Population Variation and Phylogeography of Cherry Blossom (Prunus conradinae) in China.

Prunus conradinae (subgenus Cerasus, Rosaceae) is a significant germplasm resource of wild cherry blossom in China. To ensure the comprehensiveness of this study, we used a large sample size (12 populations comprising 244 individuals) which involved the fresh leaves of P. conradinae in Eastern, Central, and Southwestern China. We combined morphological and molecular evidence (three chloroplast DNA (cpDNA) sequences and one nuclear DNA (nr DNA) sequence) to examine the population of P. conradinae variation and differentiation. Our results revealed that Central, East, and Southwest China are important regions for the conservation of P. conradinae to ensure adequate germplasm resources in the future. We also found support for a new variant, P. conradinae var. rubrum. We observed high genetic diversity within P. conradinae (haplotype diversity [Hd] = 0.830; ribotype diversity [Rd] = 0.798), with novel genetic variation and a distinct genealogical structure among populations. There was genetic variation among populations and phylogeographic structure among populations and three geographical groups (Central, East, and Southwest China). The genetic differentiation coefficient was the lowest in the Southwest region and the gene exchange was obvious, while the differentiation was obvious in Central China. In the three geographic groups, we identified two distinct lineages: an East China lineage (Central China and East China) and a Southwest China lineage ((Central China and Southwest China) and East China). These two lineages originated approximately 4.38 million years ago (Mya) in the early Pliocene due to geographic isolation. P. conradinae expanded from Central China to East China at 3.32 Mya (95% HPD: 1.12-5.17 Mya) in the Pliocene. The population of P. conradinae spread from East China to Southwest China, and the differentiation time was 2.17 Mya (95% (HPD: 0.47-4.54 Mya), suggesting that the population of P. conradinae differentiated first in Central and East China. The population of P. conradinae experienced differentiation from Central China to Southwest China around 1.10 Mya (95% HPD: 0.11-2.85 Mya) during the early Pleistocene of the Quaternary period. The southeastern region of East China, near Mount Wuyi, likely serves as a refuge for P. conradinae. This study establishes a theoretical foundation for the classification, identification, conservation, and exploitation of germplasm resources of P. conradinae.

Molecular and morphological evidence support a new species of RosaceaePrunus subg. Cerasus from Wuyishan National Park, southeast China.

Prunustongmuensis, a new species of cherry blossom, is described and illustrated from Wuyishan National Park, southeast China. This species is characterized by its tubular to nearly bottle-shaped receptacles and dark purple drupes. It can be distinguished from other wild cherry trees by its flowers and leaves, reddish brown young leaves, presence of 1-2 glands at the base of leaves, petioles densely covered with yellowish brown villi, longer pedicels (0.6-2.5 cm), villous pistil, and dark purple drupes. In the present study, we conducted a comprehensive morphological study based on specimens of the new species and its morphologically close species, field observations, and examination of pollen morphology. In addition, our phylogenetic analysis based on the complete plastid genome sequences further confirms the status of the new species and indicates that it is closely related to Prunusclarofolia, however, it notably differs in leaf shape, size, petiole villus color, gland location, timing of flower and leaf openings, and reflexed or spread sepals, as well as drupe color.

Reticulate evolution of the tertiary relict Osmanthus.

When interspecific gene flow is common, species relationships are more accurately represented by a phylogenetic network than by a bifurcating tree. This study aimed to uncover the role of introgression in the evolution of Osmanthus, the only genus of the subtribe Oleinae (Oleaceae) with its distribution center in East Asia. We built species trees, detected introgression, and constructed networks using multiple kinds of sequencing data (whole genome resequencing, transcriptome sequencing, and Sanger sequencing of nrDNA) combined with concatenation and coalescence approaches. Then, based on well-understood species relationships, historical biogeographic analyses and diversification rate estimates were employed to reveal the history of Osmanthus. Osmanthus originated in mid-Miocene Europe and dispersed to the eastern Tibetan Plateau in the late Miocene. Thereafter, it continued to spread eastwards. Phylogenetic conflict is common within the 'Core Osmanthus' clade and is seen at both early and late stages of diversification, leading to hypotheses of net-like species relationships. Incomplete lineage sorting proved ineffective in explaining phylogenetic conflicts and thus supported introgression as the main cause of conflicts. This study elucidates the diversification history of a relict genus in the subtropical regions of eastern Asia and reveals that introgression had profound effects on its evolutionary history.

Pollen Morphology in Sorbus L. (Rosaceae) and Its Taxonomic Implications.

The genus Sorbus L. in the Rosaceae family is taxonomically challenging due to its morphological variation, polyploidy, and interspecific hybridization. In this study, we used scanning electron microscopy (SEM) to observe the pollen morphology of eighty species, representing six subgenera, in order to assess the differences within the genus Sorbus and its pollen characteristics. We conducted a cluster analysis on three qualitative and four quantitative characteristics. The results demonstrated that the pollen grains of the studied Sorbus species are isopolar and tricolporate. We identified five types of pollen shapes: suboblate, spheroidal, subprolate, prolate, and perprolate. The pollen ornamentation of the investigated species could be classified into five types: striate-perforate, striate, cerebroid-perforate, cerebroid, and foveolate. Interestingly, within the same subgenera, different species exhibited multiple types of characters. The cluster analysis indicated that all 80 species could be divided into six groups, with group B consisting exclusively of species from the subgenus Sorbus. Although pollen micro-morphologies alone do not provide sufficient evidence to establish the taxonomic relationships of the subgenera within Sorbus, they do offer valuable information for species-level taxonomic treatment.

Alternative Splicing Analysis Revealed the Role of Alpha-Linolenic Acid and Carotenoids in Fruit Development of Osmanthus fragrans.

Alternative splicing refers to the process of producing different splicing isoforms from the same pre-mRNA through different alternative splicing events, which almost participates in all stages of plant growth and development. In order to understand its role in the fruit development of Osmanthus fragrans, transcriptome sequencing and alternative splicing analysis was carried out on three stages of O. fragrans fruit (O. fragrans "Zi Yingui"). The results showed that the proportion of skipping exon events was the highest in all three periods, followed by a retained intron, and the proportion of mutually exclusive exon events was the lowest and most of the alternative splicing events occurred in the first two periods. The results of enrichment analysis of differentially expressed genes and differentially expressed isoforms showed that alpha-Linolenic acid metabolism, flavonoid biosynthesis, carotenoid biosynthesis, photosynthesis, and photosynthetic-antenna protein pathways were significantly enriched, which may play an important role in the fruit development of O. fragrans. The results of this study lay the foundation for further study of the development and maturation of O. fragrans fruit and further ideas for controlling fruit color and improving fruit quality and appearance.

Scale-dependent effects of species diversity on aboveground biomass and productivity in a subtropical broadleaved forest on Mt. Huangshan.

The relationship between species diversity and biomass/productivity is a major scientific question in ecology. Exploring this relationship is essential to understanding the mechanisms underpinning the maintenance of biodiversity. Positive, negative, and neutral relationships have been identified in controlled experiments and observational research. However, increasing evidence suggests that the effects of species diversity on aboveground biomass and productivity are influenced by biotic and abiotic factors, but it remains unclear whether scale-dependent effects affect aboveground biomass and productivity. Herein, we used a generalized linear regression model and a structural equation model to explore relationships between species diversity and productivity/aboveground biomass under different scales and to investigate the effects of topographical factors and species diversity on ecosystem functioning. The results revealed a positive relationship between biodiversity and ecosystem functioning based on species diversity and aboveground biomass. Different sampling scales may impact the relationship between species diversity and ecosystem functioning. A positive relationship was found between species richness and productivity at medium and large scales; however, ambiguous relationships were found in productivity and other species diversity indices. Elevation was a key factor affecting both biomass and productivity. These results suggest that species diversity is not the only factor affecting biomass and productivity, and the positive correlation between species diversity and ecosystem functioning is mediated by abiotic factors.

Identification of miRNAs and Target Genes at Key Stages of Sexual Differentiation in Androdioecious Osmanthus fragrans.

Androdioecy is the crucial transition state in the evolutionary direction of hermaphroditism to dioecy, however, the molecular mechanisms underlying the formation of this sex system remain unclear. While popular in China for its ornamental and cultural value, Osmanthus fragrans has an extremely rare androdioecy breeding system, meaning that there are both male and hermaphroditic plants in a population. To unravel the mechanisms underlying the formation of androdioecy, we performed small RNA sequencing studies on male and hermaphroditic O. fragrans. A total of 334 miRNAs were identified, of which 59 were differentially expressed. Functional categorization revealed that the target genes of differentially expressed miRNAs were mainly involved in the biological processes of reproductive development and the hormone signal transduction pathway. We speculated that the miRNA160, miRNA167, miRNA393 and miRNA396 families may influence the sex differentiation in O. fragrans. Overall, our study is the first exploration of miRNAs in the growth and development process of O. fragrans, and is also the first study of androdioecious plants from the miRNA sequencing perspective. The analysis of miRNAs and target genes that may be involved in the sex differentiation process lay a foundation for the ultimate discovery of the androdioecious molecular mechanism in O. fragrans.

Limited accumulation of high-frequency somatic mutations in a 1700-year-old Osmanthus fragrans tree.

Lifespan varies greatly between and within species. Mutation accumulation is considered an important factor explaining this life-history trait. However, direct assessment of somatic mutations in long-lived species is still rare. In this study, we sequenced a 1700-year-old sweet olive tree and analysed the high-frequency somatic mutations accumulated in its six primary branches. We found the lowest per-year mutation accumulation rate in this oldest tree among those studied via the whole-genome sequencing approach. Investigation of mutation profiles suggests that this low rate of high-frequency mutation was unlikely to result from strong purifying selection. More intriguingly, on a per-branching scale, the high-frequency mutation accumulation rate was similar among the long-lived individuals such as oak, wild peach and sweet olive investigated here. We therefore suggest the possibility that the accumulation of high-frequency somatic mutations in very long-lived trees might have an upper boundary due to both the possible limited number of stem cell divisions and the early segregation of the stem cell lineage.

Comparative analysis of the complete plastid genomes in Prunus subgenus Cerasus (Rosaceae): Molecular structures and phylogenetic relationships.

Prunus subgenus Cerasus (cherry) is an economically important group that distributed in temperate regions of the northern hemisphere. However, shared interspecific morphological traits and variability across taxa of Cerasus are among the impediments to taxonomic efforts to correctly delimit taxa. This is further complicated by a lack of genetic information on these taxa, with no focused genomic or phylogenetic studies being done on Cerasus. In this study, we conducted comparative analysis on the complete plastid genomes (plastomes) of 20 Cerasus species to gain a greater understanding of the attributes of the plastome of these taxa while helping resolve their phylogenetic placement in Prunus sensu lato and interspecific relationships within the subgenus. Our results displayed that (1) the plastomes of the 20 Cerasus species studied exhibited a typical quadripartite structure with conversed genome arrangement, structure, and moderate divergence. (2) The average size of complete plastomes for the Cerasus taxa studied was 157,861 bp, ranging from 157,458 to 158,024 bp. A total of 134 genes were annotated, including 86 protein-coding genes, 40 tRNAs, and 8 rRNAs across all species. In simple sequence repeat analysis, we found Cerasus had a comparable number of dispersed and tandem repeats to those identified in other angiosperm taxa, with only P. pseudocerasus found to contain trinucleotide repeats. Nucleotide diversity analysis revealed that the trnG-GCC gene and rpl32-trnL region had the highest Pi value showing potential as phylogenetic markers. (3) Two phylogenetic trees of the plastomes verified the monophyletic relationship of Cerasus and provided a more resolved species-level phylogeny. Our study provides detailed plastome information for exploring the phylogeny of subg. Cerasus taxa. We identified various types of repeats and nucleotide diversity hotspots, which can be a reference for species identification and reconstruction of phylogenetic relationships.

Plastid genomes reveal evolutionary shifts in elevational range and flowering time of Osmanthus (Oleaceae).

Species of Osmanthus are economically important ornamental trees, yet information regarding their plastid genomes (plastomes) have rarely been reported, thus hindering taxonomic and evolutionary studies of this small but enigmatic genus. Here, we performed comparative genomics and evolutionary analyses on plastomes of 16 of the 28 currently accepted species, with 11 plastomes newly sequenced. Phylogenetic studies identified four main lineages within the genus that are here designated the: "Caucasian Osmanthus" (corresponding to O. decorus), "Siphosmanthus" (corresponding to O. sect. Siphosmanthus), "O. serrulatus + O. yunnanensis," and "Core Osmanthus: (corresponding to O. sect. Osmanthus + O. sect. Linocieroides). Molecular clock analysis suggested that Osmanthus split from its sister clade c. 15.83 Ma. The estimated crown ages of the lineages were the following: genus Osmanthus at 12.66 Ma; "Siphosmanthus" clade at 5.85 Ma; "O. serrulatus + O. yunnanensis" at 4.89 Ma; and "Core Osmanthus: clade at 6.2 Ma. Ancestral state reconstructions and trait mapping showed that ancestors of Osmanthus were spring flowering and originated at lower elevations. Phylogenetic principal component analysis clearly distinguished spring-flowering species from autumn-flowering species, suggesting that flowering time differentiation is related to the difference in ecological niches. Nucleotide substitution rates of 80 common genes showed slow evolutionary pace and low nucleotide variations, all genes being subjected to purifying selection.

Laboratory Characterization of Geosynthetics-Reinforced Asphalt Mixture.

In order to improve the mechanical properties of asphalt pavement, geosynthetics can be employed in asphalt mixture. This research designed 12 reinforced schemes based on the types of geosynthetics, bonding layers and reinforced position. For the relative tests carried out, reinforced specimens were prepared according to each individual scheme. Moreover, rutting tests, bending creep tests and split fatigue tests were carried out on reinforced specimens in the laboratory. The results obtained in this investigation showed that the dynamic stability, bending creep rate and fatigue life of geocell-reinforced specimens are better than those of geogrid-reinforced specimens. The bonding layer of Styrene-Butadiene-Styrene (SBS) modified asphalt is better than epoxy modified asphalt. The dynamic stability and fatigue life of middle reinforcement are better than those of the lower reinforcement, while the bending creep rate of the lower reinforcement is better than middle reinforcement. In addition, reinforced scheme (9) has the largest increase in dynamic stability and fatigue life by 103 and 137%, respectively, and reinforced scheme (12) has the largest reduction in bending creep rate by 46%. However, scheme (9) improved dynamic stability and fatigue life by 43 and 29% higher than scheme (12), while the reduction of flexural creep rate of scheme (12) is only 7% higher than that of scheme (9).

SWATH-MS based quantitive proteomics reveal regulatory metabolism and networks of androdioecy breeding system in Osmanthus fragrans.

BACKGROUND: The fragrant flower plant Osmanthus fragrans has an extremely rare androdioecious breeding system displaying the occurrence of males and hermaphrodites in a single population, which occupies a crucial intermediate stage in the evolutionary transition between hermaphroditism and dioecy. However, the molecular mechanism of androdioecy plant is very limited and still largely unknown. RESULTS: Here, we used SWATH-MS-based quantitative approach to study the proteome changes between male and hermaphroditic O. fragrans pistils. A total of 428 proteins of diverse functions were determined to show significant abundance changes including 210 up-regulated and 218 down-regulated proteins in male compared to hermaphroditic pistils. Functional categorization revealed that the differentially expressed proteins (DEPs) primarily distributed in the carbohydrate metabolism, secondary metabolism as well as signaling cascades. Further experimental analysis showed the substantial carbohydrates accumulation associated with promoted net photosynthetic rate and water use efficiency were observed in purplish red pedicel of hermaphroditic flower compared with green pedicel of male flower, implicating glucose metabolism serves as nutritional modulator for the differentiation of male and hermaphroditic flower. Meanwhile, the entire upregulation of secondary metabolism including flavonoids, isoprenoids and lignins seem to protect and maintain the male function in male flowers, well explaining important feature of androdioecy that aborted pistil of a male flower still has a male function. Furthermore, nine selected DEPs were validated via gene expression analysis, suggesting an extra layer of post-transcriptional regulation occurs during O. fragrans floral development. CONCLUSION: Taken together, our findings represent the first SWATH-MS-based proteomic report in androdioecy plant O. fragrans, which reveal carbohydrate metabolism, secondary metabolism and post-transcriptional regulation contributing to the androdioecy breeding system and ultimately extend our understanding on genetic basis as well as the industrialization development of O. fragrans.

Characterization of the complete chloroplast genome of Prunus clarofolia C.K. Schneid (Rosaceae).

Prunus clarofolia is an endemic species that widely distributed in subtropical regions of China. Here we present its complete plastome. The plastome of P. clarofolia is successfully assembled from raw reads sequenced by Illumina Hiseq 2500 platform system. The complete chloroplast of this species is 158,053 bp in length with 36.7% overall GC content, including a pair of invert repeat regions (IR) (26,393bp) that is divided by a large single copy region (LSC) (86,088bp) and a small single copy region (SSC) (19,179bp). The plastid genome contained a total of 130 genes, including 85 coding genes, 8 rRNA genes, and 37 tRNA genes. Each of rps16, atpF, rpoC1, clpP, petB, petD, rpl16, rpl2, ndhB, and ndhA contains one intron, rps12 and ycf3 contains two introns. Phylogenetic analysis indicates that P. clarofolia has a closer relationship with P. avium.

The complete plastid genome of cherry plants Prunus sargentii (Rosaceae) and its phylogenetic implication.

Prunus sargentii is an ornamental flowering cherry species, spread in Japan, Korea, Russia, and Northeast China. Little information is available regarding its genomic, with limited phylogenetic relationship study performed on P. sargentii until now. In this research, we reported the complete plastid genome of P. sargentii. The complete chloroplast of this species is 158,138 bp in length, including a pair of invert repeat regions (IR) (26,463bp) that is divided by a large single-copy region (LSC) (85,959bp) and a small single-copy region (SSC) (19,253bp). The plastid genome contained a total of 128 genes, including 84 coding genes, eight rRNA genes, and 36 tRNA genes. Phylogenetic analysis indicates that P. sargentii has a closer relationship with P. kumanoensis.

Complete chloroplast genome of a wild-type Gardenia jasminoides ellis (rubiaceae) adapted to island climate.

Gardenia jasminoides Ellis is a traditional aromatic and medicinal plant in China. Here, the complete chloroplast genome of a wild-type gardenia adapted to island climate was assembled. The assembled genome was 155,247 bp in length, with four typical regions, i.e., a large single-copy (LSC) region (85,414 bp), a small single-copy (SSC) region (18,235 bp) and two inverted repeats (IRs) regions (25,799 bp each). In total, 138 genes were predicted, including 90 protein-coding genes, 40 tRNA genes and eight rRNA genes. The overall GC content of the chloroplast genome was 37.5%. The chloroplast genome would provide more information for the phylogeography and phylogeny study of G. jasminoides.

Phylogeography and the population genetic structure of flowering cherry Cerasus serrulata (Rosaceae) in subtropical and temperate China.

Cerasus serrulata (Rosaceae) is an important flowering cherry resource which is valuable for developing new cultivars of flowering cherries. It is broadly distributed and possesses abundant variations. In this study, phylogeographic analysis was conducted to reveal the evolutionary history to better understand the genetic diversity and genetic structure of C. serrulata so as to provide more accurate molecular insights into better conservation and utilization of the germplasm resources. A total of 327 individuals from 18 wild populations were collected. Three chloroplast DNA (cpDNA) fragments (matK, trnD-E, and trnS-G) and the nuclear internal transcribed spacer (ITS) were utilized. The results showed a high genetic diversity at both species level and population level of C. serrulata. High genetic differentiation and the existence of the phylogeographic structure were detected. No significant expansion events were discovered. Two geographic lineages were inferred. One was confined to the Qinling Mountains and the Taihang Mountains. The other was from the Wuling Mountains to the Jiangnan Hilly Regions and then went northeast to the coast of Asia. In addition, some taxonomic treatments of the C. serrulata complex are discussed and reconsidered. Conservation and utilization strategies of wild C. serrulata germplasm resources were recommended.

The genome of Chinese flowering cherry (Cerasus serrulata) provides new insights into Cerasus species.

Cerasus serrulata is a flowering cherry germplasm resource for ornamental purposes. In this work, we present a de novo chromosome-scale genome assembly of C. serrulata by the use of Nanopore and Hi-C sequencing technologies. The assembled C. serrulata genome is 265.40 Mb across 304 contigs and 67 scaffolds, with a contig N50 of 1.56 Mb and a scaffold N50 of 31.12 Mb. It contains 29,094 coding genes, 27,611 (94.90%) of which are annotated in at least one functional database. Synteny analysis indicated that C. serrulata and C. avium have 333 syntenic blocks composed of 14,072 genes. Blocks on chromosome 01 of C. serrulata are distributed on all chromosomes of C. avium, implying that chromosome 01 is the most ancient or active of the chromosomes. The comparative genomic analysis confirmed that C. serrulata has 740 expanded gene families, 1031 contracted gene families, and 228 rapidly evolving gene families. By the use of 656 single-copy orthologs, a phylogenetic tree composed of 10 species was constructed. The present C. serrulata species diverged from Prunus yedoensis ~17.34 million years ago (Mya), while the divergence of C. serrulata and C. avium was estimated to have occurred ∼21.44 Mya. In addition, a total of 148 MADS-box family gene members were identified in C. serrulata, accompanying the loss of the AGL32 subfamily and the expansion of the SVP subfamily. The MYB and WRKY gene families comprising 372 and 66 genes could be divided into seven and eight subfamilies in C. serrulata, respectively, based on clustering analysis. Nine hundred forty-one plant disease-resistance genes (R-genes) were detected by searching C. serrulata within the PRGdb. This research provides high-quality genomic information about C. serrulata as well as insights into the evolutionary history of Cerasus species.

Characterization of the chloroplast genome of the Osmanthus didymopetalus.

In this study, we assembled the chloroplast genome of Osmanthus didymopetalus (Oleaceae), a rare evergreen tree native to Hainan, China. The genome of O. didymopetalus was 155,155 bp in length and contained a pair of inverted repeats (IR, 25,697-25,704 bp) regions, which were separated by the small single copy (SSC, 17,591 bp) and the large single copy (LSC, 86,225 bp) regions. The cp genome encoded 133 genes including 88 protein-coding genes, 37 tRNA genes, and eight rRNA ribosomal genes. The overall GC content of O. didymopetalus chloroplast genome is 37.8%. Phylogenetic results showed that O. didymopetalus was more closely to O. yunnanensis, O. fragrans and O. insularis. This study will be beneficial for the evolutionary study and phylogenetic reconstruction of Osmanthus.

Functional androdioecy in the ornamental shrub Osmanthus delavayi (Oleaceae).

Androdioecy is one of the rarest sexual systems among plants, characterized by males co-occurring with hermaphrodites. Osmanthus delavayi (Oleaceae), an ornamental shrub from southern China, is known to have both male and hermaphrodite individuals, but little is known regarding the breeding system of this species and whether it is functionally androdioecious, and how this potentially evolved. In this study, we explore the characteristics of the breeding system of O. delavayi through the study of phenology, sex ratio, floral organ morphology, pollen number, stigma receptivity, artificial pollination, pollinators, and gene flow within and between populations, while also discussing the evolution and maintenance of androdioecy within the genus. The proportion of males was less than 0.5 and the out-crossing index (OCI) was 5. Morphological androdioecy was observed, with hermaphrodite flowers having fertile pistils, while male flowers had degenerated pistils. Males and hermaphrodites both had large amounts of small and fertile pollen grains, although the pollen number of males was ca. 1.21 × more than that of hermaphrodites, and pollen was generally smaller. Self-pollination was found to produce a much lower fruit set than outcrossing under natural conditions. Gene flow between males and hermaphrodites within a population was greater (1.007) than that between populations (0.753). All these results indicate that O. delavayi is functionally androdioecious, which may be an intermediate state in the evolutionary transition from hermaphroditism to dioecy.

The Complete Plastid Genome of Magnolia zenii and Genetic Comparison to Magnoliaceae species.

Magnolia zenii is a critically endangered species known from only 18 trees that survive on Baohua Mountain in Jiangsu province, China. Little information is available regarding its molecular biology, with no genomic study performed on M. zenii until now. We determined the complete plastid genome of M. zenii and identified microsatellites. Whole sequence alignment and phylogenetic analysis using BI and ML methods were also conducted. The plastome of M. zenii was 160,048 bp long with 39.2% GC content and included a pair of inverted repeats (IRs) of 26,596 bp that separated a large single-copy (LSC) region of 88,098 bp and a small single-copy (SSC) region of 18,757 bp. One hundred thirty genes were identified, of which 79 were protein-coding genes, 37 were transfer RNAs, and eight were ribosomal RNAs. Thirty seven simple sequence repeats (SSRs) were also identified. Comparative analyses of genome structure and sequence data of closely-related species revealed five mutation hotspots, useful for future phylogenetic research. Magnolia zenii was placed as sister to M. biondii with strong support in all analyses. Overall, this study providing M. zenii genomic resources will be beneficial for the evolutionary study and phylogenetic reconstruction of Magnoliaceae.