Uncovering hidden biodiversity in the Cryptophyta: Clone library studies at the Helgoland Time Series Site in the Southern German Bight indentifies the cryptophycean clade potentially responsible for the majority of its genetic diversity during the spring bloom.

Cryptophyceae are important group in marine phytoplankton, but little is known about the occurrence and distribution of individual species. Recently, with use of molecular probes and microarray technology, it has been shown that species related to teleaulax spp. or Chroomonas spp. (clades 4 or 6) contributed the most to cryptophycean biomass in the north Sea. for the microarray study, the single probe (clade 4/6) recognizes members of both clades 4 and 6 and thus cannot separate them. Therefore, it was unknown as to whether the cryptophyte community was composed of clade 4, clade 6 or both of them. Here, we addressed this question and increased the genetic diversity of our investigations of cryptophycean diversity in the north Sea by sequencing 18S rRnA clone libraries made from fractionated water samples to examine specifically the picoplanktonic fraction because that fraction was studied in detail in the earlier microarray study. We focused on samples from the spring phytoplankton bloom in 2004 because the microarray signals were the strongest at this time. Excluding chimeric sequences, we detect- ed nine cryptophycean oTUs, seven of which fell into the teleaulax/Plagioselmis branch, whereas two grouped with Geminigera spp. our results indicate that these oTUs, affiliated with clade 4, may be an important component of cryptophyte community during spring bloom in the north Se

Advances in the Detection of Toxic Algae Using Electrochemical Biosensors

Harmful algal blooms (HABs) are more frequent as climate changes and tropical toxic species move northward, especially along the Iberian Peninsula, a rich aquaculture area. Monitoring programs, detecting the presence of toxic algae before they bloom, are of paramount importance to protect ecosystems, aquaculture, human health and local economies. Rapid, reliable species identification methods using molecular barcodes coupled to biosensor detection tools have received increasing attention as an alternative to the legally required but impractical microscopic counting-based techniques. Our electrochemical detection system has improved, moving from conventional sandwich hybridization protocols using di�erent redox mediators and signal probes with di�erent labels to a novel strategy involving the recognition of RNA heteroduplexes by antibodies further labelled with bacterial antibody binding proteins conjugated with multiple enzyme molecules. Each change has increased sensitivity. A 150-fold signal increase has been produced with our newest protocol using magnetic microbeads (MBs) and amperometric detection at screen-printed carbon electrodes (SPCEs) to detect the target RNA of toxic species. We can detect as few as 10 cells

MIDTAL (Microarrays for the Detection of Toxic Algae)

Microalgae in marine and brackish waters of Europe regularly cause harmful effects, considered from the human perspective, in that they cause economic damage to fisheries and tourism. Cyanobacteria cause similar problems in freshwaters. These episodes encompass a broad range of phenomena collectively referred to as harmful algal blooms (HABs). For adequate management of these phenomena, monitoring of microalgae is required. However, present day monitoring is time consuming and based on morphology as determined by light microscopy, which may be insufficient to give definitive species and toxin attribution. In the European Union (EU) FP7 project MIDTAL (microarrays for the detection of toxic algae), we will first target rapid species identification using rRNA genes. The variable regions of the rRNA genes can be used for probe design to recognize species or even strains. Second, a toxin based microarray will be developed that includes antibody reactions to specific toxins produced by these microalgae because even when cell numbers are low, toxins can be present and can accumulate in the shellfish. Microarrays are the state of the art technology in molecular biology for the processing of bulk samples for detection of target RNA/DNA sequence. Existing rRNA probes and antibodies for toxic algal species/strains and their toxins will be adapted and optimized for microarray use. The purpose of MIDTAL is to support the common fisheries policy and to aid the national monitoring agencies by providing new rapid tools for the identification of toxic algae and their toxins so they can comply with EU directive 91/1491/CEE to monitor for toxic algae, and reduce the need for the mouse bioassay

Molecular Clocks and Inferring Evolutionary Milestones and Biogeography in the Microalgae

Molecular clocks are proving to be very useful tools for unravelling the evolution of protistan taxa relative to geological change. Molecular clocks have been used to reconstruct the biogeographic history of non-fossilized microalgae from calibration of trees/clocks based on taxa with a fossil record. They have also been used to extrapolate back to the origin of microalgal lineages, to document the appearance of new morphotypes in the fossil record that should be recognised as new species, to estimate the timing of major evolutionary events, viz., endosymbioses and to infer possible explanations for selective survival during global change extinction events.International Symposium, "The Origin and Evolution of Natural Diversity". 1–5 October 2007. Sapporo, Japan