ComprehendingCornus: Puzzles and progress in the systematics of the dogwoods

Dogwoods evolved in two main lineages, a red-fruited line in which the inflorescences have basal bracts, and a blue-(or white-)fruited line in which the bracts are rudimentary or lacking. The 15 “red-line” species are mostly well marked, whereas “blue-line” species—numbering roughly 50 if some newer treatments be accepted—are mostly hard to tell apart. Red-line ovules are tenuinucellate, blue-line ovules crassinucellate. Dividing the red line separates cornelian cherries from the showy-bracted species; dividing the latter separates dwarf cornels from big-bracted dogwoods. Adding in the blue line there are thus four subgroups, which differ with regard to inflorescences— and with regard to iridoid glucosides, among other things. I downplay further subdivision (into opposite- and alternateleaved blue-line groups, for instance) and focus on few traits because I aim to trace the subgroups backward in the fossil record, to decide the direction of evolutionary changes, and to infer the causes for divergence. Unlike pollen, leaves, and wood, fossil dogwood fruits can often be assigned to one of the four subgroups and sometimes to a species group within a subgroup.Dunstania (based on fruits from England’s early Tertiary and here transferred toCornus’s cornelian cherry subgroup) or something close to the dunstanias gave rise to modernC. volkensii of Africa and four related species found today in California and Eurasia. Fossil fruit-stones of the blue line, like their modern counterparts, can be subglobose, compressed and asymmetric, fluted, or topped by a deep depression. Such a deep depression serves to link a fossil to the extant speciesC. alternifolia andC. controversa, but other features of the blue-line fruitstones, overlapping and less constant, cannot be used to prove a fossil’s tie to groups within the blue line. Evidence of several kinds makes the blue-line dogwoods oldest and connectsCornus to the nyssoids:Nyssa, Camptotheca, andDavidia. Commonly called Nyssaceae after 1910, when Wangerin got their traits wrong, the nyssoids are surer relatives ofCornus than any of Wangerin’s (“Das Pflanzenreich”) cornaceous genera.Mastixia, however, may be the actual sister group because it shares withCornus 1-celled 2-armed hairs that nyssoids lack.Cornus, nyssoids, and mastixioids (modernMastixia and its closest fossil allies) are here regarded as the true Cornaceae. Other genera once thought to be cornaceous are doubtfully to not at all related. As is now widely known, discordant features makeCorokia andKaliphora (for example) out of place in the Cornaceae, butAucuba andGarrya (for example) are neither easily excluded by a listing of their traits nor easily included by construction of branched diagrams. The novelty that brought about the radiation of true cornads was, I think, epigyny, followed fairly quickly by the hard endocarp and singleseeded locules. Mastixioid fruits of the late Cretaceous were dry or leathery and smaller than their successors. This suggests that early members of the family spread abiotically or by means of animals that ate whole plants. Bigger, fleshier fruits came later through interaction with evolving frugivores. The cause of early radiation withinCornus probably was interaction with insect predators and pollinators. One species of the blue line escaped competition for spring pollinators by switching to fall flowering and thereby kept some traits that other dogwoods lost. Though birds are now the main dispersers of the dogwoods, rodents likely play a minor role, and monkeys likely played a major role when compound fruits evolved. Though gynoecial reduction was the rule in dogwoods,C. (Dunstania) multilocularis’s multiple seed chambers reflect an evolutionary increase. I postulate that modernCornus’s 4-merous flower with 2-merous ovary has a strong developmental tie to pairing of the leaves and branches.     

Mutations in the right operator of bacteriophage lambda: physiological effects

The right operator in bacteriophage lambda vs326 has one-twentieth the in vitro binding affinity for repressor as λv⁺; for comparison λv3 has one-quarter the affinity of λv⁺. In vivo, both mutants constitutively express genes in the right operon. Both λv3 and λvs326 express gene O constitutively because they complement λimm434Oam^? in a λ lysogen, vs, more efficiently than v3. The v3 allele in cis (but not in trans) to vs326 gives significantly greater phage yields in a λ lysogen than λvs326 alone, cro gene function, measured by arrest of exonuclease synthesis, suggested the following series of increasing degree of conatitutivity: v3, vs326, v3 vs326. λv2 vs326 forms plaques on lysogens that carry λcI857, but λv2 v3 does not. These results indicate that vs326, like v3, is an operator constitutive mutation but stronger in its effects. These mutants exemplify a uniform correlation between relative weakness of repressor binding and degree of constitutive gene expression.     

Lights out: the evolution of bacterial bioluminescence in Loliginidae

Representatives of several metazoan clades engage in symbiotic interactions with bioluminescent bacteria, but the evolution and maintenance of these interactions remain poorly understood. Uroteuthis is a genus of loliginid squid (Cephalopoda: Loliginidae) characterized by paired ventral photophores (light organs) housing bioluminescent bacteria. While previous phylogenetic studies have suggested that Uroteuthis is closely related to Loliolus, a genus of non-bioluminescent species, this relationship remains unresolved. To illuminate Uroteuthis and Loliolus phylogeny and its implications for the evolution of bioluminescence in Loliginidae, we generated sequences from two mitochondrial genes from Uroteuthis specimens sampled from several sites in the Indian and western Pacific Oceans. We combined these data with data from GenBank, analyzed the concatenated data set using maximum likelihood and Bayesian methods, and reconstructed the evolution of bacterial bioluminescence on the resulting phylogenies. Our analyses support the hypothesis that Uroteuthis is paraphyletic with respect to Loliolus. Furthermore, our reconstructions suggest that the symbiosis between loliginid squid and bioluminescent bacteria evolved once in the ancestor of Loliolini (the clade comprising Uroteuthis and Loliolus), but was subsequently lost in the ancestor of Loliolus. These findings could have profound implications for our understanding of the evolution of symbiotic bioluminescence in squid.     

Characterization of the acetyltransferase used in pheromone biosynthesis in moths: Specificity for the Z isomer in tortricidae

Characterization of the acetyltransferase (acetyl-CoA: fatty alcohol acetyltransferase) that catalyzes the conversion of fatty alcohols to acetate esters was carried out in the redbanded leafroller moth, Argyrotaenia velutinana. This enzyme is one of the last enzymes used in the pheromone biosynthetic pathway of many moths. Characterization of the acetyltransferase involved an in vitro enzyme assay using a pheromone gland homogenate. The enzyme found only in the pheromone gland, was active during the photophase and at all adult ages tested (1–5-days-old). The enzyme was derermined to be microsomal and exhibited specificity for the Z isomer of all 12, 14 and 16 carbon monounsaturated fatty alcohol substrates tested. The shape of the kinetic curves for the Z and E isomers of Δ11-tetradecenol were similar for time of incubation, protein and substrate concentration, with the Z isomer always forming 1.5–2 times more product. Substrate competition assays between Z and E11-tetradecenol indicate that one enzyme is involved and it converts the Z isomer to product faster than the E isomer. Substrate preference assays conducted with the enzyme from Choristoneura rosaceana, C. fumiferana and Platynota idaeusalis (all Tortricidae) pheromone glands also indicate specificity for the Z isomer. On the other hand, assays conducted with Trichoplusia ni (Noctuidae) and three strains (Z, E and hybrid) of Ostrinia nubilalis (Pyralidae) indicate no preferences between the Z and E isomers.     

The promiscuous larvae: flexibility in the establishment of symbiosis in corals

Coral reefs thrive in part because of the symbiotic partnership between corals and Symbiodinium. While this partnership is one of the keys to the success of coral reef ecosystems, surprisingly little is known about many aspects of coral symbiosis, in particular the establishment and development of symbiosis in host species that acquire symbionts anew in each generation. More specifically, the point at which symbiosis is established (i.e., larva vs. juvenile) remains uncertain, as does the source of free-living Symbiodinium in the environment. In addition, the capacity of host and symbiont to form novel combinations is unknown. To explore patterns of initial association between host and symbiont, larvae of two species of Acropora were exposed to sediment collected from three locations on the Great Barrier Reef. A high proportion of larvae established symbiosis shortly after contact with sediments, and Acropora larvae were promiscuous, taking up multiple types of Symbiodinium. The Symbiodinium types acquired from the sediments reflected the symbiont assemblage within a wide range of cnidarian hosts at each of the three sites, suggesting potential regional differences in the free-living Symbiodinium assemblage. Coral larvae clearly have the capacity to take up Symbiodinium prior to settlement, and sediment is a likely source. Promiscuous larvae allow species to associate with Symbiodinium appropriate for potentially novel environments that may be experienced following dispersal.     

Nippostrongylus brasiliensis: feeding activity in the mouse

Nippostrongylus brasiliensis incorporated the fluorescent dye, Rhodamine B, while feeding in vivo. Uptake of dye in both sexes of helminth increased linearly from 30 to 120 min after the hosts were given dye per os. Feeding by female N. brasiliensis significantly exceeded that of the male at 4 and 5 days postinfection. Feeding declined in older helminths of both sexes. The density of helminths had no effect on their incorporation of dye in vivo. Feeding in male- and female-only groups of worms was similar to that seen in populations of mixed sexes. Feeding by helminths decreased during the first 36 h of food deprivation in the host, but increased during subsequent fasting of the host. Both sexes of N. brasiliensis resumed feeding within 15 min after the fasted hosts were fed. Growth of N. brasiliensis increased linearly from 4 to 7 days postinfection, based on dry weight. Seven-day-old and older females were significantly heavier than males.     

Mould in buildings: the air spora of domestic dwellings

Filamentous fungi in 37 genera were isolated during winter from the internal air of 62 domestic dwellings in England and Scotland. In the air spora the number of viable fungal colony-forming units (cfu), including yeasts, ranged from < 12 to 449 800 cfu m⁻³. There was considerable variation between dwellings and between weekly samples taken within the same dwelling, but in approximately one-third of all air samples there were < 200 cfu m⁻³. The filamentous fungi isolated most frequently from the air in 47 Scottish dwellings were Penicillium spp. (96% of dwellings), Cladosporium spp. (89%), Aspergillus spp. (75%, mainly A. versicolor), Ulocladium spp. (62%), Geomyces pannorum (57%) and Sistotrema brinkmannii (51%). Yeasts were isolated in 94% of these dwellings and overall comprised 13% of cfu on Andersen sampler plates. Penicillium spp., Cladosporium spp. and S. brinkmannii together accounted for > 70% of the propagules. High numbers of spores in the internal air were associated with surface mould growth and constructional work. In addition, disturbance of surface growth and vacuum cleaning of carpets caused large temporary increases in the atmospheric spore count.     

Invaders on the move: parasitism in the sexual galls of four alien gall wasps in Britain (Hymenoptera: Cynipidae)

Four alien, host-alternating cynipid gall wasps are established in the British Isles: Andricus kollari, A. lignicola, A. quercuscalicis and A. corruptrix. Their current distributions divide Britain into four zones; all four invaders are present in the south and south-east of Britain, whereas only A. kollari is present in the far north of Scotland. The rank order, according to how far north the distributions of the four invading cynipid species reach, is: A. corruptrix→ A. quercuscalicis→ A. lignicola→ A. kollari. The life cycles of all four cynipid species involve a sexual generation in spring on Quercus cerris, and an agamic generation in autumn on Q. robur. Here we studied the parasitoid attack by four pteromalid species on the sexual generations of the invaders. We collected and reared the galls of all four species at eight sites from the south to the north of the country (two sites per zone). The geographical locations of the sites reflect the invasion history and thus the residence time of the alien species in the four zones. At each site we measured the densities of all host galls on Q. cerris and of both obligatory host-tree species. We also took a series of measures, such as host-tree density and mean host-tree size, to further characterize the tree stands. These measures are referred to as local parameters. Host densities varied between sites and between years. In A. kollari, galling rates were highest in the middle of the country (zone 2) in 1994, whereas in 1995 they increased from the south to the north. In A. lignicola, galling rates in both years were lowest at the sites in zone 3 (closest to its distribution boundary). In A. quercuscalicis, galling rates were found to be lowest at the site most to the north-west in both years, again the one furthest away from the area where this species was first recorded. Mortality caused by parasitoid attack differed from less than 10% to as high as 70% and varied between host species, sites and years. In four out of six cases the historical/regional variables (north/south and east/west) correlated significantly with parasitoid attack rates that were characteristically lowest at sites close to the distribution boundaries. Of the local factors, we found parasitoid attack rates correlated negatively in one case with host density, whereas they correlated positively in four cases with the density of alternative hosts of the parasitoids. In one of the models the local density of Q. cerris trees correlated negatively with parasitoid attack on A. quercuscalicis. For all three host species the terms retained in the minimal adequate models obtained for 1994 and 1995 differed, which might indicate that these communities of native parasitoids and invading host have not yet settled in any definite structure.     

Starting from scratch: Evolution of the lichen thallus in the basidiolichen Dictyonema (Agaricales: Hygrophoraceae)

Phylogenetic studies indicate that the basidiolichen genus Dictyonema s.lat., often thought to represent only a single genus with few species, includes several well-supported genus-level clades, all of which form associations with a unique lineage of obligately lichenized cyanobacteria (Rhizonema). In an attempt to elucidate the evolution and genus- and species-level diversification in Dictyonema s.lat., we generated 68 new sequences of the nuclear large subunit rDNA (nuLSU), the internal transcribed spacer (ITS), and the RNA polymerase II subunit (RPB2), for 29 species-level lineages representing all major clades of Dictyonema s.lat. and most of the species currently known. The multilocus phylogeny obtained via maximum likelihood and Bayesian approaches indicates the presence of five genus-level groups: a basal clade, Cyphellostereum, that is sister to the rest of the species, a paraphyletic grade representing Dictyonema s.str., and three clades representing the genera Acantholichen, Cora, and Corella. To determine the evolutionary transformations of the lichenized thallus in the group, ancestral character state reconstruction was done using six characters (lichenisation, thallus type, cortex type, hyphal sheath and haustorial type, photobiont morphology, and basidiocarp type). Our analysis indicates a progressive development of the lichenized thallus from loosely organized filamentous crusts with separate, cyphelloid basidiocarps in Cyphellostereum, to filamentous crusts with derived hyphal sheath and cyphelloid–stereoid basidiocarps partially incorporated into the lichen thallus in Dictyonema, to squamulose–foliose thalli with corticioid basidiocarps entirely supported by the lichen thallus in Cora. These results indicate a remarkable evolutionary integration of lichenized and reproductive tissues in Dictyonema s.lat., supporting the hypothesis that, at least in this case, lichenized thalli may have evolved from reproductive structures in their nonlichenized ancestors.     

Comparative Phylogeography of Direct-Developing Frogs (Anura: Craugastoridae: Pristimantis) in the Southern Andes of Colombia

The Andes of South America hosts perhaps the highest amphibian species diversity in the world, and a sizable component of that diversity is comprised of direct-developing frogs of the genus Pristimantis (Anura: Craugastoridae). In order to better understand the initial stages of species formation in these frogs, this study quantified local-scale spatial genetic structuring in three species of Pristimantis. DNA sequences of two mitochondrial gene fragments (16S and COI) were obtained from P. brevifrons, P. palmeri and P. jubatus at different locations in the Cordillera Occidental. We found high levels of genetic diversity in the three species, with highly structured populations (as measured by F _ST) in P. brevifrons and P. palmeri while P. jubatus showed panmixia. Large effective population sizes, inferred from the high levels of genetic diversity, were found in the three species and two highly divergent lineages were detected within P. jubatus and P. palmeri. Estimated divergence times among populations within P. brevifrons and P. palmeri coincide with the Pleistocene, perhaps due to similar responses to climatic cycling or recent geological history. Such insights have important implications for linking alpha and beta diversity, suggesting regional scale patterns may be associated with local scale processes in promoting differentiation among populations in the Andes.     

Distal-to-proximal NO conversion in hemoproteins: the role of the proximal pocket

Hemoproteins play central roles in the formation and utilization of nitric oxide (NO) in cellular signaling, as well as in protection against nitrosative stress. Key to heme-nitrosyl function and reactivity is the Fe coordination number (5 or 6). For (five-coordinate) 5c-NO complexes, the potential for NO to bind on either heme face exists, as in the microbial cytochrome c′ from Alcaligenes xylosoxidans (AxCYTcp), which forms a stable proximal 5c-NO complex via a distal six-coordinate NO intermediate and a putative dinitrosyl species. Strong parallels between the NO-binding kinetics of AxCYTcp, the eukaryotic NO sensor soluble guanylate cyclase, and the ferrocytochrome c/cardiolipin complex have led to the suggestion that a distal-to-proximal NO switch could contribute to the selective ligand responses in gas-sensing hemoproteins. The proximal NO-binding site in AxCYTcp is close to a conserved basic (Arg124) residue that is postulated to modulate NO reactivity. We have replaced Arg124 by five different amino acids and have determined high-resolution (1.07-1.40 Å) crystallographic structures with and without NO. These, together with kinetic and resonance Raman data, provide new insights into the mechanism of distal-to-proximal heme-NO conversion, including the determinants of Fe-His bond scission. The Arg124Ala variant allowed us to determine the structure of an analog of the previously unobserved key 5c-NO distal intermediate species. The very high resolution structures combined with the extensive spectroscopic and kinetic data have allowed us to provide a fresh insight into heme reactivity towards NO, a reaction that is of wide importance in biology.     

Propagule predation in a Neotropical mangrove: the role of the Grapsid crab Goniopsis cruentata

In Neotropical mangroves the crabs Ucides cordatus and Goniopsis cruentata have been considered the most significant propagule consumers, but their relative importance has not been investigated. The aim of this study was to compare the magnitude of predation by these crabs on three mangrove species propagules: Avicennia schaueriana, Laguncularia racemosa and Rhizophora mangle. We found that G. cruentata is a more important predator than U. cordatus in both natural and restored areas. We also tested the hypothesis that Ucides and Goniopsis have antagonistic effects on propagules predation using a cage experiment where the presence/absence of these species was manipulated in a 2 × 2 factorial design. The effects of Goniopsis were stronger in the absence of Ucides due to negative interactions between these predator species. Moreover, we found that Goniopsis preference for A. schaueriana and L. racemosa can favor the dominance of R. mangle in Neotropical mangroves. This study suggests that propagule predation by Goniopsis should be controlled in mangrove restoration programs at abandoned shrimp farms and destroyed areas, if dominance by R. mangle is undesirable relative to mixed species communities.     

Facilitation and competition in the high Arctic: the importance of the experimental approach

In the last decade, plant ecologists have focussed more on the occurrence of positive plant–plant interactions than ever before. Especially in severe environments, such as the Arctic, species removal experiments tend to find facilitative rather than competitive effects, casting doubt on the importance of competition under extreme growing conditions. Two approaches to measure plant–plant interactions presented here reveal that competition affects plant growth even in the high Arctic. Luzula confusa and Salix polaris show a reduced growth in mixed stands compared with pure. This competition effect is not detected in a removal experiment, which inevitably also alters site microclimate. Indeed, in the latter experiment, facilitative effects of Luzula on Salix were found. Evidently, both facilitation and competition are acting and important. Causes and implications are discussed using a conceptual model derived from that of Brooker and Callaghan (1998).     

Xylella fastidiosa in Europe: From the Introduction to the Current Status

Xylella fastidiosa is xylem-limited bacterium capable of infecting a wide range of host plants, resulting in Pierce’s disease in grapevine, citrus variegated chlorosis, olive quick decline syndrome, peach phony disease, plum leaf scald, alfalfa dwarf, margin necrosis and leaf scorch affecting oleander, coffee, almond, pecan, mulberry, red maple, oak, and other types of cultivated and ornamental plants and forest trees. In the European Union, X. fastidiosa is listed as a quarantine organism. Since its first outbreak in the Apulia region of southern Italy in 2013 where it caused devastating disease on Olea europaea (called olive leaf scorch and quick decline), X. fastidiosa continued to spread and successfully established in some European countries (Corsica and PACA in France, Balearic Islands, Madrid and Comunitat Valenciana in Spain, and Porto in Portugal). The most recent data for Europe indicates that X. fastidiosa is present on 174 hosts, 25 of which were newly identified in 2021 (with further five hosts discovered in other parts of the world in the same year). From the six reported subspecies of X. fastidiosa worldwide, four have been recorded in European countries (fastidiosa, multiplex, pauca, and sandyi). Currently confirmed X. fastidiosa vector species are Philaenus spumarius, Neophilaenus campestris, and Philaenus italosignus, whereby only P. spumarius (which has been identified as the key vector in Apulia, Italy) is also present in Americas. X. fastidiosa control is currently based on pathogen-free propagation plant material, eradication, territory demarcation, and vector control, as well as use of resistant plant cultivars and bactericidal treatments.     

Heme-heme interaction in cytochrome c oxidase: the cooperativity of the hemes of cytochrome c oxidase as evidenced in the reaction with CO

Isolated and purified cytochrome c oxidase from beef heart muscle mitochondria (Kuboyama et al. (1972) J. Biol. Chem.247, 6375–6383) is shown to be very similar to the hemoprotein in situ with respect to its EPR absorption properties and the half-reduction potentials of the hemes and copper. The half-reduction potentials of cytochromes a and a₃ in the purified cytochrome c oxidase are 205 mV and 360 mV, respectively, and these values are the same in the presence and absence of cytochrome c.Low-temperature EPR spectra show that the binding of CO to reduced cytochrome a₃ changes the oxidized cytochrome a from high spin (g 6) to low spin (g 3). In samples at 5–8 °K the photodissociation of the reduced cytochrome a₃CO compound shifts the spectrum of the oxidized low-spin cytochrome a to a lower g value and converts approximately 5% of the low-spin form to a high-spin form. The heme-heme interaction demonstrated in this reaction is very fast as evidenced by the fact that even at 5 °K the measured change in oxidized cytochrome is complete within 5 msec.     

Structural features of the mitogenome of the leafhopper genus Cladolidia (Hemiptera: Cicadellidae: Coelidiinae) and phylogenetic implications in Cicadellidae

The first two complete mitogenomes of the leafhopper genus Cladolidia (C. biungulata and C. robusta) were sequenced and annotated to further explore the phylogeny of Cladolidia. Both the newly sequenced mitogenomes have a typical circular structure, with lengths of 15,247 and 15,376 bp and A + T contents of 78.2% and 78%, respectively. We identified a highly conserved genome organization in the two Cladolidia spp. through comparative analysis that included the following assessments: genome content, gene order, nucleotide composition, codon usage, amino acid composition, and tRNA secondary structure. Moreover, we detected the base heterogeneity of Cicadellidae mitogenomic data and constructed phylogenetic trees using the nucleotide alignments of 12 subfamilies of 58 leafhopper species. We noted a weak heterogeneity in the base composition among the Cicadellidae mitogenomes. Phylogenetic analyses showed that the monophyly of each subfamily was generally well supported in the family Cicadellidae; the main topology was as follows: (Deltocephalinae + (Treehoppers + ((Megophthalminae + (Macropsinae + (Hylicinae + (Coelidiinae +Iassinae)) + (Idiocerinae + (Cicadellinae + (Typhlocybinae + (Mileewinae + (Evacanthinae +Ledrinae)))))))))). Within Coelidiinae, phylogenetic analyses revealed that C. biungulata and C. robusta belong to Coelidiinae and the monophyly of Cladolidia is well supported. In addition, on the basis of complete mitogenome phylogenetic analysis and the comparison of morphological characteristics, we further confirm the genus Olidiana as a paraphyletic group, suggesting that the genus may need taxonomic revisions.