutionary history. We characterised the profiles of corollas using geometric morpho- metrics and compared the intraspecific shape variance of specialists and generalists in a phylogenetic context. We used three approaches that differently accounted for the high dimensionality of morphologi- cal traits, the ancestral reconstruction of pollination syndromes over time, and the error associated with the estimation of the intraspecific variance. Our findings provide partial support for the niche variation hypothesis. If considering the whole shape in the analysis corroborated this idea, decomposing the shape into principal components indicated that not all aspects of the corolla exhibit the same pattern of vari- ation. Specifically, pollination generalists tend to display greater intraspecific variation than specialists in terms of tubularity, but not of curvature. Accounting for the error in the variance estimation also reduced the support for the hypothesis, suggesting that larger sample sizes may be required to reach stronger conclusions. This study emphasises the reciprocal influence between plants and their pollinators on floral morphology at different biodiversity scales, and suggests that ecological strategies of species can affect patterns of morphological variation at macroevolutionary scales.

Cyrtandraobliquifolia (Gesneriaceae), a new species from Kaua'i, Hawaiian Islands.

Cyrtandraobliquifolia K.R. Wood & W.L. Wagner (Gesneriaceae), a new shrub species known only from Kaua'i, Hawaiian Islands, is described and illustrated with notes on its distribution, ecology, and conservation status. The new species is morphologically most similar to Cyrtandrawawrae C.B. Clarke but differs by its unique combination of oblique, non-peltate, auriculate leaf bases, more deeply divided calyx lobes, inflorescence with fewer flowers and lacking profusely umbellate cymes. Cyrtandraobliquifolia is known from only two localities which have undergone severe habitat degradation from landslides and invasive plants and animals and is determined to be Critically Endangered (CR) when evaluated under IUCN criteria.

Complete chloroplast genome of Petrocosmea qinlingensis (Gesneriaceae), a protected wild plant in the Qinling mountains.

Petrocosmea qinlingensis is a protected wild plant endemic in China, inhabiting low-light limestone cliffs but the complete chloroplast genome has not been reported. In this study, we first sequenced and assembled the complete chloroplast genome of P. qinlingensis. The total size of this genome was 153,865 bp, including a large single-copy (LSC) region (84,737 bp), a small single-copy (SSC) region (18,244 bp), and two inverted repeats (IRs) regions (25,442 bp). This genome encoded 111 uniquegenes, consisted of 77 protein-coding genes, four ribosomal RNA genes, and 30 transfer RNA genes. Phylogenomic analysis based on the chloroplast protein-coding genes and showed that the genus Petrocosmea was the closest relative to Raphiocarpus. Our results will support further phylogeographic, population genetic studies of this species.

Staurantherafloribunda, a new species of Gesneriaceae from Yunnan, China.

Staurantherafloribunda F.Su, C.Y.Hao & K.Tan, a new species of Gesneriaceae from Yunnan, China, is described and illustrated here. It is morphologically similar to S.grandifolia Benth. in the shape of corolla, stigma, leaves and the number of stamens. However, it can be readily distinguished from the compared species by its dense cymes, leaf indumentum, lack of a corolla spur, calyx colour and stamen shape. The description of the new species, photographs, detailed descriptions, notes on etymology, distribution and habitat, as well as comparisons with morphologically similar species, are provided.

A new combination and a new synonym of Gesneriaceae in China.

Didymocarpussubpalmatinervis W.T.Wang was shown to be conspecific with Petrocodonlithophilus Y.M.Shui, W.H.Chen & Mich.Möller, by checking original literature, examining specimens, tracing specimen collecting history, and conducting field surveys. The results show morphological characteristics and geographical distribution overlaps between these two species. The transfer of Didymocarpussubpalmatinervis to Petrocodon as a new combination with Petrocodonsubpalmatinervis (W.T.Wang) F.Wen & Z.L.Li is proposed here, and P.lithophilus is synonymized with P.subpalmatinervis. Color photographs and essential information are also provided, including a detailed comparison of description, distribution, habitat, and the proposed IUCN conservation status.

Two new naphthoquinone derivatives from Sinningia conspicua (Gesneriaceae).

Two new naphthoquinones, 7-hydroxy-2-O-methyldunniol (1) and 7-methoxy-2-O-methyldunniol (2) were isolated from the tubers of Sinningia conspicua (Seem.) G. Nicholson (Gesneriaceae). Nine known compounds were also obtained: 7-methoxydunniol (3), dunniol (4), 7-methoxy-α-dunnione (5), 7-hydroxydunnione (6), 2,4,4',6-tetramethoxychalcone (7), loureirin B (8), allo-calceolarioside A (9), ß-sitosterol (10) and stigmasterol (11). This is the first report of compounds 3 and 5 in Sinningia. n-Hexane, ethyl acetate and ethanol extracts, and compounds 1-5 were tested for cytotoxic activity against PC-3 (prostate) and SKMEL 103 (melanoma) human tumour cell lines and also against the 3T3 (fibroblast) non-cancer cell line. All samples were inactive.

Primulinapingnanensis, a new species of Gesneriaceae from Guangxi, China.

A new species of Primulina, P.pingnanensis, from the Guangxi Zhuangzu Autonomous Region, China, is described and illustrated here. It is morphologically similar to P.orthandra but has significant differences in the bracts, corolla tube and lobes shape, as well as in the indumentum of the outer surface of the corolla, the filaments, the staminodes and the anthers. Colorful photographs and essential information of this new taxon are also provided, including detailed taxonomic description, distribution, habitat, the comparison table, and the IUCN conservation status. We also discuss a validation of new combination P.crassifolia and Chiritacrassifolia.

Oreocharis oriolus, a new species of Gesneriaceae in a sclerophyllous oak community from Yunnan, Southwest China.

Oreocharis oriolus, a new species of Gesneriaceae in a sclerophyllous oak community from Yunnan, Southwest China, is described and illustrated. Morphologically, it resembles both O. forrestii and O. georgei, while it is distinct in combined characters of wrinkled leaves, peduncle and pedicel covered with whitish and egladular villous hairs, bract lanceolate and nearly glabrescent adaxially, and staminode absent. Molecular phylogenetic analysis based on nuclear ribosomal internal transcribed spacer (nrITS) and chloroplast DNA fragment (trnL-F) of 61 congeneric species also supported O. oriolus as a new species while it was nested with O. delavayi. It was currently assessed as 'Critically Endangered' (CR) regarding to its small-sized population and narrow distribution following the IUCN categories and criteria.

Plastome evolution and phylogenomics of Trichosporeae (Gesneriaceae) with its morphological characters appraisal.

Trichosporeae is the largest and most taxonomically difficult tribe of Gesneriaceae due to its diverse morphology. Previous studies have not clarified the phylogenetic relationships within this tribe on several DNA markers, including the generic relationships within subtribes. Recently, plastid phylogenomics have been successfully employed to resolve the phylogenetic relationships at different taxonomic levels. In this study, plastid phylogenomics were used to explore the relationships within Trichosporeae. Eleven plastomes of Hemiboea were newly reported. Comparative analyses, phylogeny and morphological character evolution within Trichosporeae were conducted on 79 species representing seven subtribes. The Hemiboea plastomes range from 152,742 bp to 153,695 bp in length. Within Trichosporeae, the sampled plastomes range from 152,196 bp to 156,614 bp and GC content from 37.2% to 37.8%. A total of 121-133 genes were annotated in each species, including 80-91 protein-coding genes, 34-37 tRNA genes, and 8 rRNA genes. The contraction and expansion of IR borders were not detected, and gene rearrangements and inversions did not occur. The 13 hypervariable regions were proposed as the potential molecular markers for species identification. A total of 24,299 SNPs and 3,378 indels were inferred, and most of the SNPs were functionally missense and silent variations. There were 1968 SSRs, 2055 tandem repeats and 2802 dispersed repeats. The RSCU and ENC values indicated that the codon usage pattern was conserved in Trichosporeae. Both the phylogenetic frameworks based on the whole plastome and 80 CDSs were basically concordant. The sister relationships between Loxocarpinae and Didymocarpinae were confirmed, and Oreocharis was a sister group of Hemiboea with high support. The morphological characters showed a complex evolutionary pattern of Trichosporeae. Our findings may contribute to future research on genetic diversity, morphological evolutionary patterns, and conservation of the tribe Trichosporeae.

A high quality, high molecular weight DNA extraction method for PacBio HiFi genome sequencing of recalcitrant plants.

BACKGROUND: PacBio HiFi sequencing provides highly accurate long-read sequencing datasets which are of great advantage for whole genome sequencing projects. One limitation of the method is the requirement for high quality, high molecular weight input DNA. This can be particularly challenging for plants that frequently contain common and species-specific secondary metabolites, which often interfere with downstream processes. Cape Primroses (genus Streptocarpus), are some of these recalcitrant plants and are selected here as material to develop a high quality, high molecular weight DNA extraction protocol for long read genome sequencing. RESULTS: We developed a DNA extraction method for PacBio HiFi sequencing for Streptocarpus grandis and Streptocarpus kentaniensis. A CTAB lysis buffer was employed to avoid guanidine, and the traditional chloroform and phenol purification steps were replaced with pre-lysis sample washes. Best cells/nucleus lysis was achieved with 4 h at 58 °C. The obtained high quality and high molecular weight DNAs were tested in PacBio SMRTBell™ library preparations, which resulted in circular consensus sequencing (CCS) reads from 17 to 27 Gb per cell, and a read length N50 from 14 to 17 kbp. To evaluate the quality of the reads for whole genome sequencing, they were assembled with HiFiasm into draft genomes, with N50 = 49 Mb and 23 Mb, and L50 = 10 and 11. The longest contigs were 95 Mb and 57 Mb respectively, showing good contiguity as these are longer than the theoretical chromosome length (genome size/chromosome number) of 78 Mb and 55 Mb, for S. grandis and S. kentaniensis respectively. CONCLUSIONS: DNA extraction is a critical step towards obtaining a complete genome assembly. Our DNA extraction method here provided the required high quality, high molecular weight DNA for successful standard-input PacBio HiFi library preparation. The contigs from those reads showed a high contiguity, providing a good starting draft assembly towards obtaining a complete genome. The results obtained here were highly promising, and demonstrated that the DNA extraction method developed here is compatible with PacBio HiFi sequencing and suitable for de novo whole genome sequencing projects of plants.

Gesneriads, a Source of Resurrection and Double-Tolerant Species: Proposal of New Desiccation- and Freezing-Tolerant Plants and Their Physiological Adaptations.

Gesneriaceae is a pantropical family of plants that, thanks to their lithophytic and epiphytic growth forms, have developed different strategies for overcoming water scarcity. Desiccation tolerance or "resurrection" ability is one of them: a rare phenomenon among angiosperms that involves surviving with very little relative water content in their tissues until water is again available. Physiological responses of desiccation tolerance are also activated during freezing temperatures, a stress that many of the resurrection gesneriads suffer due to their mountainous habitat. Therefore, research on desiccation- and freezing-tolerant gesneriads is a great opportunity for crop improvement, and some of them have become reference resurrection angiosperms (Dorcoceras hygrometrica, Haberlea rhodopensis and Ramonda myconi). However, their difficult indoor cultivation and outdoor accessibility are major obstacles for their study. Therefore, this review aims to identify phylogenetic, geoclimatic, habitat, and morphological features in order to propose new tentative resurrection gesneriads as a way of making them more reachable to the scientific community. Additionally, shared and species-specific physiological responses to desiccation and freezing stress have been gathered as a stress response metabolic basis of the family.

Three new naphthoquinones from the tubers of Sinningia mauroana.

Three new naphthoquinones, 5,6,7-trimethoxydunnione (1), 6,7-dimethoxy-α-dunnione (2), and 5,6,7-trimethoxydunniol (3) were isolated from the tubers of Sinningia mauroana Chautems, together with eleven known compounds: 5-hydroxy-6,7-dimethoxy-α-dunnione (4), 6-hydroxy-7-methoxy-α-dunnione (5), 7-hydroxy-6-methoxy-α-dunnione (6), cedrol (7), tectoquinone (8), plantainoside A (9), calceolarioside A (10), calceolarioside B (11), sanangoside (12), allo-calceolarioside A (13), and conandroside (14). The known compounds 4, 7-9 and 11-13 are being reported for the first time in this species. Hexane, ethyl acetate and ethanol extracts were tested for antioxidant activity by the ORAC-FL method, and the antioxidant capacity was measured as trolox equivalent (TE). The ethyl acetate (8808 µg TE g-1) and ethanol (7911 µg TE g-1) extracts showed high antioxidant capacity, while the hexane extract displayed weak antioxidant capacity (860 µg TE g-1).

Leaf gas exchange and water relations of the woody desiccation-tolerant Paraboea rufescens during dehydration and rehydration.

Desiccation-tolerant (DT) plants can withstand dehydration to less than 0.1 g H2O g-1 dry weight. The mechanism for whole-plant recovery from severe dehydration is still not clear, especially for woody DT plants. In the present study, we evaluated the desiccation tolerance and mechanism of recovery for a potentially new woody resurrection plant Paraboea rufescens (Gesneriaceae). We monitored the leaf water status, leaf gas exchange, chlorophyll fluorescence and root pressure of potted P. rufescens during dehydration and rehydration, and we investigated the water content and chlorophyll fluorescence of P. rufescens leaves in the field during the dry season. After re-watering from a severely dehydrated state, leaf maximum quantum yield of photosystem II of P. rufescens quickly recovered to well-watered levels. Leaf water status and leaf hydraulic conductance quickly recovered to well-watered levels after re-watering, while leaf gas exchange traits also trended to recovery, but at a slower rate. The maximum root pressure in rehydrated P. rufescens was more than twice in well-watered plants. Our study identified P. rufescens as a new DT woody plant. The whole-plant recovery of P. rufescens from extreme dehydration is potentially associated with an increase of root pressure after rehydration. These findings provide insights into the mechanisms of recovery of DT plants from dehydration.

Flavonoids and phenylethanoids from the flowers and leaves of Aeschynanthus species and cultivars (Gesneriaceae).

Forty-one flavones, each one of flavonol, chalcone and dihydroflavonol, two flavanones, and four phenylethanoids were isolated from corollas, calyces and leaves of two Aeschynanthus species, A. fulgens and A. pulcher, and six cultivars, 'Mahligai', 'Mona Lisa', SoeKa', 'Redona', 'Freshya' and 'Bravera'. Flavonoids were mainly the glucuronides and/or methylglucuronides based on hispidulin, nepetin, pectolinarigenin, 6-hydroxyluteolin, scutellarein, apigenin and luteolin, and identified by UV spectra, HR-MS, LC-MS, acid hydrolysis, NMR, and/or HPLC and TLC comparisons with authentic samples. Of these flavonoids, twelve, i.e. hispidulin 7,4'-di-O-glucuronide, 7,4'-di-O-methylglucuronide, 7-O-methylglucuronide-4'-O-glucuronide, 7-O-glucuronide-4'-O-methylglucuronide, 7-O-glucosyl-(1 â†’ 2)-glucuronide and 8-C-glucoside, nepetin 7,4'-di-O-glucuronide, 7-O-glucuronide-4'-O-methylglucuronide and 7-O-methylglucuronide-4'-O-glucuronide, pectolinarigenin 7-O-glucosyl-(1 â†’ 2)-glucuronide and 7-O-xylosyl-(1 â†’ 2)-(6″-malonylglucoside), and 6-hydroxyluteolin 7,4'-di-O-glucuronide, were previously undescribed.

The first complete chloroplast genome of Briggsia chienii W. Y. Chun and its phylogenetic position within Gesneriaceae.

Briggsia chienii W. Y. Chun 1946 is an endemic herbaceous perennial species distributed in southern China. In this study, we firstly characterized the complete chloroplast genome sequence of B. chienii and provided new molecular resources for promoting its conservation and taxonomic assignment. Its complete chloroplast genome is 154,082 bp in length and contains the typical quadripartite structure of angiosperm plastome, including two inverted repeat (IR) regions of 25,447 bp, a large single-copy (LSC) region of 85,035 bp, and a small single-copy (SSC) region of 18,153 bp. The plastome contains 114 genes, consisting of 80 protein-coding genes, 30 tRNA gene, and 4 rRNA genes. The overall GC content in the plastome of B. chienii is 37.4%, which is lower than lots of angiosperm plastome. The phylogenetic result indicated that B. chienii exhibited the closest relationship with Oreocharis cotinifolia W. T. Wang 1983, and provided new information for the phylogeny relationship of genus Briggsia.

Genetic Variability in Balkan Paleoendemic Resurrection Plants Ramonda serbica and R. nathaliae Across Their Range and in the Zone of Sympatry.

The genus Ramonda includes three Paleoendemic and Tertiary relict species that survived in refugial habitats of the Balkan Peninsula (R. nathaliae and R. serbica) and the Iberian Peninsula (R. myconi). They are all "resurrection plants," a rare phenomenon among flowering plants in Europe. Ramonda myconi and R. nathaliae are diploids (2n = 2x = 48), while R. serbica is a hexaploid (2n = 6x = 144). The two Balkan species occur in sympatry in only two localities in eastern Serbia, where tetraploid potential hybrids (2n = 4x = 96) were found. This observation raised questions about the existence of gene flow between the two species and, more generally, about the evolutionary processes shaping their genetic diversity. To address this question, genetic markers (AFLP) and an estimate of genome size variation were used in a much larger sample and at a larger geographic scale than previously. The combination of AFLP markers and genome size results suggested ongoing processes of interspecific and interploidy hybridization in the two sites of sympatry. The data also showed that interspecific gene flow was strictly confined to sympatry. Elsewhere, both Ramonda species were characterized by low genetic diversity within populations and high population differentiation. This is consistent with the fact that the two species are highly fragmented into small and isolated populations, likely a consequence of their postglacial history. Within sympatry, enormous variability in cytotypes was observed, exceeding most reported cases of mixed ploidy in complex plant species (from 2x to >8x). The AFLP profiles of non-canonical ploidy levels indicated a diversity of origin pathways and that backcrosses probably occur between tetraploid interspecific hybrids and parental species. The question arises whether this diversity of cytotypes corresponds to a transient situation. If not, the question arises as to the genetic and ecological mechanisms that allow this diversity to be maintained over time.

Oreocharis xieyongii, an unusual new species of Gesneriaceae from western Hunan, China.

A new species, Oreocharis xieyongii T. Deng, D.G. Zhang & H. Sun, from Hunan Province, central China, is described. The combination of purple zygomorphic corolla with longer adaxial lobes and exserted stamens defines the species and discriminates it from all other current Oreocharis species. Morphological traits of the new species were compared to those of two similar species, Oreocharis xiangguiensis and O. rubrostriata. Phylogenetic analysis indicates that the new species is nested within the Oreocharis. Although only half of Oreocharis species were included in our study, evolutionary character analysis indicates that the ancestral states of the genus are likely the purple corolla, longer abaxial lip and inserted stamens. The longer adaxial lip is perhaps an apomorphy and only present in O. xieyongii and O. rubrostriata. Both morphological and molecular evidence suggest that O. xieyongii is a taxon new to science.

The first genome for the Cape Primrose Streptocarpus rexii (Gesneriaceae), a model plant for studying meristem-driven shoot diversity.

Cape Primroses (Streptocarpus, Gesneriaceae) are an ideal study system for investigating the genetics underlying species diversity in angiosperms. Streptocarpus rexii has served as a model species for plant developmental research for over five decades due to its unusual extended meristem activity present in the leaves. In this study, we sequenced and assembled the complete nuclear, chloroplast, and mitochondrial genomes of S. rexii using Oxford Nanopore Technologies long read sequencing. Two flow cells of PromethION sequencing resulted in 32 billion reads and were sufficient to generate a draft assembly including the chloroplast, mitochondrial and nuclear genomes, spanning 776 Mbp. The final nuclear genome assembly contained 5,855 contigs, spanning 766 Mbp of the 929-Mbp haploid genome with an N50 of 3.7 Mbp and an L50 of 57 contigs. Over 70% of the draft genome was identified as repeats. A genome repeat library of Gesneriaceae was generated and used for genome annotation, with a total of 45,045 genes annotated in the S. rexii genome. Ks plots of the paranomes suggested a recent whole genome duplication event, shared between S. rexii and Primulina huaijiensis. A new chloroplast and mitochondrial genome assembly method, based on contig coverage and identification, was developed, and successfully used to assemble both organellar genomes of S. rexii. This method was developed into a pipeline and proved widely applicable. The nuclear genome of S. rexii and other datasets generated and reported here will be invaluable resources for further research to aid in the identification of genes involved in morphological variation underpinning plant diversification.

Monopyleglutinosa (Gesneriaceae), a new species from the western slopes of the Ecuadorian Andes.

Exploratory field expeditions to the western slopes of the Ecuadorian Andes resulted in the discovery of a new species of Monopyle (Gesneriaceae). Monopyleglutinosa J.L.Clark & Keene, sp. nov. is described as a narrow endemic from lowland forests along the border of the Reserva Ecológica Los Illinizas in the Province of Cotopaxi. The new species is unique for the presence of glutinous or sticky trichomes on the calyx lobes and outer surface of the inferior ovary. Based on IUCN guidelines, a preliminary conservation status is assigned as Critically Endangered (CR).

ResumenLas expediciones a los bosques de las laderas noroccidentales de los Andes de Ecuador dieron como resultado el descubrimiento de una nueva especie: Monopyleglutinosa J.L.Clark & Keene, sp. nov., la cual es endémica de una reducida área en el borde de la Reserva Ecológica Los Illinizas en la provincia de Cotopaxi. La nueva especie es única por la presencia de tricomas glutinosos o pegajosos en los lóbulos del cáliz y la superficie externa del ovario ínfero. Basados en los criterios de la UICN, se asigna un estado de conservación preliminar de En Peligro Crítico (CR).