Rapid evolution of post-zygotic reproductive isolation is widespread in Arctic plant lineages.

BACKGROUND AND AIMS: The Arctic tundra, with its extreme temperatures and short growing season, is evolutionarily young and harbours one of the most species-poor floras on Earth. Arctic species often show little phenotypic and genetic divergence across circumpolar ranges. However, strong intraspecific post-zygotic reproductive isolation (RI) in terms of hybrid sterility has frequently evolved within selfing Arctic species of the genus Draba. Here we assess whether incipient biological species are common in the Arctic flora. METHODS: We conducted an extensive crossing experiment including six species representing four phylogenetically distant families collected across the circumpolar Arctic. We crossed conspecific parental populations representing different spatial scales, raised 740 F1 hybrids to maturity and measured fertility under laboratory conditions. We examined genetic divergence between populations for two of these species (Cardamine bellidifolia and Ranunculus pygmaeus). KEY RESULTS: In five of the six species, we find extensive reduction in pollen fertility and seed set in F1 hybrids; 219 (46 %) of the 477 F1 hybrids generated between parents separated by ≥427 km had <20 % pollen fertility. Isolation with migration (IM) and *BEAST analyses of sequences of eight nuclear genes in C. bellidifolia suggests that reproductively isolated populations of this species diverged during, or even after, the last glaciation. Likewise, Arctic populations of R. pygmaeus were genetically very similar despite exhibiting strongly reduced fertility in crosses, suggesting that RI evolved recently also in this species. CONCLUSION: We show that post-zygotic RI has developed multiple times within taxonomically recognized Arctic species belonging to several distantly related lineages, and that RI may have developed over just a few millennia. Rapid and widespread evolution of incipient biological species in the Arctic flora might be associated with frequent bottlenecks due to glacial cycles, and/or selfing mating systems, which are common in the harsh Arctic environment where pollinators are scarce.

Allopolyploid origins of the Galeopsis tetraploids--revisiting Müntzing's classical textbook example using molecular tools.

Whole-genome duplication coupled with hybridization is of prime importance in plant evolution. Here we reinvestigate Müntzing's classical example of allopolyploid speciation; the first report of experimental synthesis of a naturally occurring allopolyploid species, Galeopsis tetrahit. Various molecular markers (cpDNA, NRPA2, amplified fragment length polymorphisms (AFLPs)) and flow cytometry were surveyed in population samples of subgenus Galeopsis, including two allopolyploid species and their potential diploid parents. The presence of two divergent copies of single-copy NRPA2 confirms the allopolyploid origins of G. tetrahit and Galeopsis bifida. However, the two allopolyploids do not share the same maternal genome, as originally suggested by Müntzing. The results support independent origins, but not recurrent formation, of the two allotetraploids. Data further indicate frequent gene flow and introgression within ploidy levels, but less so between ploidy levels. Our results confirm and elaborate on Müntzing's classical conclusion about allopolyploid origins of G. tetrahit and G. bifida. We address questions of general interest within polyploidy research, such as recurrent formation, gene flow and introgression within and between ploidy levels.

Genetics of intrinsic postzygotic isolation in a circumpolar plant species, Draba nivalis (Brassicaceae).

Sterility barriers, ranging from incomplete to fully developed, were recently demonstrated within taxonomic species of the genus Draba, suggesting the existence of numerous, cryptic biological species. Because these taxa are predominately selfers and of Pleistocene origin, it was concluded that hybrid sterility evolved quickly and possibly by genetic drift. Here we used genetic mapping and QTL analyses to determine the genetic basis of hybrid sterility between geographically distant populations of one of these taxonomic species, Draba nivalis. Fifty microsatellite loci were mapped, and QTL analyses identified five loci underlying seed fertility and two underlying pollen fertility. Four of five seed fertility QTLs reduced fertility in heterozygotes, an observation most consistent with drift-based fixation of underdominant sterility loci. However, several nuclear-nuclear interactions were also found, including two that acted like reciprocal translocations with lowest fitness in double heterozygotes, and two that had a pattern of fitness consistent with Bateson-Dobzhansky-Muller incompatibilities. In contrast, pollen fertility QTLs exhibited additive inheritance, with lowest fertility associated with the paternal allele, a pattern of inheritance suggestive of cytonuclear incompatibilities. The results imply that multiple genetic mechanisms underlie the rapid evolution of reproductive barriers in Draba.

Genetics of cryptic speciation within an Arctic mustard, Draba nivalis.

Crossing experiments indicate that hybrid sterility barriers frequently have developed within diploid, circumpolar plant species of the genus Draba. To gain insight into the rapid evolution of postzygotic reproductive isolation in this system, we augmented the linkage map of one of these species, D. nivalis, and searched for quantitative trait loci (QTLs) associated with reproductive isolation. The map adds 63 new dominant markers to a previously published dataset of 31 co-dominant microsatellites. These markers include 52 amplified fragment length polymorphisms (AFLPs) and 11 sequence-specific amplified polymorphisms (SSAPs) based on retrotransposon sequence. 22 markers displaying transmission ratio distortion were further included in the map. We resolved eight linkage groups with a total map length of 894 cM. Significant genotype-trait associations, or quantitative trait loci (QTL), were detected for reproductive phenotypes including pollen fertility (4 QTLs), seed set (3 QTLs), flowering time (3 QTLs) and number of flowers (4 QTLs). Observed patterns of inheritance were consistent with the influence of both nuclear-nuclear interactions and chromosomal changes on these traits. All seed set QTLs and one pollen fertility QTL displayed underdominant effects suggestive of the involvement of chromosomal rearrangements in hybrid sterility. Interestingly, D. nivalis is predominantly self-fertilizing, which may facilitate the establishment of underdominant loci and contribute to reproductive isolation.

High biological species diversity in the arctic flora.

The arctic flora is considered to be impoverished, but estimates of species diversity are based on morphological assessments, which may not provide accurate counts of biological species. Here we report on crossing relationships within three diploid circumpolar plant species in the genus Draba (Brassicaceae). Although 99% of parental individuals were fully fertile, the fertility of intraspecific crosses was surprisingly low. Hybrids from crosses within populations were mostly fertile (63%), but only 8% of the hybrids from crosses within and among geographic regions (Alaska, Greenland, Svalbard, and Norway) were fertile. The frequent occurrence of intraspecific crossing barriers is not accompanied by significant morphological or ecological differentiation, indicating that numerous cryptic biological species have arisen within each taxonomic species despite their recent (Pleistocene) origin.