Phylogenetic and morphological analysis of a new cliff-dwelling species reveals a remnant ancestral diversity and evolutionary parallelism in <Emphasis Type="Italic">Sonchus</Emphasis> (Asteraceae)

We describe a new cliff-dwelling species within Sonchus (Asteraceae): Sonchus boulosii and analyze its systematic position and evolutionary significance; in addition, we provide a key to the species of Sonchus in Morocco. Both morphological and ecological characteristics suggest a close relationship of S. boulosii with taxa of section Pustulati. However, ITS nrDNA and cpDNA matK markers indicate its uncertain position within the genus, but clear genetic differentiation from the remaining major clades. ITS phylogenetic trees show that likely evolutionary shifts to rocky habitat took place at least five times within genus Sonchus and that sect. Pustulati and S. boulosii clades have a clearly independent evolutionary origin. We postulate that the strong resemblance of S. boulosii to other rocky species reflects a phenomenon of homoplasy, probably driven by parallel evolutionary adaptations to the severe ecological constraints of its cliff face habitat. Therefore, a new section is also described, which includes S. boulosii as its sole representative: section Pulvinati. According to phylogenetic trees, the new clade may share its common ancestor with the clade comprising sections Maritimi and Arvenses, from which it is widely divergent in morphology and ecology, with the exception of Sonchus novae-zelandiae. However, the latter is a derived taxon, with high level of polyploidy unlike S. boulosii that shows 2n?=?18, basal chromosome number of the genus. Since sections Pulvinati and Pustulati seem to be quite old in Sonchus, we also hypothesize that some similarities, such as fruit morphology, may reflect the persistence of some primitive characteristics.     

Genetic signatures of plant resistance genes with known function within and between species

Plant disease resistance (R) genes have undergone significant evolutionary divergence to cope with rapid changes in pathogens. These highly variable evolutionary patterns may have contributed to diversity in R gene protein families or structures. Here, the evolutionary patterns of 76 identified R genes and their homologs were investigated within and between plant species. Results demonstrated that nucleotide binding sites and leucine-rich-repeat genes located in loci with complex evolutionary histories tended to evolve rapidly, have high variation in copy numbers, exhibit high levels of nucleotide variation and frequent gene conversion events, and also exhibit high non-synonymous to synonymous substitution ratios in LRR regions. However, non-NBS-LRR R genes are relatively well conserved with constrained variation and are more likely to participate in the basic defense system of hosts. In addition, both conserved and highly divergent evolutionary patterns were observed for the same R genes and were consistent with inter- and intra-specific distributions of some R genes. These results thus indicate either continuous or altered evolutionary patterns between and within species. The present investigation is the first attempt to investigate evolutionary patterns among all clearly functional R genes. The results reported here thus provide a foundation for future plant disease studies.     

Chromosome numbers in Hypericum and related genera

New or confirmatory chromosome counts for 16 taxa ofHypericum in Britain, one of northwest Africa, and 24 of North and Central America are reported. First records for any member of the genusVisma (n = 10) are also included. The counts are discussed with respect to those previously reported forHypericum and related genera. Some of this chromosomal information is incorporated in a diagram showing suggested evolutionary trends withinHypericum. There appears to be a descending series of basic numbers (x = 12, 10, 9, 8, 7), of which all but the last have been recorded in polyploid, as well as diploid, form. Observations of chromosome morphology suggest thatHypericum is cytologically relatively unspecialized. Studies of chromosome morphology, therefore, are not likely to yield much information about the evolutionary history of the genus. It is suggested that the basic number forAllanblackia andPentadesma is 7 or 14, whereas inCalophyllum andMesua it is probably 8 or 16. Known gametic chromosome numbers inMammea (n = 16, 18) andGarcinia (n = 24, ca. 27, ca. 29, ca. 38, ca. 40, and 48) do not indicate an obvious basic number for these genera, although x = 8, 9, and 16 might be involved. Ring-formation of theOenothera-type, possibly indicative of structural hybridity, is reported for the first time inHypericum mitchellianum Rydb., a close relative ofHypericum punctatum Lam., which was previously shown to possess this anomalous chromo some condition.     

Reconstruction of a composite comparative map composed of ten legume genomes

The Fabaceae (legume family) is the third largest and the second of agricultural importance among flowering plant groups. In this study, we report the reconstruction of a composite comparative map composed of ten legume genomes, including seven species from the galegoid clade (Medicago truncatula, Medicago sativa, Lens culinaris, Pisum sativum, Lotus japonicus, Cicer arietinum, Vicia faba) and three species from the phaseoloid clade (Vigna radiata, Phaseolus vulgaris, Glycine max). To accomplish this comparison, a total of 209 cross-species gene-derived markers were employed. The comparative analysis resulted in a single extensive genetic/genomic network composed of 93 chromosomes or linkage groups, from which 110 synteny blocks and other evolutionary events (e.g., 13 inversions) were identified. This comparative map also allowed us to deduce several large scale evolutionary events, such as chromosome fusion/fission, with which might explain differences in chromosome numbers among compared species or between the two clades. As a result, useful properties of cross-species genic markers were re-verified as an efficient tool for cross-species translation of genomic information, and similar approaches, combined with a high throughput bioinformatic marker design program, should be effective for applying the knowledge of trait-associated genes to other important crop species for breeding purposes. Here, we provide a basic comparative framework for the ten legume species, and expect to be usefully applied towards the crop improvement in legume breeding.     

Phylogenetic analysis reveals the origins of tetraploid and hexaploid species in the Japanese <Emphasis Type="Italic">Lepisorus thunbergianus</Emphasis> (Polypodiaceae) complex

The Japanese Lepisorus thunbergianus complex contains diploid and tetraploid races of L. thunbergianus and a hexaploid species, L. mikawanus. Here, we performed molecular phylogenetic analysis on this complex to delimit species and to elucidate the evolutionary origins of tetraploid and hexaploid species. Chloroplast DNA (cpDNA) phylogeny supported the monophyly of the complex. Based on a single-copy nuclear gene (PgiC) tree, the tetraploid L. thunbergianus samples could be classified into two variants: an allotetraploid of hybrid origin between diploid L. thunbergianus and Japanese L. angustus and another allotetraploid of hybrid origin between diploid L. thunbergianus and an unknown diploid race of L. tosaensis. These variants can be recognized morphologically and distinguished from their parent species. Hence, here we described these allopolyploids as new species, L. nigripes and L. kuratae, respectively. The hexaploid species L. mikawanus has three types of PgiC alleles, each of which was derived from diploid L. thunbergianus, L. tosaensis, and Japanese L. angustus, while cpDNA shows that it is included in Japanese L. thunbergianus clade. Based on the cpDNA phylogeny and PgiC nucleotide sequences, we therefore concluded that L. mikawanus is an allohexaploid that originated through hybridization between tetraploid species, L. nigripes and an unknown ancestral diploid race of L. tosaensis.     

Heteroplasmy due to chloroplast paternal leakage: another insight into <Emphasis Type="Italic">Phragmites</Emphasis> haplotypic diversity in North America

Chloroplasts contain several copies of their DNA, and intra-individual haplotypic variation (heteroplasmy) is common in plants, but unexplored in the cosmopolitan genus Phragmites. The aims of this study were to assess if heteroplasmy due to paternal leakage of the chloroplast occurs in Phragmites and which new insights into the evolutionary history of Phragmites australis in North America can be identified from the heteroplasmic variation. Eight non-native P. australis haplotypes occur in North America and can interbreed with P. australis ssp. americanus and P. australis var. berlandieri, creating opportunities for biparental inheritance of distinctive haplotypes. The polymorphism in the trnT-trnL sequence length revealed seventeen cases of heteroplasmy worldwide, in contact zones of distantly related haplotypes and in known hybrid populations, nine of which occurred in North America. In America, the cloned sequences, combined with nuclear markers, identified recombined haplotypes between native P. australis ssp. americanus and invasive P. australis haplotype M, and between the species P. mauritianus and P. australis, due to chloroplast paternal leakage. The occurrence of heteroplasmy and recombined haplotypes suggest a local origin for some of the rare non-native haplotypes occurring in North America, and plastid leakage events in the evolutionary histories of P. australis ssp. americanus and P. australis var. berlandieri.     

Intraspecific Variation in Mitogenomes of Five <Emphasis Type="Italic">Crassostrea</Emphasis> Species Provides Insight into Oyster Diversification and Speciation

A large number of Crassostrea oysters are found in Asia-Pacific. While analyses of interspecific variation have helped to establish historical relationships among these species, studies on intraspecific variation are necessary to understand their recent evolutionary history and current forces driving population biology. We resequenced 18 and analyzed 31 mitogenomes of five Crassostrea species from China: Crassostrea gigas, Crassostrea angulata, Crassostrea sikamea, Crassostrea ariakensis, and Crassostrea hongkongensis. Our analysis finds abundant insertions, deletions, and single-nucleotide polymorphisms in all species. Intraspecific variation varies greatly among species with polymorphic sites ranging from 54 to 293 and nucleotide diversity ranging from 0.00106 to 0.00683. In all measurements, C. hongkongensis that has the narrowest geographic distribution exhibits the least sequence diversity; C. ariakensis that has the widest distribution shows the highest diversity, and species with intermediate distribution show intermediate levels of diversity. Low sequence diversity in C. hongkongensis may reflect recent bottlenecks that are probably exacerbated by human transplantation. High diversity in C. ariakensis is likely due to divergence of northern and southern China populations that have been separated without gene flow. The significant differences in mitogenome diversity suggest that the five sister species of Crassostrea have experienced different evolutionary forces since their divergence. The recent divergence of two C. ariakensis populations and the C. gigas/angulata species complex provides evidence for continued diversification and speciation of Crassostrea species along China’s coast, which are shaped by unknown mechanisms in a north–south divide.     

Divergent Evolution of Symbiotic Bacteria: Rhizobia of the Relic Legume <Emphasis Type="Italic">Vavilovia formosa</Emphasis> Form an Isolated Group within <Emphasis Type="Italic">Rhizobium leguminosarum</Emphasis> bv. <Emphasis Type="Italic">viciae</Emphasis>

Comparative sequence analysis of symbiotic genes (nodA, nodC, nodD, nifH), which are elements of accessory component of the rhizobial genome, demonstrated that the strains of Rhizobium leguminosarum bv. viciae, isolated from the nodules of a relic legume, Vavilovia formosa, the closest relative of hypothetical common ancestor of the tribe Fabeae, represented a group separated from the strains of R. leguminosarum bv. viciae, isolated from other representatives of this tribe (Vicia, Lathyrus, Pisum, Lens). No isolation was observed relative to the genes representing the core component of the rhizobial genome (16S rDNA, ITS, glnII) or relative to host specificity of the rhizobia. The data obtained suggest that sequence divergence of symbiotic genes marks the initial stage of sympatric speciation, which can be classified as the isolation of the relic “vaviloviae” symbiotype, a possible evolutionary precursor of the “viciae” biotype.     

The intermediate sperm type and genitalia of <Emphasis Type="Italic">Zorotypus shannoni</Emphasis> Gurney: evidence supporting infraordinal lineages in Zoraptera (Insecta)

The sperm ultrastructure and the male and female genital apparatus of Zorotypus shannoni were examined and documented in detail, mainly using transmission electron microscopy micrographs. The findings suggest an evolutionary trend shared with Z. hubbardi and Z. impolitus. The three species are characterized by enlarged mitochondrial derivatives and related modifications. Giant sperm are probably a synapomorphy of Z. hubbardi and Z. impolitus, whereas an intermediate condition of this feature is found in Z. shannoni. The monophyletic origin of Z. caudelli, Z. magnicaudelli, Z. huxleyi and Z. weidneri is suggested by characteristically modified axonemes. The presence of extra-acrosomal material is also an unusual feature for Zoraptera, but this condition also occurs in the majority of polyneopteran groups. The long and convoluted female spermathecal duct with secretory and duct-forming cells is a constant feature in Zoraptera. The enlarged seminal receptacle suggests an evolutionary link between the male genital structures and the sperm size on one hand, and the size of the female spermatheca on the other. The small and otherwise uniform group Zoraptera exhibits a remarkable variation of sperm types and genital structures, suggesting the impact of different types of selection. It is likely that cryptic female choice plays a major role in shaping the genital apparatus.     

Descriptions of eight new species of <Emphasis Type="Italic">Ligophorus</Emphasis> Euzet &; Suriano, 1977 (Monogenea: Ancyrocephalidae) from Red Sea mullets

Eight new species of Ligophorus Euzet &; Suriano, 1977 (Monogenea: Ancyrocephalidae) are described from two species of mullets from the Red Sea. Ligophorus bykhowskyi n. sp. and L. zhangi n. sp. from Crenimugil crenilabris (Forsskål) differ from other species of the genus in the structure of the male copulatory organ, which has a simple accessory piece and a wide copulatory tube that arises from a large, single-chambered, expanded base. Ligophorus simpliciformis n. sp., L. bipartitus n. sp., L. campanulatus n. sp., L. mamaevi n. sp., L. lebedevi n. sp. and L. surianoae n. sp. from Liza carinata (Valenciennes) are differentiated on the basis of the morphometrics of the hard parts of the haptor and male copulatory organ. The eight species represent the first records of species directly attributed to Ligophorus from the Red Sea. Measurements of the haptoral hard-parts and the male copulatory organ of the new species are analysed with the aid of Principal Component Analysis. Three morphological types of male copulatory organ, five types of anchor, and two types of ventral and three types of dorsal bars were distinguished among these species. L. bykhowskyi and L. zhangi from C. crenilabris have the same type of male copulatory organ and anchors. Those species from Liza carinata have only one common morphological character, a thick copulatory tube, but have two types of accessory piece, four types of anchors and three types of bars. All species of Ligophorus found on mullets in the Red Sea have an accessory piece without a distal bifurcation and thus differ from most species of this genus from other regions of the world’s oceans.     

Genome-wide repeat dynamics reflect phylogenetic distance in closely related allotetraploid <Emphasis Type="Italic">Nicotiana</Emphasis> (Solanaceae)

Nicotiana sect. Repandae is a group of four allotetraploid species originating from a single allopolyploidisation event approximately 5 million years ago. Previous phylogenetic analyses support the hypothesis of N. nudicaulis as sister to the other three species. This is concordant with changes in genome size, separating those with genome downsizing (N. nudicaulis) from those with genome upsizing (N. repanda, N. nesophila, N. stocktonii). However, a recent analysis reflecting genome dynamics of different transposable element families reconstructed greater similarity between N. nudicaulis and the Revillagigedo Island taxa (N. nesophila and N. stocktonii), thereby placing N. repanda as sister to the rest of the group. This could reflect a different phylogenetic hypothesis or the unique evolutionary history of these particular elements. Here we re-examine relationships in this group and investigate genome-wide patterns in repetitive DNA, utilising high-throughput sequencing and a genome skimming approach. Repetitive DNA clusters provide support for N. nudicaulis as sister to the rest of the section, with N. repanda sister to the two Revillagigedo Island species. Clade-specific patterns in the occurrence and abundance of particular repeats confirm the original (N. nudicaulis (N. repanda (N. nesophila + N. stocktonii))) hypothesis. Furthermore, overall repeat dynamics in the island species N. nesophila and N. stocktonii confirm their similarity to N. repanda and the distinctive patterns between these three species and N. nudicaulis. Together these results suggest that broad-scale repeat dynamics do in fact reflect evolutionary history and could be predicted based on phylogenetic distance.     

Effects of Colistin and Bacteriocins Combinations on the In Vitro Growth of <Emphasis Type="Italic">Escherichia coli</Emphasis> Strains from Swine Origin

Escherichia coli strains from swine origin, either susceptible or resistant to colistin, were grown under planktonic and biofilm cultures. After which, they were treated with antibacterial agents including nisin and enterocin DD14 bacteriocins, colistin and their combinations. Importantly, the combination of colistin, enterocin DD14 and nisin eradicated the planktonic and biofilm cultures of E. coli CIP54127 and the E. coli strains with colistin-resistance phenotype such as E. coli 184 (mcr-1 ⁺) and E. coli 289 (mcr-1 ^?), suggesting therefore that bacteriocins from lactic acid bacteria could be used as agents with antibiotic augmentation capability.     

Characterization of <Emphasis Type="Italic">GA20ox</Emphasis> genes in tall and dwarf types coconut (<Emphasis Type="Italic">Cocos nucifera</Emphasis> L.)

Coconuts (Cocos nucifera L.) are divided by the height into tall and dwarf types. In many plants the short phenotype was emerged by mutation of the GA20ox gene encoding the enzyme involved in gibberellin (GA) biosynthesis. Two CnGA20ox genes, CnGA20ox1 and CnGA20ox2, were cloned from tall and dwarf types coconut. The sequences, gene structures and expressions were compared. The structure of each gene comprised three exons and two introns. The CnGA20ox1 and CnGA20ox2 genes consisted of the coding region of 1110 and 1131 bp, encoding proteins of 369 and 376 amino acids, respectively. Their amino acid sequences are highly homologous to GA20ox1 and GA20ox2 genes of Elaeis guineensis, but only 57% homologous to each other. However, the characteristic amino acids two histidines and one aspartic acid which are the two iron (Fe²⁺) binding residues, and arginine and serine which are the substrate binding residues of the dioxygenase enzyme in the 20G-FeII_Oxy domain involved in GA biosynthesis, were found in the active site of both enzymes. The evolutionary relationship of their proteins revealed three clusters in vascular plants, with two subgroups in dicots and three subgroups in monocots. This result confirmed that CnGA20ox was present as multi-copy genes, and at least two groups CnGA20ox1 and CnGA20ox2 were found in coconut. The nucleotide sequences of CnGA20ox1 gene in both coconut types were identical but its expression was about three folds higher in the leaves of tall coconut than in those of dwarf type which was in good agreement with their height. In contrast, the nucleotide sequences of CnGA20ox2 gene in the two coconut types were different, but the expression of CnGA20ox2 gene could not be detected in either coconut type. The promoter region of CnGA20ox1 gene was cloned, and the core promoter sequences and various cis-elements were found. The CnGA20ox1 gene should be responsible for the height in coconut, which is different from other plants because no mutation was present in CnGA20ox1 gene of dwarf type coconut.     

Analysis of novel high-molecular-weight prolamins from <Emphasis Type="Italic">Leymus multicaulis</Emphasis> (Kar. et Kir.) Tzvelev and <Emphasis Type="Italic">L. chinensis</Emphasis> (Trin. ex Bunge) Tzvelev

Nine novel high-molecular-weight prolamins (HMW-prolamins) were isolated from Leymus multicaulis and L. chinensis. Based on the structure of the repetitive domains, all nine genes were classified as D-hordeins but not high-molecular-weight glutenin subunits (HMW-GSs) that have been previously isolated in Leymus spp. Four genes, Lmul 1.2, 2.4, 2.7, and Lchi 2.5 were verified by bacterial expression, whereas the other five sequences (1.3 types) were classified as pseudogenes. The four Leymus D-hordein proteins had longer N-termini than those of Hordeum spp. [116/118 vs. 110 amino acid (AA) residues], whereas three (Lmul 1.2, 2.4, and 2.7) contained shorter N-termini than those of the Ps. juncea (116 vs. 118 AA residues). Furthermore, Lmul 1.2 was identified as the smallest D-hordein, and Lmul 1.2 and 2.7 had an additional cysteines. Phylogenetic analysis supported that the nine D-hordeins of Leymus formed two independent clades, with all the 1.3 types clustered with Ps. juncea Ns 1.3, whereas the others were clustered together with the D-hordeins from Hordeum and Ps. juncea and the HMW-GSs from Leymus. Within the clade of four D-hordein genes and HMW-GSs, the HMW-GSs of Leymus formed a separated branch that served as an intermediate between the D-hordeins of Ps. juncea and Leymus. These novel D-hordeins may be potentially utilized in the improvement of food processing properties particularly those relating to extra cysteine residues. The findings of the present study also provide basic information for understanding the HMW-prolamins among Triticeae species, as well as expand the sources of D-hordeins from Hordeum to Leymus.