Germination and emergence of Astrodaucus orientalis (L.) Drude populations influenced by environmental factors and seed burial depth

Astrodaucus orientalis is a weed species in cropping systems and rangelands in Iran. The effects of temperature, light, NaCl concentration, water potential, seed burial depth and crop residue cover were assessed on seed germination and seedling emergence of two populations of A. orientalis from Ardabil (Meshginshahr population) and East Azarbayjan (Tabriz population) provinces of Iran. The A. orientalis populations indicated different responses to environmental factors and burial depth. In the Tabriz population the greatest germination (88.5%) was observed in 20/12°C day/night temperature but in the Meshginshahr population (83.2%) it was obtained in 24/16°C day/night temperature. Over a broad range of light period (10–24 hr light) germination was 74–83%, but it decreased (less than 37%) under 24 hr dark in both A. orientalis populations. With respect to water potential, the C₅₀ parameters were −0.62 and − 0.49 MPa for Tabriz and Meshginshahr populations, respectively. The D₅₀ parameters (the burial depth that caused 50% decrease in emergence) for Tabriz and Meshginshahr populations were 2.42 and 3.13 cm, respectively. Generally, the results showed that emergence of both populations of A. orientalis was delayed as depth of burial increased up to 4 cm and in cropping systems a shallow tillage that locates the seeds to >4 cm of depth in soil could be used in order to suppress seedling emergence. Our findings also could be useful in integrated management of A. orientalis in winter annual crops and rangelands.     

Growth patterns of the alien perennial Bunias orientalis L. (Brassicaceae) underlying its rising dominance in some native plant assemblages

The polycarpic perennial Bunias orientalis L. (Brassicaceae) introduced to Central Europe in the 18th century recently entered a phase of rapid spread accompanied by sudden establishments of extensive dominance stands mainly on roadside locations. We studied vegetation structure and expansion rate of B. orientalis stands and performed a series of experiments to investigate key factors that underlie the colonizing and establishment of B. orientalis. Reiterated observations exposed a high current expansion rate of B. orientalis populations and vegetation surveys of B. orientalis stands showed that these stands were mainly composed of Artemisietea and Arrhenateretea species. Regeneration experiments with root fragments revealed high regeneration capacities: root fragments of 3 cm length showed 93% regeneration, varying water content (10–50% water loss) and separation into root cortex and root stele yielded regeneration of 30 to 50%. In a field study high regrowth after mowing with varying mowing intensity indicates B. orientalis to be well adapted to disturbed sites as its preferential locations for development of dominance stands. Vegetative growth parameters were studied in two controlled growth experiments with elevated nutrient availabilities. B. orientalis exhibited a high sensitivity to nutrient addition and rosette sizes of maximal 90 cm were reached. Biomass was comparable or even higher than that of native ruderals grown in the same experiment. Measurements of reproductive parameters revealed a high reproductive effort (0.2 to 0.45 g g^-1) even under intense mowing regimes, resulting in a dense seed bank with maximal values of about 400 fruits (550 seeds) l^-1 soil. With respect to colonization and establishment of B. orientalis the results of our study enable the formulation of three hypotheses.     

The co-occurrence of Artemia parthenogenetica and Branchinella spinosa (Branchiopoda: Anostraca) in a saline pond of south eastern Italy

The co-occurrence of Artemia parthenogenetica Bowen & Sterling, 1978 and Branchinella spinosa Milne-Edwards, 1840, was previously only supposed, on the basis of finding their cysts in the same lagoon, but active stages were never recorded together. In a saline lake in south eastern Italy, populations of the two species were observed for two years. A. parthenogenetica was typical of the early wet season (October–January), while B. spinosa, even if it was present before, developed in the late part of the wet season (February–May). Adult A. parthenogenetica were recorded only in the season 1999–2000. The seasonal distribution of A. parthenogenetica is probably controlled by abiotic conditions (i.e., photoperiod, temperature, conductivity, and rainfall); however, competition between specimens of similar body size (e.g., adults of A. parthenogenetica, and juveniles of B. spinosa) could explain the lower numbers of B. spinosa population in winter. The discovery of cysts of both species deep in the sediment of the lake suggests a long-lasting co-occurrence of these two Anostraca.     

On the presence of the genus Branchinecta Verrill, 1869 (Crustacea,Anostraca) in Yugoslavia

The genus Branchinecta Verrill, 1869 is represented in Yugoslavia by two species: Branchinecta ferox (Milne-Edwards, 1840) and Branchinecta orientalis G.O. Sars, 1901. The first species was collected in the steppe-like province of Banat, Pannonian Lowland, and in the lowland around the Skadar Lake, Montenegro, the second only in the province of Banat.On the basis of rich material of both species, a detailed study of the most essential morphological characters has been carried out and comparisons have been made with data in the literature. In B. ferox, significant morphological deviations are not found, whereas, in B. orientalis, essential features are clearly ascertained which were previously unknown or were not stated with sufficient precision. B. ferox is known to appear in small, temporary, natronsoda water rainpools, while B. orientalis has been found in a large temporary inundation pool with a high content of natronsoda and salt in the water.     

Phenological shifts of the alien colonizer Bunias orientalis: Image-based analysis of temporal niche separation

Abstract. Phenological development of the colonizer Bunias orientalis L. was investigated in permanent plots in herbaceous vegetation, using photography and image analysis. In all habitats but one, B. orientalis showed a two-phased rosette growth behaviour, with a first peak cover reached in spring, roughly coinciding with an early peak cover of the matrix vegetation, and a second autumn growth flush occurring in a phase of reduced matrix vegetation cover. This seasonally bimodal growth pattern of B. orientalis appears to partly compensate for its competitive inferiority in crowded herbaceous vegetation. Small or overshadowed individuals of this species particularly profit from higher above-ground performance or release from overlap in autumn. The significance of temporal niche separation for survival and growth of B. orientalis individuals varies with habitat conditions, being most apparent in occasionally disturbed habitats with a relatively low frequency of gap formation. Despite some limitations, image analysis proved to be useful for phenological investigations within herbaceous plant stands.     

Dormant egg bank characteristics and hatching pattern of the Phallocryptus spinosa (Anostraca) population in the Makgadikgadi Pans (Botswana)

The anostracan Phallocryptus spinosa has an almost exclusively palearctic distribution. The Makgadikgadi Pans area in Botswana represents the only distribution record south of the Sahara. In this ephemeral wetland, it is an important food item in the diet of migrating birds. By studying egg bank characteristics (such as depth and density) and hatching requirements, we investigated the persistence of this sub-Saharan population. At localities in the middle and north basin of Sua Pan, sediment cores were taken along a transect, and dormant eggs were isolated. Densities of the active dormant egg bank ranged from 833 to 31449 dormant eggs m⁻², indicating that this species is well established. Most of the dormant eggs were found in the top 4 cm of the sediments, and densities decreased to zero at a depth of 13 cm. Considering the expected low sedimentation rate, the presence of dormant eggs down to 13 cm indicates long-time occurrence of P. spinosa in the Makgadikgadi Pans area. We observed high hatching fractions (up to 85%) at a temperature of 22 °C and a salinity of 5 ppt. A second anostracan species, Branchinella ornata, co-occurred with P. spinosa in our study site. This population also had a large active dormant egg bank (ranging from 6634 to 50557 dormant eggs m⁻²) with dormant eggs present until a depth of about 11 cm. This pattern indicates a long-time co-occurrence of P. spinosa and B. ornata.     

Wind pollination in high-mountain populations of Hormathophylla spinosa (Cruciferae)

In this paper we study aspects of the breeding system of Hormathophylla spinosa (Crucifcrae) to identify the factors responsible for seed production in the absence of insect pollinators. The pollinator-exclusion experiments show that H. spinosa, under natural conditions, does not produce seed by apomixis or spontaneous autogamy. H. spinosa appears to be self-incompatible but slightly geitonogamous. Thus, this plant species needs pollen vectors for reproduction. The results of the wind-exclusion experiments performed during two different years in two populations of H. spinosa support the hypothesis that the wind acts as a pollen vector; flowers excluded from the wind had a lower fruit set and female fertility than flowers excluded from all pollinator insects (winged and wingless). This generalist pollination system (insects and wind) permits this species to colonize and sustain viable populations in high mountains, where this species is the only woody shrub living above 3000 m above sea level in the Sierra Nevada.     

Promotion of spinosad biosynthesis by chromosomal integration of the Vitreoscilla hemoglobin gene in Saccharopolyspora spinosa

To promote spinosad biosynthesis by improving the limited oxygen supply during high-density fermentation of Saccharopolyspora spinosa, the open reading frame of the Vitreoscilla hemoglobin gene was placed under the control of the promoter for the erythromycin resistance gene by splicing using overlapping extension PCR. This was cloned into the integrating vector pSET152, yielding the Vitreoscilla hemoglobin gene expression plasmid pSET152EVHB. This was then introduced into S. spinosa SP06081 by conjugal transfer, and integrated into the chromosome by site-specific recombination at the integration site ΦC31 on pSET152EVHB. The resultant conjugant, S. spinosa S078-1101, was genetically stable. The integration was further confirmed by PCR and Southern blotting analysis. A carbon monoxide differential spectrum assay showed that active Vitreoscilla hemoglobin was successfully expressed in S. spinosa S078-1101. Fermentation results revealed that expression of the Vitreoscilla hemoglobin gene significantly promoted spinosad biosynthesis under normal oxygen and moderately oxygen-limiting conditions (P<0.01). These findings demonstrate that integrating expression of the Vitreoscilla hemoglobin gene improves oxygen uptake and is an effective means for the genetic improvement of S. spinosa fermentation.     

A novel 53 kDa protein (BoP53) in Babesia orientalis poses the immunoreactivity using the infection serum

Babesia orientalis (B. orientalis) is responsible for water buffalo babesiosis, which caused serious economic losses in the south of China. Although the invasion process has been roughly described, there are still some unknown molecules that have not yet been identified. Recently, an invasion-related protein BOV57 has been identified in the Babesia bovis. However, there is no report available about the gene in B. orientalis. B. orientalis P53 (BoP53) sequence was obtained by blast BOV57 sequence in B. orientalis genome database, and the full length of the BoP53 gene is 1599 bp. BoP53 gene was cloned into a pGEX-6P-1 expression vector and expressed as a GST-tag fusion protein. The tertiary structure of BoP53 was predicted with the I-TASSER software. The native BoP53 was identified from of B. orientalis lysate incubation with mouse antiserum against rBoP53. BoP53 as a novel identified protein promotes the study of B. orientalis, the reaction of rBoP53 with the serum of B. orientalis-infected water buffalo but not with healthy buffalo serum indicated its good antigenicity. It may be a candidate antigen for the diagnosis of B. orientalis infection.     

Effects of ocean warming and acidification on the energy budget of an excavating sponge

Recent research efforts have demonstrated increased bioerosion rates under experimentally elevated partial pressures of seawater carbon dioxide (pCO₂) with or without increased temperatures, which may lead to net erosion on coral reefs in the future. However, this conclusion clearly depends on the ability of the investigated bioeroding organisms to survive and grow in the warmer and more acidic future environments, which remains unexplored. The excavating sponge Cliona orientalis Thiele, 1900 is a widely distributed bioeroding organism and symbiotic with dinoflagellates of the genus Symbiodinium. Using C. orientalis, an energy budget model was developed to calculate amounts of carbon directed into metabolic maintenance and growth. This model was tested under a range of CO₂ emission scenarios (temperature + pCO₂) appropriate to an Austral early summer. Under a pre‐industrial scenario, present day (control) scenario, or B1 future scenario (associated with reducing the rate of CO₂ emissions over the next few decades), C. orientalis maintained a positive energy budget, where metabolic demand was likely satisfied by autotrophic carbon provided by Symbiodinium and heterotrophic carbon via filter‐feeding, suggesting sustainability. Under B1, C. orientalis likely benefited by a greater supply of photosynthetic products from its symbionts, which increased by up to 56% per unit area, and displayed an improved condition with up to 52% increased surplus carbon available for growth. Under an A1FI future scenario (associated with ‘business‐as‐usual’ CO₂ emissions) bleached C. orientalis experienced the highest metabolic demand, but carbon acquired was insufficient to maintain the sponge, as indicated by a negative energy budget. These metabolic considerations suggest that previous observations of increased bioerosion under A1FI by C. orientalis may not last through the height of future A1FI summers, and survival of individual sponges may be dependent on the energy reserves (biomass) they have accumulated through the rest of the year.     

Enrichment of a continuous culture of <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> with the yeast <Emphasis Type="Italic">Issatchenkia orientalis</Emphasis> in the production of ethanol at increasing temperatures

A fermentation system was continuously fed with sugar-cane syrup and operated with recycling of Saccharomyces cerevisiae cells at temperatures varying from 30 to 47°C. The aim of the present work was to obtain and study the colonies of isolates showing elongated cells of yeasts which were sporadically observed at the end of this continuous process. Based on a sequence of assays involving methods of classical taxonomy and RAPD-PCR, two groups of isolates showing characteristics of non-Saccharomyces yeasts were identified in the yeast population where S. cerevisiae was the dominant yeast. The largest group of non-Saccharomyces yeasts, resulting from a slow proliferation over the 2 months, reached a final level of 29.6% at the end of the process. RAPD-PCR profiles obtained for the isolates of this dominant non-Saccharomyces yeast indicated that they were isolates of Issatchenkia orientalis. Pichia membranifaciens was the only species of non-Saccharomyces yeast detected together with I. orientalis but at a very low frequency. The optimum temperature for ethanol formation shown by the isolate 195B of I. orientalis was 42°C. This strain also showed a faster ethanol formation and biomass accumulation than the thermotolerant strain of S. cerevisiae used as the starter of this fermentation process. Some isolates of I. orientalis were also able to grow better at 40°C than at 30°C on plates containing glycerol as carbon source. Yeasts able to grow and produce ethanol at high temperatures can extend the fermentation process beyond the temperature limits tolerated by S. cerevisiae.     

The effect of the entomopathogenic fungus Conidiobolus coronatus on the composition of cuticular and internal lipids of Blatta orientalis females

The composition of cuticular and internal lipids in females of the cockroach Blatta orientalis L. exposed to the entomopathogenic fungus Conidiobolus coronatus is investigated. The compositions of the fatty acids, n‐alkanes, alcohol, sterols and methyl esters in the lipids are chemically characterized. Although contact with virulent colonies of the fungus does not induce insect mortality, significant changes in the lipid profiles, both cuticular and internal, are found. The cuticular extracts of a control group of B. orientalis females contain 24 compounds varying in carbon chain length from C6 to C22. The main cuticular fatty acids identified are: C16:1, C16:0, C18:1 and C18:0_. The cuticular lipids of B. orientalis females after exposure to C. coronatus contain only 14 free fatty acids from C8 to C20. The highest concentrations identified are C16:0, C18:2 and C18:1. Analysis by gas chromatography‐mass spectrometry identifies the presence of a homologous series of n‐alkanes containing from 25 to 31 carbon atoms. In the case of the insects after fungal exposure, the content of the n‐alkanes in the cuticular lipid is two‐fold higher compared with the controls. Of the cuticular lipids, 11 alcohols are found, ranging from C12:0 to C20:0. There is no presence of alcohols in the internal lipids of the control B. orientalis females and in all of the extracts from the B. orientalis females after fungal exposure. In the samples analyzed, the most common sterol is cholesterol. This is present in the cuticular lipids and the internal lipids of all of the insects sampled. The cuticular and internal lipids of females contain five fatty acid methyl esters, ranging in size from C15 to C19.     

Reproductive biology of twoPrunus species (Rosaceae) in the Northwest Iberian Peninsula

We studied the reproductive biology ofPrunus spinosa andPrunus mahaleb (Prunoideae, Rosaceae) in the northwest Iberian Peninsula. The two species flowered at the same time (peaking on March 9 and 11, respectively in 1990) but differ significantly in their fruit maturation times. Nectar volume peaked in the early morning in both species, and was ten times greater inP. spinosa than inP. mahaleb. Neither species shows apomixis, nor does fruit-set occur if pollinators are excluded. In both species self-pollination resulted in fewer fruits than open pollination. The principal pollinators belong to theApidae family (79% and 63% of visits toP. spinosa andP. mahaleb, respectively). Results are compared with those for other rosaceous plants with fleshy fruits.     

Testing the “Local Provenance” Paradigm: A Common Garden Experiment in Cumberland Plain Woodland,Sydney, Australia

Seed for restoration projects has traditionally been sourced locally to “preserve” the genetic integrity of the replanted site. Plants grown from locally sourced seeds are perceived to have the advantage of being adapted to local conditions, and the use of local provenance is a requirement of many restoration projects. However, the processes of climate change and habitat fragmentation, with the subsequent development of novel environments, are forcing us to reconsider this basic tenet of restoration ecology. We tested the “local provenance is best” paradigm, by comparing the performance of plants grown from local with non‐local seed sources within a common garden experiment. We selected six species representing a range of growth forms (Acacia falcata, Bursaria spinosa ssp. spinosa, Eucalyptus crebra, E. tereticornis, Hardenbergia violacea and Themeda australis) from an assemblage known as the Cumberland Plain Woodland, a threatened community in western Sydney. Multiple provenances were collected from within the range of each species and grown at two field sites on the Cumberland Plain. Growing time varied between species and ranged from 7 months to 2 years. With the exception of B. spinosa, and to a lesser extent T. australis, we found little evidence that local provenance plants were superior to distant provenances in terms of survival and establishment.     

Population studies of arthropods on Melia azedarach in Seville (Spain), with special reference to Eutetranychus orientalis (Acari: Tetranychidae) and its natural enemies

Eutetranychus orientalis has become an important pest of the ornamental tree Melia azedarach in the city of Seville (Spain). Trees suffer total defoliation at the end of summer. Studies were conducted in a regular plantation of this tree in the Miraflores Park in 2008 and 2009, to determine the arthropod faunal composition, with particular interest in the possible natural enemies of E. orientalis. Eutetranychus orientalis accounted for 98.3% of the arthropods found on the leaflets. Two species of phytoseiids were found, Euseius scutalis and Euseius stipulatus, but they only represented 0.2% of the arthropods. The most abundant insect was the predator thrips Scolothrips longicornis, which accounted for 0.9% of the arthropods found. The population of E. orientalis reached two peaks in 2008, with 325 individuals per leaflet in August, and 100 individuals per leaflet in November. Scolothrips longicornis densities closely followed E. orientalis, and predation was observed on various mite instars. Phytoseiids did not show such a response to the E. orientalis densities. Eutetranychus orientalis was more abundant in the exterior part of the plantation. No differences of arthropod densities were found between the various orientations in the plantation (north vs. south, east vs. west), although E. orientalis densities were different between rows. Distribution of E. orientalis population was highly aggregative, that of S. longicornis population was less aggregative, whereas the phytoseiid population showed a random distribution.     

酸枣果提取物在体内外对抗生素的增效作用及机制研究

作者研究团队先前从酸枣果的氯仿提取物中精制得到其低极性范围的活性组合物Fr.2a,发现Fr.2a与多种抗生素联用显示出广泛的协同抗菌作用。该研究在Fr.2a的基础上利用硅胶柱层析对酸枣果氯仿提取物中其他极性范围内的活性成分进行了分离纯化,得到精制物Fr.B,并对精制物Fr.B进行GC MS、核磁共振氢谱、红外光谱分析,以确定Fr.B的组成成分;通过抗菌谱分析和细胞通透性分析,以明确Fr.B的抗菌增效谱和抗菌增效机制;采用熔和法将精制物Fr.B制备成软膏,通过小鼠伤口感染模型评价该软膏对抗生素的增效效果。结果表明：(1)由酸枣果氯仿提取物进一步精制得到的Fr.B组分,主要包含反油酸、油酸、顺 10 十六碳烯醇、棕榈酸等脂肪酸类化合物。(2)Fr.B分别与庆大霉素、妥布霉素、氨苄青霉素、氯霉素、红霉素、夫西地酸、制霉菌素、酮康唑和两性霉素B等多种抗生素联用时显示出广泛的协同抗菌作用。(3)Fr.B可破坏细胞膜和细胞壁的完整性而增强细菌细胞的通透性。(4)在体内和体外Fr.B均能显著增强红霉素对耐甲氧西林金黄色葡萄球菌(MRSA)的杀菌作用,从而提高红霉素对MRSA菌株引起的伤口感染的治疗效果。研究表明,本研究所得到的Fr.B具有广谱的抗菌增效活性,能够增强红霉素对伤口耐药菌感染的治疗效果。该研究结果为克服微生物对抗生素的耐药性提供了新的思路和解决方案。     

Intrafloral patterns of color and scent in Capparis spinosa L. and the ghosts of its selection past

Premise

Capparis spinosa is a widespread charismatic plant, in which the nocturnal floral habit contrasts with the high visitation by diurnal bees and the pronounced scarcity of hawkmoths. To resolve this discrepancy and elucidate floral evolution of C. spinosa, we analyzed the intrafloral patterns of visual and olfactory cues in relation to the known sensory biases of the different visitor guilds (bees, butterflies, and hawkmoths).

Methods

We measured the intrafloral variation of scent, reflectance spectra, and colorimetric properties according to three guilds of known visitors of C. spinosa. Additionally, we sampled visitation rates using a motion-activated camera.

Results

Carpenter bees visited the flowers eight times more frequently than nocturnal hawkmoths, at dusk and in the following morning. Yet, the floral headspace of C. spinosa contained a typical sphingophilous scent with high emission rates of certain monoterpenes and amino-acid derived compounds. Visual cues included a special case of multisensory nectar guide and color patterns conspicuous to the visual systems of both hawkmoths and bees.

Conclusions

The intrafloral patterns of sensory stimuli suggest that hawkmoths have exerted strong historical selection on C. spinosa. Our study revealed two interesting paradoxes: (a) the flowers phenotypically biased towards the more inconsistent pollinator; and (b) floral display demands an abundance of resources that seems maladaptive in the habitats of C. spinosa. The transition to a binary pollination system accommodating large bees has not required phenotypic changes, owing to specific eco-physiological adaptations, unrelated to pollination, which make this plant an unusual case in pollination ecology.     

Karyomorphological studies onWoodwardia sensu lato of Japan

Karyomorphological comparisons were made of five species of JapaneseWoodwardia. There were no marked differences at interphase and prophase among the five species.Woodwardia japonica, W. prolifera, andW. unigemmata were diploid with 2n=68 and the formulas of their metaphase karyotypes uniformly 4m(median centromeric chromosomes)+12sm(submedian)+52(st+t)(subterminal and terminal).Woodwardia orientalis was tetraploid with 2n=136 and 8m+24sm+104(st+t), and the ratio of each chromosomal type to total complement was identical to that of three diploid species. These four species had several characteristics in common:x=34, the longest chromosome of sm, and a mean chromosome length over 3.0 μm. AlthoughWoodwardia orientalis showed some similarity toW. prolifera, it seems to be an allotetraploid which originated by chromosome doubling of a hybrid ofW. prolifera and a diploid species as yet karyomorphologically unknown.Woodwardia kempii was tetraploid with 2n=124 and 8m+24sm+92(st+t), and differed from the others in havingx=31, the longest chromosomes of t, and a mean chromosome length under 3.0 μm. This species has been classified as an independent genus,Chieniopteris, and our karyomorphological study supports this treatment.     

Analyzing the radiation of the proenkephalin gene in tetrapods: cloning of a Bombina orientalis proenkephalin cDNA

Analyzing the Radiation of the Proenkephalin Gene in Tetrapods: Cloning of a Bombina orientalis Proenkephalin cDNA: A proenkephalin cDNA was cloned from the brain of the anuran amphibian, Bombina orientalis (Family: Discoglossidae). This cDNA is 1358 nucleotides in length, and contains an open reading frame that codes for 251 amino acids. Within the open reading frame there are seven opioid (YGGF) sequences. There were five Met-enkephalin (YGGFM) sequences that are flanked by sets of paired basic amino acid proteolytic cleavage sites and two C-terminally extended Met-enkephalin sequences: YGGFMRGY and YGGFMRF. No Leu-enkephalin sequences were found in B. orientalis proenkephalin. It was possible to align the amino acid sequences of proenkephalin from several vertebrate taxa (human, Australian lungfish, B. orientalis, Xenopus laevis, Spea multiplicatus) by inserting a minimum of nine gaps. This alignment was then used to analyze the corresponding nucleotides for each proenkephalin sequence using maximum likelihood. This analysis yielded a single tree. In this tree, the Australian lungfish sequence was the outgroup or the tetrapod ingroup. The amphibian sequences form a clade separate from the human sequence. The bootstrap value for the amphibian clade was 100%. Within the amphibian clade the Bombina sequence was the sister group to a clade composed of the X. laevis and S. multiplicatus sequences. The bootstrap value for the X. laevis/S. multiplicatus clade was 94%. Collectively, these data indicate that the sequence of Bombina proenkephalin may be more similar to the proposed ancestral anuran proenkephalin sequence, than either X. laevis or S. multiplicatus proenkephalin.     

Cryptic variation and the tragedy of unrecognized taxa: the case of international trade in the spiny turtle Heosemys spinosa (Testudines: Geoemydidae)

Loss of habitat and human exploitation have driven many turtle species to the brink of extinction, particularly in many parts of southern Asia. The spiny turtle (Heosemys spinosa) is a terrestrial species distributed throughout the Sundaland region of South‐East Asia. Despite international legislative protection, H. spinosa continues to be illegally collected for the food and traditional medicine markets of China. Given its widespread distribution, taxonomists have reasonably questioned whether H. spinosa truly represents a single evolutionary lineage or multiple undiagnosed species. Recently, a large and illegal shipment of rare, wild‐caught H. spinosa was confiscated in Hong Kong, China, and the turtles were eventually distributed to several zoos and academic collections. Based on analyses of these individuals, along with additional individuals from the pet trade and museum collections, we found concordant genetic and phenotypic variation, indicating that two distinct types of H. spinosa exist in this collection of turtles. Further characterization of this variation will require field surveys and the collection of additional morphological and genetic data from specimens of known geographic provenance. However, our data indicate that this highly exploited, endangered species may contain additional cryptic taxa, and emphasize the critical need for systematic evaluation of species before unrecognized variation is lost forever. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 164 , 811–824.