Extreme Multiple Reticulate Origins of the Pteris cadieri Complex (Pteridaceae)

The Pteris cadieri complex displays extensive morphological variation and seems to have originated through hybridization. However, the members of this complex reproduce by apogamy, which usually limits genetic variation. To evaluate the hypotheses of hybrid origins, the pattern of evolution in this species complex is reconstructed. Multiple methodologies were used. Diploids, triploids, and tetraploids were identified by chromosome counts and flow cytometry. Nuclear DNA markers (cytosolic phosphoglucose isomerase gene, PgiC) were used, together with chloroplast DNA markers (atpB-rbcL spacer and rbcL gene) to infer the biparental and maternal lineages of the Pteris cadieri complex. The three cpDNA haplotype groups and five PgiC alleles found in this study indicate that the evolution of the Pteris cadieri complex has been extremely reticulate. Up to 11 taxa belonging to eight morphs were identified. By comparing genetic variation in the Pteris cadieri in two independent areas, Hainan and Taiwan, we inferred that hybridization has occurred independently in different areas. Furthermore, we found evidence for phenological divergence (evergreen and deciduous) within Taiwan. We propose that the Pteris cadieri complex originated from different genetic lineages through multiple hybridizations in different geographical areas, leading to its present morphological diversity.     

Admixture Analysis Using Genotyping-by-Sequencing Reveals Genetic Relatedness and Parental Lineage Distribution in Highbush Blueberry Genotypes and Cross Derivatives

Blueberries (Vaccinium section Cyanococcus) are perennial shrubs widely cultivated for their edible fruits. In this study, we performed admixture and genetic relatedness analysis of northern highbush (NHB, primarily V. corymbosum) and southern highbush (SHB, V. corymbosum introgressed with V. darrowii, V. virgatum, or V. tenellum) blueberry genotypes, and progenies of the BNJ16-5 cross (V. corymbosum × V. darrowii). Using genotyping-by-sequencing (GBS), we generated more than 334 million reads (75 bp). The GBS reads were aligned to the V. corymbosum cv. Draper v1.0 reference genome sequence, and ~2.8 million reads were successfully mapped. From the alignments, we identified 2,244,039 single-nucleotide polymorphisms, which were used for principal component, haplotype, and admixture analysis. Principal component analysis revealed three main groups: (1) NHB cultivars, (2) SHB cultivars, and (3) BNJ16-5 progenies. The overall fixation index (F_ST) and nucleotide diversity for NHB and SHB cultivars indicated wide genetic differentiation, and haplotype analysis revealed that SHB cultivars are more genetically diverse than NHB cultivars. The admixture analysis identified a mixture of various lineages of parental genomic introgression. This study demonstrated the effectiveness of GBS-derived single-nucleotide polymorphism markers in genetic and admixture analyses to reveal genetic relatedness and to examine parental lineages in blueberry, which may be useful for future breeding plans.     

Molecular and Cytogenetic Analysis of rDNA Evolution in Crepis Sensu Lato

Although Crepis was the first model plant group in which chromosomal changes were considered to play an important role in speciation, their chromosome structure and evolution have been barely investigated using molecular cytogenetic methods. The aim of the study was to provide a better understanding of the patterns and directions of Crepis chromosome evolution, using comparative analyses of rDNA loci number and localisation. The chromosome base number and chromosomal organisation of 5S and 35S rDNA loci were analysed in the phylogenetic background for 39 species of Crepis, which represent the evolutionary lineages of Crepis sensu stricto and Lagoseris, including Lapsana communis. The phylogenetic relationships among all the species were inferred from nrITS and newly obtained 5S rDNA NTS sequences. Despite high variations in rDNA loci chromosomal organisation, most species had a chromosome with both rDNA loci within the same (usually short) chromosomal arm. The comparative analyses revealed several independent rDNA loci number gains and loci repositioning that accompanied diversification and speciation in Crepis. Some of the changes in rDNA loci patterns were reconstructed for the same evolutionary lineages as descending dysploidy.     

Allelopathic effects of hackberry in a bottomland forest community

Hackberry was previously found to produce toxins that inhibit the growth of associated herbaceous species in a grassland community. Nevertheless, it was hypothesized that bare areas under hackberry trees in a bottomland forest community may not be caused by allelopathy, since inhibitory compounds may be leached or metabolized faster than in a grassland community. Investigations indicated that the relatively bare areas under hackberry were not due to competition for minerals, light, or water, or to differences in texture or pH. The percent of soil moisture was always higher under hackberry trees than under bur oak trees where herbaceous species were prominent. Decaying hackberry leaves, leaf leachate, and soil collected from under hackberry trees significantly reduce seed germination and seedling growth of test species. Ferulic, caffeic, gentisic, andp-coumaric acids, and scopolin, and scopoletin were identified as phenolic phytotoxins produced in hackberry leaves. Thus it appears that the reduced growth of herbaceous vegetation associated with hackberry trees in both grassland and forest communities is due primarily to allelopathy, with the initial inhibition being accentuated by competition.     

Phenols in Leaves and Bark of Fagus sylvatica as Determinants of Insect Occurrences

Beech forests play an important role in temperate and north Mediterranean ecosystems in Greece since they occupy infertile montane soils. In the last glacial maximum, Fagus sylvatica (beech) was confined to Southern Europe where it was dominant and in the last thousand years has expanded its range to dominate central Europe. We sampled four different beech forest types. We found 298 insect species associated with beech trees and dead beech wood. While F. sylvatica and Quercus (oak) are confamilial, there are great differences in richness of the associated entomofauna. Insect species that inhabit beech forests are less than one fifth of those species living in oak dominated forests despite the fact that beech is the most abundant central and north European tree. There is a distinct paucity of monophagous species on beech trees and most insect species are shared between co-occurring deciduous tree species and beech. This lack of species is attributed to the vegetation history and secondary plant chemistry. Bark and leaf biophenols from beech indicate that differences in plant secondary metabolites may be responsible for the differences in the richness of entomofauna in communities dominated by beech and other deciduous trees.     

Evolutionary Analysis of Dipeptidyl Peptidase I

Human dipeptidyl peptidase I (DPPI) belongs to the family of papain-like cysteine peptidases. Its distinctive features are the unique exclusion domain which enables the eponymous activity and homotetramerization of DPPI, and its dependence on chloride ions for enzymatic activity. The oligomeric state of DPPI is unique in this family of predominantly monomeric peptidases. However, a distant DPPI ortholog from Plasmodium falciparum has been shown to be monomeric, indicating that the oligomeric state of DPPI varies between lineages. The aim of this work was to study the evolution of DPPI, with particular attention to the structural features that determine its characteristic enzymatic activity and preferences, and to reconstruct the evolution of its oligomerization. We analyzed fifty-seven selected sequences of DPPI and confirmed its presence in three lineages, namely, Amorphea (including animals and Amoebozoa), Alveolates and the metamonad Giardia. The amino acid residues that bind the chloride ion are highly conserved in all species, indicating that the dependence on chloride ions for activity is an evolutionarily conserved feature of DPPI. The number of N-glycosylation sites is significantly increased in animals, particularly vertebrates. Analysis of homology models and subunit contacts suggests that oligomerization is likely restricted to DPPIs in the Amorphea group.     

Genetic Distance and Age Affect the Cuticular Chemical Profiles of the Clonal Ant Cerapachys biroi

Although cuticular hydrocarbons (CHCs) have received much attention from biologists because of their important role in insect communication, few studies have addressed the chemical ecology of clonal species of eusocial insects. In this study we investigated whether and how differences in CHCs relate to the genetics and reproductive dynamics of the parthenogenetic ant Cerapachys biroi. We collected individuals of different ages and subcastes from several colonies belonging to four clonal lineages, and analyzed their cuticular chemical signature. CHCs varied according to colonies and clonal lineages in two independent data sets, and correlations were found between genetic and chemical distances between colonies. This supports the results of previous research showing that C. biroi workers discriminate between nestmates and non-nestmates, especially when they belong to different clonal lineages. In C. biroi, the production of individuals of a morphological subcaste specialized in reproduction is inversely proportional to colony-level fertility. As chemical signatures usually correlate with fertility and reproductive activity in social Hymenoptera, we asked whether CHCs could function as fertility-signaling primer pheromones determining larval subcaste fate in C. biroi. Interestingly, and contrary to findings for several other ant species, fertility and reproductive activity showed no correlation with chemical signatures, suggesting the absence of fertility related CHCs. This implies that other cues are responsible for subcaste differentiation in this species.     

Antiherbivore Chemistry of Eucalyptus--Cues and Deterrents for Marsupial Folivores

Formylated phloroglucinol compounds (FPCs) are the single most important factor determining the amount of foliage that marsupial folivores eat from individual Eucalyptus trees. Folivores need to recognize which trees contain FPCs if they are to avoid them and forage efficiently, they are challenged by great diversity in the types and quantities of FPCs present, even within eucalypt species. We investigated the relationship between FPCs and terpenoids in species with both simple and complex FPC profiles and found strong positive correlations between terpenes generally, and several monoterpenes in particular, and FPCs. Terpene cues also indicated qualitative differences in trees' FPC profiles. We describe significant qualitative and quantitative variation in FPCs in several species that are important food sources for marsupial folivores. New discoveries include the fact that macrocarpals occur as two major, distinct groups and several new dimeric acylphloroglucinols from Eucalyptus strzeleckii. These patterns add to the chemical complexity of the foraging environment for folivores.     

Species-specific Leaf Volatile Compounds of Obligate <Emphasis Type="Italic">Macaranga</Emphasis> Myrmecophytes and Host-specific Aggressiveness of Symbiotic <Emphasis Type="Italic">Crematogaster</Emphasis> Ants

Macaranga myrmecophytes harbor species-specific Crematogaster ants that defend host trees from herbivores. We examined ant aggressive behaviors when artificially damaged leaf pieces from another tree were offered to four sympatric species of obligate Macaranga myrmecophytes. The ants showed aggressive behavior in response to leaf pieces regardless of the leaf species; however, aggressiveness was higher when conspecific leaf pieces were offered than when nonhost species were offered. Thus, ants can recognize leaf damage and distinguish among damaged leaf species. Chemical analyses of volatile compounds emitted from damaged leaves that may induce ant defense showed that the composition of the minor compounds differed among the four Macaranga species, although there were many compounds in common.     

Phylogeography and Genetic Differentiation among Populations of the Moon Turban Snail Lunella granulata Gmelin, 1791 (Gastropoda: Turbinidae)

We examined the genetic variation and phylogeographic relationships among 10 populations of Lunella granulata from mainland China, Penghu Archipelago, Taiwan Island, and Japan using mitochondrial COI and 16S markers. A total of 45 haplotypes were obtained in 112 specimens, and relatively high levels of haplotype diversity (h = 0.903) and low levels of nucleotide diversity (π = 0.0046) were detected. Four major phylogenetic lineage clusters were revealed and were concordant with their geographic distribution, agreeing with the haplotype network. These results suggested that geographic barrier isolating effects were occurring among the populations. This hypothesis was also supported by a significant genetic differentiation index (F_ST = 0.709) and by a spatial analysis of molecular variance (SAMOVA) analysis. A mismatch distribution analysis, neutrality tests and Bayesian skyline plots found a single significant population expansion. This expansion occurred on the coast of mainland China before 20–17 ka. Consequently, although the dispersal ability of the planktonic stage and the circulation of ocean currents generally promote genetic exchanges among populations, L. granulata has tended to maintain distinct genetic groups that reflect the respective geographic origins of the constituent lineages. Although the circulation of ocean currents, in principle, may still play a role in determining the genetic composition of populations, long-distance migration between regions is difficult even at the planktonic stage.     

Effects of large mammalian herbivores and ant symbionts on condensed tannins of Acacia drepanolobium in Kenya

Condensed tannins have been considered to be important inducible defenses against mammalian herbivory. We tested for differences in condensed tannin defenses in Acacia drepanolobium in Kenya over two years among different large mammalian herbivore treatments [total exclusion, antelope only, and megaherbivore (elephants and giraffes) + antelope] and with four different ant symbiont species on the trees. We predicted that (1) condensed tannin concentrations would be lowest in the mammal treatment with the lowest level of herbivory (total exclusion), (2) trees occupied by mutualist ants that protect the trees most aggressively would have lower levels of tannins, and (3) if chemical defense production is costly, there would be a trade-off between tannin concentrations, growth, and mechanical defenses. Mean tannin concentrations increased from total exclusion treatments to wildlife-only treatments to megaherbivore + antelope treatments. In 1997, condensed tannin concentrations were significantly lower in trees occupied by the ant Crematogaster nigriceps, the only ant species that actively removed axillary buds. Contrary to our prediction, trees occupied by ant species that protect the trees more aggressively against mammalian herbivores did not have lower overall levels of condensed tannins. There was no consistent evidence of a trade-off between tannin concentrations and growth rate, but there was a positive correlation between mean thorn length and mean tannin concentrations across species of ant inhabitants and across herbivore treatments in 1997. Contrary to our expectation, trees had higher tannin concentrations in the upper parts of the canopy where there is little herbivory by mammals.     

Effector Profiles of Endophytic Fusarium Associated with Asymptomatic Banana (Musa sp.) Hosts

During the infection of a host, plant pathogenic fungi secrete small proteins called effectors, which then modulate the defence response of the host. In the Fusarium oxysporum species complex (FOSC), the secreted in xylem (SIX) gene effectors are important for host-specific pathogenicity, and are also useful markers for identifying the various host-specific lineages. While the presence and diversity of the SIX genes has been explored in many of the pathogenic lineages of F. oxysporum, there is a limited understanding of these genes in non-pathogenic, endophytic isolates of F. oxysporum. In this study, universal primers for each of the known SIX genes are designed and used to screen a panel of endophytically-associated Fusarium species isolated from healthy, asymptomatic banana tissue. SIX gene orthologues are identified in the majority of the Fusarium isolates screened in this study. Furthermore, the SIX gene profiles of these endophytic isolates do not overlap with the SIX genes present in the pathogenic lineages of F. oxysporum that are assessed in this study. SIX gene orthologues have not been commonly identified in Fusarium species outside of the FOSC nor in non-pathogenic isolates of F. oxysporum. The results of this study indicate that the SIX gene effectors may be more broadly distributed throughout the Fusarium genus than previously thought. This has important implications for understanding the evolution of pathogenicity in the FOSC.     

Ruffed grouse feeding behavior and its relationship to secondary metabolites of quaking aspen flower buds

Quaking aspen (Populus tremuloides Michx.) staminate flower buds and the extended catkins are primary food resources for ruffed grouse (Bonasa umbellus). Winter feeding observations indicate that ruffed grouse select specific trees or clones of quaking aspen to feed in. Flower buds and catkins of quaking aspen were analyzed for secondary compounds (tannins, alkaloids, and phenolics) that might cause ruffed grouse to avoid trees with high levels of these compounds. Coniferyl benzoate, a compound that has not been previously found in quaking aspen, exists in significantly higher concentrations in buds from trees with no feeding history as compared to ruffed grouse feeding trees. Aspen catkins were also significantly lower in coniferyl benzoate than buds from the same tree. Ruffed grouse feeding preference was not related to the tannin or total phenolic levels found in buds or catkins. Buds from feeding trees had higher protein levels than trees with no feeding history; however, catkins did not differ from buds in protein concentration. The high use of extended catkins in the spring by ruffed grouse is probably due to a lower percentage of bud scale material in the catkin as opposed to the dormant bud. Bud scales contain almost all of the nontannin phenolics in catkins and dormant buds. A feeding strategy where bud scales are avoided may exist for other bird species that feed on quaking aspen. Dormant flower buds are significantly lower in protein-precipitable tannins than catkins and differ in secondary metabolite composition from other aspen foliage.Minnesota Agriculture Experiment Station Journal Series No. 16,953.     

Fundamental aspects of wood as a component of thermoplastic composites

In order to utilize wood‐based particles and fibers effectively as fillers or reinforcements in thermoplastic composites, a fundamental understanding of the structural and chemical characteristics of wood is required. Wood is the secondary xylem of trees, shrubs, and woody lianas (vines). Although the physiological function of wood is similar among different groupings of trees, significant differences are found in the basic anatomical structure of the broadest groupings of gymnosperms (coniferous trees or “softwoods”) and angiosperms (broad‐leaved trees or “hardwoods”). In addition, there are anatomical differences in wood structure among the various species of trees. These structural differences may have effects on the use of these materials in composites. Wood cell walls comprise three major organic constituents, namely, cellulose hemicelluloses, and lignin. In addition to these structural polymers, numerous other organic materials (“extractives”) may be present within the wood. The chemical composition of wood varies between species. The basic characteristics of anatomy and structure combine to impart variations in permeability, bulk chemistry, and surface chemistry. Characterization of particle size and shape, as well as surface tension characteristics as indicators of wettability, becomes important as we try to understand how these biopolymeric materials will behave when introduced into synthetic polymer systems.     

Genetic Isolation among the Northwestern,Southwestern and Central-Eastern Indian Ocean Populations of the Pronghorn Spiny Lobster Panulirus penicillatus

The pronghorn spiny lobster Panulirus penicillatus is a highly valuable species which is widely distributed in Indo-West Pacific and Eastern Pacific regions. Mitochondrial DNA control region sequences (566–571 bp) were determined to investigate the population genetic structure of this species in the Indian Ocean. In total, 236 adult individuals of Panulirus penicillatus were collected from five locations in the Indian Ocean region. Almost all individuals had a unique haplotype. Intrapopulation haplotype (h) and nucleotide (π) diversities were high for each locality, ranging from h = 0.9986–1.0000 and π = 0.031593–0.043441. We observed distinct genetic isolation of population located at the northwestern and southwestern edge of the species range. Gene flow was found within localities in the central and eastern region of the Indian Ocean, probably resulting from an extended planktonic larval stage and prevailing ocean currents.     

Amino Acid Fluctuations in Young and Old Orange Trees and Their Influence on Glassy-Winged Sharpshooter (<Emphasis Type="Italic">Homalodisca vitripennis</Emphasis>) Population Densities

The glassy-winged sharpshooter (GWSS) is an invasive insect pest in California that proliferated in citrus while incorporating many other plant species in its host range. Field studies were conducted from September 2002 to September 2003 in adjacent young and old orange groves to determine the influence of amino acid concentrations in the xylem fluid on relative densities of the xylophagous GWSS. Beginning in early September 2002, higher densities of adult GWSS were observed on young compared to old trees. This difference became increasingly pronounced through late October in conjunction with increasing concentrations of essential amino acids in xylem fluid of the young trees. Greater densities of adult GWSS remained on young trees through early February 2003. Thereafter, the population crashed unexpectedly, leaving only negligible numbers of GWSS on either young or old trees. Mean concentrations of the essential amino acids for insect growth and development were higher in young compared to old trees. Besides these essential amino acids, asparagine, serine, glutamine, and tyrosine were also significantly higher in young trees during the September–February period when GWSS was present. The pattern of elevated amino acid concentrations in young trees continued through the entire year irrespective of the presence or absence of GWSS. Principal component analysis followed by factor analysis revealed three factors for young or old orange trees that accounted for 81.6 and 78.9% of the total variation in the young and old tree analyses, respectively. Various groups of amino acids in different factors displayed peak or elevated levels in young trees corresponding to the increased densities of GWSS. The potential roles of these amino acids in GWSS host selection are discussed.     

Allelopathic effects ofCitrus aurantium L.

Field observations on undisturbed stands of sour orange revealed thatCynodon dactylon, Chenopodium album, Avena sativa, andAmaranthus retroflexus were not able to grow normally and complete their life cycles under its canopies, although the same species grow well under adjacent trees of date palm. Investigations revealed that the failure of the test species to grow normally under sour orange was not due to competition for light, moisture and minerals or to differences in soil texture or pH. Soil under sour orange trees drastically reduced seed germination and/or seedling growth of test species. Aqueous extracts, decaying materials, and volatile compounds of senescent and nonsenescent sour orange leaves were found to inhibit seed germination and/or seedling growth of test species. Therefore, allelopathy appeared to be the basic factor responsible for the reduction in plant growth with competition propably accentuating its effects.     

Volatile Profile Differences and the Associated Sirex noctilio Activity in Two Host Tree Species in the Northeastern United States

Sirex noctilio females are known to be attracted to stem sections of stressed pine trees for oviposition. The volatile profiles and attractiveness of Eastern white pine (Pinus strobus) and two chemotypes of Scots pine (P. sylvestris) were compared after stem injection with herbicide. In general, trap captures on herbicide-treated trees were higher than on controls. The high-carene chemotype of Scots pine captured the highest numbers of females, followed by the low-carene chemotype, and finally the Eastern white pine. Herbicide-treated trees of both species emitted larger quantities of volatiles than the controls. The herbicide treatment induced higher volatile emission rates in the Scots pine chemotypes than in white pine, although there was no difference between the two chemotypes. However, qualitative differences were found between the volatile profiles of the two species as well as between the two Scots pine chemotypes, which could account for the differential attractiveness of the species and chemotypes tested.     

Long-term monitoring of post-fire aboveground biomass recovery in a lowland dipterocarp forest in East Kalimantan,Indonesia

Between 1988 and 2000, changes in the above-ground biomass (AGB) of trees in an East Kalimantan lowland forest, damaged by fires in 1982–83 and 1998, were estimated using allometric functions and an annual inventory of stem diameter. The original vegetation of the study site was lowland dipterocarp forest which has since been affected by selective logging and wild fires. The 1982–83 fire killed large trees of primary species and the opened sites became dominated by a few pioneer species. Between 1988 and 1997, a few pioneer tree species, namely Macaranga spp., dominated a heavily disturbed stand (HDS). Primary tree species that survived the 1982–83 fire dominated a lightly disturbed stand (LDS). A moderately disturbed stand (MDS) contained vegetation intermediate between the HDS and the LDS. In 1997, there were 553, 499 and 356 trees ha-1 in the HDS, MDS and LDS, respectively. Macaranga trees accounted for 70, 40 and 11 of the total number of trees in the HDS, MDS, and LDS, respectively. In 1997, the AGB of trees in the HDS, MDS, and LDS was 117, 280, and 315 Mg ha-1, respectively. The proportion of biomass accounted for by Macaranga trees for the HDS, MDS, and LDS was 34, 8 and 1, respectively. The pioneer trees did not compensate for the loss of aboveground biomass resulting from the death of large primary trees. The fire in 1998 again decreased AGB of the stands. In 2000, the AGB of trees in the HDS, MDS, and LDS was 27, 106, and 219 Mg ha-1, respectively. The sites opened up by the 1998 fire were covered with the pioneer seedlings and seemingly dominated by the pioneer trees with a larger number of stems per ha, but lower biomass as compared to the original forest (> 400 Mg ha-1).