Chromosome-scale genome assembly and annotation of Cotoneaster glaucophyllus.

Cotoneaster glaucophyllus is a semi-evergreen plant that blossoms in late summer, producing dense, attractive, fragrant white flowers with significant ornamental and ecological value. Here, a chromosome-scale genome assembly was obtained by integrating PacBio and Illumina sequencing data with the aid of Hi-C technology. The genome assembly was 563.3 Mb in length, with contig N50 and scaffold N50 values of ~6 Mb and ~31 Mb, respectively. Most (95.59%) of the sequences were anchored onto 17 pseudochromosomes (538.4 Mb). We predicted 35,856 protein-coding genes, 1,401 miRNAs, 655 tRNAs, 425 rRNAs, and 795 snRNAs. The functions of 34,967 genes (97.52%) were predicted. The availability of this chromosome-level genome will provide valuable resources for molecular studies of this species, facilitating future research on speciation, functional genomics, and comparative genomics within the Rosaceae family.

High-quality chromosome-level genome assembly and multi-omics analysis of rosemary (Salvia rosmarinus) reveals new insights into the environmental and genome adaptation.

High-quality genome of rosemary (Salvia rosmarinus) represents a valuable resource and tool for understanding genome evolution and environmental adaptation as well as its genetic improvement. However, the existing rosemary genome did not provide insights into the relationship between antioxidant components and environmental adaptability. In this study, by employing Nanopore sequencing and Hi-C technologies, a total of 1.17 Gb (97.96%) genome sequences were mapped to 12 chromosomes with 46 121 protein-coding genes and 1265 non-coding RNA genes. Comparative genome analysis reveals that rosemary had a closely genetic relationship with Salvia splendens and Salvia miltiorrhiza, and it diverged from them approximately 33.7 million years ago (MYA), and one whole-genome duplication occurred around 28.3 MYA in rosemary genome. Among all identified rosemary genes, 1918 gene families were expanded, 35 of which are involved in the biosynthesis of antioxidant components. These expanded gene families enhance the ability of rosemary adaptation to adverse environments. Multi-omics (integrated transcriptome and metabolome) analysis showed the tissue-specific distribution of antioxidant components related to environmental adaptation. During the drought, heat and salt stress treatments, 36 genes in the biosynthesis pathways of carnosic acid, rosmarinic acid and flavonoids were up-regulated, illustrating the important role of these antioxidant components in responding to abiotic stresses by adjusting ROS homeostasis. Moreover, cooperating with the photosynthesis, substance and energy metabolism, protein and ion balance, the collaborative system maintained cell stability and improved the ability of rosemary against harsh environment. This study provides a genomic data platform for gene discovery and precision breeding in rosemary. Our results also provide new insights into the adaptive evolution of rosemary and the contribution of antioxidant components in resistance to harsh environments.

A new species of Cotoneaster (Rosaceae) from western Sichuan, China.

Cotoneasterdensiflorus, a new species of Rosaceae from western Sichuan, China, is described and illustrated. Morphologically, we inferred that the new species belongs to CotoneasterSer.Salicifolii sensu Yü et al. (1974) in the Flora of China and Fryer and Hylmö (2009). This species is most similar to C.salicifolius, but differs in its leaf blade of ovate-lanceolate to obovate shape (vs. elliptic-oblong to ovate-lanceolate), smaller length-width ratio of 2.37 ± 0.31 (vs. 3.17 ± 0.32), slightly conduplicate (vs. not conduplicate), less lateral veins of 6-8 pairs (vs. 12-16 pairs), upper surface slightly rugose (vs. rugose), leaf margin plane (vs. revolute), lower surface densely grey tomentose (vs. grey tomentose, with bloom), greater corolla diameter of 7-9 mm (vs. 5-6 mm), styles 2 (vs. 2-3), pyrenes 2 (vs. 2-3), larger pollen grains P/E values of 2.05 ± 0.12 (vs. 1.19 ± 0.05) and leaf epidermis type W (vs. type I). Based on phylogenetic analysis of the whole chloroplast genome, C.densiflorus is sister to C.rhytidophyllus, but distantly related to C.salicifolius.

Contribution of Constituents in Rosemary-Magnolia Compound Essential Oil to Antibacterial, Antioxidant and Cytotoxicity Bioactivities Revealed by Grey Correlation Analysis.

The compound of essential oils (EOs) is a key approach to achieving the superimposed efficacy of plant EOs. In this article, grey correlation analysis was applied for the first time to explore the compound ratios and contribution between constituents and the bioactivity of the compound EOs. There were 12 active constituents shared in rosemary and magnolia EOs prepared by negative pressure distillation. With different proportions, these two EOs were blended and analyzed for the antioxidant, bacteriostatic and antitumor effects. According to the results of the inhibition circle, minimum bactericidal and inhibitory concentration, the most obvious inhibition effect of the compound EOs on different strains of bacteria was shown in Staphylococcus aureus. The results of antioxidant test showed that single EO from rosemary had the best antioxidant effect, and its EO content was directly proportional to the antioxidant effect. The cytotoxicity results showed that, there was a significant difference in the lethality of the compound EOs between tumor cells Mcf-7 (human breast cancer cells) and SGC-7901 cells (human gastric cancer cells). Furthermore, single EO from magnolia had an obvious inhibitory effect on the growth of Mcf-7 cells and SGC-7901 cells, and the cell lethality rate was as high as 95.19 % and 97.96 %, respectively. As the results of grey correlation analysis, the constituents with the maximal correlation of inhibitory effects on bacteria were as follows: S. aureus - Terpinolene (0.893), E. coli - Eucalyptol (0.901), B. subtilis - α-Pinene (0.823), B. cereus - Terpinolene (0.913) and Salmonella - α-Phellandrene (0.855). For the ABTS and DPPH scavenging effects, the constituents with the maximal correlation were (-)-Camphor (0.860) and ß-Pinene (0.780), respectively. In terms of the effects of the active constituents of compound EOs on the inhibitory activities of tumor cells Mcf-7 and SGC-7901, the three active constituents of γ-Terpinene, (R)-(+)-ß-Citronellol and (-)-Camphor were in the top three, and their correlation were Mcf-7 (0.833, 0.820, 0.795) and SGC-7901 (0.797, 0.766, 0.740). Our study determined the contribution degree of active constituents in the antibacterial, antioxidant, and antitumor bioactivities of rosemary-magnolia compound EOs, and also provided new insights for the research of EOs combination formulations.

Biotransformation, metabolic response, and toxicity of UV-234 and UV-326 in larval zebrafish (Danio rerio).

Benzotriazole ultraviolet stabilizers (BUVSs) are emerging pollutants that are widely detected in aquatic ecosystems. While structure-dependent effects of BUVSs are reported, the relationship between biotransformation and toxicity outcomes remains unclear. In this study, zebrafish embryos were exposed to two common BUVSs (UV-234 and UV-326) at 1, 10, and 100 µg/L for up to 7 days. Comparison of their uptake and biotransformation revealed that the bioaccumulation capacity of UV-234 was higher than that of UV-326, while UV-326 was more extensively biotransformed with additional conjugation reactions. However, UV-326 showed low metabolism due to inhibited phase II enzymes, which may result in the comparable internal concentrations of both BUVSs in larval zebrafish. Both BUVSs induced oxidative stress while decreased MDA, suggesting the disturbance of lipid metabolism. The subsequent metabolomic profiling revealed that UV-234 and UV-326 exerted different effects on arachidonic acid, lipid, and energy metabolism. However, both BUVSs negatively impacted the cyclic guanosine monophosphate / protein kinase G pathway. This converged metabolic change resulted in comparable toxicity of UV-234 and UV-326, which was confirmed by the induction of downstream apoptosis, neuroinflammation, and abnormal locomotion behavior. These data have important implications for understanding the metabolism, disposition, and toxicology of BUVSs in aquatic organisms.

Comparative analysis of cold-responsive genes under short-term cold stimulation and cold-adaptive genes under long-term heterogeneous environments reveals a cold adaptation mechanism in weeping forsythia.

Identifying cold-related genes can provide insights into the cold adaptation mechanism of weeping forsythia. In this study, we compared the changes in gene expressions and physiological and biochemical indices under short-term cold stimulation with the changes in gene sequences under a long-term heterogeneous environment to investigate the cold adaptation mechanism in weeping forsythia. The data of adaptive gene sequence changes, e.g., single nucleotide polymorphisms, were obtained from previous landscape genomics studies. The physiological and biochemical indicators and transcriptome results showed that weeping forsythia initiated a series of programs, including increasing cell osmotic pressures, scavenging ROS, activating the defense mechanism that crosses with pathogen infection, and upregulating CBF/DREB1 transcription factor 1, to cope with short-term cold stress. A reanalysis of landscape genomic data suggested that weeping forsythia responded to long-term heterogeneous cold stress by the differentiation of genes related to synthesis of aromatic substances and adenosine triphosphate. Our results supported the hypothesis that the adaptation mechanisms of species to short-term environmental stimulation and long-term stress in heterogeneous environments are different. The differences in cold tolerance among populations are not necessarily obtained by changing cold-responsive gene sequences. This study provides new insights into the cold adaptation mechanisms of plants.

Diversity Analysis of Leaf Nutrient Endophytes and Metabolites in Dioecious Idesia polycarpa Maxim Leaves during Reproductive Stages.

Leaves are essential vegetative organs of plants. Studying the variations in leaf nutrient content and microbial communities of male and female plants at reproductive stages helps us understand allocation and adaptation strategies. This study aimed to determine the nutrient characteristics and microbial differences in the leaves of male and female Idesia polycarpa at reproductive stages. Seven-year-old female and male plants were used as test materials in this experiment. The samples were collected at three stages: flowering (May), fruit matter accumulation (July), and fruit ripening (October). The nitrogen (TN), phosphorus (TP), potassium (TK), carbon (TC), and the pH of the female and male leaves were analyzed. In addition, the leaf microbial diversity and differential metabolites were determined using the Illumina high-throughput sequencing method and the ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method at the reproductive developmental stages. This study found that male and female plant leaves had different TN and TK contents over time but no difference in TC and TP content. The significant differences in bacterial diversity between male and female plants and the richness of the fungi of male plants at the flowering and fruit maturity stages were observed. Proteobacteria, Pseudomonadaceae, Ascomycota, and Aspergillus were the dominant bacteria and fungi in the Idesia polycarpa leaves. The presence of microorganisms differed in the two sexes in different periods. Alphaproteobacteria and Sordariomycetes were the indicator groups for male leaves, and Pseudomonas and Sordariomycetes were the indicator groups for female leaves. Significant differences in phenolic acid were found between male and female leaves. A KEGG enrichment analysis revealed that differential metabolites were enriched in metabolic pathways, amino acid biosynthesis, and the nucleotide metabolism. According to a correlation analysis, leaf TK and TP were strongly correlated with endophytic bacteria abundance and differential metabolite composition. This study revealed the changes in substances and microorganisms in the leaves of male and female plants in their reproductive stages. It provides a theoretical basis for developing and utilizing the leaves of Idesia polycarpa and for field management.

Soil Quality Assessment in Tourism-Disturbed Subtropical Mountain Meadow Areas of Wugong Mountain, Central Southeast China.

Meadow soil is a vital ecosystem component and can be influenced by meadow vegetation. Evaluating soil quality in mountain meadows subjected to different levels of tourism disturbance is essential for scientific research, ecological restoration, and sustainable management. This study aimed to evaluate meadow soil quality at different tourism-disturbance levels and attempted to establish a minimum data set (MDS) with compatible indicators for soil quality assessment of subtropical mountain meadows. We analyzed fifteen soil physical, chemical, and biological indicators in control check (CK), light disturbance (LD), medium disturbance (MD), and severe disturbance (SD) meadow areas in Wugong Mountain, west of Jiangxi, China. In addition, a soil quality index (SQI) was determined using the established MDS based on the integrated soil quality index. Average soil permeability, soil pH, available nitrogen (AN), available phosphorus (AP), and number of fungal OTUs were finally introduced into the MDS to evaluate meadow soil quality at different tourism-disturbance levels. The study found that the soil of the Wugong Mountain meadow was acidic, the bulk density was loose, and the nutrient content was rich. Additionally, SQI decreased with increase in tourism-disturbance level. The mean SQI values of the Wugong Mountain meadow areas were: CK, 0.612; LD, 0.493; MD, 0.448; and SD, 0.416. Our results demonstrate that the SQI based on the MDS method could be a valuable tool with which to indicate the soil quality of mountain meadow areas, and the SQI can be regarded as a primary indicator of ecological restoration and sustainable management.

Zero waste multistage utilization of Ginkgo biloba branches.

Zero waste multistage utilization of biomass from Ginkgo biloba branches (GBBs) was achieved through extraction of bioactive components, analysis of antioxidant and antibacterial activities, preparation and composition of pyrolyzate, adsorption and reuse of modified biochar. The results showed that GBBs had abundant bioactive components for potential application in the industry of food, chemical raw materials and biomedicine. Especially, the bioactive compounds in acetone extract (10 mg/mL) of GBBs identified by DPPH and ABTS had free radical scavenging abilities of 92.28% and 98.18%, respectively, which are equivalent to Vitamin C used as an antioxidant in food additives. Fourier Transform Infrared and X-Ray Diffraction analysis showed that carboxymethyl cellulose (CMC) and magnetic Fe3O4 were successfully incorporated into raw biochar (RB) to form CMC-Fe3O4-RB nanomaterial. Scanning electron microscopy and X-Ray Diffraction spectroscopy displayed Fe, C, and O existed on the surface of CMC-Fe3O4-RB. Compared with RB, CMC-Fe3O4-RB had a larger specific surface area, pore volume and pore size. Meanwhile, nanomagnetic CMC-Fe3O4-RB solved the problem of agglomeration in traditional magnetized biochar production, and improved the adsorption capacity of Pb2+, which was 29.90% higher than that of RB by ICP-OES. Further, the Pb2+ (10 mg/L) adsorption capacity of CMC-Fe3O4-RB reached the highest level in 2 h at the dosage of 0.01 g/L, and remained stable at 52.987 mg/g after five cycles of adsorption and desorption. This research aided in the creation of a strategy for GBBs zero waste multistage usage and a circular economic model for GBBs industry development, which can be promoted and applied to the fields of food industry and environment improvement.

Physiological and transcriptional changes provide insights into the effect of root waterlogging on the aboveground part of Pterocarya stenoptera.

Pterocarya stenoptera is a tree species that occurs along rivers and has high tolerance to waterlogging. Identification of waterlogging response genes in the aboveground part of P. stenoptera will increase understanding of tolerance mechanisms under root waterlogging conditions. In this study, we employed four physiological indicators and comparative transcriptome sequencing to investigate the waterlogging tolerance mechanism in P. stenoptera. The physiological results showed that the aboveground part of P. stenoptera was not obviously affected by waterlogging. P. stenoptera enhanced waterlogging tolerance by increasing the synthesis of alpha-Linolenic acids and flavonoids and activating the jasmonic acid, ethylene, and auxin signaling pathways. Our results confirmed our hypothesis that P. stenoptera, a species that is widely distributed along rivers, has evolved a range of mechanisms in response to waterlogging. Our research will provide new insights for understanding the tolerance mechanism of species to waterlogging.

Phylogenomic analyses based on genome-skimming data reveal cyto-nuclear discordance in the evolutionary history of Cotoneaster (Rosaceae).

As a consequence of hybridization, polyploidization, and apomixis, the genus Cotoneaster (Rosaceae) represents one of the most complicated and controversial lineages in Rosaceae, with ca. 370 species which have been classified into two subgenera and several sections, and is notorious for its taxonomic difficulty. The infrageneric relationships and taxonomy of Cotoneaster have remained poorly understood. Previous studies have focused mainly on natural hybridization involving only several species, and phylogeny based on very limited markers. In the present study, the sequences of complete chloroplast genomes and 204 low-copy nuclear genes of 72 accessions, representing 69 species as ingroups, were used to conduct the most comprehensive phylogenetic analysis so far for Cotoneaster. Based on the sequences of complete chloroplast genomes and many nuclear genes, our analyses yield two robust phylogenetic trees respectively. Chloroplast genome and nuclear data confidently resolved relationships of this genus into two major clades which largely supported current classification based on morphological evidence. However, conflicts between the chloroplast genome and low-copy nuclear phylogenies were observed in both the species level and clade level. Cyto-nuclear discordance in the phylogeny could be caused by frequent hybridization events and incomplete sorting lineage (ILS). In addition, our divergence-time analysis revealed an evolutionary radiation of the genus from late Miocene to date.

Characterization of the complete chloroplast genome of Hovenia acerba (Rhamnaceae).

Hovenia acerba is a widely distributed species with economic, ornamental, and medicinal value in China. In this study, we assembled and characterized the complete chloroplast genome of H. acerba for the first time. The circular genome has a quadripartite structure with 161,651 bp in length and contains a pair of 26,619 bp inverted repeat (IR) regions, separated by the large single-copy (LSC, 89,443 bp) region and small single-copy (SSC, 18,970 bp) region. The plastid genome harbours 104 unique genes, including 72 protein-coding genes, 28 tRNAs, and four rRNAs. The overall GC content of the whole genome was 36.7%. Further, the phylogenetic analysis showed that H. acerba clustered together with Ziziphus genus. The complete chloroplast genome of H. acerba will provide important information for phylogenetic and evolutionary studies in Rhamnaceae, as well as the other closely related family.

Molecular Phylogeography Analysis Reveals Population Dynamics and Genetic Divergence of a Widespread Tree Pterocarya stenoptera in China.

The geological events, past climatic fluctuations, and river systems played key roles in the spatial distribution, population dynamics, and genetic differentiation of species. In this work, we selected Pterocarya stenoptera, a widespread tree species in China, to test the roles of these factors. Four noncoding spacers, eight microsatellite (simple sequence repeat) markers, and species distribution modeling were used to examine the phylogeographical pattern of P. stenoptera. Based on chloroplast DNA data, populations of P. stenoptera were clearly clustered into three groups. The divergence time of these groups fell into the stage of the Qinghai-Tibet Movement, 1.7-2.6 Ma. For simple sequence repeat data, only one western marginal population YNYB could be separated from other populations, whereas other populations were mixed together. Our results indicated that the environmental heterogeneity resulting from the Qinghai-Tibet movement might be response for this genetic divergence. The climatic fluctuations in the Pleistocene did not cause the substantial range shift of P. stenoptera, while the fluctuations affected its population size. Moreover, we also confirmed the river systems did not act as channels or barrier of dispersal for P. stenoptera.

Functional nano-catalyzed pyrolyzates from branch of Cinnamomum camphora.

Cinnamomum camphora is an excellent tree species for construction of forest construction of Henan Province, China. The diverse bioactive components of nano-catalyzed pyrolyzates form cold-acclimated C. camphora branch (CCB) in North China were explored. The raw powder of CCB treated with nano-catalyst (Ag, NiO, 1/2Ag + 1/2NiO) were pyrolyzed at two temperatures (550 °C and 700 °C), respectively. The main pyrolyzates are bioactive components of bioenergy, biomedicines, food additive, spices, cosmetics and chemical, whose total relative contents at 550 °C pyrolyzates are higher than those at 700 °C pyrolyzates. There are abundant components of spices and biomedicine at 550 °C pyrolyzates, while more spices and food additive at 700 °C pyrolyzates. At 550 °C, the content of biomedicine components reaches the highest by 1/2Ag + 1/2NiO nanocatalysis, while the contents of spices and food additive components reach the highest by NiO nanocatalysis. At 700 °C, the content of bioenergy components reaches the highest by 1/2Ag + 1/2NiO nanocatalysis, and the content of cosmetics components reaches the highest by Ag nanocatalysis. The findings suggested that the branch of the cold-acclimated C. camphora have the potential to develop into valued-added products of bioenergy, biomedicine, cosmetics, spices and food additive by nanocatalysis.

Molecular Evidence for Natural Hybridization between Cotoneaster dielsianus and C. glaucophyllus.

Hybridization accompanied by polyploidization and apomixis has been demonstrated as a driving force in the evolution and speciation of many plants. A good example to study the evolutionary process of hybridization associated with polyploidy and apomixis is the genus Cotoneaster (Rosaceae), which includes approximately 150 species, most of which are polyploid apomicts. In this study, we investigated all Cotoneaster taxa distributed in a small region of Malipo, Yunnan, China. Based on the morphological characteristics, four Cotoneaster taxa were identified and sampled: C. dielsianus, C. glaucophyllus, C. franchetii, and a putative hybrid. Flow cytometry analyses showed that C. glaucophyllus was diploid, while the other three taxa were tetraploid. A total of five low-copy nuclear genes and six chloroplast regions were sequenced to validate the status of the putative hybrid. Sequence analyses showed that C. dielsianus and C. glaucophyllus are distantly related and they could be well separated using totally 50 fixed nucleotide substitutions and four fixed indels at the 11 investigated genes. All individuals of the putative hybrid harbored identical sequences: they showed chromatogram additivity for all fixed differences between C. dielsianus and C. glaucophyllus at the five nuclear genes, and were identical with C. glaucophyllus at the six chloroplast regions. Haplotype analysis revealed that C. dielsianus possessed nine haplotypes for the 11 genes, while C. glaucophyllus had ten, and there were no shared haplotypes between the two species. The putative hybrid harbored two haplotypes for each nuclear gene: one shared with C. dielsianus and the other with C. glaucophyllus. They possessed the same chloroplast haplotype with C. glaucophyllus. Our study provided convincing evidence for natural hybridization between C. dielsianus and C. glaucophyllus, and revealed that all hybrid individuals were derivatives of one initial F1 via apomixes. C. glaucophyllus served as the maternal parent at the initial hybridization event. We proposed that anthropological disturbance provided an opportunity for hybridization between C. dielsianus and C. glaucophyllus, and a tetraploid F1 successfully bred many identical progenies via apomixis. Under this situation, species integrity could be maintained for these Cotoneaster species, but attentions should be kept for this new-born hybrid.

Isolation and identification of EST-SSR markers in Chunia bucklandioides (Hamamelidaceae).

PREMISE OF THE STUDY: Chunia bucklandioides (Hamamelidaceae), endemic to Hainan, China, is listed as threatened in the IUCN Red List and is now only found on Mt. Diaoluo and Mt. Jianfeng. Thus, microsatellite markers were developed for future conservation genetic studies of this species. METHODS AND RESULTS: A total of 115 primers were designed on the basis of the transcriptome data of C. bucklandioides. Of them, 59 successfully amplified in C. bucklandioides and polymorphisms were detected in 11; the number of alleles per locus varied from two to five, the observed heterozygosity ranged from 0.000 to 0.941, and the expected heterozygosity ranged from 0.000 to 0.699. A total of 13 primers amplified in Mytilaria laosensis, and five primers amplified in Exbucklandia tonkinensis and E. populnea. CONCLUSIONS: The markers screened here provide a basis to assess genetic structure and further establish conservation strategies for C. bucklandioides.

A luminescent ultrathin film with reversible sensing toward pressure.

A flexible ultrathin film based on alternate assembly of a sodium polyacrylate (PAA) modified styrylbiphenyl derivative (BTBS) and layered double hydroxide nanosheets is fabricated, which exhibits pressure-responsive photoluminescence with a high sensitivity and good reversibility.

De Novo Transcriptome Assembly in Firmiana danxiaensis, a Tree Species Endemic to the Danxia Landform.

Many Firmiana species are locally endemic, providing an interesting system for studying adaptation and speciation. Among these species, F. danxiaensis is a tree species endemic to Mount Danxia in Guangdong, China, which is an area known for presenting the Danxia landform. How F. danxiaensis could have adapted to the stressful environment of rocky cliffs covered with barren soils in the Danxia landform is still unknown. In this study, we performed de novo assembly of the transcriptome of F. danxiaensis, obtaining 47,221 unigenes with an N50 value of 987 bp. Homology analysis showed that 32,318 of the unigenes presented hits in the NCBI non-redundant database, and 31,857 exhibited significant matches with the protein database of Theobroma cacao. Gene Ontology (GO) annotation showed that hundreds of unigenes participated in responses to various stresses or nutritional starvation, which may help us to understand the adaptation of F. danxiaensis to Danxia landform. Additionally, we found 263 genes related to responses to Cd, partially explaining the high accumulation of Cd observed in Firmiana species. The EuKaryotic Orthologous Groups (KOG) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotations revealed many genes playing roles in the biosynthesis of secondary metabolites and environmental adaptation, which may also contribute to the survivor and success of Firmiana species in extreme environments. Based on the obtained transcriptome, we further identified a Firmiana-specific whole-genome duplication event that occurred approximately 20 Mya, which may have provided raw materials for the diversification of Firmiana species.

Molecular evidence for natural hybridization between wild loquat (Eriobotrya japonica) and its relative E. prinoides.

BACKGROUND: Interspecific hybridization has long been recognized as a pivotal process in plant evolution and speciation. It occurs fairly common in the genera of the subtribe Pyrinae. In Eriobotrya, a small tree genus of Pyrinae, E. prinoides var. daduheensis has been recognized as either a variety of E. prinoides, a natural hybrid between E. prinoides and E. japonica, or a variety of E. japonica. However, to date, there has been no convincing evidence on its status. RESULTS: Four nuclear genes and two chloroplast regions were sequenced in 89 individuals of these three Eriobotrya taxa from two locations where they coexist. A few fixed nucleotide substitutions or gaps were found in each of the investigated nuclear and chloroplast loci between E. japonica and E. prinoides. Of the 35 individuals of E. prinoides var. daduheensis, 33 showed nucleotide additivity of E. japonica and E. prinoides in at least one nuclear gene, and 10 of them harboured nucleotide additivity at all the four nuclear genes. Most haplotypes of E. prinoides var. daduheensis were also shared with those of E. japonica and E. prinoides. In the two chloroplast regions, 28 and 7 individuals were identical with E. japonica and E. prinoides, respectively. CONCLUSIONS: Our study provides compelling evidence for a hybrid status for E. prinoides var. daduheensis. Most hybrid individuals are later-generation hybrids. Both E. japonica and E. prinoides can serve as female parent. Differential adaptation might maintain the species boundary of E. prinoides and E. japonica in the face of hybridization and potential introgression.

Development and characterization of microsatellite markers from the transcriptome of Firmiana danxiaensis (Malvaceae s.l.).

PREMISE OF THE STUDY: Firmiana consists of 12-16 species, many of which are narrow endemics. Expressed sequence tag (EST)-simple sequence repeat (SSR) markers were developed and characterized for size polymorphism in four Firmiana species. • METHODS AND RESULTS: A total of 102 EST-SSR primer pairs were designed based on the transcriptome sequences of F. danxiaensis; these were then characterized in four Firmiana species-F. danxiaensis, F. kwangsiensis, F. hainanensis, and F. simplex. In these four species, 17 primer pairs were successfully amplified, and 14 were polymorphic in at least one species. The number of alleles ranged from one to 13, and the observed and expected heterozygosities ranged from 0 to 1 and 0 to 0.925, respectively. The lowest level of polymorphism was observed in F. danxiaensis. • CONCLUSIONS: These polymorphic EST-SSR markers are valuable for conservation genetics studies in the endangered Firmiana species.