Phylogenetic position of Rickettsia tsutsugamushi and the relationship among its antigenic variants by analyses of 16S rRNA gene sequences

Abstract The 16S rRNA gene sequences of Rickettsia tsutsugamushi and Rickettsia sibirica were determined by PCR and DNA sequencing. Phylogenetic analysis revealed that R. sibirica is positioned in a cluster of the genus Rickettsia with a similarity value of 98.1–99.6%, whereas R. tsutsugamushi is located apart from the cluster with a similarity value of 90.2–90.6%. This evidence suggests that R. tsutsugamushi should be excluded taxonomically from the genus Rickettsia . The phylogenetic classification of six antigenic variants in R. tsutsugamushi moderately reflected their antigenic relationship known in closely and distantly related strains.     

Thecamoeba foliovenanda n. sp. (Amoebozoa,Discosea, Thecamoebida) – One more case of sibling species among amoebae of the genus Thecamoeba

Until recently it seemed that amoebae of the genus Thecamoeba can be reliably identified using light-microscopic characters, like the size and shape of the cell, a characteristic pattern of folds and ridges and structure of the nucleus. However, recent molecular studies show the presence of sibling species that can be reliably distinguished based on the gene sequence data only. Here we describe a new terrestrial species, Thecamoeba foliovenanda n. sp. This species is almost identical with Thecamoeba similis in light-microscopy, which has minor differences in the ultrastructure but considerable differences in the SSU rRNA gene sequence. We investigated the light-microscopic data, as well as transmission electron-microscopic images and videorecords on the type strain of T. similis 1583/8 and performed its comparison with the new species. This study further supports the conclusion that species of the genus Thecamoeba nowadays require gene sequencing for reliable identification and species distinction.     

Structural and developmental studies of Chlamydomyzium oviparasiticum from Rhabditis nematodes and in culture

The oomycete (Peronosporomycete) Chlamydomyzium oviparasiticum, previously recorded as a parasite of rotifer eggs, was found infecting Rhabditis nematodes in a sample of rotting garden compost. For the first time C. oviparasiticum was cultured in liquid media, which enabled more detailed studies of zoospore behaviour and facilitated the use of confocal microscopy. Rhabditis nematodes were successfully re-infected from liquid-cultured inoculum. Light (including video) microscopy and transmission electron microscopy were used to document details of thallus development, zoospore release and resting spore morphology to enable comparison with other oomycete species. This species showed several significant saprolegnialian characters such as the ‘achlyoid’ pattern of spore formation, centrifugal cleavage and structured encystment vesicles. In contrast, spore release into a transient vesicle was a peronosporalean characteristic. The thick-walled resting spores showed relatively poor cytoplasmic preservation and had a thick multi-layered wall. It was still not possible to unequivocably decide whether these were chlamydospores or parthenogenically formed oospores. The phylogenetic significance of these observations is discussed.     

Jsowerbya, new genus of Muricidae (Mollusca: Gastropoda) from the Eocene of the Paris (France) and Hampshire (England) basins with a phylogenetic assessment of its Ocenebrine versus Ergalataxine affinities

Jsowerbya, nov. gen. (Gastropoda: Muricidae) includes three species from the Eocene of the Paris and Hampshire basins. It increases the number of extinct muricid genera, which curiously represent a very small fraction of the described genera. The species of Jsowerbya, often mistaken for the muricopsine genus Muricopsis, possess a unique combination of characters shared with the subfamilies Ocenebrinae and Ergalataxinae. A cladistic analysis, based on structural homologies of the spiral sculpture, however, suggests that Jsowerbya is closely related to the Ocenebrinae. Thus, Jsowerbya is here regarded as one the most basal Ocenebrinae.     

Phosphorylase activity in relation to starch synthesis in developing Hordeum distichum grain

Activity, control and primer requirements of starch phosphorylase in developing barley endosperm were investigated. Phosphorylase was detected in endosperm extracts from 3 days after anthesis. Unprimed activity was predominant between 2 and 10 days after anthesis, when it constituted 70–80% of total activity, but this proportion declined rapidly as the grain developed. The existence of at least 2 isoenzymes was indicated by studies of pH dependence and phosphate inhibition, and was further supported by acrylamide gel electrophoresis and column chromatography using DEAE-cellulose. The two isoenzymes which ere possibly both glyco proteins, appear in barley endosperm soon after anthesis. One appears capable of unprimed activity, and may be associated with the initiation of a-1,2 glucans, which then serve as primers for starch synthetase. This disappears by 13–15 days after anthesis. The other isoenzyme is capable of some unprimed activity but undergoes modification between 15 and 20 days after anthesis, resulting in the loss of unprimed activity. The relevance of the results to initiation of starch synthesis and to starch synthetase in amyloplasts is discussed.     

Les Pseudholaster (Echinoidea,Holasteroida) du Crétacé de France

In the order of Holasteroida, the fossil record highlights a contradiction between the genus Pseudholaster that appears in the Aptian, whose plastron is prostostern close to the Jurassic ancestors and the genus Holaster, which appears in the Valanginian, whose meridostern plastron appears more derived. This inconsistency can be explained by the ignorance of the plastronal architecture on the part of the early authors. A review of the species of Pseudholaster from the Cretaceous period of France was therefore carried out. The objective was to statistically determine the discriminating morphological characters, and to study the modifications of the architecture of the interambulacrum 5 of the French species belonging to this genus, as well as to the species included in the genus Holaster incorrectly by earlier authors. This review of the species of the genus Pseudholaster begins with a study of the ontogeny of the species Holaster intermedius Münster in Goldfuss, 1826–1833, first representative of the genus Pseudholaster, which appears in the Hauterivian in the Parisian and Rhodano-vocontian basins. The modifications during growth concern the overall shape, but also the plastron architecture: the number of plastron plates increases while the number of plates located between the peristome and the periproct remains fixed. The plastron of this species is protosternal and not meridosternal as Lambert pointed out. The labrum is cupuliform in contact with the second sternal 5a2 by a narrow digitation. However, this arrangement differs from that observed on a protosternal breastplate. This apomorphism of the plastron plate pattern, called “labrotaxienne”, is found in all the Pseudholaster studied, and the study of the architecture of the interambulacrum 5 also reveals a gradual decrease in the number of preanal plates between the oldest (Hauterivian) and the younger (Cenomanian-Lower Turonian) species studied. Most of the French species have been revised, with some synonyms. A new species, P. neraudeaui, is the last known Pseudholaster dated from the upper Cenomanian and lower Turonian of southwestern France. Our study illustrates the evolution of the genus Pseudholaster between the Hauterivian and the early Turonian in France. The interest of the study is to show that the appearance of the genus Pseudholaster is older than that of the genus Holaster. Pseuholaster intermedius, of Hauterivian age, possesses a derived protostern plastron called here “labrotaxien” and not meridostern as defined historically by Lambert, and to reveal that the number of preanals decreases over geological time. This data is essential for future phylogenetic studies. On a palaeobiogeographical level, the study reveals the expansion of the genus Pseudholaster during early Cretaceous in western Europe, with diversification during the Albian, its disappearance during late Cenomanian in the Paris basin while it still persists in the Aquitain basin, its predilection for circalitoral environments.     

Cloning of genes for bacteriophage T5 stable RNAs

One EcoRI-generated fragment (440 basepairs) and two EcoRI/HindIII fragments (220 and 960 basepairs) from the deletion region of T5 phage have been inserted into the phage λ XIII and the plasmid pBR322 as vectors. Recombinant DNA molecules were studied by hybridization with in vivo ³²P-labeled T5 4–5 S RNAs on nitrocellulose filters. Two-dimensional polyacrylamide gel electrophoretic fractionation and fingerprint analysis of the RNAs eluted from the filters were carried out to identify RNAs coded by cloned fragments. For the accurate localization of the genes for these RNAs, RNA-DNA hybrids were treated with T1 and pancreatic RNAases, and the eluted RNA fragments stable against RNAase action were electrophoresed. It was shown that the EcoRI¹440 fragment contains the gene for tRNA 10 (tRNA^Asp), the EcoRI/HindIII¹220 fragment contains the gene for RNA III (107 bases) and parts of the genes for RNA I (107 bases) and tRNA 12 (tRNA^His), and the EcoRI/HindIII¹960 fragment contains only a part of the gene for tRNA 9 (tRNA^Gln). The arrangement of these genes on the physical map of T5 phage was as follows: -tRNA^Gln-tRNA^His-RNA III-RNA I-…-tRNA^Asp.