Paraphyly and budding speciation in the hairy snail (Pulmonata,Hygromiidae)

Delimitation of species is often complicated by discordance of morphological and genetic data. This may be caused by the existence of cryptic or polymorphic species. The latter case is particularly true for certain snail species showing an exceptionally high intraspecific genetic diversity. The present investigation deals with the Trochulus hispidus complex, which has a complicated taxonomy. Our analyses of the COI sequence revealed that individuals showing a T. hispidus phenotype are distributed in nine highly differentiated mitochondrial clades (showing p‐distances up to 19%). The results of a parallel morphometric investigation did not reveal any differentiation between these clades, although the overall variability is quite high. The phylogenetic analyses based on 12S, 16S and COI sequences show that the T. hispidus complex is paraphyletic with respect to several other morphologically well‐defined Trochulus species (T. clandestinus, T. villosus, T. villosulus and T. striolatus) which form well‐supported monophyletic groups. The nc marker sequence (5.8S–ITS2–28S) shows only a clear separation of T. o. oreinos and T. o. scheerpeltzi, and a weakly supported separation of T. clandestinus, whereas all other species and the clades of the T. hispidus complex appear within one homogeneous group. The paraphyly of the T. hispidus complex reflects its complicated history, which was probably driven by geographic isolation in different glacial refugia and budding speciation. At our present state of knowledge, it cannot be excluded that several cryptic species are embedded within the T. hispidus complex. However, the lack of morphological differentiation of the T. hispidus mitochondrial clades does not provide any hints in this direction. Thus, we currently do not recommend any taxonomic changes. The results of the current investigation exemplify the limitations of barcoding attempts in highly diverse species such as T. hispidus.     

Multiple mode of binding of phencyclidines: high affinity association between phencyclidine receptors in rat brain and a monovalent ion-sensitive polypeptide

Two populations of phencyclidine (PCP) binding sites are shown to exist in the rat brain: a high-affinity monovalent ion-sensitive site (Kd of 10-14 nM for [3H]TCP, [3H]N-[1-(2-thienyl)cyclohexyl]piperidine), which exists in both the frontal cortex and the hippocampus, and a lower affinity site (Kd of 80-130 nM for [3H]TCP) which is found in the hippocampus but not in the frontal cortex. The nature of the interactions between the ion-binding sites and the high affinity PCP receptors depend on both ligand structure (PCP or TCP) and the ion involved (K' or Na'). The high-affinity sites are associated with an Mr 90,000 polypeptide whose labeling by [3H]azido phencyclidine is selectively inhibited by monovalent ions.     

Geranyl diphosphate synthase is required for biosynthesis of gibberellins

Geranyl diphosphate synthase (GPS) is generally considered to be responsible for the biosynthesis of monoterpene precursors only. However, reduction of LeGPS expression in tomato (Lycopersicon esculentum) by virus-induced gene silencing resulted in severely dwarfed plants. Further analysis of these dwarfed plants revealed a decreased gibberellin content, whereas carotenoid and chlorophyll levels were unaltered. Accordingly, the phenotype could be rescued by application of gibberellic acid. The dwarfed phenotype was also obtained in Arabidopsis thaliana plants transformed with RNAi constructs of AtGPS. These results link geranyl diphosphate (GPP) to the gibberellin biosynthesis pathway. They also demand a re-evaluation of the role of GPS in precursor synthesis for other di-, tri-, tetra- and/or polyterpenes and their derivatives.     

New roads towards chloroplast transformation in higher plants

The attempts to manipulate organelle DNA of higher plants have not yet been succesfull. Although some Agrobacterium mediated and direct gene transfer experi-ments suggested that chloroplasts of Nicotiana tabacum could be transformed, these experiments could not be reproduced. New transformation techniques have emerged: laser-injection, microinjection and high velocity microprojectiles. The biolistic ex-periments with Chlamydomonas made it clear that organelle DNA can support introduced DNA fragments by homologous recombination. In view of these devel-opments, we describe the parameters and new possibilities for chloroplast trans-formation of higher plants.     

Range‐constrained co‐occurrence simulation reveals little niche partitioning among rock‐dwelling Montenegrina land snails (Gastropoda: Clausiliidae)

Aim

Taxon co‐occurrence analysis is commonly used in ecology, but it has not been applied to range‐wide distribution data of partly allopatric taxa because existing methods cannot differentiate between distribution‐related effects and taxon interactions. Our first aim was to develop a taxon co‐occurrence analysis method that is also capable of taking into account the effect of species ranges and can handle faunistic records from museum databases or biodiversity inventories. Our second aim was to test the independence of taxon co‐occurrences of rock‐dwelling gastropods at different taxonomic levels, with a special focus on the Clausiliidae subfamily Alopiinae, and in particular the genus Montenegrina.

Location

Balkan Peninsula in south‐eastern Europe (46N–36N, 13.5E–28E).

Methods

We introduced a taxon‐specific metric that characterizes the occurrence probability at a given location. This probability was calculated as a distance‐weighted mean of the taxon's presence and absence records at all sites. We applied corrections to account for the biases introduced by varying sampling intensity in our dataset. Then we used probabilistic null‐models to simulate taxon distributions under the null hypothesis of no taxon interactions and calculated pairwise and cumulated co‐occurrences. Independence of taxon occurrences was tested by comparing observed co‐occurrences to simulated values.

Results

We observed significantly fewer co‐occurrences among species and intra‐generic lineages of Montenegrina than expected under the assumption of no taxon interaction.

Main conclusions

Fewer than expected co‐occurrences among species and intra‐generic clades indicate that species divergence preceded niche partitioning. This suggests a primary role of non‐adaptive processes in the speciation of rock‐dwelling gastropods. The method can account for the effects of distributional constraints in range‐wide datasets, making it suitable for testing ecological, biogeographical, or evolutionary hypotheses where interactions of partly allopatric taxa are in question.     

Characterization of Blue Light Signal Transduction Chains That Control Development and Maintenance of Sexual Competence in Chlamydomonas reinhardtii

Blue light induces the differentiation of Chlamydomonas reinhardtii pregametes to gametes. The light-induced conversion of pregametes to gametes is protein synthesis dependent and proceeds only after a lag phase. Upon incubation in the dark, gametes lost their mating ability, resulting in dark-inactivated gametes. Reillumination rapidly restored mating competence and this was shown to be independent of protein synthesis. Apparently, differentiation and maintenance of gametic competence are both regulated by light. Whether one or two light-activated signal pathways are involved was investigated using pharmacological compounds that affect signal transduction. Compounds that affected pregamete-to-gamete conversion affected the expression of a gamete-specific gene in a similar fashion. Other drugs affected only dark-inactivated gametes, suggesting that reactivating gametes requires a separate signaling pathway. Combined treatments provided evidence for the consecutive action of a phosphatase and a protein kinase C-like kinase in the light-induced reactivation process.     

Characterisation and cloning of a calmodulin-like domain protein kinase from Chlamydomonas moewusii (Gerloff )

Calcium-stimulated protein kinase activity in the flagella of the green alga Chlamydomonas moewusii (Gerloff) was characterised. Using SDS-PAGE and an on-blot phosphorylation assay, a 65-kDa protein was identified as the major calcium-stimulated protein kinase. Its activity was directly stimulated by calcium, a characteristic of the calmodulin-like domain protein kinases (CDPKs). Monoclonal antibodies raised against the CDPKα from soybean cross-reacted with the 65-kDa protein in the flagella, and also with other proteins in the flagellum and cell body. The same monoclonal antibodies were used to screen a C. moewusii cDNA expression library in order to isolate CDPK cDNAs from C. moewusii. The CCK1 cDNA encodes a protein with a kinase and calmodulin-like domain linked by a junction domain typical of CDPKs. From Southern analyses, evidence was obtained for a CDPK gene family in C. moewusii and C. reinhardtii. Received: 9 July 1996 / Accepted: 13 November 1996