Insights into comparative genomics, structural features, and phylogenetic relationship of species from Eurasian Aster and its related genera (Asteraceae: Astereae) based on complete chloroplast genome.

Aster L. is an economically and phylogenetically important genus in the tribe Astereae. Here, the complete plastomes of the eight Aster species were assembled and characterized using next-generation sequencing datasets. The results indicated the complete plastomes of Aster had a quadripartite structure. These genomes were 152,045-152,729 bp in length and contained 132-133 genes, including 87 protein-coding genes, 37-38 tRNA genes, and eight rRNA genes. Expansion or contraction of inverted repeat regions and forward, palindromic, complement, and reverse repeats were detected in the eight Aster species. Additionally, our analyses showed the richest type of simple sequence repeats was A/T mononucleotides, and 14 highly variable regions were discovered by analyzing the border regions, sequence divergence, and hotspots. Phylogenetic analyses indicated that 27 species in Astereae were clustered into six clades, i.e., A to D, North American, and outgroup clades, and supported that the genera Heteropappus, Kalimeris, and Heteroplexis are nested within Aster. The results indicated the clades B to D might be considered as genera. Divergence time estimate showed the clades A, B, C, and D diverged at 23.15 Mya, 15.13 Mya, 24.29 Mya, and 21.66 Mya, respectively. These results shed light on the phylogenetic relationships of Aster and provided new information on species identification of Aster and its related genera.

A new species, Aster yaoshanensis (Asteracae, Astereae), from Guangxi (China), based on morphology and molecular phylogenetic data.

Aster yaoshanensis sp. nov., a new species of the family Asteraceae is here described and illustrated. The species is presently known only from rock crevices of mountain valleys in Dayaoshan National Nature Reserve, Guangxi autonomous region, China. Phylogenetic analyses based on ITS sequences and complete plastome data have shown that this new species is a member of genus Aster with high support. Morphologically, it mostly resembles A. jishouensis, but it can be easily distinguished from the latter by bract indumentum (glabrous except margin ciliate vs. villous especially on veins abaxially, glabrous adaxially) and color (green vs. purple), shorter corolla (3.2-3.5 mm vs. 4.5-5.3 mm), bract stalk (obvious, ca.1.2 mm vs. sessile), and different distribution (Guangxi vs. Hunan). The detailed description, distribution map, and photos are provided. This study further elucidates the species identification, phylogeny and characteristic evolution of Aster.

Intraspecific differentiation of Lindera obtusiloba as revealed by comparative plastomic and evolutionary analyses.

Lindera obtusiloba Blume is the northernmost tree species in the family Lauraceae, and it is a key species in understanding the evolutionary history of this family. The species of L. obtusiloba in East Asia has diverged into the Northern and Southern populations, which are geographically separated by an arid belt. Though the morphological differences between populations have been observed and well documented, intraspecific variations at the plastomic level have not been systematically investigated to date. Here, ten chloroplast genomes of L. obtusiloba individuals were sequenced and analyzed along with three publicly available plastomes. Comparative plastomic analysis suggests that both the Northern and the Southern populations share similar overall structure, gene order, and GC content in their plastomes although the size of the plasome and the level of intraspecific variability do vary between the two populations. The Northern have relatively larger plastomes while the Southern population possesses higher intraspecific variability, which could be attributed to the complexity of the geological environments in the South. Phylogenomic analyses also support the split of the Northern and Southern clades among L. obtusiloba individuals. However, there is no obvious species boundary between var. obtusiloba and var. heterophylla in the Southern population, indicating that gene flow could still occur between these two varieties, and this could be used as a good example of reticulate evolution. It is also found that a few photosynthesis-related genes are under positive selection, which is mainly related to the geological and environmental differences between the Northern and the Southern regions. Our results provide a reference for phylogenetic analysis within species and suggest that phylogenomic analyses with a sufficient number of nuclear and chloroplast genomic target loci from widely distributed individuals could provide a deeper understanding of the population evolution of the widespread species.

Characterization and phylogenetic analysis of the chloroplast genome of Duhaldea cappa (Buch.-Ham. ex D.Don) Pruski & Anderb. (Asteraceae).

Duhaldea cappa, a valuable medicinal plant of genus Duhaldea in the tribe Inuleae, is predominantly found in China, Bhutan, India, Malaysia, Nepal, Pakistan, Thailand, and Vietnam. However, the genomic studies of Duhaldea cappa are limited. In this study, we successfully sequenced and assembled the complete chloroplast genome of Duhaldea cappa. The chloroplast genome is 150,819 bp in length with a 37.73% GC content. The chloroplast genome has a quadripartite structure, consisting of a large single-copy region of 82,731 bp, a small single-copy region of 18,168 bp, and a pair of inverted repeat sequences of 24,960 bp. The genome contains 133 genes. Among these genes, there are 88 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The phylogeny reconstructed from data of the complete chloroplast genome indicated that Duhaldea cappa is closely related to Pluchea indica in the tribe Inuleae. Analyzing and reporting the chloroplast genome of Duhaldea cappa will establish a solid theoretical and data foundation for the efficient development, conservation, and utilization of this plant species.

Primulaxilingensis (Primulaceae), a new species from Sichuan, China.

A new species, Primulaxilingensis K.Huang & Z.X.Fu, sp. nov. (Primulaceae), is described and illustrated. In gross morphology, it is clearly allied to section Minutissimae on account of having stolons, being glabrous, leaf rosette less than or equal to corolla, flower solitary and bract not swollen at base. The new species is easily distinguished by the combination of scape densely yellow farinose, leaf apex acute, rarely broadly obtuse, corolla pale purplish blue and style 3.0-6.0 mm above base of corolla tube, stamens reaching the corolla tube mouth in thrum flower. In addition, the distribution map, morphological comparison of related species and conservation status of the new species are also provided.

Characterization and phylogenetic analysis of the complete chloroplast genome of Saussurea sagittifolia (Asteraceae, Cardueae).

The species of Saussurea sagittifolia Y. S. Chen & S. R. Yi belongs to the family Asteraceae (Cardueae). The complete chloroplast genome of S. sagittifolia was assembled and annotated for the first time in this study. The complete chloroplast genome of S. sagittifolia was 152,535 bp, including a large single-copy (LSC) region of 83,511 bp, a small single-copy (SSC) region of 18,632 bp, and a pair of inverted repeats (IRs) of 25,196 bp. The overall GC content of the chloroplast genome was 37.7%. The chloroplast genome encoded 131 genes, including 87 protein-coding genes, 36 tRNA genes, and eight rRNA genes. Phylogenetic analysis based on complete chloroplast sequences revealed that it related closely to Saussurea medusa.

Improved assembly and annotation of the sesame genome.

Sesame (Sesamum indicum L.) is an important oilseed crop that produces abundant seed oil and has a pleasant flavor and high nutritional value. To date, several Illumina-based genome assemblies corresponding to different sesame genotypes have been published and widely used in genetic and genomic studies of sesame. However, these assemblies consistently showed low continuity with numerous gaps. Here, we reported a high-quality, reference-level sesame genome assembly by integrating PacBio high-fidelity sequencing and Hi-C technology. Our updated sesame assembly was 309.35 Mb in size with a high chromosome anchoring rate (97.54%) and contig N50 size (13.48 Mb), which were better than previously published genomes. We identified 163.38 Mb repetitive elements and 24,345 high-confidence protein-coding genes in the updated sesame assembly. Comparative genomic analysis showed that sesame shared an ancient whole-genome duplication event with two Lamiales species. A total of 2,782 genes were tandemly duplicated. We also identified several genes that were likely involved in fatty acid and triacylglycerol biosynthesis. Our improved sesame assembly and annotation will facilitate future genetic studies and genomics-assisted breeding of sesame.

Evolution of the CBL and CIPK gene families in Medicago: genome-wide characterization, pervasive duplication, and expression pattern under salt and drought stress.

BACKGROUND: Calcineurin B-like proteins (CBLs) are ubiquitous Ca2+ sensors that mediate plant responses to various stress and developmental processes by interacting with CBL-interacting protein kinases (CIPKs). CBLs and CIPKs play essential roles in acclimatization of crop plants. However, evolution of these two gene families in the genus Medicago is poorly understood. RESULTS: A total of 68 CBL and 135 CIPK genes have been identified in five genomes from Medicago. Among these genomes, the gene number of CBLs and CIPKs shows no significant difference at the haploid genome level. Phylogenetic and comprehensive characteristic analyses reveal that CBLs and CIPKs are classified into four clades respectively, which is validated by distribution of conserved motifs. The synteny analysis indicates that the whole genome duplication events (WGDs) have contributed to the expansion of both families. Expression analysis demonstrates that two MsCBLs and three MsCIPKs are specifically expressed in roots, mature leaves, developing flowers and nitrogen fixing nodules of Medicago sativa spp. sativa, the widely grown tetraploid species. In particular, the expression of these five genes was highly up-regulated in roots when exposed to salt and drought stress, indicating crucial roles in stress responses. CONCLUSIONS: Our study leads to a comprehensive understanding of evolution of CBL and CIPK gene families in Medicago, but also provides a rich resource to further address the functions of CBL-CIPK complexes in cultivated species and their closely related wild relatives.

Experimental Study of the Influence of Inorganic Salts on Foam Stability.

The aqueous film-forming foam (AFFF) extinguishing agent is suitable for fighting various hydrocarbon fuel fires due to its dual fire-fighting effect of foam and liquid film. Because the action law and microscopic mechanism of inorganic salts on the stabilization process of surfactant-generated AFFF are not perfect, this paper employs an experimental approach to investigate the effects of inorganic salt types and concentrations on sodium dodecyl sulfate-containing foam systems (SDS). Prior to critical micelle concentration (CMC), increasing inorganic salt concentration decreased solution surface tension, but the opposite was true after CMC. The CMC value of an SDS solution decreases as inorganic salt concentration increases, and the "salting effect" of inorganic salt cations also has an effect on the CMC value. Based on the resistance of the liquid film at the gas-liquid interface affecting gas transport, the foam evolution was divided into three stages: foam generation, liquid drainage, and gas transfer. The effect of inorganic salts on these three stages was studied at the molecular level, and it was discovered that the addition of NH4Cl and MgCl2 could improve the saturation adsorption at the gas-liquid interface, reduce the surface tension of the surfactant solution, and improve foam stability. Meanwhile, inorganic salts can change the force of gas molecules, so the equilibrium force of gas across the liquid membrane increases as inorganic salt concentration increases. Additionally, the addition of inorganic salts raises the diffusive drainage coefficient, but the gravity drainage coefficient still reigns supreme in the predecay period.

Characterization of the complete chloroplast genome sequence of Chinese endemic species of Nouelia insignis (Hyalideae, Asteraceae) and its phylogenetic implications.

This study was the first report complete chloroplast genome of Nouelia insignis (Asteraceae, Hyalideae), the large shrubs to small trees endemic to China. The circular whole cp genome of N. insignis was 151,524 bp in length, containing a large single-copy (LSC) region of 83,145 bp and a small single-copy (SSC) region of 18,261 bp. These two regions were separated by a pair of inverted repeat regions (IRa and IRb), each of them 25,060 bp in length. A total of 135 functional genes were encoded, consisting of 89 protein-coding genes, 38 tRNA genes, and eight rRNA genes. The overall GC content of the chloroplast genome sequence was 37.8%, and the GC contents of the LSC, SSC, and IR regions were 35.9, 31.5, and 43.2%, respectively. The phylogenetic analysis by the Bayesian analysis showed that the species of N. insignis was sister group with Gerbera jamesonii by strong support values, and thus was closely related to members of subfamilies of Cichorioideae and Pertyoideae. These results will be useful for the future studies of Asteraceae in the worldwide.

Chromosome-level genome assembly of a xerophytic plant, Haloxylon ammodendron.

Haloxylon ammodendron is a xerophytic perennial shrub or small tree that has a high ecological value in anti-desertification due to its high tolerance to drought and salt stress. Here, we report a high-quality, chromosome-level genome assembly of H. ammodendron by integrating PacBio's high-fidelity sequencing and Hi-C technology. The assembled genome size was 685.4 Mb, of which 99.6% was assigned to nine pseudochromosomes with a contig N50 value of 23.6 Mb. Evolutionary analysis showed that both the recent substantial amplification of long terminal repeat retrotransposons and tandem gene duplication may have contributed to its genome size expansion and arid adaptation. An ample amount of low-GC genes was closely related to functions that may contribute to the desert adaptation of H. ammodendron. Gene family clustering together with gene expression analysis identified differentially expressed genes that may play important roles in the direct response of H. ammodendron to water-deficit stress. We also identified several genes possibly related to the degraded scaly leaves and well-developed root system of H. ammodendron. The reference-level genome assembly presented here will provide a valuable genomic resource for studying the genome evolution of xerophytic plants, as well as for further genetic breeding studies of H. ammodendron.

Unprecedented organelle genomic variations in morning glories reveal independent evolutionary scenarios of parasitic plants and the diversification of plant mitochondrial complexes.

BACKGROUND: The morning glories (Convolvulaceae) are distributed worldwide and produce economically important crops, medicinal herbs, and ornamentals. Members of this family are diverse in morphological characteristics and trophic modes, including the leafless parasitic Cuscuta (dodders). Organelle genomes were generally used for studying plant phylogeny and genomic variations. Notably, plastomes in parasitic plants always show non-canonical features, such as reduced size and accelerated rates. However, few organelle genomes of this group have been sequenced, hindering our understanding of their evolution, and dodder mitogenome in particular. RESULTS: We assembled 22 new mitogenomes and 12 new plastomes in Convolvulaceae. Alongside previously known ones, we totally analyzed organelle genomes of 23 species in the family. Our sampling includes 16 leafy autotrophic species and 7 leafless parasitic dodders, covering 8 of the 12 tribes. Both the plastid and mitochondrial genomes of these plants have encountered variations that were rarely observed in other angiosperms. All of the plastomes possessed atypical IR boundaries. Besides the gene and IR losses in dodders, some leafy species also showed gene and intron losses, duplications, structural variations, and insertions of foreign DNAs. The phylogeny reconstructed by plastid protein coding sequences confirmed the previous relationship of the tribes. However, the monophyly of 'Merremieae' and the sister group of Cuscuta remained uncertain. The mitogenome was significantly inflated in Cuscuta japonica, which has exceeded over 800 kb and integrated massive DNAs from other species. In other dodders, mitogenomes were maintained in small size, revealing divergent evolutionary strategies. Mutations unique to plants were detected in the mitochondrial gene ccmFc, which has broken into three fragments through gene fission and splicing shift. The unusual changes likely initially happened to the common ancestor of the family and were caused by a foreign insertion from rosids followed by double-strand breaks and imprecise DNA repairs. The coding regions of ccmFc expanded at both sides after the fission, which may have altered the protein structure. CONCLUSIONS: Our family-scale analyses uncovered unusual scenarios for both organelle genomes in Convolvulaceae, especially in parasitic plants. The data provided valuable genetic resources for studying the evolution of Convolvulaceae and plant parasitism.

Elsholtziazhongyangii (Lamiaceae), a new species from Sichuan, China.

Elsholtziazhongyangii (Lamiaceae), a new species from Sichuan Province, China, is described and illustrated. The new species is morphologically similar to E.feddeif.feddei, but it can be easily distinguished from E.feddeif.feddei by smaller corolla (3.2-3.5 mm vs. 4.5-5.3 mm), bract indumentum (glabrous, except margin ciliate vs. villous, especially on veins abaxially, glabrous adaxially) and bract stalked (ca. 1.2 mm vs. sessile). Phylogenetic analyses, based on two nuclear ribosomal (ETS, ITS) and five plastid (rbcL, matK, trnL-F, ycf1, ycf1-rps15) regions, confirmed that the new species formed a monophyletic clade with robust support. The new species is currently known from western Sichuan.

Chrysanthemumdabieshanense, a new name for Chrysanthemumvestitum var. latifolium (Asteraceae, Anthemideae).

Recent phylogenetic analyses have revealed that Chrysanthemumvestitumvar.latifolium and C.vestitumvar.vestitum were placed in different clades based on their chloroplast genomes and nuclear LFAFY gene sequences. Accordingly, based on previous morphological analysis, molecular phylogenetic results, fieldwork, and herbarium studies, Chrysanthemumvestitumvar.latifolium should be raised to the species level. Considering the condition of the material found and Articles 6.9, 6.11, 41.2, 58.1 of the International Code of Nomenclature for Algae, Fungi, and Plants (Shenzhen Code) that is currently in force, Chrysanthemumdabieshanense Z.X.Fu, A.G.Zhen, & Y.P.Ma, nom. nov. is proposed as the new name for Chrysanthemumvestitumvar.latifolium J.Zhou & Jun Y.Chen. The detailed emended description, distribution map, insights into its habitat, and an updated comparative morphological study are presented in this study.

Mitochondrial Phylogenomics of Fagales Provides Insights Into Plant Mitogenome Mosaic Evolution.

Fagales are an order of woody plants and comprise more than 1,100 species, most of which produce economically important timbers, nuts, and fruits. Their nuclear and plastid genomes are well-sequenced and provided valuable resources to study their phylogeny, breeding, resistance, etc. However, little is known about the mitochondrial genomes (mitogenomes), which hinder a full understanding of their genome evolution. In this study, we assembled complete mitogenomes of 23 species, covering five of the seven families of Fagales. These mitogenomes had similar gene sets but varied 2.4 times in size. The mitochondrial genes were highly conserved, and their capacity in phylogeny was challenging. The mitogenomic structure was extremely dynamic, and synteny among species was poor. Further analyses of the Fagales mitogenomes revealed extremely mosaic characteristics, with horizontal transfer (HGT)-like sequences from almost all seed plant taxa and even mitoviruses. The largest mitogenome, Carpinus cordata, did not have large amounts of specific sequences but instead contained a high proportion of sequences homologous to other Fagales. Independent and unequal transfers of third-party DNA, including nuclear genome and other resources, may partially account for the HGT-like fragments and unbalanced size expansions observed in Fagales mitogenomes. Supporting this, a mitochondrial plasmid-like of nuclear origin was found in Carpinus. Overall, we deciphered the last genetic materials of Fagales, and our large-scale analyses provide new insights into plant mitogenome evolution and size variation.

Characterization of the complete chloroplast genome sequence of Chinese endemic species of Aster batangensis (Asteraceae: Astereae) and its phylogenetic implications.

This study was the first report complete chloroplast genome of Aster batangensis (Astereae: Asteraceae), the perennial herb endemic to China. The plastid genome of Aster batangensis include 132 unique genes, with 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. Among these genes, 21 duplicate genes, including10 protein-coding genes, 7 tRNA genes, and 4 rRNA genes were detected. The complete genome size of Aster batangensis has a typical quadripartite circular structure with 152,605 bp in total length, consisting a large single copy (LSC) of 84,351 bp and a small single copy (SSC) of 18,212 bp, separated by a pair of invested repeats (IR) of 25,021 bp. The average GC content of whole plastome sequence is 37.3%, and the LSC, SSC and IR regions is 35.3%, 31.3%, and 43.0%, respectively. The phylogenetic analysis by the maximum likelihood method showed that A. batangensis was closely related to the other members of Astereae (e.g. Aztecaster matudae, Conyza bonariensis, Lagenophora cuchumatanica, Baccharis tricuneata, Baccharis genistelloides).

High-quality genome assembly of an important biodiesel plant, Euphorbia lathyris L.

Caper spurge, Euphorbia lathyris L., is an important energy crop and medicinal crop. Here, we generated a high-quality, chromosome-level genome assembly of caper spurge using Oxford Nanopore sequencing, Illumina sequencing, and Hi-C technology. The final genome assembly was ∼988.9 Mb in size, 99.8% of which could be grouped into 10 pseudochromosomes, with contig and scaffold N50 values of 32.6 and 95.7 Mb, respectively. A total of 651.4 Mb repetitive sequences and 36,342 protein-coding genes were predicted in the genome assembly. Comparative genomic analysis showed that caper spurge and castor bean clustered together. We found that no independent whole-genome duplication event had occurred in caper spurge after its split from the castor bean, and recent substantial amplification of long terminal repeat retrotransposons has contributed significantly to its genome expansion. Furthermore, based on gene homology searching, we identified a number of candidate genes involved in the biosynthesis of fatty acids and triacylglycerols. The reference genome presented here will be highly useful for the further study of the genetics, genomics, and breeding of this high-value crop, as well as for evolutionary studies of spurge family and angiosperms.

Above- and Belowground Plant Functional Composition Show Similar Changes During Temperate Forest Swamp Succession.

Plant functional composition, defined by both community-weighted mean (CWM) traits and functional diversity, can provide insights into plant ecological strategies and community assembly. However, our understanding of plant functional composition during succession is largely based on aboveground traits. Here we investigated community-level traits and functional diversity for six pairs of analogous leaf and fine root traits of understory plants in a temperate forest swamp during succession with a decrease in soil pH and nutrient availability. CWMs of traits related to resource acquisition (including specific leaf area, specific root length, leaf N, leaf P, root N, and root P) decreased with succession, whereas those related to resource conservation (leaf dry matter content, root dry matter content, leaf tissue density, leaf C, and root C) increased along the forest swamp successional gradient. Multi-trait functional dispersion (FDis) of both leaf and fine root traits tended to decrease along the successional gradient, but functional richness and evenness were highest at the middle successional stage. Moreover, FDis of individual plant traits except N showed the same pattern as multi-trait FDis. Soil pH and nutrient availability were the main drivers of successional changes in both CWM traits and FDis. The changes of community-level traits along succession indicated a shift from acquisitive to conservative strategy of understory plants during forest swamp succession. Similar trends in leaf and fine root functional diversity along succession may indicate above- and belowground functional diversity are coordinated during the processes of plant community assembly. These findings of linkages between above- and belowground plant functional composition have important implications for plant community dynamics and assembly rules.

The complete chloroplast genome of Pertya phylicoides (Asteraceae, Pertyeae): a shurby endemic species from China.

This study was the first report about complete chloroplast genome of Pertya phylicoides (Asteraceae: Pertyeae), a critically shurby plant. The complete chloroplast genome of Pertya phylicoides was 153,379 bp in length and a typical circular structure, which comprises a pair of inverted repeat (IR) region of 25,191 bp divided by a large single-copy (LSC) region of 84,535 bp and a small single-copy (SSC) region of 18,462 bp. The general GC content was 37.6% in the whole sequence, whereas corresponding values of 35.6%, 31.2%, and 43.1% in the LSC, SSC, and IR regions, respectively. The whole genome contained 113 unique genes, including 79 protein-coding genes, 4 ribosomal RNA genes, and 30 tRNA genes. The phylogenetic hypotheses obtained based on the analyses of 21 cp genomes indicates Pertya phylicoides was supported as a sister group to the tribe Carduoideae.

The complete chloroplast genome of Aster hypoleucus (Asteraceae: Astereae): an endemic species from China.

This study was the first report about complete chloroplast genome of Aster hypoleucus (Asteraceae, Astereae), an endemic species in Xizang (China). The circular whole cp genome of A. hypoleucus was 152,300 bp in length, contained a large single-copy (LSC) region of 84,031 bp and a small single-copy (SSC) region of 18,269 bp. These two regions were separated by a pair of inverted repeat regions (IRa and IRb), each of them 25,000 bp in length. A total of 134 functional genes were encoded, consisted of 89 protein-coding genes, 37 tRNA genes, and eight rRNA genes. The overall GC content of the chloroplast genome sequence was 37.3%, and the GC contents of the LSC, SSC, and IR regions were 35.2%, 31.2%, and 43.0%, respectively.