Low genetic variability in the rare,recently differentiated Aquilegia paui (Ranunculaceae)

Aquilegia paui is a narrow endemic species from the north-eastern Iberian Peninsula. It has a reduced number of populations in the limestone mountain massif of Els Ports. Genetic diversity was evaluated through allozyme electrophoresis in this rare columbine, together with its widespread congener Aquilegia vulgaris. Low levels of genetic diversity were detected in both taxa, especially for A. paui. Genetic identity between the two columbines was particularly high (I = 0.990), and the allele pool of A. paui is a subset of that of its widespread congener (with the exception of a low-frequency allele that was only detected in one population of A. paui). A. paui may have recently evolved from nearby populations of A. vulgaris. Speciation of this taxon, which may still be ongoing, seems to be driven by ecological differentiation, as generally occurred for the European columbines as well as in many Mediterranean endemic species.     

Limitations on reproductive success in endemic <Emphasis Type="Italic">Aquilegia viscosa</Emphasis> (Ranunculaceae) relative to its widespread congener <Emphasis Type="Italic">Aquilegia vulgaris</Emphasis>: the interplay of herbivory and pollination

Plant reproduction can be strongly affected by herbivory and different features of pollination ecology, such as pollinator visitation rates and capacity for self-pollination. The purpose of this study is to compare the relative impact of herbivory and pollination on maternal reproductive success in endemic Aquilegia viscosa and its widespread congener Aquilegia vulgaris. We conducted herbivore exclusion experiments in two populations of each species in 2 different years and showed that the maternal fertility of A. viscosa was significantly more limited by floral predation and pre-dispersal seed predation than its widespread congener. In the absence of herbivory, A. viscosa retained significantly lower maternal fertility than A. vulgaris. Experimental pollinations in an insect-free glasshouse showed that the two species have an equal seed/ovule ratio both in the absence of pollinators and in the presence of non-limiting outcross pollination. Pollinator visitation rates were significantly higher in populations of A. vulgaris than in populations of A. viscosa. In addition, path analyses showed that spur length, an important trait for pollinator attraction in Aquilegia, and, indirectly sepal and petal width, contribute positively to the seed/ovule ratio in A. vulgaris, but not in A. viscosa. These results indicate that maternal fertility of endemic A. viscosa is strongly reduced by flower and seed predation despite low rates of pollinator visitation, and that pollen or resource limitation in the wild may further reduce maternal fertility. Finally, floral trait variation appears to be decoupled from fertility variation in endemic A. viscosa, which possibly constrains the evolution of reproductive traits in this species.     

Glandular trichomes as an inflorescence defence mechanism against insect herbivores in Iberian columbines

Glandular trichomes play a defensive role against herbivores in the leaves of many plant species. However, their functional role in inflorescences has not been studied, even though theory suggests that tissues with a higher fitness value, such as inflorescences, should be better defended. Using manipulative experiments, we analysed the defensive role of glandular trichomes against herbivorous insects in the inflorescence of Iberian columbines (genus Aquilegia), and its inter-population and inter-taxa variation in relation to herbivore abundance and potential selective pressure. The experiments were conducted in eight populations belonging to four subspecies of two columbines (Aquilegia vulgaris and Aquilegia pyrenaica). For each population, we estimated the density of glandular trichomes in the inflorescences, the abundance of insects stuck in the inflorescences, the abundance of small herbivorous insects, the incidence of damage on flowers and fruits, and the fruit set. The density of glandular trichomes on the inflorescence of A. vulgaris and A. pyrenaica was higher in regions of higher herbivore abundance. We also found that when the plants lose the protection of glandular trichomes, small insects have better access to flowers and fruits, causing more damage and reducing plant fitness. This study concludes that glandular trichomes are part of an adaptive response against phytophagous insect herbivory. The observed variation in herbivore pressure between taxa, likely caused by habitat differentiation, might have played a role in trait differentiation through divergent selection. This result adds evidence to the differentiation of the Iberian columbines through habitat specialization.     

Floral isolation between Aquilegia formosa and Aquilegia pubescens

The acquisition of floral nectar spurs is correlated with increased species diversity across multiple clades. We tested whether variation in nectar spurs influences reproductive isolation and, thus, can potentially promote species diversity using two species of Aquilegia, Aquilegia formosa and Aquilegia pubescens, which form narrow hybrid zones. Floral visitors strongly discriminated between the two species both in natural populations and at mixed-species arrays of individual flowers. Bees and hummingbirds visited flowers of A. formosa at a much greater rate than flowers of A. pubescens. Hawkmoths, however, nearly exclusively visited flowers of A. pubescens. We found that altering the orientation of A. pubescens flowers from upright to pendent, like the flowers of A. formosa, reduced hawkmoth visitation by an order of magnitude. In contrast, shortening the length of the nectar spurs of A. pubescens flowers to a length similar to A. formosa flowers did not affect hawkmoth visitation. However, pollen removal was significantly reduced in flowers with shortened nectar spurs. These data indicate that floral traits promote floral isolation between these species and that specific floral traits affect floral isolation via ethological isolation while others affect floral isolation via mechanical isolation.     

The nature of polyploidy inReseda sect.Leucoreseda (Resedaceae)

We have found two chromosome levels (n = 10 and n = 20) in Spanish species ofReseda sect.Leucoreseda, four species on the first (R. undata, R. paui, R. suffruticosa, R. barrelieri), only one on the second level (R. alba). As already the species with n = 10 apparently behave as polyploids, we propose x = 5 as the original basic chromosome number for this section. The seed protein profile ofR. alba (2n = 40) reveals close relationships withR. undata (2n = 20), while the esterase isozymes suggest affinities withR. paui (2n = 20). Thus,R. alba can be regarded either as an autopolyploid fromR. paui or more likely as an allopolyploid fromR. paui andR. undata. Finally, the DNA values suggest a diploidization process inR. alba since its origin.     

Antibacterial activity and mode of action of plant flavonoids against Proteus vulgaris and Staphylococcus aureus

(−)-Epigallocatechin, an antibiotic found in Elaeagnus glabra, and 28 other related plant flavonoids were tested for their antibacterial activity against Proteus vulgaris and Staphylococcus aureus. A free 3′,4′,5′-trihydroxy B-ring and a free 3-OH were necessary for antibacterial activity. DNA synthesis was predominantly inhibited by the active flavonoids in P. vulgaris, whereas RNA synthesis was inhibited in S. aureus.     

Gonadal development and reproductive strategies of the tropical octopus (Octopus insularis) in northeast Brazil

Octopus insularis inhabits shallow waters along the coast and oceanic islands of northeastern Brazil, where it is the main target of commercial fishing of octopuses. This study aims to investigate the hypothesis that the tropical O. insularis has a distinct gonadal development and reproductive features when compare to its congener O. vulgaris from the subtropical regions. In order to describe its reproductive development, 545 octopuses were collected in the Northeastern Brazil. A good correspondence was observed between the gonad morphology and its histological structure. Oocytes in different development stages were observed in mature females. Most female in early maturity stages had sperm stored in the spermathecae, indicating that females copulate when still immature. There was no correlation between testis weight and the Needham complex, suggesting a protracted period of spermatophore production. Octopus insularis has a general gonadal development pattern similar to O. vulgaris, however, some differences were observed, as maturation at a smaller size, probably associated to a shorter life, and lower fecundity. The distinct reproductive features of O. insularis seem to be related to less variable conditions in the tropical environments. Management should take into account the differences and establish specific rules for the Northeast Brazil octopus fisheries.     

Evidence for two types of Aquilegia ecalcarata and its implications for adaptation to new environmentsOA

Spurs have played an important role in the radiation of the genus Aquilegia,but little is known about how the spurless state arose in A.ecalcarata.Here we aim to characterize the genetic divergence within A.ecalcarata and gain insights into the origin of this species.A total of 19 populations from A.ecalcarata and 23 populations from three of its closest relatives(Aquilegia kansuensis,Aquilegia rockii and Aquilegia yabeana) were sampled in this study.We sequenced fifteen nuclear gene fragments a...     

Effect of extremely low frequency magnetic fields on the seedlings of wild plants growing in Central Yakutia

It was shown that permanent (B = 50 μT, horizontal plane, direction to the north) and alternating magnetic fields (North–South direction) exerted influences on seed germination as well as on cytological and biochemical features of seedlings characteristic of investigated species (Lepidium apetalum, Artemisia vulgaris, A. jacutica, and A. dracunculus) of wild plants growing in Central Yakutia. Under the effect of permanent magnetic field (MF), germinating capacity of seeds decreased (except for A. vulgaris), whereas alternating MF of different frequencies improved their germinating capacity, except for L. apetalum and A. jacutica at frequencies of 200 and 300 Hz, respectively. Under permanent MF, the rate of lipid peroxidation in the tissues of the seedlings decreased, whereas the content of low molecular weight antioxidants rose; when the plants were exposed to an alternating magnetic field, the content of MDA and peroxidase activity increased, and the content of low molecular weight antioxidants followed an ambiguous pattern.     

Comparison of the effects of Artemisia vulgaris and Artemisia absinthium growing in western Anatolia against trichinellosis (Trichinella spiralis) in rats

Trichinellosis often causing diarrhea and more rarely fever, periorbital edema and myositis in human, is commonly treated with benzimidazole derivatives. The Artemisia genus has been found to be effective against a variety of parasites. In the present study, the efficacy against trichinellosis (Trichinella spiralis) of Artemisia vulgaris and Artemisia absinthium was examined for the first time in rats. The results of trichinoscopy and artificial digestion, during the enteral (adult) phase of the illness show that 300 mg/kg doses of methanol extracts of the aerial parts of A. vulgaris and A. absinthium reduced the larval rate by 75.6% and 63.5% in tongue, 53.4% and 37.7% in diaphragm, 67.8% and 46.2% in quadriceps, and 66.7% and 60.5% in biceps-triceps muscles of rats, respectively. Furthermore, during the parenteral (encapsulated larvae) phase, 600 mg/kg doses of A. vulgaris and A. absinthium extracts decreased the larval rate by 66.4% and 59.9% in tongue, 57.4% and 50.0% in diaphragm, 47.6% and 43.7% in quadriceps, 60.2% and 46.4% in biceps-triceps muscles of rats, respectively. Analysis of antibody also showed that A. vulgaris significantly reduced the antibody response (P < 0.05) during the enteral and parenteral phases. Thus, the results of the present study revealed that A. vulgaris could be an alternative drug against trichinellosis.     

Can Newts Cope with the Heat? Disparate Thermoregulatory Strategies of Two Sympatric Species in Water

Many ectotherms effectively reduce their exposure to low or high environmental temperatures using behavioral thermoregulation. In terrestrial ectotherms, thermoregulatory strategies range from accurate thermoregulation to thermoconformity according to the costs and limits of thermoregulation, while in aquatic taxa the quantification of behavioral thermoregulation have received limited attention. We examined thermoregulation in two sympatric newt species, Ichthyosaura alpestris and Lissotriton vulgaris, exposed to elevated water temperatures under semi-natural conditions. According to a recent theory, we predicted that species for which elevated water temperatures pose a lower thermal quality habitat, would thermoregulate more effectively than species in thermally benign conditions. In the laboratory thermal gradient, L. vulgaris maintained higher body temperatures than I. alpestris. Semi-natural thermal conditions provided better thermal quality of habitat for L. vulgaris than for I. alpestris. Thermoregulatory indices indicated that I. alpestris actively thermoregulated its body temperature, whereas L. vulgaris remained passive to the thermal heterogeneity of aquatic environment. In the face of elevated water temperatures, sympatric newt species employed disparate thermoregulatory strategies according to the species-specific quality of the thermal habitat. Both strategies reduced newt exposure to suboptimal water temperatures with the same accuracy but with or without the costs of thermoregulation. The quantification of behavioral thermoregulation proves to be an important conceptual and methodological tool for thermal ecology studies not only in terrestrial but also in aquatic ectotherms.     

Decrease of sexual organ reciprocity between heterostylous primrose species,with possible functional and evolutionary implications

Background and Aims

Heterostyly is a floral polymorphism that has fascinated evolutionary biologists since Darwin''s seminal studies on primroses. The main morphological characteristic of heterostyly is the reciprocal placement of anthers and stigmas in two distinct (distyly) floral morphs. Variation in the degree of intermorph sexual reciprocity is relatively common and known to affect patterns of pollen transfer within species. However, the partitioning of sexual organ reciprocity within and between closely related species remains unknown. This study aimed at testing whether intermorph sexual reciprocity differs within vs. between primrose species that hybridize in nature and whether the positions of sexual organs are correlated with other floral traits.

Methods

Six floral traits were measured in both floral morphs of 15 allopatric populations of Primula elatior, P. veris and P. vulgaris, and anther–stigma reciprocity was estimated within and between species. A combination of univariate and multivariate approaches was used to test whether positions of reproductive organs were less reciprocal between than within species, to assess correlations between sexual organ positions and other corolla traits, and to quantify differences between morphs and species.

Key Results

The three species were morphologically well differentiated in most floral traits, except that P. veris and P. vulgaris did not differ significantly in sexual organ positions. Overall, lower interspecific than intraspecific sexual organ reciprocity was detected. This decrease was marked between P. elatior and P. vulgaris, intermediate and variable between P. elatior and P. veris, but negligible between P. veris and P. vulgaris.

Conclusions

Differences in anther and stigma heights between the analysed primrose species were of the same magnitude or larger than intraspecific differences that altered pollen flow within other heterostylous systems. Therefore, it is possible to suggest that considerable reductions of sexual organ reciprocity between species may lower interspecific pollen flow, with potential effects on reproductive isolation.     

Population genetic diversity of the clonal self‐incompatible herbaceous plant Linaria vulgaris along an urbanization gradient

How increasing urbanization affects biodiversity is one of the most understudied aspects of global change biology. It is, however, known that it may negatively affect plant population genetic diversity in numerous ways, for example through its negative effects on plant population size, between‐population connectivity, and reproductive success. Therefore, it is important to investigate to what extent different levels of urbanization result in these negative phenomena. Here we used microsatellite markers to investigate urbanization effects on the population genetic structure of 23 populations of the self‐incompatible, partially clonal herb Linaria vulgaris which were sampled across a gradient of urbanization. Clonal diversity as measured by the Pareto‐parameter varied between 1.11 and 2.97 and was negatively correlated to both the degree of urbanization and to population size. Urbanization and population size were not interrelated. The least clonally rich populations also experienced significantly reduced seed set. Irrespective of the degree of urbanization, L. vulgaris populations exhibited strong genetic differentiation (F_ST = 0.33) and there was no significant correlation between genetic and geographic distances, suggesting low gene flow among populations. In conclusion, we showed that urbanization negatively affected fitness of L. vulgaris populations through decreasing their clonal diversity and reproductive success, an effect that may be exacerbated by the low gene flow between populations. Although the effect was modest, the results could probably be extrapolated to bigger cities where it would be considerably more pronounced.     

Habitat-Dependent Species Recognition in Hybridizing Newts

Many species breed in heterogeneous environments where conditions affecting signaling fidelity may vary. Species recognition may be impaired under particular environmental conditions enhancing the hybridization risk. We investigated the influence of habitat on species recognition efficiency in two hybridizing newts, Lissotriton vulgaris and L. helveticus. The former is considered to be an open habitat species where the two species are in sympatry, whereas the latter also breeds in forest ponds where dissolved humic acids attenuate wavelength transmission, especially in the UV range. Because UV sexual signalling occurs in L. vulgaris, we predicted that species recognition would be reduced in water stained by humic acids. We conducted two-choice preference tests in males and females in stained and clear water. Females of both species preferred the conspecific male in clear water but not in stained water. Males did not show species recognition in either treatment. In both newts, visual species recognition is likely to depend upon habitat. Resorting to chemical communication may offset the loss of visual information but the same environmental factors that affect the transmission of visual signal can also affect the transmission of chemical signals. This environmental disruption of species recognition may account for the general avoidance of forest ponds by L. vulgaris in sympatry with L. helveticus. Stochastic variations of visual conditions in ponds may also explain the ongoing hybridization between two long diverged species that exhibit many and well differentiated sexual ornaments, and more generally between taxa naturally experiencing strong variations of their sensory environment.     

Arbuscular mycorrhizas in phosphate-polluted soil: interrelations between root colonization and nitrogen

To investigate whether arbuscular mycorrhizal fungi (AMF) – abundant in a phosphate-polluted but nitrogen-poor field site – improve plant N nutrition, we carried out a two-factorial experiment, including N fertilization and fungicide treatment. Percentage of root length colonized (% RLC) by AMF and tissue element concentrations were determined for four resident plant species. Furthermore, soil nutrient levels and N effects on aboveground biomass of individual species were measured. Nitrogen fertilization lowered % RLC by AMF of Artemisia vulgaris L., Picris hieracioides L. and Poa compressa L., but not of Bromus japonicus Thunb. This – together with positive N addition effects on N status, N:P-ratio and aboveground biomass of most species – suggested that plants are mycorrhizal because of N deficiency. Fungicide treatment, which reduced % RLC in all species, resulted in lower N concentrations in A. vulgaris and P. hieracioides, a higher N concentration in P. compressa, and did not consistently affect N status of B. japonicus. Evidently, AMF had an influence on the N nutrition of plants in this P-rich soil; however – potentially due to differences in their mycorrhizal responsiveness – not all species seemed to benefit from a mycorrhiza-mediated N uptake and accordingly, N distribution.     

Reproductive interference between two serious pests,oriental fruit flies <Emphasis Type="Italic">Bactrocera carambolae</Emphasis> and <Emphasis Type="Italic">B. dorsalis</Emphasis> (Diptera: Tephritidae), with very wide but partially overlapping host ranges

Bactrocera carambolae and B. dorsalis (Diptera: Tephritidae) are extremely destructive pests of fruits and vegetables in the Asia-Pacific region. Earlier reports have described that B. carambolae and B. dorsalis, respectively, use mainly star fruit and mango, suggesting a certain level of host partitioning which can be ascribed neither to differences in larval food qualities nor host-specific parasitoid mortality. This study specifically examined reproductive interference (antagonistic sexual interaction) between B. carambolae and B. dorsalis as a potential factor strongly affecting their host partitioning. We observed mating behaviors, especially interspecific courtships and mating, by cohabiting the conspecific and heterospecific pairs together. Consequently, we quantified their effects on the reproductive success of females. Males of both species frequently courted their own females, but they also courted females of other species. Courtship refusal by females was not selective in males of either species. This incomplete discrimination of both sexes led to frequent occurrences of interspecific sexual interactions in both species, but only B. carambolae females showed reduced reproductive success. These results suggest that B. dorsalis, superior in reproductive interference, can occupy high-quality mango, whereas B. carambolae, inferior in reproductive interference, must use low-quality star fruit.     

Genomic diversity in two related plant species with and without sex chromosomes--Silene latifolia and S. vulgaris

Background

Genome size evolution is a complex process influenced by polyploidization, satellite DNA accumulation, and expansion of retroelements. How this process could be affected by different reproductive strategies is still poorly understood.

Methodology/Principal Findings

We analyzed differences in the number and distribution of major repetitive DNA elements in two closely related species, Silene latifolia and S. vulgaris. Both species are diploid and possess the same chromosome number (2n = 24), but differ in their genome size and mode of reproduction. The dioecious S. latifolia (1C = 2.70 pg DNA) possesses sex chromosomes and its genome is 2.5× larger than that of the gynodioecious S. vulgaris (1C = 1.13 pg DNA), which does not possess sex chromosomes. We discovered that the genome of S. latifolia is larger mainly due to the expansion of Ogre retrotransposons. Surprisingly, the centromeric STAR-C and TR1 tandem repeats were found to be more abundant in S. vulgaris, the species with the smaller genome. We further examined the distribution of major repetitive sequences in related species in the Caryophyllaceae family. The results of FISH (fluorescence in situ hybridization) on mitotic chromosomes with the Retand element indicate that large rearrangements occurred during the evolution of the Caryophyllaceae family.

Conclusions/Significance

Our data demonstrate that the evolution of genome size in the genus Silene is accompanied by the expansion of different repetitive elements with specific patterns in the dioecious species possessing the sex chromosomes.     

Enhanced immune function does not depress reproductive output

Costs of reproduction might be mediated by a physiological (resource allocation) trade-off between immune function and reproductive effort, and several recent studies have shown that an experimental increase in reproductive effort is associated with decreased immune function. Here we test the complementary prediction of this hypothesis: that increased immune function (specific antibody production) depresses reproductive output. Female European starlings (Sturnus vulgaris) were injected with a non-pathogenic antigen (sheep red blood cells) following completion of laying of their first clutch, to stimulate an in vivo humoral immune response (primary antibody production). We induced laying of a second clutch by removing the first clutch, and assessed changes in reproductive performance in individual females pre- and post-treatment. Injection of sheep red blood cells produced a significant antibody response in 96% (n=29) of treated females, with titres comparable to previous studies (range 1 to 7). However, increased antibody production did not decrease primary or secondary female reproductive effort (re-laying interval, egg size, clutch size, chick growth or fledging success), compared with control, saline-injected birds (n=22). These data do not support a simple resource allocation model for the cost of reproduction, based on a reciprocal, negative relationship between resources allocated to immune function and reproduction.     

Polyamine Biosynthetic Enzymes in the Cell Cycle of Chlorella: Correlation between Ornithine Decarboxylase and DNA Synthesis at Different Light Intensities

During the life cycle of Chlorella vulgaris Beijerinck var vulgaris fa. vulgaris growing synchronously, the specific activity of ornithine decarboxylase peaked at the 2nd hour of the cycle, whereas that of arginine decarboxylase changed only slightly, increasing towards the end of the cycle. The endogenous level of putrescine and spermidine on a per cell basis increased gradually up to the 8th hour of the cycle, and declined thereafter. Thus, the peak of ornithine decarboxylase activity and the polyamine increase preceded both DNA replication (which took place between the 6th and 8th hours of the cycle) and autospore release (which started at the 8th hour). A 2-fold increase in the light intensity caused doubling of the DNA content, resulting in doubling of the number of autospores per mother cell. It also brought about a 2-fold increase in the specific activity of ornithine decarboxylase and polyamine content, the peaks being at the same hour of the cycle under high and low light intensities. The increase in cell number and polyamine content in a Chlorella culture grown under high light intensity was inhibited by α-difluoromethyl ornithine, a specific inhibitor of ornithine decarboxylase, this inhibition being partially reversed by putrescine.

It is suggested that in C. vulgaris the sequence of events which relates polyamine biosynthesis to cell division is as follows: increased ornithine decarboxylase activity, accumulation of polyamines, DNA replication, and autospore release.

     

Does the surrounding matrix influence corridor effectiveness for pollen dispersal in farmland?

Recent decades have seen a shift in agricultural land use from pasture to arable combined with increased use of fertilisers and pesticides. In this quite hostile landscape matrix, pollinator movements between native vegetation remnants may be impeded. Linear landscape elements (LLEs) in farmland can function as biological corridors by facilitating pollinator movements and pollen flow between fragmented plant populations. The type of landscape matrix surrounding LLEs and plant populations, and LLE habitat quality may influence the effectiveness of LLEs as corridors for pollen dispersal through the availability of floral resources and nesting opportunities for pollinators. Using fluorescent dyes as pollen analogues, we investigated dye dispersal patterns between fragmented populations of the rare insect-pollinated Primula vulgaris connected by existing LLEs. We examined how dye deposition on P. vulgaris and within LLEs (on other co-flowering insect-pollinated species) could be influenced by the surrounding matrix type (pasture, arable field), the recipient population traits of P. vulgaris (flowering population size, flower display, flowering plant density and co-flowering floral resources) and by LLE traits (LLE length and co-flowering floral resources). Dye dispersal through corridors was significantly higher when the landscape matrix surrounding P. vulgaris recipient populations and LLEs consisted of pastures rather than arable fields (or a mix of both), even after accounting for differences in floral resources. A higher cover and diversity of insect-pollinated plants increased dye deposition when co-flowering within small P. vulgaris populations, but led to dye loss within LLEs. Large P. vulgaris populations appeared more attractive thereby increasing heterospecific dye deposition in the LLEs. Our study shows that farming practices shifting from pastures to maize arable fields have a negative impact on dye dispersal patterns, and so possibly affect pollen dispersal of P. vulgaris, likely through a reduced pollinator service. Corridor effectiveness depends on the type of surrounding matrix, and the abundance of floral resources directly influences dye dispersal patterns. Preserving the remaining pastureland is essential, but restoring rich flowering vegetation at field boundaries and along LLEs may also promote corridor effectiveness for pollen dispersal.