Discrimination Amongst Leishmania by Polymerase Chain Reaction and Hybridization with Small Subunit Ribosomal DNA Derived Oligonucleotides

ABSTRACT. A method for discriminating among Leishmania is described, based upon small subunit ribosomal DNA sequence differences. The method was to amplify the entire 2.2 kb small subunit rDNA by polymerase chain reaction using conserved primers specific for the 5' and 3' termini of the small subunit ribosomal RNA, and then hybridize the product dotted onto nylon membranes with labeled oligonucleotides. The design of the hybridization probes was based upon complete small subunit rDNA sequences from L. amazonensis, L. major and L. guyanensis and partial sequences of L. mexicana, L. braziliensis, L. tropica and L. chagasi. A high degree of sequence similarity (> 99%) among species was found. However, sufficient sequence divergence occurred to permit the design of internal oligonucleotide probes specific for species complexes. This procedure successfully discriminated amongst a wide range of Leishmania isolates. The method detected as few as 10 cultured organisms and detected parasites in tissue samples from experimentally infected animals. Non-radioactive labeling showed the same specificity and sensitivity as radioactive probes.     

Anatomy of extrafloral nectaries in Fabaceae from dry‐seasonal forest in Brazil

Extrafloral nectaries (EFNs) are found in many species of Fabaceae. The aim of this work is to describe the internal morphology of the EFNs from species of Fabaceae found in areas of dry‐seasonal forest in north‐eastern Brazil. All species of Fabaceae with EFNs found were collected and samples were submitted to conventional techniques for anatomical and scanning electronic microscopy analysis. EFNs were found in 35 species, of which 32 were examined anatomically. All types have epidermal cells, secretory tissues and vascular bundles in the EFNs. Sclerenchymatous cells were found between the secretory tissues and the vascular tissues, with a few exceptions. The function of these cells is not clear; however, a role in the transportation of the sap in the nectary or with the support of the secretory tissue is possible. The nectar is released through glandular trichomes, secretory pores or even by breaking the epidermal cells and cuticle. The internal patterns found in the EFNs from different species and genera can provide important information for taxonomic and evolutionary studies in the family. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 163 , 87–98.     

New microsatellite resources for groupers (Serranidae)

We characterized nine microsatellite loci and identified an additional 60 genomic regions containing microsatellites in the red hind grouper, Epinephelus guttatus. The nine loci were highly polymorphic, and primers designed from red hind genomic DNA produced a strong amplification product in a test panel of 16 other groupers in the genera Epinephelus and Mycteroperca collected from across the world. Most of the amplified regions were homologous to the red hind locus and a well‐defined microsatellite repeat was generally evident. The nine loci, together with the 60 uncharacterized microsatellite‐containing regions, provide a powerful tool for ecological and evolutionary studies in groupers.     

Where and when the earliest coccolithophores?

Gardin, S., Krystyn, L., Richoz, S., Bartolini, A. & Galbrun, B. 2012: Where and when the earliest coccolithophores? Lethaia, Vol. 45, pp. 507–523. New calcareous nannofossil analyses from the Northern Calcareous Alps of Austria are herein used to update and improve the state of knowledge about the oldest occurrence of coccolithophores reported in the literature. Previously reported Norian occurrences of coccoliths were based on an obsolete Triassic chronostratigraphy, in which the Rhaetian stage was subsumed into the Norian (‘Sevatian 2’). The oldest stratigraphical record of coccoliths spp. lies just below the Norian‐Rhaetian boundary and the first coccolith species, Crucirhabdus minutus, is recorded from the base of Rhaetian stage. The latter bio‐event is located just above the First Occurrence of the conodont Misikella posthersteni and the first occurrence of the ammonoid Paracochloceras suessi in the Steinbergkogel section (Austria), Global Stratotype Section and Point (GSSP) candidate for the Norian–Rhaetian boundary. The appearance of the coccolith Crucirhabdus minutus is seen as a robust biochronological datum that will provide useful constraints for Triassic biostratigraphy, palaeoclimatic modelling and phylogenetic reconstructions. The new calcareous nannofossil biochronology of Steinbergkogel that we present herein completes the existing biostratigraphic characterization of the Norian‐Rhaetian transition based on conodonts and ammonoids and strengthens the position of Steinbergkogel as the best GSSP proposed section for the base of the Rhaetian. The record of coccolithophores across the Norian–Rhaetian boundary at Steinbergkogel takes place along with a discernible increase in abundance of Prinsiosphaera triassica, as well as the appearance of Euconusphaera zlambachensis, which are the two most important Rhaetian pelagic carbonate producers. This succession of bio‐events is interpreted as the initiation of the pelagic carbonate production driven by the successful spreading of calcareous nannofossils in the Western Tethys during the Rhaetian. □Austria, Calcareous Alps, coccolithophores, Triassic.