Multilocus genetic analyses provide insight into speciation and hybridization in aquatic grasses, genus Ruppia

Aquatic plants of the genus Ruppia inhabit some of the most threatened habitats in the world, such as coastal lagoons and inland saline to brackish waters where their meadows play several key roles. The evolutionary history of this genus has been affected by the processes of hybridization, polyploidization, and vicariance, which have resulted in uncertainty regarding the number of species. In the present study, we apply microsatellite markers for the identification, genetic characterization, and detection of hybridization events among populations of putative Ruppia species found in the southern Iberian Peninsula, with the exception of a clearly distinct species, the diploid Ruppia maritima. Microsatellite markers group the populations into genetically distinct entities that are not coincident with geographical location and contain unique diagnostic alleles. These results support the interpretation of these entities as distinct species: designated here as (1) Ruppia drepanensis, (2) Ruppia cf. maritima, and (3) Ruppia cirrhosa. A fourth distinct genetic entity was identified as a putative hybrid between R. cf. maritima and R. cirrhosa because it contained a mixture of microsatellite alleles that are otherwise unique to these putative species. Hence, our analyses were able to discriminate among different genetic entities of Ruppia and, by adding multilocus nuclear markers, we confirm hybridization as an important process of speciation within the genus. In addition, careful taxonomic curation of the samples enabled us to determine the genotypic and genetic diversity and differentiation among populations of each putative Ruppia species. This will be important for identifying diversity hotspots and evaluating patterns of population genetic connectivity. © 2015 The Linnean Society of London, Biological Journal of the Linnean Society, 2015, 00, 000–000.

云南中新世和山西上新世果实和种子的研究

研究化石果实和种子的古果实学在欧洲和北美开展较早，近年来发展迅速，我国在这个领域的研究相对薄弱，迄今为止尚没有专项的研究工作。本论文首次对我国云南和山西晚第三纪的果实和种子进行专门研究，并根据它们对应的现存最近亲缘植物的分布及生态特征分析和重建了当时当地的吉植被、古环境和古气候。 1．云南省西部陇川盆地芒旦地区早一中中新世被子植物果实和种子植物群，包括果实和种子29种，详细研究了14种，隶属于ll科12属：壳斗科的石栎属、金缕梅科的蜡瓣花属、金丝桃科的金丝桃属、樟科的木姜子属、木兰科的木兰属和鹅掌楸属、桑科的榕属、杨梅科的杨梅属、蓝果树科的蓝果树属、芸香科的花椒属、清风藤科的清风藤属以及山矾科的山矾属。所有这些果实和种子的化石在中国均为首次报道，其中蜡瓣花属、鹅掌楸属、蓝果树属、清风藤属和山矾属还是该属在中国的首次大化石记录。在植物群组成中，石栎属占绝对优势，其他主要类群有蜡瓣花属、木姜子属、木兰属、花椒属和山矾属等。芒旦果实和种子植物群反映当时当地为亚热带常绿阔叶林的森林面貌和温暖湿润的亚热带气候条件。 2．山西省榆社盆地张村地区晚上新世( 2.3 Ma-3.5 Ma)水生植物川蔓藻属(Ruppia)果实和种子的大量发现及其与川蔓藻属2个化石种和5个现代种的比较表明，这些果实和种子属于一个新种—榆社川蔓藻Ruppia yushensis sp. nov。该新种的主要特征为内果皮卵形、表面光滑、侧面具两个显著的窄椭圆形凹陷、顶端具一个明显的小尖突，以及种子顶端具显著的球形膨大的种脐。榆社川蔓藻将川蔓藻属的第三纪地理分布范围从欧洲延伸到了东亚，并且填补了它在上新世地层的空缺。榆社川蔓藻作为榆社盆地晚新生代水生植物的发现，指示了当时当地的咸水环境。根据川蔓藻属植物的现代生态资料并结合其它矿物学和生物学指标，可以推测晚上新世张村地区为一个浅而平静的微咸水湖。榆社川蔓藻果实和种子的大量出现说明该植物可能在该水域形成了单一的优势类群，其光滑的内果皮表面指示当时当地为暖温带或温带气候。

Assessing the sensitivity of seagrass bed biotopes to pressures associated with marine activities.

This project was commissioned to generate an improved understanding of the sensitivities of seagrass habitats to pressures associated with human activities in the marine environment - to provide an evidence base to facilitate and support management advice for Marine Protected Areas; development of UK marine monitoring and assessment, and conservation advice to offshore marine industries. Seagrass bed habitats are identified as a Priority Marine Feature (PMF) under the Marine (Scotland) Act 2010, they are also included on the OSPAR list of threatened and declining species and habitats, and are a Habitat of Principle Importance (HPI) under the Natural Environment and Rural Communities (NERC) Act 2006, in England and Wales. The purpose of this project was to produce sensitivity assessments with supporting evidence for the HPI, OSPAR and PMF seagrass/Zostera bed habitat definitions, clearly documenting the evidence behind the assessments and any differences between assessments. Nineteen pressures, falling in five categories - biological, hydrological, physical damage, physical loss, and pollution and other chemical changes - were assessed in this report. Assessments were based on the three British seagrasses Zostera marina, Z. noltei and Ruppia maritima. Z. marina var. angustifolia was considered to be a subspecies of Z. marina but it was specified where studies had considered it as a species in its own rights. Where possible other components of the community were investigated but the basis of the assessment focused on seagrass species. To develop each sensitivity assessment, the resistance and resilience of the key elements were assessed against the pressure benchmark using the available evidence. The benchmarks were designed to provide a ‘standard’ level of pressure against which to assess sensitivity. Overall, seagrass beds were highly sensitive to a number of human activities: • penetration or disturbance of the substratum below the surface; • habitat structure changes – removal of substratum; • physical change to another sediment type; • physical loss of habitat; • siltation rate changes including and smothering; and • changes in suspended solids. High sensitivity was recorded for pressures which directly impacted the factors that limit seagrass growth and health such as light availability. Physical pressures that caused mechanical modification of the sediment, and hence damage to roots and leaves, also resulted in high sensitivity. Seagrass beds were assessed as ‘not sensitive’ to microbial pathogens or ‘removal of target species’. These assessments were based on the benchmarks used. Z. marina is known to be sensitive to Labyrinthula zosterae but this was not included in the benchmark used. Similarly, ‘removal of target species’ addresses only the biological effects of removal and not the physical effects of the process used. For example, seagrass beds are probably not sensitive to the removal of scallops found within the bed but are highly sensitive to the effects of dredging for scallops, as assessed under the pressure penetration or disturbance of the substratum below the surface‘. This is also an example of a synergistic effect Assessing the sensitivity of seagrass bed biotopes to pressures associated with marine activities between pressures. Where possible, synergistic effects were highlighted but synergistic and cumulative effects are outside the scope off this study. The report found that no distinct differences in sensitivity exist between the HPI, PMF and OSPAR definitions. Individual biotopes do however have different sensitivities to pressures. These differences were determined by the species affected, the position of the habitat on the shore and the sediment type. For instance evidence showed that beds growing in soft and muddy sand were more vulnerable to physical damage than beds on harder, more compact substratum. Temporal effects can also influence the sensitivity of seagrass beds. On a seasonal time frame, physical damage to roots and leaves occurring in the reproductive season (summer months) will have a greater impact than damage in winter. On a daily basis, the tidal regime could accentuate or attenuate the effects of pressures depending on high and low tide. A variety of factors must therefore be taken into account in order to assess the sensitivity of a particular seagrass habitat at any location. No clear difference in resilience was established across the three seagrass definitions assessed in this report. The resilience of seagrass beds and the ability to recover from human induced pressures is a combination of the environmental conditions of the site, growth rates of the seagrass, the frequency and the intensity of the disturbance. This highlights the importance of considering the species affected as well as the ecology of the seagrass bed, the environmental conditions and the types and nature of activities giving rise to the pressure and the effects of that pressure. For example, pressures that result in sediment modification (e.g. pitting or erosion), sediment change or removal, prolong recovery. Therefore, the resilience of each biotope and habitat definitions is discussed for each pressure. Using a clearly documented, evidence based approach to create sensitivity assessments allows the assessment and any subsequent decision making or management plans to be readily communicated, transparent and justifiable. The assessments can be replicated and updated where new evidence becomes available ensuring the longevity of the sensitivity assessment tool. The evidence review has reduced the uncertainty around assessments previously undertaken in the MB0102 project (Tillin et al 2010) by assigning a single sensitivity score to the pressures as opposed to a range. Finally, as seagrass habitats may also contribute to ecosystem function and the delivery of ecosystem services, understanding the sensitivity of these biotopes may also support assessment and management in regard to these. Whatever objective measures are applied to data to assess sensitivity, the final sensitivity assessment is indicative. The evidence, the benchmarks, the confidence in the assessments and the limitations of the process, require a sense-check by experienced marine ecologists before the outcome is used in management decisions.

Padrões espaciais e temporais da comunidade de invertebrados bentônicos no estuário Tramandaí-Armazém, RS, e a resposta da macro e meiofauna a um derrame experimental de óleo bruto

A distribuição espaço-temporal dos invertebrados bentônicos, no estuário Tramandaí-Armazém, localizado no Litoral Norte do Rio Grande do Sul, Brasil, foi analisada em três sub-ambientes: laguna Armazém (3 áreas amostrais), laguna Tramandaí (3 áreas amostrais) e Canal de ligação com o Oceano Atlântico (1 área amostral). Na laguna Tramandaí, ocorre um aporte de água doce pelo rio Tramandaí, na laguna Armazém, ocorre a entrada mais freqüente da cunha salina e no Canal verifica-se uma maior influência do Oceano Atlântico. No ARTIGO I, foram analisados os organismos da meiofauna, obtidos a partir de amostragens sazonais em 2000. Em cada área foram tomadas seis amostras, até a profundidade de 5 cm no interior do sedimento, com um corer de 2,7cm de diâmetro. Nematoda foi o grupo taxonômico dominante em todos os ambientes. As análises uni e multivariadas mostraram que a estrutura da meiofauna na laguna Armazém, caracterizada pelas densidades mais elevadas, difere significativamente dos demais ambientes amostrados. A similaridade das amostras coletadas na laguna Tramandaí e no Canal ocorreu em quase todas as estações, com exceção da primavera. Picos de densidade da meiofauna foram encontrados no verão. Os resultados sugerem que os fatores abióticos, como salinidade, hidrodinâmica e temperatura, são importantes condicionantes desta variabilidade espaço-temporal detectada. No ARTIGO II, detalhou-se a identificação de Nematoda, em gênero, e sua classificação em grupo trófico. As análises multivariadas mostraram que a associação de Nematoda é distinta entre os três ambientes analisados. Os atributos densidade, diversidade e riqueza de gêneros foram significativamente mais elevados na laguna Armazém. Nesta laguna, os nematódeos comedores de epistrato e de bactérias foram os dominantes, enquanto que no Canal e Tramandaí prevaleceram os comedores de depósito e predadores facultativos. Uma variabilidade temporal foi detectada tanto para os gêneros, como para os grupos tróficos, com densidades mais elevadas no verão. A identificação de Nematoda e de seu comportamento alimentar mostraram que além da salinidade, hidrodinâmica e temperatura, a disponibilidade de alimento e a forma como está disponível, determinam a estrutura desta associação. No ARTIGO III, foi analisada a distribuição espaço-temporal da macrofauna, a partir de amostragens sazonais entre o outono de 2002 e o verão de 2003. Em cada área foram obtidas 6 amostras com um corer de 10cm de diâmetro enterrado até a profundidade de 20 cm no interior do sedimento. No Canal, a comunidade da macrofauna, foi caracterizada pelas menores densidades e riqueza específica, em todas as estações do ano. Diferente do verificado para Nematoda, a estrutura da macrofauna, encontrada na laguna Armazém e laguna Tramandaí foi similar. Este resultado se deve a baixa riqueza específica da macrofauna e a dominância de espécies oportunistas. Em situações de maior oferta de alimento, oriunda da decomposição de detritos (no inverno) e ao aparecimento da macrófita Ruppia maritima (na primavera), observou-se uma diferenciação da estrutura da macrofauna encontrada na laguna Armazém da laguna Tramandaí. Na laguna Armazém densidades mais elevadas ocorreram no inverno, nos demais ambientes não foram constatadas diferenças significativas entre as estações. Provavelmente as características geomorfológicas do estuário e a hidrodinâmica atuante promovam uma instabilidade em todo o estuário, que para a macrofauna se sobrepõem às diferenças ambientais encontradas entre as lagunas. No ARTIGO IV, avaliou-se a macro e meiofauna, frente a um derrame experimental de óleo, através de análises univariadas. No experimento foram considerados dois fatores: tratamento (controle, corer sem óleo, corer com óleo) e tempo (4 e 9 horas após a adição do óleo). Cada tratamento constou de cinco réplicas, nos dois períodos de coleta. Não foram detectados efeitos do óleo sobre a macrofauna, no tempo de 4 horas e 9 horas de exposição. Para a meiofauna e gêneros de Nematoda, não foram encontradas variações significativas de densidade, com exceção do gênero Theristus, que apresentou um maior número de organismos mortos na presença do óleo.

Axenic cell culture of the seagrass Cymodocea nodosa from cotyledonary tissue

[EN] Plant Tissue Culture, also called “micropropagation”, is the propagation of plants from different tissues (or explants) in a shorter time than conventional propagation, making use of the ability that many plant cells have to regenerate a whole plant (totipotency).There are two alternative mechanisms by which an explant can regenerate an entire plant, namely organogenesis and somatic embryogenesis. Since the last decades, the number of higher terrestrial plants species from which these techniques have been successfully applied has continually increased. However, few attempts have been carried out in marine plants. Previous seagrasses authors have focused their studies on i) vegetative propagation of rhizome fragments as explants in Ruppia maritima, Halophila engelmannii, Cymodocea nodosa and Posidonia oceanica; ii) culture of meristems in Heterozostera tasmanica, C. nodosa or P. oceanica; and iii) culture of germinated seeds on aseptic conditions, in Thalassia testudinum, H. ovalis, P. coriacea, P. oceanica, and H. decipiens. All these studies determine the most adequate culture medium for each species (seawater, nutrients, vitamins, carbon sources, etc...), often supplemented with different plant growth regulators and the necessary conditions for the culture maintenance, such as light and temperature. On the other hand, several studies have previously established protocols for cell or protoplast isolation in the species Zostera marina, Z. muelleri, P. oceanica, and C. nodosa, using shoots collected from natural meadows as original vegetal source, but further cell growth was never accomplished. Due to the absence of somatic embryogenesis or organogenetic studies in seagrasses we wonder: IS THE SUCCESSFUL APPLICATION OF TISSUE CULTURE TECHNIQUES POSSIBLE IN SEAGRASSES?

Age determination and pollen profile of sediment core Voulkaria 1966, Greece

Lake Voulkaria is situated in northwestern Greece in the Prefecture of Etoloakarnania, 6 km SW of the city of Vonitsa and 10 km east of the northern tip of the island of Levkás (Leukás, Lefkada). The lake is separated from the Ionian Sea on the West by a narrow limestone ridge ca 10 m high and has a size of 940 ha. An almost continuous fringe of Phragmites surrounds the open water. This reed bank is up to 500 m wide along the southern shore of the lake. Water depth is low, predominantly less than 2 m. In the south-eastern part of the lake a maximum depth of 3.1 m was measured in September 1997.

Biostratigraphical investigations of Lago d'Averno, near Naples

Remains of diatoms, molluscs, ostracods, foraminifera and pollen exines preserved in the sediments of Lago d'Averno, a volcanic lake in the Phlegrean Fields west of Naples, allowed us to reconstruct the changes in the ecological conditions of the lake and of the vegetation around it for the period from 800 BC to 800 AD. Lago d'Averno was at first a freshwater lake, temporarily influenced by volcanic springs. Salinity increased slowly during Greek times as a result of subsidence of the surrounding land. Saline conditions developed only after the lake was connected with the sea by a canal, when Portus Julius was built in 37 BC. The first post-Roman period of uplift ended with a short freshwater phase during the 7th century after Christ. Deciduous oakwoods around the lake was transformed into a forest of evergreen oaks in Greek times and thrived there - apparently almost uninfluenced by man - until it was felled, when the Avernus was incorporated into the new Roman harbour in 37 BC, to construct a shipyard and other military buildings there. Land-use was never more intense than during Roman times and weakest in Greek and Early Roman times, when the Avernus was considered a holy place, the entrance to the underworld.