Flavonoid chemistry of Coreopsis grandiflora (Compositae)

The leaf flavonoid chemistryof Coreopsis grandiflora, which includes var.harveyana, var.longipes, var.saxicola and the typical var.grandiflora, is quite uniform with 6-hy-droxyquercetin 7-O-glucoside, luteolin 7-O-glucoside, marein-maritimein chal-cone-aurone pair and lanceolin-leptosin chalcone-aurone pair as consistent com-ponents. Flavonoid data lend support to the hypothesis that the hexaploid var.longipes originated from parents which would be included withinC. grandiflora, i.e., there is no evidence that other species were involved in its formation. One population of var.grandiflora and several collections of var.saxicola contain additional flavonoid components in the form of flavonol 3-O-glycosides. In nearly all instances the additional compounds are attributable to hybridization withC. lanceolata orC. pubescens because these flavonols are characteristic of these two species and morphological considerations also suggest it. Flavonoid chemistry supports the treatment of var.saxicola as a variety ofC. grandiflora rather than as a distinct species.     

Mating behavior and the evolutionary significance of mate guarding in three species of crane flies (Diptera: Tipulidae)

Three species of crane flies-Dactylolabis montana, Limonia simulans,and Antocha saxicola-gather near streams to mate and oviposit. All species are polygamous and sex ratios at these sites are male-biased. After a short mating bout, males guard females by standing over them during oviposition. Sperm competition appears to be intense and to follow last-male advantage, based on the packing of sperm within the two elongate spermathecae. Males of A. saxicolasuccessfully defend against rivals over 85% of the time. In contrast, defending males of D. montanaand L. simulanslose the female over 65% of the time during interactions with rivals. Despite the high frequency of loss, defending males gain additional oviposition time by engaging rivals in combat while the female continues to oviposit. Thus, a guarding male does not have to retain the female for guarding to be adaptive. Legs and claws of all species are sexually dimorphic and play an important role in guarding and defending.     

Molecular genetic relationships and some issues of systematics of rock lizards of the genus Darevskia (Squamata: Lacertidae) based on locus analysis of SINE-type repeats (Squam1)

To study the molecular genetic relationships and correlate them with the taxonomy within the complex of lacertid lizards of the genus Darevskia, the locus analysis of the copies of the SINE-type repeat (Squam1) specific for the order Squamata was used. It was demonstrated that one of the loci (No. 34) contained the Squam1 copy insert in all species and subspecies of the examined genus. SINE allelic copies in some of the loci contained large indels and specific sets of mutations. The allelic variant M (medium, about 340 bp) was found most frequently; it was detected in all subspecies of D. saxicola (saxicola, darevskii, szczerbaki, lindholmi) and in most of the other species of the genus. Two species, D. derjugini and D. praticola, differed from the other species in the presence of long (L) and short (S) alleles. The longest allele was characteristic of the D. derjugini population from the Northern Caucasus (L, 379 bp, ssp. silvatica), while the shortest allele (97 bp) united the derjugini and barani subspecies. The second allele S (279 bp) characterizes the subspecies D. praticola praticola, some individuals of which also carry allele M. The second subspecies, D. p. pontica, contains allele L2, which differs from all other medium alleles in the presence of strictly specific short indel. In addition to apomorphic indels, the specificity and mutation distribution patterns among the Squam1 alleles were also examined. An analysis of the NJ tree indicated the concordance between morphological and molecular genetic characters of the species derjugini, praticola, and saxicola. Furthermore, four subspecies of D. saxicola were much closer to each other than the subspecies within the first two species; D. d. silvatica and the group of D. d. derjugini + barani were clearly separated. It cannot be excluded that populations from Azerbaijan and Serbia can be treated as the independent subspecies of D. praticola.     

Divergent Receiver Responses to Components of Multimodal Signals in Two Foot-Flagging Frog Species

Multimodal communication of acoustic and visual signals serves a vital role in the mating system of anuran amphibians. To understand signal evolution and function in multimodal signal design it is critical to test receiver responses to unimodal signal components versus multimodal composite signals. We investigated two anuran species displaying a conspicuous foot-flagging behavior in addition to or in combination with advertisement calls while announcing their signaling sites to conspecifics. To investigate the conspicuousness of the foot-flagging signals, we measured and compared spectral reflectance of foot webbings of Micrixalus saxicola and Staurois parvus using a spectrophotometer. We performed behavioral field experiments using a model frog including an extendable leg combined with acoustic playbacks to test receiver responses to acoustic, visual and combined audio-visual stimuli. Our results indicated that the foot webbings of S. parvus achieved a 13 times higher contrast against their visual background than feet of M. saxicola. The main response to all experimental stimuli in S. parvus was foot flagging, whereas M. saxicola responded primarily with calls but never foot flagged. Together these across-species differences suggest that in S. parvus foot-flagging behavior is applied as a salient and frequently used communicative signal during agonistic behavior, whereas we propose it constitutes an evolutionary nascent state in ritualization of the current fighting behavior in M. saxicola.     

FENESTROCONIDIA CALOPLACAE,A NEW SPORODOCHIAL LICHENICOLOUS FUNGUS WITH CATENATE CONIDIOGENOUS CELLS

The genus Fenestroconidia is described to accommodate the new species Fenestroconidia caloplacae. This lichenicolous fungus produces black sporodochia, has conidiogenous cells arranged in chains, and simple, pale brown conidia with a subhyaline guttule-like zone. It is known only from central Spain (Cuenca province), growing in the apothecia of Caloplaca saxicola s.lat.     

Pollination and reproductive system of synchronopatric species of Cactaceae (Cactoideae) subject to interspecific flow of pollen: an example of ecological adaptation in the Brazilian Chaco

• Three synchronopatric Cactaceae species, Echinopsis rhodotricha, Harrisia balansae and Praecereus saxicola, have mostly nocturnal anthesis and similar flowers, characteristics that motivated us to perform a comparative study of reproductive ecology.
• Reproductive phenology was sampled monthly from December 2014 to November 2015. We describe floral biology, breeding system via pollination treatments and evaluate floral visitors from focal and filming observations. Pollen grains found on moth proboscis were compared among cactus species under light microscopy. We used fluorescent dye particles to test intra‐ and interspecific pollen flow.
• These three species have extended flowering with greater intensity in the wet season, causing high overlap. They have white and hypocrateriformis flowers that open at twilight or nightfall and last about 15 h. H. balansae seems to be self‐incompatible, while E. rhodotricha presented self‐compatibility. P. saxicola presented self‐fertility, but most of the population seems to be self‐incompatible. We suggest sphingophily for the three species, but only P. saxicola was visited by Manduca rustica (Sphingidae). However, we observed pollen grains of all three species on the proboscis of moths, especially M. rustica and M. sexta. Prolonged anthesis allowed bees (herein considered as secondary pollinators) to visit flowers of E. rhodotricha and P. saxicola.
• It can be concluded that the studied species share nocturnal and diurnal pollinators, suggesting interspecific pollen flow, which, however, could not be detected with fluorescent dye particles. In view of the low frequency of primary pollinators, it appears that these three species have different reproductive strategies, ensuring the fruiting and production of viable seeds.

     

Aster saxicola sp. nov. (Asteraceae,Astereae), a new species from Guizhou,China: evidence from morphological,molecular and cytological studies

Aster saxicola W. P. Li & Z. Li, a new species of Asteraceae from southeastern Guizhou province, China, is described and illustrated based on morphological, molecular and cytological data. Morphological comparisons showed that A. saxicola is similar to A. oliganthus, but can be distinguished from the latter by its purple abaxial surface of the lower and middle cauline leaves, cordate‐ovate basal leaves with strigose hairs, outer three‐seriate phyllaries with purple or purplish apex, 9–14 ray florets and 10–18 disk florets. Phylogenetic analyses based on nrDNA ITS, ETS and plastid trnL–F DNA sequence data support that A. saxicola is a distinct species and belongs to Aster subgen. Aster sect. Ageratoides. Cytological observation shows that the new species is diploid with a karyotype formula 2n = 2x = 18 = 16m + 2sm. The new species should be considered endangered (EN) based on the IUCN red list criteria because of its restricted geographic range.     

New distributional data of the rare endemic species Eokochia saxicola (Guss.) Freitag and G. Kadereit (Chenopodiaceae): Effects on biogeography and conservation

Eokochia saxicola (Guss.) Freitag & G. Kadereit is a very rare endemic species of southern Italy and one of the rarest plants in Europe, included as priority species in the Annex II of the Council Directive 92/43/EEC. During the floristic surveys in the Cilento e Vallo di Diano National Park (Campania region, southern Italy), with the aim to assess the conservation status of rare and/or endemic species, new sites of occurrence of E. saxicola have been recorded representing the first known mainland sites; in fact, this endemic species was previously recorded only in the insular sites of Ischia (where it is extinct) and Capri in Campania region and Strombolicchio in Sicily region. Moreover, a further site was found in Capri island. So, three new sites of occurrence of E. saxicola have been recorded, increasing the total number from the two previously known to the actual five; in Campania, the total number increased from one to four. The research also increased significantly the global population size, currently made up of about 270 individuals compared with the 150 previously known. Consequences for the biogeography of the species and practical issues for conservation are discussed.     

Taxonomic and functional diversity in <Emphasis Type="Italic">Calogaya</Emphasis> (lichenised Ascomycota) in dry continental Asia

The genus Calogaya (Teloschistaceae, Xanthorioideae) was established to accommodate mainly epilithic lichens with lobate thalli, previously regarded as the “Caloplaca saxicola group.” Data supporting the recognition of this new genus came from European lichens, and although the genus is soundly based, we have found in Asia numerous epiphytic lineages and lineages with reduced, non-lobate thallus in dry continental areas. The taxonomic and functional diversity of Calogaya is distinctly higher in steppe and desert areas of Asia than in the less arid regions of Europe. We sampled 238 specimens, mostly from arid regions of north-western China, Iran, southern Siberia and Turkey. Three nuclear DNA loci were analysed separately and jointly by Bayesian inference, maximum likelihood and *BEAST approaches. Delimitations of 28 putative species were tested by BP&P multispecies coalescent model with joint analysis of species delimitation and species-tree estimation. Finally, we recognised 22 taxonomic units: 16 are at species rank, 3 are treated as subspecies and 3 are complexes, treated here as a single entity, but in reality probably including more than one species. Calogaya altynis, C. biatorina subsp. asiatica, C. decipiens subsp. esorediata, C. haloxylonis, C. orientalis, C. xanthoriella and C. xinjiangis are newly described. Caloplaca zoroasteriorum is combined into Calogaya, and Calogaya persica is reduced to a subspecies. The taxonomic status of Calogaya saxicola is unclear, and the name is employed here “sensu lato” for several non-monophyletic epilithic lineages with short-lobed thalli. Calogaya biatorina and C. ferrugineoides are the two other heterogeneous taxonomic units probably including more species.     

<Emphasis Type="Italic">Galerina saxicola</Emphasis> (Fungi,Agaricales) is conspecific with <Emphasis Type="Italic">G. stordalii</Emphasis> and new data on ecology of the latter species

The holotype of Galerina saxicola Svr?ek, a never revised species described from the Czech Republic in 1994, was studied in detail. Due to its poor state and the impossibility to obtain DNA data, we fixed its taxonomic position by designating an epitype using recent material from the type locality. The species proved to be conspecific with Norwegian and Czech collections of Galerina stordalii A.H.Sm., both morphologically and molecularly. Full synonymy and diagnostic characters of G. stordalii are provided, its morphological and ecological variability is discussed, and information on type specimens is corrected. Our collections document that G. stordalii has a broader ecological amplitude than thought before, living not only on Sphagnum and peat in boreal and arctic-alpine habitats like bogs and snow beds, but also among mosses in boggy spruce forests, on decaying conifer trunks in old-growth forests and on moist sandstone rocks in a river canyon under the influence of climatic inversion.     

Sex Allocation by a Parasitoid Wasp (Hymenoptera: Ichneumonidae) to Different Host Species: A Question for the Mechanism of Host Size Estimation

Sex allocation by the polyphagous solitary pupal parasitoid wasp Pimpla luctuosa Smith to a small host species, Galleria mellonella (L.), and a large host species, Mamestra brassicae L., was investigated to test whether female wasps responded to hosts of different sizes across different host species. In the experiments, both host species were presented to each test female wasp. Primary and secondary sex ratio experiments revealed that female wasps laid more female eggs in larger pupae of each host species, indicating that female wasps recognized size differences within host species. The wasp sex ratio (male ratio) from M. brassicae, however, was much higher than that expected on the basis of the sex ratio curve from different-sized G. mellonella. Larger hosts of each host species yielded larger wasps, indicating that the host size estimation by female wasps across different host species was incomplete or was not simple. These results suggested that P. luctuosa evaluated host size not only by physical measures such as dimension but also by other unknown measures. A possible explanation for the adaptiveness of different sex ratio responses by Pimpla luctuosa to different host species was discussed.     

The lichen genus caloplaca in polar regions

Extensive material of Caloplaca from Arctic and Antarctic regions has been critically examined. A list of 49 species is presented for Arctic regions. They are presumed to have a more or less circumpolar distribution. Twenty-two species are listed from the Antarctic region, but about ten more, probably undescribed species, are present there. About one-third of the species in the Antarctic region are bipolar or widespread in cold regions; these include mainly terricolous and muscicolous species and none of them are maritime. It is assumed that migration of the bipolar or cosmopolitan species has taken place along the Andean mountain chain, whereas the maritime polar species have evolved separately in the two hemispheres. The Caloplaca species of the Antarctic region are provisionally assigned to the following distribution types: continental Antarctic, western Antarctic, insul-Antarctic and sub-Antarctic. Caloplaca exsecuta, C. saxicola and C. phaeocarpella are recorded as new to the Antarctic region. Caloplaca johnstonii (Dodge) Søchting & Olech, comb, nov., is established as the correct name of C. tenuis Øvstedal.     

Chemosystematische Untersuchungen zur Phylogenese der Sammelgattung Lacerta (Reptilia: Sauria: Lacertidae)

Chemosystematic investigations concerning the phylogeny of the collective genus Lacerta (Reptilia: Sauria: Lacertidae) Investigated the albumins from 31 species and subspecies of the genus Lacerta s.1. by means of the immunoloical techniue Micro-Complement-fixation. Dendrograms were computed by means of the ktch-MargJiash algorithm. Moreover the taxa of the Lacerta saxicola and the Lacerta danfordi comlexes were analyzed by electrophoresis concerning 14 genetic loci. The results were discussel together with karyological and genitalmorphological data so far known yielding the following relationships: Lacerta graeca shows a closer relationshi to the subgenus Podarcis as does the quite closely related species air L. dugesii and L. perspicikata. Lacerta Iaevis probabl is connected closely to Podarcis, too. Lacerta parva shows a closer relationship to the “small izards” than to the sub-genus Lacerta s. str. A clear decision concerning the position of the Lacerta danfordi complex according to chemosystematical data was not possible. It is isolated from the other species probably about as long as L. puma. Biochemical differences between its “species” are too small to confirm their taxonomical revalorisation by Eiselt and Schmidtler Within the Lacerta saxicola complex the investigated taxa show a close relationship. Lacerta praticola and L. derjugini were definitly assigned to this group.     

Highly Differentiated ZW Sex Microchromosomes in the Australian Varanus Species Evolved through Rapid Amplification of Repetitive Sequences

Transitions between sex determination systems have occurred in many lineages of squamates and it follows that novel sex chromosomes will also have arisen multiple times. The formation of sex chromosomes may be reinforced by inhibition of recombination and the accumulation of repetitive DNA sequences. The karyotypes of monitor lizards are known to be highly conserved yet the sex chromosomes in this family have not been fully investigated. Here, we compare male and female karyotypes of three Australian monitor lizards, Varanus acanthurus, V. gouldii and V. rosenbergi, from two different clades. V. acanthurus belongs to the acanthurus clade and the other two belong to the gouldii clade. We applied C-banding and comparative genomic hybridization to reveal that these species have ZZ/ZW sex micro-chromosomes in which the W chromosome is highly differentiated from the Z chromosome. In combination with previous reports, all six Varanus species in which sex chromosomes have been identified have ZZ/ZW sex chromosomes, spanning several clades on the varanid phylogeny, making it likely that the ZZ/ZW sex chromosome is ancestral for this family. However, repetitive sequences of these ZW chromosome pairs differed among species. In particular, an (AAT)n microsatellite repeat motif mapped by fluorescence in situ hybridization on part of W chromosome in V. acanthurus only, whereas a (CGG)n motif mapped onto the W chromosomes of V. gouldii and V. rosenbergi. Furthermore, the W chromosome probe for V. acanthurus produced hybridization signals only on the centromeric regions of W chromosomes of the other two species. These results suggest that the W chromosome sequences were not conserved between gouldii and acanthurus clades and that these repetitive sequences have been amplified rapidly and independently on the W chromosome of the two clades after their divergence.     

Development of PCR‐Based Markers to Determine the Sex of Kelps

Sex discriminating genetic markers are commonly used to facilitate breeding programs in economically and ecologically important animal and plant species. However, despite their considerable economic and ecological value, the development of sex markers for kelp species has been very limited. In this study, we used the recently described sequence of the sex determining region (SDR) of the brown algal model Ectocarpus to develop novel DNA-based sex-markers for three commercially relevant kelps: Laminaria digitata, Undaria pinnatifida and Macrocystis pyrifera. Markers were designed within nine protein coding genes of Ectocarpus male and female (U/V) sex chromosomes and tested on gametophytes of the three kelp species. Seven primer pairs corresponding to three loci in the Ectocarpus SDR amplified sex-specific bands in the three kelp species, yielding at least one male and one female marker for each species. Our work has generated the first male sex-specific markers for L. digitata and U. pinnatifida, as well as the first sex markers developed for the genus Macrocystis. The markers and methodology presented here will not only facilitate seaweed breeding programs but also represent useful tools for population and demography studies and provide a means to investigate the evolution of sex determination across this largely understudied eukaryotic group.     

Application of microwave‐assisted extraction coupled with high‐speed counter‐current chromatography for separation and purification of Dehydrocavidine from Corydalis saxicola Bunting

Introduction – Dehydrocavidine is a major component of Corydalis saxicola Bunting with sedative, analgesic, anticonvulsive and antibacterial activities. Conventional methods have disadvantages in extracting, separating and purifying dehydrocavidine from C. saxicola. Hence, an efficient method should be established. Objective – To develop a suitable preparative method in order to isolate dehydrocavidine from a complex C. saxicola extract by preparative HSCCC. Methodology – The methanol extract of C. saxicola was prepared by optimised microwave‐assisted extraction (MAE). The analytical HSCCC was used for the exploration of suitable solvent systems and the preparative HSCCC was used for larger scale separation and purification. Dehydrocavidine was analysed by high‐performance liquid chromatography (HPLC) and further identified by ESI‐MS and 1H NMR. Results – The optimised MAE experimental conditions were as follows: extraction temperature, 60°C; ratio of liquid to solid, 20; extraction time, 15 min; and microwave power, 700 W. In less than 4 h, 42.1 mg of dehydrocavidine (98.9% purity) was obtained from 900 mg crude extract in a one‐step separation, using a two‐phase solvent system composed of chloroform–methanol–0.3 m hydrochloric acid (4 : 0.5 : 2, v/v/v). Conclusion – Microwave‐assisted extraction coupled with high‐speed counter‐current chromatography is a powerful tool for extraction, separation and purification of dehydrocavidine from C. saxicola. Copyright © 2009 John Wiley & Sons, Ltd.     

Efficacy of a biocide tested on selected lichens and its effects on their substrata

The efficacy of Koretrel^©, a biocide used in south-eastern Asia to remove epilithic photoautotrophic organisms, was tested in the field and laboratory using lichens as target organisms. In situ observations were carried out at increasing time intervals for 6 month on 15 vegetation plots and 21 saxicolous species representative of the lichen flora of Italian monuments, selected according to their different ecology, growth-form and distribution. Change in chlorophyll fluorescence was monitored in three species for 32 day after biocide treatment. The effects of the biocide on the substratum was studied by monitoring changes in capillary water absorption and surface colorimetry of two CaCO₃-containing rocks (Aurisina limestone and Muggia sandstone), and a grey granite. Rock crystal integrity was monitored by SEM. The results show that Koretrel^© is very efficient in killing the lichens. Pronounced effects were observed in all species after 1–2 weeks, with chlorophyll fluorescence reduced to zero in 4 day in the case of Lecanora muralis and Neofuscelia loxodes and in 8 days for Caloplaca saxicola. The biocide considerably decreases capillary water absorption, and alters the surface colour of the substrata, particularly for Aurisina limestone. However, no corrosion features were observed in the rock minerals.     

Seasonal changes in situ grazing of the mesoherbivores Idotea baltica and Gammarus oceanicus on the brown algae Fucus vesiculosus and Pylaiella littoralis in the central Gulf of Finland, Baltic Sea

The in situ grazing experiments were performed in the shallow water rocky habitat of the northern Baltic Sea during ice-free season 2002. In the experiments the effects of algal species and choice on the grazing of the mesoherbivores Idotea baltica (Pallas) and Gammarus oceanicus Segerstråle were tested. Salinity, temperature, concentration of nutrients in water and macroalgae and net production of macroalgae were considered as random effects in the analysis. The invertebrate feeding rate was mainly a function of the net photosynthetic activity of Pylaiella littoralis (L.) Kjellman and Fucus vesiculosus L. Feeding rate increased significantly with decreasing algal photosynthetic activity. When the two algal species were incubated together invertebrates fed primarily on P. littoralis. Low selectivity towards P. littoralis coincided with its high photosynthetic activity. The presence of F. vesiculosus did not modify the invertebrate feeding on P. littoralis. The results indicated that (1) the grazing on F. vesiculosus depended on the availability of P. littoralis, (2) the photosynthetic activity of algae explained the best the variation in grazing rate and (3) the grazers are not likely to control the early outbreak of filamentous algae in the northern Baltic Sea by avoiding young and photosynthetically active algae. The likely mechanism behind the relationship is that the increased photosynthetic activity of macroalgae coincides with their higher resistance to herbivory.