Studying Whole-Genome Duplication Using Experimental Evolution of Chlamydomonas.

In this chapter, we present the use of Chlamydomonas reinhardtii in experiments designed to study the evolutionary impacts of whole genome duplication. We shortly introduce the algal species and depict why it is an excellent model for experimental evolution. Subsequently, we discuss the most relevant steps and methods in the design of a ploidy-related Chlamydomonas experiment. These steps include strain selection, ploidy determination, different methods of making diplo- and polyploid Chlamydomonas cells, replication, culturing conditions, preservation, and the ways to quantify phenotypic and genotypic change.

Limits to gene flow in a cosmopolitan marine planktonic diatom.

The role of geographic isolation in marine microbial speciation is hotly debated because of the high dispersal potential and large population sizes of planktonic microorganisms and the apparent lack of strong dispersal barriers in the open sea. Here, we show that gene flow between distant populations of the globally distributed, bloom-forming diatom species Pseudo-nitzschia pungens (clade I) is limited and follows a strong isolation by distance pattern. Furthermore, phylogenetic analysis implies that under appropriate geographic and environmental circumstances, like the pronounced climatic changes in the Pleistocene, population structuring may lead to speciation and hence may play an important role in diversification of marine planktonic microorganisms. A better understanding of the factors that control population structuring is thus essential to reveal the role of allopatric speciation in marine microorganisms.

Natural hybrids in the marine diatom Pseudo-nitzschia pungens (Bacillariophyceae): genetic and morphological evidence.

Hybridization between genetically distinguishable taxa provides opportunities for investigating speciation. While hybridization is a common phenomenon in various macro-organisms, natural hybridization among micro-eukaryotes is barely studied. Here we used a nuclear and a chloroplast molecular marker and morphology to demonstrate the presence of natural hybrids between two genetically and morphologically distinct varieties of the marine planktonic diatom Pseudo-nitzschia pungens (vars. pungens and cingulata) in a contact zone in the northeast Pacific. Cloning and sequencing of the rDNA internal transcribed spacer region revealed strains containing ribotypes from both varieties, indicating hybridization. Both varieties were found to also have different chloroplast-encoded rbcL sequences. Hybrid strains were either hetero- or homoplastidial, as demonstrated by denaturing gradient gel electrophoresis, which is in accordance with expectations based on the mode of chloroplast inheritance in Pseudo-nitzschia. While most hybrids are probably first generation, there are also indications for further hybridization. Morphologically, the hybrids resembled var. pungens for most characters rather than having an intermediate morphology. Further research should focus on the hybridization frequency, by assessing the spatial and temporal extent of the contact zone, and hybrid fitness, to determine the amount of gene flow between the two varieties and its evolutionary consequences.